Phenological Synchrony, Parental Contribution, and Pollen Dispersal Pattern affects the Genetic Composition of Cupressus funebris Dwarf Seed Orchard Progenies

Phenological Synchrony, Parental Contribution, and Pollen Dispersal Pattern affects the Genetic Composition of Cupressus funebris Dwarf Seed Orchard Progenies | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Phenological Synchrony, Parental Contribution, and Pollen Dispersal Pattern affects the Genetic Composition of Cupressus funebris Dwarf Seed Orchard Progenies Hao-bo Zhao, sen Cao, zhen Zhang, zhi-chun Zhou, guo-qing Jin, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7253675/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Context Cupressus funebris is a high - value conifer, is important for timber and ecological protection. In its seed production, optimizing parental composition in seed orchards and using “dwarfed” trees after topping are key to achieving high - yield and high - gain seeds. However, compared to traditional seed orchards, the parental contributions and pollen dispersal patterns in dwarfed seed orchards remain unclear. Aims By analyzing flowering synchrony, parental contributions, and pollen dispersal patterns in a dwarfed Cupressus funebris seed orchard, we investigated their effects on offspring genetics. Methods Using three years of flowering-phenology observations, we quantified flowering synchrony and identified the principal climatic drivers governing its variation. We further characterised the mating system within the dwarf seed orchard, estimated the gametic contribution of each parent to the offspring cohort, and finally compared pollen dispersal patterns between dwarf and non-dwarf orchards. Results Three years of monitoring revealed significant interclonal variation among parental ortets in the seed orchard with respect to both strobilus production and flowering phenology, indicating asymmetrical gametic contribution within the orchard. The low within-year synchrony index of male and female strobili is predominantly governed by clonal variation, as well as by significant effects of mean daily maximum temperature, mean daily minimum temperature, and cumulative precipitation during the flowering period. The offspring population in the dwarfed seed orchard maintained high genetic diversity but had inbreeding risk. Compared to the non - dwarfed orchard, spatial distance was a key factor affecting pollination success in the dwarfed one. After dwarfing, the spatial distribution of female and male cone flowers shifted downward, shortening the average pollen dispersal distance, reducing outcrossing rates, and increasing self - pollination probability. Conclusion Branch management and plant spacing in Cupressus funebris dwarfed seed orchards should be emphasized. Improving ventilation and light penetration, combined with flowering management, are essential management strategies. Cupressus funebris dwarfed seed orchards flowering synchrony mating system paternity analysis pollen spread Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Key Message This study demonstrates that the low flowering synchrony in dwarf seed orchards is primarily attributable to inter-clonal variation and to temperature and precipitation conditions during the anthesis period. Although the progeny population of dwarf orchards retains the high genetic diversity of the parental clones, unbalanced gametic contributions within the orchard predispose the offspring to increased self- or inbreeding. Moreover, compared with non-dwarf orchards, pollen dispersal distances in dwarf orchards are significantly reduced. These findings underscore the necessity for improving clonal selection, spatial arrangement, and shoot growth management in dwarf seed orchards. 1 Introduction Seed orchards, special - purpose tree populations for seed production, are the "output delivery system" of breeding populations (Allender 2011; Li et al. 2012; Heuchel et al. 2022). For conifers lacking clonal propagation materials, seed orchards link genetic improvement with artificial forest establishment. They contain the best individuals (breeding values), and theoretically, their offspring should have high genetic gain and diversity to enhance major economic traits or adaptability. This is crucial for global seed production programs (El-Kassaby and Reynolds 1990; Ruotsalainen 2014). To produce seeds in a predictable manner, in open - pollinated seed orchards, balanced mating probabilities between all parents, low - level inbreeding, and random mating are expected. Each parent should contribute equally to the offspring, ensuring a broad genetic base and the expected genetic value (Lindgren and Prescher 2005; Chen et al. 2018). This requires several conditions: (a) all parents provide equal numbers of male and female gametes for seed production; (b) female flower fertilization timing is synchronized with pollen