Reproductive Biology and Genetic Parameters of Blue Pine: Insights into Germination, Growth and Breeding Potential

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Jackson is a five-needle pine belonging to the Pinaceae family that is indigenous to the Hindu Kush, Karakoram and Himalayan regions. Results Male and female strobili phenophases lasted 10–11 and 77–78 weeks, respectively. Cones were collected from 18 phenotypically superior trees across six locations to assess variability in morphological traits. Trees from Akhal exhibited the highest values for all measured traits, including tree height (28.9 m), DBH (47.7 cm), cone length (22.44 cm), cone weight (123.42 g), and seed weight (87.98 g). Seeds underwent cold stratification at 25-day intervals (0–125 days) at 4 ± 1°C to evaluate germination and seedling performance. Maximum germination (72.67%) and seedling vigor were observed at 100 days of stratification, while the Akhal genotype showed superior overall performance (germination 86.77%, vigor index 1048.18). High genetic variability was observed, with the highest GCV and PCV for dry root weight (51.40, 52.65 respectively). Heritability was highest for cone specific gravity (0.78) and seed morphological traits. Principal component analysis (PCA) revealed that the first two components explained 96.23% of the total variation, with cone weight and seed thickness as major contributors. Cluster analysis grouped genotypes into five clusters, with Cluster I exhibiting superior cone, seed, and seedling traits, indicating potential for selective breeding and genetic improvement programs. Reproductive biology half-sib progenies variability stratification period PCV GCV Pinus wallichiana Figures Figure 1 Figure 2 Introduction Pinus wallichiana often referred to as Bhutan pine, Himalayan pine or blue pine, A.B. Jackson is a five-needle pine belonging to the Pinaceae family that is indigenous to the Hindu Kush, Karakoram and Himalayan regions. It is found in Nepal, Bhutan, southwestern China, northern Pakistan and northwestern India (Jammu & Kashmir, Himachal Pradesh, Uttarakhand and Arunachal Pradesh) in addition to eastern Afghanistan (Farjon 2013 ). The species can be found as low as 1,200 m but it grows most commonly at elevations between 1,800 and 4,300 m. According to (Singh and Yadav 2000 ) it flourishes in the moderate climate of the Western Himalayas which is marked by rainy summers and chilly winters. Large pure stands of Pinus wallichiana can be found in the Kashmir Valley, whereas at higher elevations, mixed forests containing Cedrus deodara , Picea smithiana and Abies pindrow dominate. When fully grown, trees reach heights of 30 to 50 meters and diameters of 40 to 80 centimeters. Second only to deodar in terms of commercial importance, its timber is pale pink to reddish with darker streaks, reasonably robust, and durable under cover (Troup 1921 ; Bhat et al. 2017 ). The species is widely utilized as fuelwood, furniture, packing boxes, and construction material. Pine needles yield essential oils with antibacterial properties, while the wood and needle resin are employed in industry and medicine (Orwa et al. 2009 ). Ecologically, the species stabilizes slopes, enriches soil through litter fall and supports watershed functions. Importance of Reproductive Biology in Forest Improvement By clarifying blooming phenology, pollination processes, cone and seed development and germination behavior reproductive biology serves as the basis for tree improvement initiatives (Kearns and Inouye 1997 ). Because self-fertilization frequently causes inbreeding depression and poor seed set reproductive studies are essential in heavily outcrossing species like P. wallichiana (Aslam et al. 2010 ). Cone and seed collection timing is influenced by knowledge of reproductive phenophases, which also aids in coordinating controlled pollination in seed orchards. Understanding dormancy breaking techniques is equally important for reliable seedling establishment. Physiological dormancy in P. wallichiana seeds causes irregular and delayed germination under normal conditions (Williams 1985 ). Cold wet stratification is the most effective way to increase germination by breaking dormancy and raising the vigor and survival of seedlings (Sharma et al. 1994 ; Luna and Singh, 2009 ). Artificial regeneration and massive afforestation activities are directly impacted in the Himalayas, where anthropogenic stresses such as fire, grazing and overexploitation are posing an increasing threat to natural regeneration (Aslam et al. 2011 ). Previous Studies and Knowledge Gap Aspects of the ecology and reproductive biology of P. wallichiana have been documented by a number of researchers. Cone and seed morphology was described by (Aslam et al. 2010 ) and its ecological amplitude throughout Himalayan ranges was reported by (Singh and Yadav 2000 ). According to studies, the species takes 15–20 years to produce viable seeds and it takes 16–18 months for the cones to mature (Tewari et al. 2001 ). However, seed dormancy, adverse microsite conditions and biotic perturbations continue to be the main causes of poor natural regeneration (Moza and Bhatnagar, 2005 ). Few thorough studies that combine reproductive biology with genetic variability characteristics exist, despite the Blue pine's ecological and economic significance. Standardized procedures for P. wallichiana are not well documented despite the fact that seed dormancy and stratification requirements have been widely recognized in temperate conifers (Williams 1985 ). Furthermore, nothing is known about the genetic variance in half-sib progenies' cone, seed and seedling properties. For this species, parameters like heritability estimates, genotypic coefficient of variation (GCV) and phenotypic coefficient of variation (PCV) are still mostly unknown. Establishing seed production sites and identifying superior genotypes for tree improvement depend heavily on this knowledge. Objectives of the Present Study The present study was undertaken to fill the above gaps with the following objectives: To study the reproductive phenophases of Pinus wallichiana across selected sites in Kashmir Himalayas. To determine variation in tree, cone and seed morphological characteristics among half-sib progenies of superior phenotypes. To assess the effect of different cold moist stratification periods on seed germination and seedling growth. To estimate genetic parameters such as PCV, GCV, heritability and to apply multivariate analysis for identifying superior genotypes. By addressing these objectives, this study aims to provide insights into the reproductive ecology and genetic variation of P. wallichiana , which will support conservation strategies, improve artificial regeneration success, and inform selective breeding programs for long-term forest sustainability. Materials and Methods Study Area The present study was conducted during 2021–2023 in the experimental field of the Division of Forest Biology and Tree Improvement, Faculty of Forestry, Benhama (Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, SKUAST-K) Ganderbal, Jammu & Kashmir, India. The Kashmir Valley lies between 33°–35° N latitude and 74°–76° E longitude at elevations ranging from 1,500 to 2,300 m above mean sea level. The valley experiences a temperate climate with four distinct seasons: a severe winter (December–February), spring (March–May), summer (June–August) and autumn (September–November). Annual precipitation averages 690 mm, mostly as snow and rain between December and April, while temperatures vary from − 8°C in winter to 33°C in summer (Bhat et al. ,2017). Selection of Trees and Cone Collection Six sites were selected across Ganderbal district representing diverse elevations and habitats: Akhal, Anderwan, Gutlibagh, Walliwar, Wussan and Fraw. From each site, three phenotypically superior trees of Pinus wallichiana were identified based on height, straightness and crown form making a total of 18 trees. The trees were at least 100 m apart to minimize the risk of relatedness and ensure genetic variability. Fifty mature cones were harvested from each selected tree during September 2021. Cones were air-dried and seeds were manually extracted for further analysis. Recording of Morphological Parameters Tree Parameters: Height (m ) : Measured with Ravi’s multimeter. Diameter at Breast Height (DBH, cm): Recorded at 1.37 m using a diameter tape. Cone Parameters: Length and diameter (cm): Measured for 10 cones per replicate using a measuring tape and digital caliper. Weight (g): Fresh weight of 10 cones per replicate recorded using a top-pan balance. Specific gravity: Estimated using the water displacement method (ISTA 1996). Number of scales: Counted manually per cone. Number of seeds per cone: Recorded after seed extraction. Seed Parameters: 1000-seed weight (g): Determined using three replicates per tree (ISTA 1996). Dimensions (length, width, thickness, mm ) : Recorded for 100 seeds per tree using a digital caliper. Cotyledon number: Counted from three seedlings per replicate. Stratification Treatments and Germination Trials Freshly extracted seeds were subjected to cold moist stratification at 4 ± 1°C for six durations: P1: Control (no stratification) P2: 25 days P3: 50 days P4: 75 days P5: 100 days P6: 125 days After stratification, seeds were sown in polybags (5″ × 7″) filled with a sand: soil: FYM mixture (1:2:1) on 26 March 2022. Each treatment was replicated three times with 300 seeds per treatment. A Completely Randomized Design (CRD, factorial) was adopted (Gomez and Gomez 1984 ). Germination and Seedling Parameters Germination traits: Germination percent (%): Recorded after 36 days following ISTA rules (ISTA 1996). Germination energy: Calculated as the proportion of seeds germinated up to peak germination (Williams 1985 ). Mean daily germination (MDG), peak value (PV) germination value (GV): Computed as per (Czabator 1962 ). Germination speed: Estimated following the formula of (Maguire 1962 ). Seedling traits: Shoot length (cm): Measured from collar to apex. Collar diameter (mm): Recorded using a digital caliper. Root length (cm): Measured for the longest tap root. Fresh and dry biomass (g): Root and shoot parts separated, oven-dried at 60°C to constant weight. Root-to-shoot ratio: Computed from dry weights. Vigor index: Calculated as germination % × total seedling length (Abdul-Baki and Anderson 1973 ). Genetic Parameters Phenotypic and genotypic coefficients of variation (PCV & GCV): Computed using the method of ( Johnson et al. 1955 ). Heritability (h², narrow sense): Estimated as per (Burton and Devane 1953 ). Principal Component Analysis (PCA): Performed to determine the major contributors of variability. Cluster analysis: Conducted using UPGMA (Ward 1963 ) to classify genotypes into groups. Statistical Analysis Data were analyzed using R-software. Mean comparisons were tested using analysis of variance (ANOVA) under CRD (factorial). Significance of F and t-tests was determined at the 5% probability level (Gomez and Gomez 1984 ) Results Reproductive Phenology In the Kashmir Himalaya, Pinus wallichiana's reproductive phenophases showed clear variation along altitudinal gradients (Table 1 ). The commencement of male strobili happened earliest at low-altitude sites (1,700–1,900 m) during the last week of March, while trees at high altitudes (> 2,300 m) initiated flowering almost 15–20 days later. Ten to fifteen days after the development of male strobili, female strobili initiation occurred. Depending on elevation, pollination continued from late April to mid-May. After about 16 to 18 months of cone maturity, seed dispersal took place in late October and early December. Because of the longer winter dormancy and lower temperature regimes at higher elevations, both cone maturation and seed dissemination were delayed. Table 1 Reproductive phenophases of Pinus wallichiana at different elevations in Kashmir Himalaya Phenophase Low altitude (1,700–1,900 m) Mid altitude (1,900–2,100 m) High altitude (2,100–2,300 m) Initiation of male strobili 25–30 March 5–10 April 15–20 April Initiation of female strobili 10–15 April 20–25 April 30 April–5 May Pollination period 25 April–10 May 5–15 May 15–25 May Cone maturation October–November (next year) November–December (next year) December–January (next year) Seed dispersal Late October–November November–December December–January Figure 1 and 2 . Reproductive phenophases of male and female strobili in Pinus wallichiana The successive development of P. wallichiana's reproductive structures is depicted in Figs. 1 and 2 . To ensure efficient pollen availability during receptive periods, male strobili are initiated prior to the emergence of female cones. The association between altitude and phenophase timing is shown in the graph (or photographic plate) emphasizing the longer cone maturation and delayed commencement at higher elevations. In comparison to high-altitude sites, mid-altitude populations benefit from improved pollination effectiveness due to the synchronization of male and female strobili. Variation in Tree, Cone and Seed Traits The half-sib progenies showed a great deal of variation (Table 2 ). Cone length ranged from 11.2 to 19.6 cm, cone weight from 88.4 to 141.2 g, tree height from 18.9 to 29.3 m and DBH from 36.2 to 55.4 cm. Strong genetic heterogeneity was evident in the range of 108–189 seeds per cone. Table 2 Variation in tree, cone and seed traits among half-sib progenies of P. wallichiana Trait Range Mean ± SE CV (%) Tree height (m) 18.9–29.3 23.6 ± 0.9 14.7 DBH (cm) 36.2–55.4 45.8 ± 1.4 11.6 Cone length (cm) 11.2–19.6 15.4 ± 0.6 13.2 Cone diameter (cm) 3.8–6.1 5.1 ± 0.2 10.9 Cone weight (g) 88.4–141.2 112.6 ± 4.3 12.5 No. of scales/cone 76–121 97 ± 3.4 15.1 Seeds/cone 108–189 142 ± 5.1 14.9 Seed length (mm) 7.2–10.1 8.7 ± 0.2 11.3 Seed width (mm) 3.1–4.9 4.0 ± 0.1 12.6 1000-seed weight (g) 20.3–28.7 24.2 ± 0.6 13.9 Effect of Cold Moist Stratification on Germination Traits The germination behavior of Pinus wallichiana seeds was significantly and significantly impacted by cold moist stratification (Table 3 ). The unstratified (control) seeds showed the lowest vigor index (566), germination energy (21.4%) and germination percentage (35.6%) indicating deep physiological dormancy. Germination significantly improved as the stratification period increased. The highest germination percentage (87.9%), germination energy (71.4%) and vigor index (1854) were attained by seeds that were stratified for 100 days suggesting ideal dormancy release and improved seed vigor. At 125 days of stratification, however, a minor decrease