dispersal; (c) the seed orchard parents have sufficient genetic diversity to pass on to the offspring; and (d) the seed orchard is isolated from natural stands of the same species to prevent external pollen contamination. As observed in many tree breeding programs, these expectations for seed orchards are rarely fully met (Funda et al. 2011; Torimaru et al. 2012; Lisa and Keith 2017, Song et al. 2018; Bruna et al. 2020). Parental phenology, pollen quantity, and seed yield in seed orchards can vary significantly among parent sources and genotypes. Parental contributions to the genetic pool of the offspring are often unbalanced, potentially leading to a loss of genetic diversity (Heike and Dagmar 2022; Chatterjee et al. 2025). When selecting parents for seed orchards, it is essential to optimize a large number of parental materials and evaluate offspring genetic traits to maximize gains while maintaining high diversity levels (Yang et al. 2020; Chaloupková and Lstibůrek 2022; Lee et al. 2022). Phenology is a key part of a plant's life history and a major fitness factor that affects reproductive success (Primack 2003; Paul et al. 2014; Du et al. 2024; Martins et al. 2025). Differences in phenological variation and flowering synchrony can impact the frequency of gene exchange between parents and the genetic composition of seeds in seed orchards. These differences can also reduce effective population size, cause year - to - year changes in mating systems, increase self - pollination probabilities, and raise the risk of external pollen contamination (Barrett et al. 2011; Darlin et al. 2024). These issues are particularly evident in conifer seed orchards where wind is the main pollen dispersal agent (Li et al. 2012; Heuchel et al. 2022; Wang et al. 2023). In conifers, 34% - 52% of parents typically contribute to 80% of the offspring in seed orchards, showing a clear imbalance (Wang et al. 2023; Martins et al. 2025). Pollen dispersal and female cone flower pollination usually occur within two to three weeks. During this time, factors such as rainfall, wind direction, and wind speed can become critical, alter random mating in seed orchards, and increase pollen contamination (Pearse et al. 2015; Tania et al. 2024). In recent years, tree dwarfing has been used to address the problem of tall trees in seed orchards and difficulties in seed collection. It's an important part of modern intensive seed orchard management (Han et al. 2013; Wu et al. 2015; Wang et al. 2025). Methods like topping, pruning, girdling, and hormone induction are used to suppress apical dominance and control upward growth. This makes it easier to harvest cones and conduct artificial pollination. However, it also causes female and male cone flowers to shift downward (Kolpak et al. 2015). But tree - breeding programs rarely focus on flower - related management in dwarfed seed orchards. It's still unclear whether pollen and mating patterns change in these orchards. This information is crucial for making genetic management decisions in seed orchards, such as dwarfing, thinning, artificial pollination, and clone arrangement (Chen et al. 2022; Tania et al. 2024). Cupressus funebris is a key afforestation and ecological tree species in southern China. It has excellent wood quality, strong adaptability, and can tolerate drought and poor soils, making it highly effective for afforestation and ecological restoration in barren mountains (Yang et al. 2023). However, significant clonal differences in reproductive capacity and flowering phenology have been observed in Cupressus funebris . Factors such as multiple branches, poor ventilation and light penetration in the inner canopy, and adverse conditions like temperature drops and rainfall during the flowering period may affect mating probabilities among parents in dwarfed seed orchards (Huntsman and Leslie 2023; Martins et al. 2021). To ensure seed quality for afforestation and develop rational genetic management strategies for dwarfed seed orchards, long - term monitoring of reproductive phenology and flowering synchrony within seed orchards is essential. It is also crucial to evaluate the impact of these factors on seed production, as well as pollen dispersal patterns and mating systems among parents. The objectives of our study are to: 1) Characterize differences in flowering synchrony and identify major climatic factors in dwarfed Cupressus funebris seed orchards. 2) Assess parental contributions to the genetic pool of offspring. 3) Examine changes in mating mechanisms and pollen dispersal patterns following tree management practices. Our findings offer new insights into the mating mechanisms of dwarfed seed orchards and provide valuable references for flower - related management practices. 