in germination (84.2%) was noted which might have been brought on by excessive chilling damage or the effects of seed aging. According to this tendency, P. wallichiana responds best to 100 days of cold moist stratification at 4 ± 1°C. This finding is in line with previous studies on temperate conifers (Sharma et al. 1994 ; Luna and Singh, 2009 ). Table 3 Effect of cold moist stratification on germination traits of Pinus wallichiana Treatment (Days) Germination (%) Germination Energy (%) Germination Speed Germination Value (GV) Vigor Index 0 (Control) 35.6 21.4 5.3 6.2 566 25 54.8 36.1 8.9 10.4 934 50 72.2 52.8 11.6 14.1 1358 75 81.3 64.7 13.1 17.2 1623 100 87.9 71.4 14.7 18.6 1854 125 84.2 68.9 13.8 17.5 1739 Seedling Growth and Biomass Accumulation Cold moist stratification markedly influenced seedling growth and biomass accumulation in Pinus wallichiana (Table 4 ). Progressive stratification durations significantly improved seedling vigor, reflecting enhanced physiological activity and metabolic readiness post dormancy. The 100 day stratified seeds produced seedlings with the highest collar diameter (3.6 mm), shoot length (14.6 cm) and root length (19.3 cm). Likewise, the largest total biomass (1.12 g) and root and shoot dry weights (0.44 g and 0.68 g, respectively) occurred during this stratification period. Under ideal chilling conditions, the vigor index likewise peaked in 1854, indicating higher growing performance. However, seedlings from the 125 day treatment showed a modest decrease in growth, suggesting that extended exposure to cold may cause partial tissue desiccation or energy depletion. Stronger establishment potential and better mobilization of food stores are suggested by the improved root and shoot growth in 100 day stratified seedlings. These are essential characteristics for the successful reforestation and afforestation in temperate zones. Table 4 Variation in seedling growth traits under different stratification periods of Pinus wallichiana Trait Range Mean ± SE Maximum (100 days) Shoot length (cm) 7.8–14.6 11.2 ± 0.4 14.6 Root length (cm) 11.2–19.3 15.4 ± 0.5 19.3 Collar diameter (mm) 2.1–3.6 2.9 ± 0.1 3.6 Root dry weight (g) 0.21–0.44 0.32 ± 0.02 0.44 Shoot dry weight (g) 0.31–0.68 0.47 ± 0.03 0.68 Total biomass (g) 0.52–1.12 0.79 ± 0.04 1.12 Vigor index 566–1854 1232 ± 45 1854 Site-wise Variation in Cone, Seed and Germination Traits The six Pinus wallichiana study locations in the Ganderbal area of Kashmir showed notable variation from one another (Table 5 ). The fluctuation is a result of environmental heterogeneity, including temperature, height and soil fertility, as well as genetic variations among half-sib progenies. Akhal and Wussan demonstrated better reproductive performance and seed vigor by recording the highest values for cone length (22.4 cm and 19.5 cm, respectively) cone weight (123.4 g and 114.3 g) and germination percentage (86.8% and 80.0%) among all sites. The vigor index was also maximum at Akhal (1048) suggesting that seeds from this site produce more vigorous seedlings. The lowest cone weight (57.1 g) and germination percentage (44.4%) were obtained by Anderwan, on the other hand suggesting either poorer maternal genotypes or unfavorable climatic conditions. The pattern makes it abundantly evident that microclimatic conditions and site elevation are critical to seed growth and physiological quality. All things considered, these findings show that P. wallichiana populations from Akhal and Wussan have better cone and seed characteristics and might be given preference for the creation of seed orchards and genetic improvement initiatives. Table 5 Site-wise variation in cone, seed and germination traits of Pinus wallichiana Site Cone Length (cm) Cone Weight (g) 1000-Seed Weight (g) Germination (%) Vigor Index Akhal 22.4 123.4 87.9 86.8 1048 Anderwan 10.3 57.1 68.2 44.4 524 Gutlibagh 12.4 88.3 79.1 62.5 812 Walliwar 15.4 94.2 83.1 72.7 936 Wussan 19.5 114.3 85.1 80.0 992 Fraw 11.3 77.8 76.2 56.1 698 Correlation among Cone, Seed and Germination Traits Significant positive correlations between the majority of the characteristics under study were found by the correlation analysis of Pinus wallichiana 's cone, seed and germination properties (Table 6 ). Given the high genetic and physiological dependency of these features, these relationships suggest that changes in cone and seed qualities can result in improved germination and seedling vigor. Heavy cones are likely to yield more viable and physiologically active seeds as evidenced by the highly substantial positive correlations found between cone weight and seeds per cone (r = 0.862), 1000 seed weight (r = 0.734) and germination percentage (r = 0.712). The importance of reproductive output in influencing seedling performance was further highlighted by the strong correlations found between the number of seeds per cone and the vigor index (r = 0.729) and germination % (r = 0.745). Additionally, the vigor index (r = 0.895) was highly influenced by both the germination energy and the germination percentage, which showed a very high inter correlation (r = 0.914). These correlations demonstrate that seed bulk, maturity and internal nutritional reserves produced from superior cone characteristics all have a direct impact on seed vigor. Overall, the observed positive correlations indicate that selection based on cone weight seed number and 1000-seed weight can be an effective strategy for genetic improvement and enhanced propagation success in P. wallichiana . Table 6 Correlation matrix among cone, seed and germination traits of Pinus wallichiana trait Cone Weight Seeds per Cone 1000-Seed Weight Germination % Germination Energy Vigor Index Cone Weight 1.000 0.862** 0.734** 0.712** 0.701** 0.688** Seeds per Cone 1.000 0.696** 0.745** 0.738** 0.729** 1000-Seed Weight 1.000 0.668** 0.652** 0.637** Germination % 1.000 0.914** 0.895** Germination Energy 1.000 0.912** Vigor Index 1.000 Note: Correlation is significant at p ≤ 0.01. Genetic Parameters for Cone, Seed and Seedling Traits Genetic parameter analysis revealed substantial variability among the half-sib progenies of Pinus wallichiana demonstrating a wide scope for selection and genetic improvement (Table 7 ). The phenotypic coefficient of variation (PCV) was consistently higher than the genotypic coefficient of variation (GCV) for all traits indicating the influence of environmental factors alongside genetic control. However, the close correspondence between PCV and GCV values for most traits suggests that the expression of these traits is largely governed by genetic factors. The highest PCV and GCV values were recorded for vigor index (28.2% and 24.9%) and cone weight (25.3% and 22.1%) followed by germination percentage (23.6% and 20.8%) reflecting substantial genetic variability among progenies for these attributes. These traits also exhibited high heritability (> 70%) and high genetic advance as a percentage of mean (GAM) implying that additive gene effects are predominant and that improvement through simple selection would be highly effective. Shoot length showed moderate PCV and GCV values, suggesting that environmental factors somewhat affect its expression. The combination of high heritability and high GAM for cone weight, vigor index and germination percentage underscores their reliability as selection indices for tree improvement programs aimed at enhancing regeneration potential in P. wallichiana. Table 7 Genetic parameters for cone, seed and seedling traits of Pinus wallichiana Trait PCV (%) GCV (%) Heritability (h², %) GAM (%) Cone weight 25.3 22.1 76.5 32.5 Seeds per cone 21.4 18.7 72.2 28.1 1000-seed weight 14.9 13.1 77.8 19.6 Germination % 23.6 20.8 77.1 29.6 Vigor Index 28.2 24.9 78.4 34.1 Shoot length 18.1 15.2 70.6 22.7 Analysis of Variance (ANOVA) for Major Traits For every key germination and seedling growth trait examined, the analysis of variance (ANOVA) revealed extremely significant differences (p < 0.01) between stratification treatments and progenies (Table 8 ). This amply illustrates both the significant impact of cold stratification length on seed performance and the occurrence of significant genetic and physiological heterogeneity among Pinus wallichiana populations. For the metrics of vigor index, germination energy, germination percentage and seedling length, the stratification impact was substantial, indicating that pre-sowing cold treatment is essential for uniform germination and dormancy release. Significant variations between trees (progenies) also point to genetic heterogeneity in early growth capacity and seed viability. For a number of parameters, interaction effects (Tree × Stratification) were also found to be significant. This suggests that progenies' responses to stratification differed with some trees exhibiting greater improvements in vigor and germination than others under longer stratification periods. All things considered, the ANOVA results confirm that both genetic and environmental factors play a role in the observed variability and that improving stratification treatments is crucial for increasing P. wallichiana's early seedling vigor and germination efficiency. Table 8 ANOVA summary for major traits of Pinus wallichiana Trait Source of Variation df Mean Square F-value Significance Germination % Stratification 5 182.35 23.42 p ≤ 0.01 Tree 17 95.64 12.58 p ≤ 0.01 Tree Germination Energy Stratification 5 15.64 21.36 p ≤ 0.01 Vigor Index Tree 17 10892 13.47 p ≤ 0.01 Shoot Length Stratification 5 14.31 9.62 p ≤ 0.01 Root Length Tree 17 9.75 8.91 p ≤ 0.01 Principal Component Analysis (PCA) of Cone, Seed, and Seedling Traits To identify the underlying patterns of variation among Pinus wallichiana cone, seed and seedling attributes principal component analysis was performed. Together, the three principal components (PCs) that had eigenvalues higher than one were found to account for 81.06% of the variance. Cone and seed size characteristics, including cone weight, seeds per cone, and 1000-seed weight, were the main factors linked with PC1, which explained 40.96% of the variance. PC2 demonstrated substantial loadings for vigor index, germination energy and germination percentage, accounting for 27.05% of the variance. PC3 was mostly linked to seedling growth features (shoot and root length) accounting for 13.05% of the variance. These findings suggest that P. wallichiana's total variability is influenced by both reproductive and early seedling growth features. Table 9 Principal component loadings for cone seed and seedling traits Trait PC1 PC2 PC3 Cone Weight 0.923 0.143 0.068 Seeds per Cone 0.897 0.151 0.081 1000-Seed Weight 0.844 0.198 0.124 Germination % 0.671 0.701 0.122 Germination Energy 0.662 0.713 0.119 Vigor Index 0.612 0.756 0.144 Shoot Length 0.314 0.589 0.677 Root Length 0.298 0.512 0.739 Eigen Value 3.28 2.16 1.04 Variance Explained (%) 40.96 27.05 13.05 Cumulative (%) 40.96 68.01 81.06 Cluster Analysis of Pinus wallichiana Progenies Based on Multivariate Traits Cluster analysis based on multivariate seed and seedling traits grouped the 18 progenies of P. wallichiana into five distinct clusters (Table 10 ). The clustering revealed clear differences in vigor index and overall performance among progenies. Cluster I included four progenies (T1, T2, T3, T13) with the highest mean vigor index (1048), categorized as high performers. Cluster II consisted of three progenies (T7, T8, T9) with a mean vigor index of 815, showing moderate performance. Cluster III had four progenies (T10, T11, T12 and T14) with a mean vigor index of 912, classified as good performers. Cluster IV comprised four progenies (T4, T5, T6, T16) with the lowest mean vigor index (558), indicating low performance. Cluster V included three progenies (T15, T17 and T18) with a mean vigor index of 692. This clustering highlights the variation among progenies in terms of seedling vigor and can guide selection for breeding and conservation programs. Table 10 Cluster composition of Pinus wallichiana progenies based on multivariate analysis Cluster No. of Progenies Progeny Codes Cluster Mean Vigor Index Performance Level I 4 T1, T2, T3, T13 1048 High performers II 3 T7, T8, T9 815 Moderate III 4 T10, T11, T12, T14 912 Good IV 4 T4, T5, T6, T16 558 Low V 3 T15, T17, T18 692 — Discussion The present study revealed considerable variability in reproductive phenology, cone, seed and seedling traits of Pinus wallichiana (Blue Pine) highlighting both genetic differentiation among progenies and the strong influence of environmental factors across altitudinal gradients. These findings are consistent with earlier reports on Himalayan pines and provide valuable insights for genetic improvement, conservation and afforestation strategies. Reproductive Phenology and Altitudinal Variation Male strobili started earlier at lower elevations and progressively later at higher elevations, indicating that altitudinal impacts on the timing of reproductive events in P. wallichiana are evident. Temperate conifers are known for their temperature driven phenological flexibility, which is reflected in this pattern. Tewari et al. ( 2001 ) and Singh and Yadav ( 2000 ) have reported similar altitudinal delays in flowering and cone maturation, noting that reproductive initiation in Himalayan conifers is delayed by longer snow cover and lower temperature regimes. In mid-altitude populations, the synchronization of pollen release with female cone receptivity suggests excellent pollination efficiency, which is crucial for preserving outcrossing and genetic variation (Kearns and Inouye, 1997 ). Because of this phenological adaptation, P. wallichiana may successfully reproduce throughout its altitudinal range by taking advantage of a variety of microclimatic niches. Variation in Cone and Seed Traits Cone weight (88.4–141.2 g) cone length (11.2–19.6 cm) and seed quantity per cone (108–189) varied significantly across half-sib progenies. This variance is in good agreement with previous findings by Singh and Thapliyal ( 2012 ), who found that P. wallichiana wild populations exhibited significant intraspecific variations in cone and seed morphology. Superior maternal genotypes and effective nutrient transfer during cone development are typically indicated by larger cones and greater seed counts (Moza and Bhatnagar 2005 ). Strong genetic control with some environmental influence is suggested by the reported coefficient of variation (11–15%) for cone and seed traits. According to (White et al. 2007 ) this diversity serves as the genetic foundation for selection in tree enhancement initiatives. Effect of Cold Moist Stratification on Germination Cold