2 Material and methods 2.1 Study Area and Material Sources The study materials were obtained from a Cupressus funebris seed orchard located in Chun'an County, Zhejiang Province (119°03'57″ E, 29°32'24″N). The orchard covers an area of 3.7 hm², with an altitude ranging from 250 to 300 m. It is situated on a gentle slope with a gradient of less than 15°, facing south - southeast. The soil conditions of the seed orchard were determined, with mountain red soil (pH 4.74) containing 20.08 g/kg organic matter, 1.12 g/kg total nitrogen, 0.49 g/kg total phosphorus, 95.87 mg/kg alkaline - hydrolyzed nitrogen, 283.92 mg/kg available phosphorus, 93.36 mg/kg available potassium, 212.90 mg/kg exchangeable calcium, and 108.55 mg/kg total boron. The improved seed orchard was established in 2015. After genetic testing of the parental materials from the first - generation seed orchard (established in 1990) with 45 parents, 27 genotypes were retained to form this improved clonal seed orchard. The seed orchard is divided into five blocks, with a total of 1,680 trees planted. The clones are arranged in a random spatial arrangement within the orchard. Each clone is repeated 10 - 15 times within a plot, and the distance between ramets of the same clone is over 20 m, with a tree spacing of 4 m × 4 m. A pollen - free zone of over 300 m surrounds the seed orchard. The seed orchard first flowered and produced cones in 2018. In 2020, the mother trees in blocks 2 - 5 underwent topping to facilitate cone harvest and pollination management, forming the Dwarfed Seed Orchard (DSO). Specifically, the top 1 - 2 whorls were cut, with the cutting height controlled at 2.5 m, with no further treatment (Fig. 1) . Block 1 was undwarfized seed orchards (USO). The management of both seed orchards remained consistent. 2.2 Phenological Observation Phenological observations were conducted during the flowering stage in 2021, 2024, and 2025. For each clone, five ramets were selected. The following method was used: one first - order branch in the middle - upper part of the canopy was chosen in each of the four directions (east, south, west, and north). These branches were observed daily until the end of the flowering period (Li et al. 2012) .The flowering phases were determined as follows: (1) When 0% - 30% of female cone scales crack or 0% - 30% of male cones shed pollen, it's the onset of flowering. (2) When 30% - 75% of female cone scales crack or 30% - 75% of male cones shed pollen, it's full flowering. (3) The end of flowering for female cones is when 75% - 100% of female cone scales close, thicken, and turn dark purple - red. For male cones, it's when 75% - 100% of pollen is shed, and the flower branches turn yellowish - brown and wilt. The entire flowering period, which included the beginning, full, and end of flowering phases, was recorded. At the end of the flowering period, the number of female cone flowers and male cone flower spikes on each observed branch were recorded (Beren et al. 2020). 2.3 Sampl es of mating systems Sample collection was carried out in two parts. In the first part, in May 2023, we collected the current year's needles from 27 parents in the dwarfed seed orchard and stored them in a -80 ℃ freezer for DNA extraction. During the cone maturation period, we collected cones from all clones and processed the seeds for sowing and seedling cultivation. When the seedlings reached a height of 20 cm, we randomly selected 20 seedlings from each clones, forming a total of 540 offspring individuals for genetic diversity analysis between parental and offspring generations in the seed orchard. The second part involved selecting five identical clones from both non - dwarfed and dwarfed plots. We marked the locations of the mother trees and collected current - year needles. Upon cone maturation, we collected cones from the middle - upper part of the tree, air - dried them in the laboratory, and gathered over 1,000 seeds per tree. After sowing and seedling cultivation, 30 offspring seedlings were randomly chosen from each mother tree, forming 150 offspring individuals. Young tissues were collected and stored in a -80 ℃ freezer for DNA extraction. These samples were used for paternal analysis and pollen dispersal pattern research in dwarfed and non - dwarfed seed orchards. 2.4 DNA Extraction, Primer Screening, and SSR-PCR Amplification DNA was extracted from parent trees and seedling progeny in the seed orchard using an improved CTAB plant genomic DNA extraction kit (Aidlab, Beijing, China), and DNA quality and concentration were measured using a NanoDrop 2000 spectrophotometer. For parent - offspring relationship analysis and genetic diversity assessment, 12 SSR markers were used, with primer information listed in Table S1. PCR amplification and detection followed previously described methods (Yang et al. 2016). Parent and offspring DNA samples were amplified with primers, and the amplified products were separated by electrophoresis on a Qsep100 TM automatic nucleic acid protein analysis system (BIOptic, Taiwan). 