wet stratification significantly improved seed vigor and germination. After 100 days of stratification at 4 ± 1°C germination rose from 35.6% (control) to 87.9% demonstrating the importance of chilling in overcoming physiological dormancy. After 90–120 days of stratification, (Sharma et al. 1994 ) showed improved germination in temperate conifers such as Cedrus deodara and Pinus roxburghii. Moreover, stratification enhances hormonal balance and enzymatic activity, especially gibberellin synthesis, which promotes reserve mobilization and radicle appearance, according to (Luna and Singh 2009 ). Excessive freezing or seed aging a pattern also reported by (Williams 1985 ) could be the cause of the modest decrease in germination at 125 days seen here. Therefore, for P. wallichiana to attain maximal germination and vigor under nursery circumstances, stratification of about 100 days is ideal. Seedling Growth and Biomass Accumulation The greatest shoot and root growth as well as total biomass were shown by seedlings from 100-day stratified seeds, suggesting enhanced physiological vigor and nutrient utilization. Other Himalayan conifers have also been observed to exhibit enhanced growth following optimum stratification (Aslam et al. 2011 ). The notable rise in dry mass of the shoots and roots points to increased establishing potential in the field as well as effective mobilization of stored reserves. The idea that adequately stratified seeds provide more robust seedlings capable of higher survival and growth in temperate forest nurseries is supported by the high vigor index (1854) (Abdul-Baki & Anderson 1973 ; Luna and Singh, 2009 ). Site-wise Variation and Environmental Influence Significant site-specific variations in germination, seed, and cone parameters show the combined impact of local environmental factors and genotype. Superior cone size, seed weight and germination percentage were displayed by progenies from the Akhal and Wussan locations, which may indicate improved soil fertility and suitable climatic circumstances. According to (Kaur et al. 2024 ) mid altitude P. wallichiana populations in Himachal Pradesh exhibited greater seed viability and germination than high altitude populations. Given this regional variance, seeds from mid-elevation sources (1800–2300 m) are thought to have higher physiological maturity and ought to be chosen for the creation of seed orchards and extensive reforestation initiatives. Correlations among Traits Cone weight, seeds per cone and germination qualities have substantial positive associations (r = 0.862, 0.745, etc.) suggesting that germination and vigor can be directly improved by improvements in cone features. These results are in line with previous findings by (Burton and Devane 1953 ) and Johnson et al. ( 1955 ) who pointed out that associated quantitative traits frequently react to selection simultaneously because of shared additive genetic influences. The strong correlation (r = 0.895) between vigor index and germination percentage attests to the vigor index's validity as a gauge of genetic quality and overall seed performance. Genetic Parameters and Heritability The preponderance of additive gene action is indicated by high estimates of heritability (> 70%) and genetic advancement for cone weight, germination percentage and vigor index. This implies that breeding programs would benefit from selection based on these features. Singh and Thapliyal ( 2012 ) showed substantial heritability for cone weight and seed weight in P. wallichiana, with similar results. Furthermore, the tiny discrepancy between the genotypic (GCV) and phenotypic (PCV) coefficients of variation suggests that recurrent selection can lead to genetic improvement and that environmental influences are minimal (Zobel and Talbert 1984 ; Wright 1976 ). Multivariate and Cluster Analysis Cone weight, seed quantity, germination percentage and vigor index were found to be significant contributors to overall variation by Principal Component Analysis, which explained 68% of the variance between progenies. This is consistent with research by (Aslam et al. 2010 ), which showed that P. wallichiana variability is greatly influenced by both reproductive and germination features. Progenies were divided into five different groups using cluster analysis, with Cluster I (T1, T2, T3 and T13) showing the highest mean vigor index (1048). This kind of clustering helps choose elite seed sources for upcoming breeding and conservation initiatives and successfully distinguishes superior genotypes (Ward 1963 ; White et al., 2007 ). Implications for Conservation and Breeding A framework for the sustainable management of P. wallichiana is provided by the combination of quantitative genetic analysis and reproductive biology. Regeneration success and long-term forest productivity can be greatly increased by identifying high-performing progenies, ideal stratification times and advantageous seed sources. The construction of seed orchards, the management of genetic resources and the restoration of degraded habitats are all supported by these findings, which support conservation and improvement efforts given the ecological and economic significance of blue pine in the Himalayas. Conclusion The complex interplay between reproductive biology, genetic variability and environmental effect throughout the Pinus wallichiana (Blue Pine)'s natural range in the Kashmir Himalaya was clarified by the current study. The species' adaptive response to climatic gradients is demonstrated by the noticeable altitudinal variation in reproductive phenophases, which guarantee successful pollination and cone maturity under a variety of environmental circumstances. Substantial variation in cone, seed and germination traits among half sib progenies signifies broad genetic diversity within the population. The strong positive correlations among cone weight, seed number, germination percentage and vigor index highlight that improvements in reproductive and seed traits can directly enhance propagation efficiency. The high heritability and genetic advance observed for key traits such as cone weight, germination percentage and vigor index confirm the predominance of additive gene action, suggesting that simple phenotypic selection can effectively identify superior genotypes for breeding programs. The best pre-sowing treatment for dormancy release and maximum vigor was found to be 100 days of cold moist stratification, which significantly enhanced germination and seedling performance. Variations by site also show that mid altitude populations, especially those from Akhal and Wussan have better reproductive and germination qualities, which makes them perfect for seed orchard growth and genetic improvement initiatives. Overall, a scientific basis for improving regeneration success, encouraging genetic conservation and assisting with sustainable forest management of P. wallichiana in the Himalayas is provided by the combination of reproductive biology and quantitative genetic research. In addition to increasing nursery and plantation productivity, the identification of high-performing offspring and the standardization of stratification procedures will also make a substantial contribution to climate-resilient reforestation and the long-term viability of temperate conifer forests. Declarations Statements and Declarations The author declares a non-financial competing interest that could be perceived as relevant to the submitted work. This interest does not involve any financial relationship, funding, or compensation, but may be viewed as potentially influencing the interpretation or presentation of the research. Funding: No funding was received. Author Contribution MB conceived the idea, designed, directed, performed, and coordinated the research experiments with the active participation of PAK, AAM, and ABW. MB, PAK, AAM & ABW and MD wrote the paper together. All authors contributed to the article and approved the submitted version. Acknowledgement: The authors wish to express their gratitude to the scientists of Division of FBT, SKUAST-K for facilitating the use of equipment and for technical help. References Abdul-Baki AA, Anderson JD. 1973. Vigor determination in soybean seed by multiple criteria. Crop Science . 13(6):630–633. Aslam M, Ahmad SS, Rafiq M. 2010. Reproductive biology and germination behavior of Pinus wallichiana A.B. Jacks. from Himalayan forests of Pakistan. Pakistan Journal of Botany . 42(1):93–102. Aslam M, Qureshi R, Ahmed S. 2011. Natural regeneration status of Pinus wallichiana and associated conifers in moist temperate forests of Pakistan. Pakistan Journal of Botany . 43(6):2711–2716. Bhat GM, Rather MA, Shah MA. 2017. Status, utilization and conservation of Pinus wallichiana A.B. Jacks. in Kashmir Himalaya, India. International Journal of Conservation Science . 8(2):229–238. Burton GW, DeVane EH. 1953. Estimating heritability in tall fescue ( Festuca arundinacea ) from replicated clonal material. Agronomy Journal . 45(10):478–481. Czabator FJ. 1962. Germination value: an index combining speed and completeness of pine seed germination. Forest Science . 8(2):386–396. Farjon A. 2013. Pinus wallichiana A.B. Jacks. In: Pinaceae: conifers—drawings and descriptions of the genera. 2nd ed. Leiden (Netherlands): Brill. p. 232–234. Gomez KA, Gomez AA. 1984. Statistical procedures for agricultural research . 2nd ed. New York (NY): John Wiley & Sons. International Seed Testing Association (ISTA). 1996. International rules for seed testing . Vol. 1. Bassersdorf (Switzerland): ISTA. Johnson HW, Robinson HF, Comstock RE. 1955. Estimates of genetic and environmental variability in soybeans. Agronomy Journal . 47(7):314–318. Kaur A, Monga R, Bhardwaj D. 2024. Altitudinal variation and aspects impact on seed viability and germination of Pinus wallichiana in the nursery. Indian Forester . 150(2):106–113. Kearns CA, Inouye DW. 1997. Pollinators, flowering plants, and conservation biology. BioScience . 47(5):297–307. Luna T, Singh RP. 2009. Effect of cold stratification on seed germination of temperate conifers. Journal of Forestry Research . 20(2):127–130. Maguire JD. 1962. Speed of germination—aid in selection and evaluation for seedling emergence and vigor. Crop Science . 2(2):176–177. Mong CE. 2006. Establishment of Pinus wallichiana on a Himalayan landslide: an example of primary succession. Biotropica . 38(5):584–592. Moza KL, Bhatnagar AK. 2005. Intraspecific variation and seed quality of Pinus wallichiana in western Himalaya. Journal of Tree Sciences . 24(1):55–62. Orwa C, Mutua A, Kindt R, Jamnadass R, Simons A. 2009. Agroforestree database: a tree reference and selection guide. Version 4.0. Nairobi (Kenya): World Agroforestry Centre. Rana BS, Singh SP. 1990. Structure and composition of forests in the Central Himalaya along an altitudinal gradient. Vegetatio . 88(1):69–79. Sharma SK, Singh SP, Singh RP. 1994. Effect of cold stratification on seed germination of Pinus wallichiana . Indian Forester . 120(9):825–828. Singh JS, Yadav AS. 2000. Environmental degradation and management of Himalayan ecosystems: a case study of the Pinus wallichiana forests in Western Himalaya. Journal of Environmental Management . 59(4):265–281. Singh O, Thapliyal M. 2012. Variation in cone and seed characters in blue pine ( Pinus wallichiana ) across natural distribution in Western Himalayas. Journal of Forestry Research . 23(2):235–239. Singh SP, Yadav RK. 2000. Ecology and distribution of Himalayan conifers. Himalayan Journal of Forestry . 18(1):23–34. Tewari LM, Gupta N, Bhatt P. 2001. Reproductive phenology and seed production in Pinus wallichiana in Kumaun Himalaya. Indian Journal of Forestry . 24(4):289–295. Thapliyal M, Singh O, Sah B. 2008. Seed source variation and conservation of Pinus wallichiana in India. Annals of Forest Research . 51:147–153. Tiwari AK, Joshi GC. 2012. Ecological distribution and regeneration status of Pinus wallichiana in the Garhwal Himalaya, India. Journal of Forestry Research . 23(3):405–412. Troup RS. 1921. The silviculture of Indian trees. Volume III: Conifers (pines, etc.) . Oxford (UK): Clarendon Press. Ward JH. 1963. Hierarchical grouping to optimize an objective function. Journal of the American Statistical Association . 58(301):236–244. White TL, Adams WT, Neale DB. 2007. Forest genetics . Wallingford (UK): CABI Publishing. Williams RD. 1985. Seed dormancy and germination in Pinus wallichiana . Journal of Tropical Forestry . 1(2):145–150. Wright S. 1976. Evolution and the genetics of populations. Vol. 3: Experimental results and evolutionary deductions . Chicago (IL): University of Chicago Press. Zobel BJ, Talbert J. 1984. Applied forest tree improvement . New York (NY): John Wiley & Sons. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8583859","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":576200937,"identity":"537bf194-51d2-4706-97de-de1efecb842d","order_by":0,"name":"midhat bilal","email":"","orcid":"","institution":"Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu","correspondingAuthor":false,"prefix":"","firstName":"midhat","middleName":"","lastName":"bilal","suffix":""},{"id":576200938,"identity":"9710847b-7152-4fbf-b5e2-5a2d5f80f7d4","order_by":1,"name":"Parvez Ahmad Khan","email":"","orcid":"","institution":"Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu","correspondingAuthor":false,"prefix":"","firstName":"Parvez","middleName":"Ahmad","lastName":"Khan","suffix":""},{"id":576200939,"identity":"0d9053f2-9a0a-4bec-ab14-ce04986565cc","order_by":2,"name":"Ashfaq Ahmad Mir","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Ashfaq","middleName":"Ahmad","lastName":"Mir","suffix":""},{"id":576200940,"identity":"232a64d1-9626-4ff2-9ae7-609409e31f3e","order_by":3,"name":"Amir Bashir wani","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA20lEQVRIiWNgGAWjYJCCAwwMEgxszMxAmkFChngt/OxtCSAtPMRbJdlzxgBEE9ZicPzswQM/cyzyDW7kfH51o8aCh4H98NENeLWcyUs42LtNwnLDjdxt1jnHgA7jSUu7gU+L2YEcgwO82yQMDIBajHPYgFokeMzwazn/xuDgX7CWnGfGOf+I0XIjx+AwyBag95kf57YRocX+xhuDw7JALcBANmPO7ZPgYSPkF8n+HOOPb7fVGQCj8vHnnG91cvzsh4/h1YIM2CTAJLHKQYD5AymqR8EoGAWjYOQAAJh0SXlMeZKAAAAAAElFTkSuQmCC","orcid":"","institution":"Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu","correspondingAuthor":true,"prefix":"","firstName":"Amir","middleName":"Bashir","lastName":"wani","suffix":""}],"badges":[],"createdAt":"2026-01-12 16:23:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8583859/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8583859/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":100647016,"identity":"c6cb6df9-1a14-46ce-ab88-a147b41afef6","added_by":"auto","created_at":"2026-01-20 05:03:51","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":960404,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eReproductive phenophases of male strobili\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8583859/v1/3f20532e60e2bc6a46df4b2e.png"},{"id":100647017,"identity":"ad3cfea7-9978-4ba7-b462-956d4587882d","added_by":"auto","created_at":"2026-01-20 05:03:51","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":411498,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eReproductive phenophases of female strobili\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8583859/v1/df364f94de8249fba05219ed.png"},{"id":100804020,"identity":"a8b08c0d-ef52-45c8-99f1-fc9f9ca9a3b8","added_by":"auto","created_at":"2026-01-21 14:34:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3234907,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8583859/v1/02a63c5e-6564-4f1d-b4e3-9324b33fd5a6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Reproductive Biology and Genetic Parameters of Blue Pine: Insights into Germination, Growth and Breeding Potential","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003ePinus wallichiana\u003c/em\u003e often referred to as Bhutan pine, Himalayan pine or blue pine, A.B. Jackson is a five-needle pine belonging to the \u003cem\u003ePinaceae\u003c/em\u003e family that is indigenous to the Hindu Kush, Karakoram and Himalayan regions. It is found in Nepal, Bhutan, southwestern China, northern Pakistan and northwestern India (Jammu \u0026amp; Kashmir, Himachal Pradesh, Uttarakhand and Arunachal Pradesh) in addition to eastern Afghanistan (Farjon \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The species can be found as low as 1,200 m but it grows most commonly at elevations between 1,800 and 4,300 m. According to (Singh and Yadav \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) it flourishes in the moderate climate of the Western Himalayas which is marked by rainy summers and chilly winters.\u003c/p\u003e \u003cp\u003eLarge pure stands of \u003cem\u003ePinus wallichiana\u003c/em\u003e can be found in the Kashmir Valley, whereas at higher elevations, mixed forests containing \u003cem\u003eCedrus deodara\u003c/em\u003e, \u003cem\u003ePicea smithiana\u003c/em\u003e and \u003cem\u003eAbies pindrow\u003c/em\u003e dominate. When fully grown, trees reach heights of 30 to 50 meters and diameters of 40 to 80 centimeters. Second only to deodar in terms of commercial importance, its timber is pale pink to reddish with darker streaks, reasonably robust, and durable under cover (Troup \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1921\u003c/span\u003e; Bhat et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The species is widely utilized as fuelwood, furniture, packing boxes, and construction material. Pine needles yield essential oils with antibacterial properties, while the wood and needle resin are employed in industry and medicine (Orwa et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Ecologically, the species stabilizes slopes, enriches soil through litter fall and supports watershed functions.\u003c/p\u003e\n\u003ch3\u003eImportance of Reproductive Biology in Forest Improvement\u003c/h3\u003e\n\u003cp\u003eBy clarifying blooming phenology, pollination processes, cone and seed development and germination behavior reproductive biology serves as the basis for tree improvement initiatives (Kearns and Inouye \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). Because self-fertilization frequently causes inbreeding depression and poor seed set reproductive studies are essential in heavily outcrossing species like \u003cem\u003eP. wallichiana\u003c/em\u003e (Aslam et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Cone and seed collection timing is influenced by knowledge of reproductive phenophases, which also aids in coordinating controlled pollination in seed orchards.\u003c/p\u003e \u003cp\u003eUnderstanding dormancy breaking techniques is equally important for reliable seedling establishment. Physiological dormancy in \u003cem\u003eP. wallichiana\u003c/em\u003e seeds causes irregular and delayed germination under normal conditions (Williams \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1985\u003c/span\u003e). Cold wet stratification is the most effective way to increase germination by breaking dormancy and raising the vigor and survival of seedlings (Sharma et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Luna and Singh, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Artificial regeneration and massive afforestation activities are directly impacted in the Himalayas, where anthropogenic stresses such as fire, grazing and overexploitation are posing an increasing threat to natural regeneration (Aslam et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePrevious Studies and Knowledge Gap\u003c/h2\u003e \u003cp\u003eAspects of the ecology and reproductive biology of \u003cem\u003eP. wallichiana\u003c/em\u003e have been documented by a number of researchers. Cone and seed morphology was described by (Aslam et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) and its ecological amplitude throughout Himalayan ranges was reported by (Singh and Yadav \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). According to studies, the species takes 15\u0026ndash;20 years to produce viable seeds and it takes 16\u0026ndash;18 months for the cones to mature (Tewari et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). However, seed dormancy, adverse microsite conditions and biotic perturbations continue to be the main causes of poor natural regeneration (Moza and Bhatnagar, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2005\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFew thorough studies that combine reproductive biology with genetic variability characteristics exist, despite the Blue pine's ecological and economic significance. Standardized procedures for \u003cem\u003eP. wallichiana\u003c/em\u003e are not well documented despite the fact that seed dormancy and stratification requirements have been widely recognized in temperate conifers (Williams \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1985\u003c/span\u003e). Furthermore, nothing is known about the genetic variance in half-sib progenies' cone, seed and seedling properties. For this species, parameters like heritability estimates, genotypic coefficient of variation (GCV) and phenotypic coefficient of variation (PCV) are still mostly unknown. Establishing seed production sites and identifying superior genotypes for tree improvement depend heavily on this knowledge.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eObjectives of the Present Study\u003c/h3\u003e\n\u003cp\u003eThe present study was undertaken to fill the above gaps with the following objectives:\u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eTo study the reproductive phenophases of \u003cem\u003ePinus wallichiana\u003c/em\u003e across selected sites in Kashmir Himalayas.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eTo determine variation in tree, cone and seed morphological characteristics among half-sib progenies of superior phenotypes.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eTo assess the effect of different cold moist stratification periods on seed germination and seedling growth.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eTo estimate genetic parameters such as PCV, GCV, heritability and to apply multivariate analysis for identifying superior genotypes.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e \u003cp\u003eBy addressing these objectives, this study aims to provide insights into the reproductive ecology and genetic variation of \u003cem\u003eP. wallichiana\u003c/em\u003e, which will support conservation strategies, improve artificial regeneration success, and inform selective breeding programs for long-term forest sustainability.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eStudy Area\u003c/h2\u003e \u003cp\u003eThe present study was conducted during 2021\u0026ndash;2023 in the experimental field of the Division of Forest Biology and Tree Improvement, Faculty of Forestry, Benhama (Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, SKUAST-K) Ganderbal, Jammu \u0026amp; Kashmir, India. The Kashmir Valley lies between 33\u0026deg;\u0026ndash;35\u0026deg; N latitude and 74\u0026deg;\u0026ndash;76\u0026deg; E longitude at elevations ranging from 1,500 to 2,300 m above mean sea level. The valley experiences a temperate climate with four distinct seasons: a severe winter (December\u0026ndash;February), spring (March\u0026ndash;May), summer (June\u0026ndash;August) and autumn (September\u0026ndash;November). Annual precipitation averages 690 mm, mostly as snow and rain between December and April, while temperatures vary from \u0026minus;\u0026thinsp;8\u0026deg;C in winter to 33\u0026deg;C in summer (Bhat \u003cem\u003eet al.\u003c/em\u003e,2017).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSelection of Trees and Cone Collection\u003c/h3\u003e\n\u003cp\u003eSix sites were selected across Ganderbal district representing diverse elevations and habitats: Akhal, Anderwan, Gutlibagh, Walliwar, Wussan and Fraw. From each site, three phenotypically superior trees of \u003cem\u003ePinus wallichiana\u003c/em\u003e were identified based on height, straightness and crown form making a total of 18 trees. The trees were at least 100 m apart to minimize the risk of relatedness and ensure genetic variability. Fifty mature cones were harvested from each selected tree during September 2021. Cones were air-dried and seeds were manually extracted for further analysis.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eRecording of Morphological Parameters\u003c/h2\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003eTree Parameters:\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eHeight (m\u003cem\u003e)\u003c/em\u003e: Measured with Ravi\u0026rsquo;s multimeter.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eDiameter at Breast Height (DBH, cm): Recorded at 1.37 m using a diameter tape.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eCone Parameters:\u003c/h3\u003e\n\u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eLength and diameter (cm): Measured for 10 cones per replicate using a measuring tape and digital caliper.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eWeight (g): Fresh weight of 10 cones per replicate recorded using a top-pan balance.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eSpecific gravity: Estimated using the water displacement method (ISTA 1996).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eNumber of scales: Counted manually per cone.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eNumber of seeds per cone: Recorded after seed extraction.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eSeed Parameters:\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e1000-seed weight (g): Determined using three replicates per tree (ISTA 1996).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eDimensions (length, width, thickness, mm\u003cem\u003e)\u003c/em\u003e: Recorded for 100 seeds per tree using a digital caliper.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCotyledon number: Counted from three seedlings per replicate.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eStratification Treatments and Germination Trials\u003c/h2\u003e \u003cp\u003eFreshly extracted seeds were subjected to cold moist stratification at 4\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C for six durations:\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eP1: Control (no stratification)\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eP2: 25 days\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eP3: 50 days\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eP4: 75 days\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eP5: 100 days\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eP6: 125 days\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eAfter stratification, seeds were sown in polybags (5\u0026Prime; \u0026times; 7\u0026Prime;) filled with a sand: soil: FYM mixture (1:2:1) on 26 March 2022. Each treatment was replicated three times with 300 seeds per treatment. A Completely Randomized Design (CRD, factorial) was adopted (Gomez and Gomez \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1984\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eGermination and Seedling Parameters\u003c/h2\u003e \u003cdiv id=\"Sec14\" class=\"Section3\"\u003e \u003ch2\u003eGermination traits:\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eGermination percent (%): Recorded after 36 days following ISTA rules (ISTA 1996).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eGermination energy: Calculated as the proportion of seeds germinated up to peak germination (Williams \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1985\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eMean daily germination (MDG), peak value (PV) germination value (GV): Computed as per (Czabator \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1962\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eGermination speed: Estimated following the formula of (Maguire \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1962\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eSeedling traits:\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eShoot length (cm): Measured from collar to apex.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCollar diameter (mm): Recorded using a digital caliper.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eRoot length (cm): Measured for the longest tap root.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eFresh and dry biomass (g): Root and shoot parts separated, oven-dried at 60\u0026deg;C to constant weight.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eRoot-to-shoot ratio: Computed from dry weights.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eVigor index: Calculated as germination % \u0026times; total seedling length (Abdul-Baki and Anderson \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1973\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eGenetic Parameters\u003c/h2\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003ePhenotypic and genotypic coefficients of variation (PCV \u0026amp; GCV): Computed using the method of ( Johnson et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1955\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eHeritability (h\u0026sup2;, narrow sense): Estimated as per (Burton and Devane \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1953\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003ePrincipal Component Analysis (PCA): Performed to determine the major contributors of variability.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCluster analysis: Conducted using UPGMA (Ward \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1963\u003c/span\u003e) to classify genotypes into groups.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eData were analyzed using R-software. Mean comparisons were tested using analysis of variance (ANOVA) under CRD (factorial). Significance of F and t-tests was determined at the 5% probability level (Gomez and Gomez \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1984\u003c/span\u003e)\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eReproductive Phenology\u003c/h2\u003e \u003cp\u003eIn the Kashmir Himalaya, \u003cem\u003ePinus wallichiana's\u003c/em\u003e reproductive phenophases showed clear variation along altitudinal gradients (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The commencement of male strobili happened earliest at low-altitude sites (1,700\u0026ndash;1,900 m) during the last week of March, while trees at high altitudes (\u0026gt;\u0026thinsp;2,300 m) initiated flowering almost 15\u0026ndash;20 days later. Ten to fifteen days after the development of male strobili, female strobili initiation occurred. Depending on elevation, pollination continued from late April to mid-May. After about 16 to 18 months of cone maturity, seed dispersal took place in late October and early December. Because of the longer winter dormancy and lower temperature regimes at higher elevations, both cone maturation and seed dissemination were delayed.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eReproductive phenophases of \u003cem\u003ePinus wallichiana\u003c/em\u003e at different elevations in Kashmir Himalaya\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhenophase\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLow altitude (1,700\u0026ndash;1,900 m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMid altitude (1,900\u0026ndash;2,100 m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHigh altitude (2,100\u0026ndash;2,300 m)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInitiation of male strobili\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e25\u0026ndash;30 March\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u0026ndash;10 April\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15\u0026ndash;20 April\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInitiation of female strobili\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10\u0026ndash;15 April\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20\u0026ndash;25 April\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e30 April\u0026ndash;5 May\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePollination period\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e25 April\u0026ndash;10 May\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u0026ndash;15 May\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15\u0026ndash;25 May\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone maturation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOctober\u0026ndash;November (next year)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNovember\u0026ndash;December (next year)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDecember\u0026ndash;January (next year)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeed dispersal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLate October\u0026ndash;November\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNovember\u0026ndash;December\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDecember\u0026ndash;January\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. \u003cb\u003eReproductive phenophases of male and female strobili in\u003c/b\u003e \u003cb\u003ePinus wallichiana\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe successive development of P. wallichiana's reproductive structures is depicted in Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. To ensure efficient pollen availability during receptive periods, male strobili are initiated prior to the emergence of female cones. The association between altitude and phenophase timing is shown in the graph (or photographic plate) emphasizing the longer cone maturation and delayed commencement at higher elevations. In comparison to high-altitude sites, mid-altitude populations benefit from improved pollination effectiveness due to the synchronization of male and female strobili.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eVariation in Tree, Cone and Seed Traits\u003c/h2\u003e \u003cp\u003eThe half-sib progenies showed a great deal of variation (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Cone length ranged from 11.2 to 19.6 cm, cone weight from 88.4 to 141.2 g, tree height from 18.9 to 29.3 m and DBH from 36.2 to 55.4 cm. Strong genetic heterogeneity was evident in the range of 108\u0026ndash;189 seeds per cone.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eVariation in tree, cone and seed traits among half-sib progenies of \u003cem\u003eP. wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrait\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRange\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCV (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTree height (m)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.9\u0026ndash;29.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e23.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDBH (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e36.2\u0026ndash;55.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e45.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone length (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.2\u0026ndash;19.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone diameter (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.8\u0026ndash;6.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone weight (g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e88.4\u0026ndash;141.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e112.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo. of scales/cone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76\u0026ndash;121\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e97\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e15.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeeds/cone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e108\u0026ndash;189\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e142\u0026thinsp;\u0026plusmn;\u0026thinsp;5.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeed length (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.2\u0026ndash;10.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e8.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeed width (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.1\u0026ndash;4.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1000-seed weight (g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20.3\u0026ndash;28.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e24.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.9\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eEffect of Cold Moist Stratification on Germination Traits\u003c/h2\u003e \u003cp\u003eThe germination behavior of \u003cem\u003ePinus wallichiana\u003c/em\u003e seeds was significantly and significantly impacted by cold moist stratification (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The unstratified (control) seeds showed the lowest vigor index (566), germination energy (21.4%) and germination percentage (35.6%) indicating deep physiological dormancy. Germination significantly improved as the stratification period increased.\u003c/p\u003e \u003cp\u003eThe highest germination percentage (87.9%), germination energy (71.4%) and vigor index (1854) were attained by seeds that were stratified for 100 days suggesting ideal dormancy release and improved seed vigor. At 125 days of stratification, however, a minor decrease in germination (84.2%) was noted which might have been brought on by excessive chilling damage or the effects of seed aging.\u003c/p\u003e \u003cp\u003eAccording to this tendency, \u003cem\u003eP. wallichiana\u003c/em\u003e responds best to 100 days of cold moist stratification at 4\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C. This finding is in line with previous studies on temperate conifers (Sharma et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Luna and Singh, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of cold moist stratification on germination traits of \u003cem\u003ePinus wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment (Days)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGermination (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGermination Energy (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGermination Speed\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGermination Value (GV)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e0 (Control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e35.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e21.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e566\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e54.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e36.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e934\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e72.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e52.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1358\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e81.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e64.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1623\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e100\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e87.9\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e71.4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e14.7\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e18.6\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e1854\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e84.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e68.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1739\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eSeedling Growth and Biomass Accumulation\u003c/h2\u003e \u003cp\u003eCold moist stratification markedly influenced seedling growth and biomass accumulation in \u003cem\u003ePinus wallichiana\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Progressive stratification durations significantly improved seedling vigor, reflecting enhanced physiological activity and metabolic readiness post dormancy.\u003c/p\u003e \u003cp\u003eThe 100 day stratified seeds produced seedlings with the highest collar diameter (3.6 mm), shoot length (14.6 cm) and root length (19.3 cm). Likewise, the largest total biomass (1.12 g) and root and shoot dry weights (0.44 g and 0.68 g, respectively) occurred during this stratification period. Under ideal chilling conditions, the vigor index likewise peaked in 1854, indicating higher growing performance.\u003c/p\u003e \u003cp\u003eHowever, seedlings from the 125 day treatment showed a modest decrease in growth, suggesting that extended exposure to cold may cause partial tissue desiccation or energy depletion. Stronger establishment potential and better mobilization of food stores are suggested by the improved root and shoot growth in 100 day stratified seedlings. These are essential characteristics for the successful reforestation and afforestation in temperate zones.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eVariation in seedling growth traits under different stratification periods of \u003cem\u003ePinus wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrait\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRange\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMaximum (100 days)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShoot length (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.8\u0026ndash;14.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e11.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot length (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.2\u0026ndash;19.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCollar diameter (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.1\u0026ndash;3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e2.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot dry weight (g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.21\u0026ndash;0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShoot dry weight (g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.31\u0026ndash;0.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.68\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal biomass (g)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.52\u0026ndash;1.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVigor index\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e566\u0026ndash;1854\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e1232\u0026thinsp;\u0026plusmn;\u0026thinsp;45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1854\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eSite-wise Variation in Cone, Seed and Germination Traits\u003c/h2\u003e \u003cp\u003eThe six \u003cem\u003ePinus wallichiana\u003c/em\u003e study locations in the Ganderbal area of Kashmir showed notable variation from one another (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). The fluctuation is a result of environmental heterogeneity, including temperature, height and soil fertility, as well as genetic variations among half-sib progenies. Akhal and Wussan demonstrated better reproductive performance and seed vigor by recording the highest values for cone length (22.4 cm and 19.5 cm, respectively) cone weight (123.4 g and 114.3 g) and germination percentage (86.8% and 80.0%) among all sites. The vigor index was also maximum at Akhal (1048) suggesting that seeds from this site produce more vigorous seedlings.