2.5 Genetic Diversity Parameter Calculation After exporting and converting the genotyping data from the Qsep100TM system, genetic diversity parameters were calculated using GenAlex 6.5. These parameters included the number of alleles ( Na ), effective number of alleles ( Ne ), Shannon’s diversity index ( I ), observed heterozygosity ( Ho ), expected heterozygosity ( He ), fixation index ( F). Polymorphism information content ( PIC ) for microsatellite loci were assessed using Cervus 3.0 (Zoe et al. 2023). 2.6 Paternity Analysis CERVUS 3.0 was used to conduct parentage analysis of free - pollinated offspring within a 70% - 80% confidence interval, determining paternal contributions. Paternity Identification Rate ( PIR ), outcrossing rate ( OCR ), selfing rate ( SR ), Number of Unique Alleles in Progeny ( NUAIP ) and pollen contamination rate by foreign pollen ( PCFP ) were calculated for non - dwarfed and dwarfed plots. Pollen dispersal distance was also calculated based on the actual distances between parents in the seed orchard (Heike and Dagmar 2022). 2.7 Statistical Analysis of Flowering Synchrony The flowering synchrony among parents as male, female, and self - pollination was analyzed using the flowering synchrony index ( W ij ) proposed by Li et al. (Li et al.2012). In the formula, W ij is the flowering synchrony index between the male cones of the i -th clone and the female cones of the j -th clone; M ki is the flowering frequency of the male cones of the i -th clone on day k ; P kj is the flowering frequency of the female cones of the j -th clone on day k ; and n is the number of days from the earliest flowering start to the latest flowering end between clones i and j . When i = j , the index represents the self - pollination flowering synchrony index W i . A W ij value of 1 means complete overlap of male and female cone flowering periods, while a value of 0 means no overlap. 2.8 Collection and Analysis of Climatic Data A micro - automatic weather station was used to record weather factors during the flowering period. The average high temperature (AHT) was the mean of the highest daily temperatures. The average low temperature (ALT) was the mean of the lowest daily temperatures. DMA stands for daily average temperature. Rainfall amount (RA) was the sum of daily rainfall. The number of rainy days (NRD) was counted. Average wind speed (AWS) was the mean of wind speeds, and average humidity (AH) was the mean of humidity levels during the flowering period. 3 Results 3.1 Reproductive Capacity During the three observation years, in the dwarfed seed orchard, 51.85% to 66.67% of clones contributed over 85% of female cones, and 44.44% to 51.85% of clones contributed over 85% of male cones. The parental contribution curves for female and male cones deviated from the ideal (Fig. 2-a), showing unbalanced gamete contributions. Paternal contribution analysis (Fig. 2-b) showed that in the dwarfed plot, paternal contribution ranged from 0.99% to 14.85%, with 23.81% of fathers producing over 50% of the offspring. In the non - dwarfed plot, paternal contribution was between 0.87% and 13.04%, with 24% of fathers producing over 50% of the offspring. Compared to the non - dwarfed plot, the dwarfed plot had fewer effective pollen - producing fathers, reducing gene exchange. Overall, a small number of clones contributed more pollen during reproduction. 3.2 Mating Pattern The dwarfed seed orchard is mainly characterized by outcrossing, with an outcrossing rate of 72.19%. Among the offspring, 101 individuals (70.89% identification rate) matched with parental clones, and 8 offspring-specific alleles ( Na ) were found. The pollen contamination rate was 5.80%. In contrast, the non-dwarfed seed orchard had a higher outcrossing rate of 81.70%, with 115 offspring (76.76% identification rate) matching the parentals and 12 offspring-specific alleles detected. Its pollen contamination rate was 8.00% (Table 1). 3.3 Pollen Dispersal Patterns Pollen propagation among seed orchard clones is random without a specific pollination direction. The highest proportion of offspring occurs when effective pollen dispersal is 30 - 40 m. In non - dwarfed plots, the average distance between mother trees and pollen donors was 35.03 m, with a maximum pollination distance of 68.82 m and 64.86% of effective pollination within 40 m. In dwarfed plots, the maximum pollination distance was 60.00 m, the average pollination distance was 33.35 m, and 75.76% of effective pollination occurred within 40 m (Fig. 3). Dwarfing reduces the effective pollen dissemination distance. 