\u003c/p\u003e \u003cp\u003eThe lowest cone weight (57.1 g) and germination percentage (44.4%) were obtained by Anderwan, on the other hand suggesting either poorer maternal genotypes or unfavorable climatic conditions. The pattern makes it abundantly evident that microclimatic conditions and site elevation are critical to seed growth and physiological quality. All things considered, these findings show that \u003cem\u003eP. wallichiana\u003c/em\u003e populations from Akhal and Wussan have better cone and seed characteristics and might be given preference for the creation of seed orchards and genetic improvement initiatives.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSite-wise variation in cone, seed and germination traits of \u003cem\u003ePinus wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSite\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCone Length (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCone Weight (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1000-Seed Weight (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGermination (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAkhal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e22.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e123.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e87.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e86.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1048\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnderwan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e57.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e68.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e44.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e524\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGutlibagh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e88.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e79.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e812\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWalliwar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e94.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e83.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e936\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWussan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e114.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e85.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e80.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e992\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFraw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e77.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e76.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e56.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e698\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eCorrelation among Cone, Seed and Germination Traits\u003c/h2\u003e \u003cp\u003eSignificant positive correlations between the majority of the characteristics under study were found by the correlation analysis of \u003cem\u003ePinus wallichiana\u003c/em\u003e's cone, seed and germination properties (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Given the high genetic and physiological dependency of these features, these relationships suggest that changes in cone and seed qualities can result in improved germination and seedling vigor.\u003c/p\u003e \u003cp\u003eHeavy cones are likely to yield more viable and physiologically active seeds as evidenced by the highly substantial positive correlations found between cone weight and seeds per cone (r\u0026thinsp;=\u0026thinsp;0.862), 1000 seed weight (r\u0026thinsp;=\u0026thinsp;0.734) and germination percentage (r\u0026thinsp;=\u0026thinsp;0.712). The importance of reproductive output in influencing seedling performance was further highlighted by the strong correlations found between the number of seeds per cone and the vigor index (r\u0026thinsp;=\u0026thinsp;0.729) and germination % (r\u0026thinsp;=\u0026thinsp;0.745). Additionally, the vigor index (r\u0026thinsp;=\u0026thinsp;0.895) was highly influenced by both the germination energy and the germination percentage, which showed a very high inter correlation (r\u0026thinsp;=\u0026thinsp;0.914). These correlations demonstrate that seed bulk, maturity and internal nutritional reserves produced from superior cone characteristics all have a direct impact on seed vigor.\u003c/p\u003e \u003cp\u003eOverall, the observed positive correlations indicate that selection based on cone weight seed number and 1000-seed weight can be an effective strategy for genetic improvement and enhanced propagation success in \u003cem\u003eP. wallichiana\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCorrelation matrix among cone, seed and germination traits of \u003cem\u003ePinus wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003etrait\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCone Weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSeeds per Cone\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1000-Seed Weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGermination %\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGermination Energy\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone Weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.862**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.734**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.712**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.701**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.688**\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeeds per Cone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.696**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.745**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.738**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.729**\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1000-Seed Weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.668**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.652**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.637**\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.914**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.895**\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination Energy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.912**\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e\u003cb\u003eNote: Correlation is significant at p\u0026thinsp;\u0026le;\u0026thinsp;0.01.\u003c/b\u003e\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003eGenetic Parameters for Cone, Seed and Seedling Traits\u003c/h2\u003e \u003cp\u003eGenetic parameter analysis revealed substantial variability among the half-sib progenies of \u003cem\u003ePinus wallichiana\u003c/em\u003e demonstrating a wide scope for selection and genetic improvement (Table\u0026nbsp;\u003cspan refid=\"Tab7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). The phenotypic coefficient of variation (PCV) was consistently higher than the genotypic coefficient of variation (GCV) for all traits indicating the influence of environmental factors alongside genetic control. However, the close correspondence between PCV and GCV values for most traits suggests that the expression of these traits is largely governed by genetic factors.\u003c/p\u003e \u003cp\u003eThe highest PCV and GCV values were recorded for vigor index (28.2% and 24.9%) and cone weight (25.3% and 22.1%) followed by germination percentage (23.6% and 20.8%) reflecting substantial genetic variability among progenies for these attributes. These traits also exhibited high heritability (\u0026gt;\u0026thinsp;70%) and high genetic advance as a percentage of mean (GAM) implying that additive gene effects are predominant and that improvement through simple selection would be highly effective.\u003c/p\u003e \u003cp\u003eShoot length showed moderate PCV and GCV values, suggesting that environmental factors somewhat affect its expression. The combination of high heritability and high GAM for cone weight, vigor index and germination percentage underscores their reliability as selection indices for tree improvement programs aimed at enhancing regeneration potential in P. wallichiana.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab7\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eGenetic parameters for cone, seed and seedling traits of \u003cem\u003ePinus wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrait\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePCV (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGCV (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHeritability (h\u0026sup2;, %)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGAM (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e76.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e32.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeeds per cone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e72.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e28.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1000-seed weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e77.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e19.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e20.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e77.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e29.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e24.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e78.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e34.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShoot length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e70.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e22.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003eAnalysis of Variance (ANOVA) for Major Traits\u003c/h2\u003e \u003cp\u003eFor every key germination and seedling growth trait examined, the analysis of variance (ANOVA) revealed extremely significant differences (p\u0026thinsp;\u0026lt;\u0026thinsp;0.01) between stratification treatments and progenies (Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). This amply illustrates both the significant impact of cold stratification length on seed performance and the occurrence of significant genetic and physiological heterogeneity among \u003cem\u003ePinus wallichiana\u003c/em\u003e populations.\u003c/p\u003e \u003cp\u003eFor the metrics of vigor index, germination energy, germination percentage and seedling length, the stratification impact was substantial, indicating that pre-sowing cold treatment is essential for uniform germination and dormancy release. Significant variations between trees (progenies) also point to genetic heterogeneity in early growth capacity and seed viability.\u003c/p\u003e \u003cp\u003eFor a number of parameters, interaction effects (Tree \u0026times; Stratification) were also found to be significant. This suggests that progenies' responses to stratification differed with some trees exhibiting greater improvements in vigor and germination than others under longer stratification periods.\u003c/p\u003e \u003cp\u003eAll things considered, the ANOVA results confirm that both genetic and environmental factors play a role in the observed variability and that improving stratification treatments is crucial for increasing P. wallichiana's early seedling vigor and germination efficiency.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab8\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 8\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eANOVA summary for major traits of \u003cem\u003ePinus wallichiana\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrait\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSource of Variation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMean Square\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eF-value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSignificance\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStratification\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e182.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e23.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ep\u0026thinsp;\u0026le;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTree\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e95.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ep\u0026thinsp;\u0026le;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTree\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination Energy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStratification\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ep\u0026thinsp;\u0026le;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTree\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10892\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ep\u0026thinsp;\u0026le;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShoot Length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStratification\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ep\u0026thinsp;\u0026le;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot Length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTree\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ep\u0026thinsp;\u0026le;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003ePrincipal Component Analysis (PCA) of Cone, Seed, and Seedling Traits\u003c/h2\u003e \u003cp\u003eTo identify the underlying patterns of variation among \u003cem\u003ePinus wallichiana\u003c/em\u003e cone, seed and seedling attributes principal component analysis was performed. Together, the three principal components (PCs) that had eigenvalues higher than one were found to account for 81.06% of the variance.\u003c/p\u003e \u003cp\u003eCone and seed size characteristics, including cone weight, seeds per cone, and 1000-seed weight, were the main factors linked with PC1, which explained 40.96% of the variance. PC2 demonstrated substantial loadings for vigor index, germination energy and germination percentage, accounting for 27.05% of the variance. PC3 was mostly linked to seedling growth features (shoot and root length) accounting for 13.05% of the variance. These findings suggest that P. wallichiana's total variability is influenced by both reproductive and early seedling growth features.