3.4 Parental Contribution to the Offspring Gene Pool In the parental population of the dwarfed seed orchard, the number of observed alleles ( Na ) ranged from 3 to 11, with an average of 6.000, and the average number of effective alleles ( Ne ) was 3.529. Among 540 offspring, Na ranged from 4 to 11, averaging 6.583, which is 9.717% higher than the parental average. Effective allele numbers were 9.493% higher. Genetic diversity was comparable to the parental population, indicating no reduction in offspring genetic diversity (Table 2). Offspring largely retained parental genetic diversity. However, offspring heterozygosity ( Ho ) was lower (0.536) than parental Ho (0.620), showing a decrease in offspring heterozygosity. Both parental and offspring populations had positive fixation indices ( F ), 0.026 and 0.201, respectively. This suggests potential excess homozygosity and inbreeding, but with relatively low heterozygote deficiency. 3.5 Flowering Synchrony Index Due to clonal differences, male and female cone flowering durations vary. Over three observation years, pollen shedding lasted 5 - 11, 9 - 14, and 10 - 12 days, respectively. The female cone pollination period was 5 - 12, 9 - 13, and 9 - 11 days (Fig. 3a). The full flowering period accounted for the largest proportion, with male cone full flowering comprising 36.36% - 66.67% of the total flowering period and female cone full flowering comprising 37.50% - 71.42% (Fig. 4). In 2021, 2024, and 2025, the flowering synchrony index ( W i j ) for clone combinations ranged from 0 to 0.89, 0 to 0.72, and 0 to 0.69, respectively, with averages of 0.26, 0.39, and 0.42. Overall, the flowering synchrony index was low. In 2021, 41.54% of clone combinations had a flowering synchrony index above 0.3, while 58.46% had an index below 0.3, and only 23.84% exceeded 0.4. In 2024, 74.36% of clone combinations had an index above 0.3, 25.64% below 0.3, and 48.72% exceeded 0.4. In 2025, 83.76% of clone combinations had an index above 0.3, 16.24% below 0.3, and 62.11% exceeded 0.4. (Figure 5 - b). Within the same year, the flowering synchrony indices between clones as male and female parents were similar, with variations within 5%. The self - pollination flowering synchrony indices for each clone were 0.341, 0.472, and 0.492 over the three years, higher than when they served as male or female parents. This indicates increasing self - pollination risk with orchard age (Fig. 6). 3.6 Climatic Impact Factors Factorial analysis of major meteorological factors during the flowering period and flowering phenology shows that rainfall and temperature are key drivers of the flowering synchrony index (Fig. 7). Interestingly, in years with good weather, the shorter full - flowering period leads to a lower synchrony index. Conversely, in years with rainy and cold conditions, the synchrony index rises. Among the three observed years, two with low temperatures and rainfall showed similar increases in the flowering synchrony index (Fig. 8). During the flowering period, the frequency of rainfall events is independent of floral duration; however, cumulative precipitation can prolong anthesis. Daily mean temperature inversely (DMT) modulates floral longevity, with elevated thermal regimes shortening the flowering window in clonal genotypes. Genetically mediated clonal effects consequently diminish phenological synchrony. 4 Discussion 4.1 Imbalanced Parental Contribution to the Offspring Gene Pool Compared to other conifer seed orchards, the genetic diversity of Cupressus funebris parental populations is slightly higher or similar, indicating a proper selection intensity of high - gain parents from the first - generation seed orchard for the improved one. Despite fewer parents than the breeding population ( Ho , 0.567) (Yang et al. 2016), the seed orchard maintains comparable genetic diversity ( Ho , 0.620), showing a broad genetic base. However, significant gamete contribution imbalance exists: 51.85% - 66.67% of clones contribute over 85% of female cones, and 44.44% - 51.85% contribute over 85% of male cones (Gömöry et al. 2003). This may lead to a few parents dominating the offspring gene pool, reducing genetic diversity. Similar cases have been reported in other conifer seed orchards (Chen et al. 2022; Wang et al. 2023). This study also reveals that the offspring's observed heterozygosity ( Ho = 0.536) is lower than the parents' ( Ho = 0.620), and the inbreeding coefficient ( F ) is positive for both, suggesting excess homozygosity and inbreeding risk. The decrease in offspring heterozygosity mainly stems from the imbalanced parental contribution to the offspring gene pool (Nyabera et al. 2021; Heuchel et al. 2022). 