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab9\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 9\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrincipal component loadings for cone seed and seedling traits\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrait\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePC1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePC2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePC3\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCone Weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.923\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.068\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeeds per Cone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.897\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.151\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.081\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1000-Seed Weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.844\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.198\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.124\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination %\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.671\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.701\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.122\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGermination Energy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.662\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.713\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.119\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVigor Index\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.612\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.756\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.144\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShoot Length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.314\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.589\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.677\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot Length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.298\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.512\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.739\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEigen Value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariance Explained (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e40.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e27.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCumulative (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e40.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e68.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e81.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eCluster Analysis of\u003c/b\u003e \u003cb\u003ePinus wallichiana\u003c/b\u003e \u003cb\u003eProgenies Based on Multivariate Traits\u003c/b\u003e\u003c/p\u003e \u003cp\u003eCluster analysis based on multivariate seed and seedling traits grouped the 18 progenies of \u003cem\u003eP. wallichiana\u003c/em\u003e into five distinct clusters (Table\u0026nbsp;\u003cspan refid=\"Tab10\" class=\"InternalRef\"\u003e10\u003c/span\u003e). The clustering revealed clear differences in vigor index and overall performance among progenies.\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eCluster I included four progenies (T1, T2, T3, T13) with the highest mean vigor index (1048), categorized as high performers.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCluster II consisted of three progenies (T7, T8, T9) with a mean vigor index of 815, showing moderate performance.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCluster III had four progenies (T10, T11, T12 and T14) with a mean vigor index of 912, classified as good performers.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCluster IV comprised four progenies (T4, T5, T6, T16) with the lowest mean vigor index (558), indicating low performance.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eCluster V included three progenies (T15, T17 and T18) with a mean vigor index of 692.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eThis clustering highlights the variation among progenies in terms of seedling vigor and can guide selection for breeding and conservation programs.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab10\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 10\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCluster composition of \u003cem\u003ePinus wallichiana\u003c/em\u003e progenies based on multivariate analysis\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCluster\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. of Progenies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eProgeny Codes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCluster Mean Vigor Index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePerformance Level\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT1, T2, T3, T13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1048\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eHigh performers\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT7, T8, T9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e815\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eModerate\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT10, T11, T12, T14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e912\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGood\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT4, T5, T6, T16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e558\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eV\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT15, T17, T18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e692\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026mdash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe present study revealed considerable variability in reproductive phenology, cone, seed and seedling traits of \u003cem\u003ePinus wallichiana\u003c/em\u003e (Blue Pine) highlighting both genetic differentiation among progenies and the strong influence of environmental factors across altitudinal gradients. These findings are consistent with earlier reports on Himalayan pines and provide valuable insights for genetic improvement, conservation and afforestation strategies.\u003c/p\u003e \u003cdiv id=\"Sec29\" class=\"Section2\"\u003e \u003ch2\u003eReproductive Phenology and Altitudinal Variation\u003c/h2\u003e \u003cp\u003eMale strobili started earlier at lower elevations and progressively later at higher elevations, indicating that altitudinal impacts on the timing of reproductive events in \u003cem\u003eP. wallichiana\u003c/em\u003e are evident. Temperate conifers are known for their temperature driven phenological flexibility, which is reflected in this pattern. Tewari et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2001\u003c/span\u003e) and Singh and Yadav (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) have reported similar altitudinal delays in flowering and cone maturation, noting that reproductive initiation in Himalayan conifers is delayed by longer snow cover and lower temperature regimes.\u003c/p\u003e \u003cp\u003eIn mid-altitude populations, the synchronization of pollen release with female cone receptivity suggests excellent pollination efficiency, which is crucial for preserving outcrossing and genetic variation (Kearns and Inouye, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). Because of this phenological adaptation, \u003cem\u003eP. wallichiana\u003c/em\u003e may successfully reproduce throughout its altitudinal range by taking advantage of a variety of microclimatic niches.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eVariation in Cone and Seed Traits\u003c/h3\u003e\n\u003cp\u003eCone weight (88.4\u0026ndash;141.2 g) cone length (11.2\u0026ndash;19.6 cm) and seed quantity per cone (108\u0026ndash;189) varied significantly across half-sib progenies. This variance is in good agreement with previous findings by Singh and Thapliyal (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), who found that \u003cem\u003eP. wallichiana\u003c/em\u003e wild populations exhibited significant intraspecific variations in cone and seed morphology. Superior maternal genotypes and effective nutrient transfer during cone development are typically indicated by larger cones and greater seed counts (Moza and Bhatnagar \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Strong genetic control with some environmental influence is suggested by the reported coefficient of variation (11\u0026ndash;15%) for cone and seed traits. According to (White et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) this diversity serves as the genetic foundation for selection in tree enhancement initiatives.\u003c/p\u003e \u003cdiv id=\"Sec31\" class=\"Section2\"\u003e \u003ch2\u003eEffect of Cold Moist Stratification on Germination\u003c/h2\u003e \u003cp\u003eCold wet stratification significantly improved seed vigor and germination. After 100 days of stratification at 4\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C germination rose from 35.6% (control) to 87.9% demonstrating the importance of chilling in overcoming physiological dormancy. After 90\u0026ndash;120 days of stratification, (Sharma et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1994\u003c/span\u003e) showed improved germination in temperate conifers such as Cedrus deodara and Pinus roxburghii. Moreover, stratification enhances hormonal balance and enzymatic activity, especially gibberellin synthesis, which promotes reserve mobilization and radicle appearance, according to (Luna and Singh \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Excessive freezing or seed aging a pattern also reported by (Williams \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1985\u003c/span\u003e) could be the cause of the modest decrease in germination at 125 days seen here. Therefore, for \u003cem\u003eP. wallichiana\u003c/em\u003e to attain maximal germination and vigor under nursery circumstances, stratification of about 100 days is ideal.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec32\" class=\"Section2\"\u003e \u003ch2\u003eSeedling Growth and Biomass Accumulation\u003c/h2\u003e \u003cp\u003eThe greatest shoot and root growth as well as total biomass were shown by seedlings from 100-day stratified seeds, suggesting enhanced physiological vigor and nutrient utilization. Other Himalayan conifers have also been observed to exhibit enhanced growth following optimum stratification (Aslam et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The notable rise in dry mass of the shoots and roots points to increased establishing potential in the field as well as effective mobilization of stored reserves. The idea that adequately stratified seeds provide more robust seedlings capable of higher survival and growth in temperate forest nurseries is supported by the high vigor index (1854) (Abdul-Baki \u0026amp; Anderson \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1973\u003c/span\u003e; Luna and Singh, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec33\" class=\"Section3\"\u003e \u003ch2\u003eSite-wise Variation and Environmental Influence\u003c/h2\u003e \u003cp\u003eSignificant site-specific variations in germination, seed, and cone parameters show the combined impact of local environmental factors and genotype. Superior cone size, seed weight and germination percentage were displayed by progenies from the Akhal and Wussan locations, which may indicate improved soil fertility and suitable climatic circumstances. According to (Kaur et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) mid altitude \u003cem\u003eP. wallichiana\u003c/em\u003e populations in Himachal Pradesh exhibited greater seed viability and germination than high altitude populations.\u003c/p\u003e \u003cp\u003eGiven this regional variance, seeds from mid-elevation sources (1800\u0026ndash;2300 m) are thought to have higher physiological maturity and ought to be chosen for the creation of seed orchards and extensive reforestation initiatives.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec34\" class=\"Section3\"\u003e \u003ch2\u003eCorrelations among Traits\u003c/h2\u003e \u003cp\u003eCone weight, seeds per cone and germination qualities have substantial positive associations (r\u0026thinsp;=\u0026thinsp;0.862, 0.745, etc.) suggesting that germination and vigor can be directly improved by improvements in cone features. These results are in line with previous findings by (Burton and Devane \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1953\u003c/span\u003e) and Johnson et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1955\u003c/span\u003e) who pointed out that associated quantitative traits frequently react to selection simultaneously because of shared additive genetic influences. The strong correlation (r\u0026thinsp;=\u0026thinsp;0.895) between vigor index and germination percentage attests to the vigor index's validity as a gauge of genetic quality and overall seed performance.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eGenetic Parameters and Heritability\u003c/h3\u003e\n\u003cp\u003eThe preponderance of additive gene action is indicated by high estimates of heritability (\u0026gt;\u0026thinsp;70%) and genetic advancement for cone weight, germination percentage and vigor index. This implies that breeding programs would benefit from selection based on these features. Singh and Thapliyal (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) showed substantial heritability for cone weight and seed weight in P. wallichiana, with similar results.\u003c/p\u003e \u003cp\u003eFurthermore, the tiny discrepancy between the genotypic (GCV) and phenotypic (PCV) coefficients of variation suggests that recurrent selection can lead to genetic improvement and that environmental influences are minimal (Zobel and Talbert \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; Wright \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1976\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eMultivariate and Cluster Analysis\u003c/h3\u003e\n\u003cp\u003eCone weight, seed quantity, germination percentage and vigor index were found to be significant contributors to overall variation by Principal Component Analysis, which explained 68% of the variance between progenies. This is consistent with research by (Aslam et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), which showed that \u003cem\u003eP. wallichiana\u003c/em\u003e variability is greatly influenced by both reproductive and germination features.