4.2 Pollen Dispersal Patterns and Paternal Reproductive Success The mating system affects the reproductive level and genetic structure of parent trees in seed orchards. The extent of effective pollen dispersal within seed orchards is limited, and pollen from external unknown sources has a wide dispersion range, leading to imbalanced paternal contributions (Lisa and Keith 2017). Funda and El-Kassaby indicated that 20% of parental clones in seed orchards can produce up to 80% of the cones (Funda et al. 2012). In this study, over three years of investigation in the seed orchard, a smaller number of clones were found to contribute a larger amount of pollen during reproduction. The parental balance curves for female and male cone quantities deviated significantly from the ideal state (Fig. 1). The offspring parentage analysis results showed that in the dwarfed plots, the pollen source could be determined for 70.89% of the offspring, and in the non-dwarfed plots, the pollen source could be determined for 76.67% of the offspring. The range of the number of offspring produced by the mother tree and the candidate father tree was 1 to 11, with the majority producing only one offspring. The parental reproductive contribution rate ranged from 3.57% to 42.31%, indicating an imbalance in paternal contributions within the seed orchard, especially in the dwarfed plots, where the frequency of gene exchange was reduced. The contribution of the paternal candidate trees decreased with the increase of the distance between the mother tree and the candidate tree. The wind direction between the mother tree and the pollen donor was also an important factor determining the paternal contribution. This is because in wind-pollinated tree species, pollen wind direction is affected by the prevailing wind direction (Heuchel et al. 2022). This phenomenon also occurs in the western larch experimental forest (Ben et al. 2008), but the influencing factors are different. The low pollination efficiency and asynchronous flowering among clones are the main reasons for the imbalance in paternal reproductive output within the seed orchard. Gene flow through the dynamic dispersal of pollen and seeds helps the genetic variation of plant populations (Huang et al. 2018; Chen et al. 2018; Bruna et al. 2020). The pollen dispersal range in Cupressus funebris seed orchards is relatively wide, with a pollination pattern similar to that of other wind-pollinated tree species, such as Pinus massoniana, Pinus tabuliformis , and Abies . This is different from the traditional pollination pattern of tall trees (Armstrong et al. 2024; Yang et al. 2025). In dwarfed seed orchards, the widespread pattern of pollen dispersal is likely due to the significantly higher plant density in the seed orchard compared to natural populations, and the lower position of male and female cones on dwarfed trees compared to non-dwarfed trees. Studies show that the outcrossing rate in Cupressus funebris seed orchards is lower than in other conifers. This suggests that the genetic diversity of parental clones in the seed orchard isn’t being effectively and stably transmitted. Even with a fixed - clone design, there's a risk of inbreeding and pollen contamination. The flowering asynchrony among parents exacerbates this (Li et al. 2012; Luc 2019). Also, the low density of flowering plants in the seed orchard is a significant factor increasing the inbreeding rate (Tania et al. 2024). Parental flowering synchrony, flower quantity, and selective fertilization may cause non - random mating and inbreeding. Importantly, the seed orchard has been subjected to topping and pruning. The maximum and average pollen dispersal distances in non - dwarfed plots are greater than those in dwarfed plots. Thus, dwarfing reduces effective pollen dispersal and increases self - pollination risk. This implies that more attention should be paid to parental clone arrangement and spacing in dwarfed seed orchards. Pollen dispersal and paternal reproductive success are key for seed orchard management (Wang et al. 2023; Martins et al. 2025). To boost seed yield and quality, a "wide - row, close - plant" spacing is more suitable for conifers (El-Kassaby et al. 2010). After dwarfing, the fruit - bearing layer shifts downward, and male/female cones enter a "relatively dense zone." So, more pollination space is needed to reduce inbreeding risk. 4.3 Effects of Flowering Synchrony and Climatic Factors During flowering, synchrony among individuals impacts gene flow and may be influenced by changing climatic conditions (Pearse et al. 2015; Tania et al. 2024). Low synchrony may reduce resource competition among clones but decreases pollination efficiency and affects offspring genetic gain (Wang et al. 2023). High synchrony can enhance pollination efficiency through resource sharing (Li et al. 2012). However, over the three - year observation period, overall synchrony in the seed orchard was low. Differences in flowering synchrony among clones and between years were observed, indicating that varying flowering times among clones is common in forest tree seed orchards. This asynchrony can reduce effective parental mating combinations, increase self - pollination and external pollen contamination, and thus lower the genetic gain of seeds (Wang et al. 2010). The flowering synchrony index is closely related to environmental factors such as temperature, rainfall, and altitude. In years with low temperatures and high