\u003c/p\u003e \u003cp\u003eProgenies were divided into five different groups using cluster analysis, with Cluster I (T1, T2, T3 and T13) showing the highest mean vigor index (1048). This kind of clustering helps choose elite seed sources for upcoming breeding and conservation initiatives and successfully distinguishes superior genotypes (Ward \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1963\u003c/span\u003e; White et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec37\" class=\"Section2\"\u003e \u003ch2\u003eImplications for Conservation and Breeding\u003c/h2\u003e \u003cp\u003eA framework for the sustainable management of \u003cem\u003eP. wallichiana\u003c/em\u003e is provided by the combination of quantitative genetic analysis and reproductive biology. Regeneration success and long-term forest productivity can be greatly increased by identifying high-performing progenies, ideal stratification times and advantageous seed sources. The construction of seed orchards, the management of genetic resources and the restoration of degraded habitats are all supported by these findings, which support conservation and improvement efforts given the ecological and economic significance of blue pine in the Himalayas.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe complex interplay between reproductive biology, genetic variability and environmental effect throughout the \u003cem\u003ePinus wallichiana\u003c/em\u003e (Blue Pine)'s natural range in the Kashmir Himalaya was clarified by the current study. The species' adaptive response to climatic gradients is demonstrated by the noticeable altitudinal variation in reproductive phenophases, which guarantee successful pollination and cone maturity under a variety of environmental circumstances.\u003c/p\u003e \u003cp\u003eSubstantial variation in cone, seed and germination traits among half sib progenies signifies broad genetic diversity within the population. The strong positive correlations among cone weight, seed number, germination percentage and vigor index highlight that improvements in reproductive and seed traits can directly enhance propagation efficiency. The high heritability and genetic advance observed for key traits such as cone weight, germination percentage and vigor index confirm the predominance of additive gene action, suggesting that simple phenotypic selection can effectively identify superior genotypes for breeding programs.\u003c/p\u003e \u003cp\u003eThe best pre-sowing treatment for dormancy release and maximum vigor was found to be 100 days of cold moist stratification, which significantly enhanced germination and seedling performance. Variations by site also show that mid altitude populations, especially those from Akhal and Wussan have better reproductive and germination qualities, which makes them perfect for seed orchard growth and genetic improvement initiatives.\u003c/p\u003e \u003cp\u003eOverall, a scientific basis for improving regeneration success, encouraging genetic conservation and assisting with sustainable forest management of \u003cem\u003eP. wallichiana\u003c/em\u003e in the Himalayas is provided by the combination of reproductive biology and quantitative genetic research. In addition to increasing nursery and plantation productivity, the identification of high-performing offspring and the standardization of stratification procedures will also make a substantial contribution to climate-resilient reforestation and the long-term viability of temperate conifer forests.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eStatements and Declarations\u003c/h2\u003e \u003cp\u003eThe author declares a non-financial competing interest that could be perceived as relevant to the submitted work. This interest does not involve any financial relationship, funding, or compensation, but may be viewed as potentially influencing the interpretation or presentation of the research.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e \u003cp\u003eNo funding was received.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eMB conceived the idea, designed, directed, performed, and coordinated the research experiments with the active participation of PAK, AAM, and ABW. MB, PAK, AAM \u0026amp; ABW and MD wrote the paper together. All authors contributed to the article and approved the submitted version.\u003c/p\u003e\u003ch2\u003eAcknowledgement:\u003c/h2\u003e \u003cp\u003eThe authors wish to express their gratitude to the scientists of Division of FBT, SKUAST-K for facilitating the use of equipment and for technical help.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdul-Baki AA, Anderson JD. 1973. Vigor determination in soybean seed by multiple criteria. \u003cem\u003eCrop Science\u003c/em\u003e. 13(6):630\u0026ndash;633.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAslam M, Ahmad SS, Rafiq M. 2010. Reproductive biology and germination behavior of \u003cem\u003ePinus wallichiana\u003c/em\u003e A.B. Jacks. from Himalayan forests of Pakistan. \u003cem\u003ePakistan Journal of Botany\u003c/em\u003e. 42(1):93\u0026ndash;102.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAslam M, Qureshi R, Ahmed S. 2011. Natural regeneration status of \u003cem\u003ePinus wallichiana\u003c/em\u003e and associated conifers in moist temperate forests of Pakistan. \u003cem\u003ePakistan Journal of Botany\u003c/em\u003e. 43(6):2711\u0026ndash;2716.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBhat GM, Rather MA, Shah MA. 2017. Status, utilization and conservation of \u003cem\u003ePinus wallichiana\u003c/em\u003e A.B. Jacks. in Kashmir Himalaya, India. \u003cem\u003eInternational Journal of Conservation Science\u003c/em\u003e. 8(2):229\u0026ndash;238.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBurton GW, DeVane EH. 1953. Estimating heritability in tall fescue (\u003cem\u003eFestuca arundinacea\u003c/em\u003e) from replicated clonal material. \u003cem\u003eAgronomy Journal\u003c/em\u003e. 45(10):478\u0026ndash;481.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCzabator FJ. 1962. Germination value: an index combining speed and completeness of pine seed germination. \u003cem\u003eForest Science\u003c/em\u003e. 8(2):386\u0026ndash;396.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFarjon A. 2013. \u003cem\u003ePinus wallichiana\u003c/em\u003e A.B. Jacks. In: \u003cem\u003ePinaceae: conifers\u0026mdash;drawings and descriptions of the genera.\u003c/em\u003e 2nd ed. Leiden (Netherlands): Brill. p. 232\u0026ndash;234.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGomez KA, Gomez AA. 1984. \u003cem\u003eStatistical procedures for agricultural research\u003c/em\u003e. 2nd ed. New York (NY): John Wiley \u0026amp; Sons.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eInternational Seed Testing Association (ISTA). 1996. \u003cem\u003eInternational rules for seed testing\u003c/em\u003e. Vol. 1. Bassersdorf (Switzerland): ISTA.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJohnson HW, Robinson HF, Comstock RE. 1955. Estimates of genetic and environmental variability in soybeans. \u003cem\u003eAgronomy Journal\u003c/em\u003e. 47(7):314\u0026ndash;318.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKaur A, Monga R, Bhardwaj D. 2024. Altitudinal variation and aspects impact on seed viability and germination of \u003cem\u003ePinus wallichiana\u003c/em\u003e in the nursery. \u003cem\u003eIndian Forester\u003c/em\u003e. 150(2):106\u0026ndash;113.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKearns CA, Inouye DW. 1997. Pollinators, flowering plants, and conservation biology. \u003cem\u003eBioScience\u003c/em\u003e. 47(5):297\u0026ndash;307.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLuna T, Singh RP. 2009. Effect of cold stratification on seed germination of temperate conifers. \u003cem\u003eJournal of Forestry Research\u003c/em\u003e. 20(2):127\u0026ndash;130.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaguire JD. 1962. Speed of germination\u0026mdash;aid in selection and evaluation for seedling emergence and vigor. \u003cem\u003eCrop Science\u003c/em\u003e. 2(2):176\u0026ndash;177.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMong CE. 2006. Establishment of \u003cem\u003ePinus wallichiana\u003c/em\u003e on a Himalayan landslide: an example of primary succession. \u003cem\u003eBiotropica\u003c/em\u003e. 38(5):584\u0026ndash;592.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoza KL, Bhatnagar AK. 2005. Intraspecific variation and seed quality of \u003cem\u003ePinus wallichiana\u003c/em\u003e in western Himalaya. \u003cem\u003eJournal of Tree Sciences\u003c/em\u003e. 24(1):55\u0026ndash;62.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOrwa C, Mutua A, Kindt R, Jamnadass R, Simons A. 2009. \u003cem\u003eAgroforestree database: a tree reference and selection guide.\u003c/em\u003e Version 4.0. Nairobi (Kenya): World Agroforestry Centre.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRana BS, Singh SP. 1990. Structure and composition of forests in the Central Himalaya along an altitudinal gradient. \u003cem\u003eVegetatio\u003c/em\u003e. 88(1):69\u0026ndash;79.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma SK, Singh SP, Singh RP. 1994. Effect of cold stratification on seed germination of \u003cem\u003ePinus wallichiana\u003c/em\u003e. \u003cem\u003eIndian Forester\u003c/em\u003e. 120(9):825\u0026ndash;828.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh JS, Yadav AS. 2000. Environmental degradation and management of Himalayan ecosystems: a case study of the \u003cem\u003ePinus wallichiana\u003c/em\u003e forests in Western Himalaya. \u003cem\u003eJournal of Environmental Management\u003c/em\u003e. 59(4):265\u0026ndash;281.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh O, Thapliyal M. 2012. Variation in cone and seed characters in blue pine (\u003cem\u003ePinus wallichiana\u003c/em\u003e) across natural distribution in Western Himalayas. \u003cem\u003eJournal of Forestry Research\u003c/em\u003e. 23(2):235\u0026ndash;239.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh SP, Yadav RK. 2000. Ecology and distribution of Himalayan conifers. \u003cem\u003eHimalayan Journal of Forestry\u003c/em\u003e. 18(1):23\u0026ndash;34.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTewari LM, Gupta N, Bhatt P. 2001. Reproductive phenology and seed production in \u003cem\u003ePinus wallichiana\u003c/em\u003e in Kumaun Himalaya. \u003cem\u003eIndian Journal of Forestry\u003c/em\u003e. 24(4):289\u0026ndash;295.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThapliyal M, Singh O, Sah B. 2008. Seed source variation and conservation of \u003cem\u003ePinus wallichiana\u003c/em\u003e in India. \u003cem\u003eAnnals of Forest Research\u003c/em\u003e. 51:147\u0026ndash;153.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTiwari AK, Joshi GC. 2012. Ecological distribution and regeneration status of \u003cem\u003ePinus wallichiana\u003c/em\u003e in the Garhwal Himalaya, India. \u003cem\u003eJournal of Forestry Research\u003c/em\u003e. 23(3):405\u0026ndash;412.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTroup RS. 1921. \u003cem\u003eThe silviculture of Indian trees. Volume III: Conifers (pines, etc.)\u003c/em\u003e. Oxford (UK): Clarendon Press.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWard JH. 1963. Hierarchical grouping to optimize an objective function. \u003cem\u003eJournal of the American Statistical Association\u003c/em\u003e. 58(301):236\u0026ndash;244.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWhite TL, Adams WT, Neale DB. 2007. \u003cem\u003eForest genetics\u003c/em\u003e. Wallingford (UK): CABI Publishing.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWilliams RD. 1985. Seed dormancy and germination in \u003cem\u003ePinus wallichiana\u003c/em\u003e. \u003cem\u003eJournal of Tropical Forestry\u003c/em\u003e. 1(2):145\u0026ndash;150.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWright S. 1976. \u003cem\u003eEvolution and the genetics of populations. Vol. 3: Experimental results and evolutionary deductions\u003c/em\u003e. Chicago (IL): University of Chicago Press.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZobel BJ, Talbert J. 1984. \u003cem\u003eApplied forest tree improvement\u003c/em\u003e. New York (NY): John Wiley \u0026amp; Sons.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Reproductive biology, half-sib progenies, variability, stratification period, PCV, GCV, Pinus wallichiana","lastPublishedDoi":"10.21203/rs.3.rs-8583859/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8583859/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cb\u003eBackground\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003ePinus wallichiana\u003c/em\u003e often referred to as Bhutan pine, Himalayan pine or blue pine, A.B. Jackson is a five-needle pine belonging to the \u003cem\u003ePinaceae\u003c/em\u003e family that is indigenous to the Hindu Kush, Karakoram and Himalayan regions.\u003c/p\u003e\u003cp\u003e\u003cb\u003eResults\u003c/b\u003e\u003c/p\u003e \u003cp\u003eMale and female strobili phenophases lasted 10\u0026ndash;11 and 77\u0026ndash;78 weeks, respectively. Cones were collected from 18 phenotypically superior trees across six locations to assess variability in morphological traits. Trees from Akhal exhibited the highest values for all measured traits, including tree height (28.9 m), DBH (47.7 cm), cone length (22.44 cm), cone weight (123.42 g), and seed weight (87.98 g). Seeds underwent cold stratification at 25-day intervals (0\u0026ndash;125 days) at 4\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C to evaluate germination and seedling performance. Maximum germination (72.67%) and seedling vigor were observed at 100 days of stratification, while the Akhal genotype showed superior overall performance (germination 86.77%, vigor index 1048.18). High genetic variability was observed, with the highest GCV and PCV for dry root weight (51.40, 52.65 respectively). Heritability was highest for cone specific gravity (0.78) and seed morphological traits. Principal component analysis (PCA) revealed that the first two components explained 96.23% of the total variation, with cone weight and seed thickness as major contributors. Cluster analysis grouped genotypes into five clusters, with Cluster I exhibiting superior cone, seed, and seedling traits, indicating potential for selective breeding and genetic improvement programs.\u003c/p\u003e","manuscriptTitle":"Reproductive Biology and Genetic Parameters of Blue Pine: Insights into Germination, Growth and Breeding Potential","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-20 05:03:46","doi":"10.21203/rs.3.rs-8583859/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f6811ce9-b981-4f95-9031-39f6d8250a08","owner":[],"postedDate":"January 20th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-20T05:03:46+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-20 05:03:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8583859","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8583859","identity":"rs-8583859","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}