rainfall (e.g., 2024 and 2025), the flowering synchrony index was higher. This is because adverse weather conditions slowed the flowering process of the clones. Those clones that were ready or already in the flowering stage experienced a longer flowering period due to the impact of the weather. The cold and rainy weather typically lasted for 5 - 8 days. Once the weather improved, the male and female strobili of the clones opened in a more concentrated manner, leading to a relatively higher flowering synchrony index. In contrast, in sunny years (e.g., 2021), the flowering synchrony was lower. This is attributed to the fact that under favorable weather conditions, clones capable of pollen shedding or female flower pollination completed their flowering process in a shorter time frame, making clonal differences more pronounced. This finding differs from studies on trees like Pinus radiata , which generally suggest that high temperatures and humidity reduce synchrony (Pederick and Brown 2013; Beren et al. 2020). Therefore, the flowering management of Cupressus funebris seed orchards should be tailored to the local climate. Monitoring meteorological data during the flowering period, forecasting changes in synchrony, and implementing measures such as assisted pollination or flowering induction can enhance pollination efficiency (Codesido and Merlo 2005). 5 Conclusions In Cupressus funebris dwarfed seed orchards, gamete contributions are unbalanced. The genetic composition of offspring is jointly influenced by flowering synchrony, parental contributions, and pollen dispersal patterns, with clonal differences playing a leading role. Climatic factors also have complex effects on flowering synchrony. Although dwarfing does not significantly alter pollen dispersal distances, it lowers the fruit - bearing layer, causing male and female cones to enter a "relatively dense zone." This requires more extensive pollen cloud dispersal to reduce inbreeding risks. Consequently, in Cupressus funebris dwarfed seed orchards, clone selection should be optimized based on cone quantities, phenology, and parent origins. Additionally, greater attention should be paid to branch management and plant spacing to enhance ventilation and light penetration. Combined with flowering - period management, such as assisted pollination and phenological regulation, these measures can improve the genetic gain and stability of seed orchards. Declarations Acknowledgments The authors are deeply grateful to the Research Group of Forest Tree Genetic Breeding and Cultivation of the Research Institute of Subtropical Forestry, Chinese Academy of Forestry for their great assistance in laboratory aspects. Author contributions The roles and contributions of each author must be described in the subsequent manner: The authors confirm contribution to the paper as follows: study conception and design: Z Z; data collection: H-B Z, S C, G-Q J and Z-P F; analysis and interpretation of results: Z Z, H-B Z, S C, G-Q J and Z-C Z; draft manuscript preparation: Z Z, H-B Z, S C. All authors reviewed the results and approved the final version of the manuscript. An author name can appear multiple times, and each author name must appear at least once. Funding This study was funded by the project “Research on intelligent and high-yield management technology for seed orchards of southern coniferous tree species” of the National Key R&D Program of China during the 14th Five-year Plan period (2023YFD2200601-4), and the project “Breeding of new high-carbon-sequestration and high-quality timber tree varieties” of the Zhejiang Provincial Forest Tree New Variety Breeding Program during the 14th Five-year Plan period(2021C02070-8). Data availability The data and materials used in this study are available upon reasonable request from the corresponding authors. Ethics approval and consent to participate Not applicable. Consent for publication All authors gave their informed consent to this publication and its content. Competing interests The authors declare that they have no competing interests. References Allender C (2011) The Second Report on the State of the World's Plant Genetic Resources for Food and Agriculture. Exp Agr 47:574. https://doi.org/10.1017/s0014479711000275 Armstrong C, Ganasamurthy S, Wigley K, et al. (2024) The microorganisms and metabolome of Pinus radiata Pollen. Environ microbiome 19: 103. https://doi.org/10.1186/s40793-024-00656-4 Bruna IA, Miguel LMF, Antonio SZ, et al. (2020) The effects of pollen dispersal and mating pattern on inbreeding depression and hybrid vigor in Balfourodendron riedelianum (Engl.) Engl. (Rutaceae). Conserv Genet 21:305–317. https://doi.org/10.1007/s10592-020-01250-0 Barrett JW, Knowles P, Cheliak WM. (2011) The mating system in a black spruce clonal seed orchard. Can J Forest Res 17:379-382. https://doi.org/10.1139/x87-065 Beren S, Richard M, Amir A, et al . (2020) Flowering phenology in a Eucalyptus loxophleba seed orchard, heritability and genetic correlation with biomass production and cineole: breeding strategy implications. Sci Rep-UK 10: 15303. https://doi.org/10.1038/s41598-020-72346-3 Ben SK, Funda T, Chen C, et al. (2008) Pollination dynamics in a Douglas-fir seed orchard as revealed by pedigree reconstruction. Ann Forest Sci 65: 808. https://doi.org/10.1051/forest/2010044 Chatterjee T, Chakrabarty S, Mitra B, et al. (2025) Unveiling the association of mangrove flowering phenology on the dynamics of insect abundance. Acta Oecologica 126:104052. https://doi.org/10.1016/j.actao.2024.104052 Chen XB, Sun X, Dong L, et al. (2018) Mating patterns and pollen dispersal in a Japanese larch (Larix kaempferi) clonal seed orchard: a case study. Sci China Life Sci 61:1011-1023. https://doi.org/10.1007/s11427-018-9305-7 Chen SF, Wataru I, Yamato U, et al. (2022) Pollen dispersal patterns and male reproductive success based on pedigree reconstruction in a hybrid larch ( Larix gmelinii var. Japonica × L. kaempferi ) open‑pollinated seed orchard. Tree Genet Genomes 18:33. https://doi.org/10.1007/s11295-022-01566-7 Chaloupková K, Lstibůrek M. (2022) Spatial optimization of genetic thinning in seed orchards. Ann Forest Sci 79:37. https://doi.org/10.1186/s13595-022-01158-4 Codesido V, Merlo EFJ. (2005) Variation in Reproductive Phenology in a Pinus radiata D. Don Seed Orchard in Northern Spain. Silvae Genet 54:246-256. https://doi.org/10.1515/sg-2005-0035 Du YJ, Zhang RC, Tang XR et al. (2024) The mid-domain effect in flowering phenology. Plant Diversity 46:502-509. https://doi.org/10.1016/j.pld.2024.05.005 Darlin UGZ, Alexandre MS, Bruno CR. (2024) Mating system and inbreeding depression in Hymenaea stigonocarpa . Tree Genet Genomes 20: 6. https://doi.org/10.1007/s11295-024-01639-9 El-Kassaby YA, Funda T, Lai BSK (2010) Female reproductive success variation in a Pseudotsuga menziesii seed orchard as revealed by pedigree reconstruction from a bulk seed collection. J Hered 101:164-168. https://doi.org/10.1093/jhered/esp126 Funda FT, Liewlaksaneeyanawin L C, Fundova F I, et al. (2011) Congruence between parental reproductive investment and success determined by DNA-based pedigree reconstruction in conifer seed orchards. Can J Forest Res 41:380-389. https://doi.org/10.1139/x10-190 Funda T. (2012) Seed orchard genetics. Forest Sci 59: 1-12. https://doi.org/10.1079/pavsnnr20127013 Gömöry D, Bruchánik R, Longauer R. (2003) Fertility variation and flowering asynchrony in Pinus sylvestris: consequences for the genetic structure of progeny in seed orchards. For Ecol Manage 174:117-126. https:// doi. org/ 10. 1016/ S0378- 1127(02) 00031-2 Heuchel A, Hall D, Zhao W, et al. (2022) Genetic diversity and background pollen contamination in Norway spruce and Scots pine seed orchard crops. Forestry research 2: 67-78. https://doi.org/10.48130/fr-2022-0008. Heike L, Dagmar S. (2022) Flowering behavior of clones in a Norway maple ( Acer platanoides ) seed orchard and mating system analysis using nuclear SSR markers. Eur J Forest Res 141:561-569. https://doi.org/10.1007/s10342-022-01459-3 Han S, Kang K, Kim C, et al. (2013) Effect of top-pruning in a clonal seed orchard of Pinus koraiensis . Ann Forest Sci 51:155-156. https://doi.org/10.15287/afr.2008.157 Huntsman SV, Leslie AB .The ontogeny of disparity in Cupressaceae seed cones. New Phytol 242:2301-2311. https://doi.org/10.1111/nph.19482 Huang L, Song J, Sun Y, et al. (2018) Pollination dynamics in a Platycladus orientalis seed orchard as revealed by partial pedigree reconstruction. Can J Forest Res 48:952-957. https://doi.org/10.1186/s40793-024-00656-4 Kolpak ES, Smith J, Albrecht JM, et al. (2015) High-density miniaturized seed orchards of Douglas-fir. New For 46:121-140. https://doi.org/10.1007/s11056-014-9452-6 Li W, Wang X R, Li Y. (2012) Variation in Floral Phenological Synchronization in a Clonal Seed Orchard of Pinus tabuliformis in Northeast of China. Silvae Genet 61:133-142. https://doi.org/10.1515/sg-2012-0017 Lindgren D, Prescher F. (2005) Optimal clone number for seed orchards with tested clones. Silvae Genet 54:80-92. https://doi.org/10.1515/sg-2005-0013 Lisa A, Keith W. (2017) Pollen gene flow, male reproductive success, and genetic correlations among offspring in a northern red oak ( Quercus rubra L.) seed orchard. PLoS ONE 12:e0171598. https://doi.org/10.1371/journal.pone.0171598 Lee K, Kim IS, Kang KS (2022) Pedigree reconstruction and spatial analysis for genetic testing and selection in a Larix kaempferi (Lamb.) Carrière plantation. BMC Plant Biol 22:152. https://doi.org/10.1186/s12870-022-03530-y Luc EP. (2019) Hybrid breakdown and inbreeding depression for seed yield and early growth in 2nd-generation interspecific hybrids of larch ( Larix. × eurolepis × L. × eurolepis ). Ann Forest Sci 76:1-10. https://doi.org/10.1007/s13595-019-0853-8 Martins AE, Loiola PdP, Pareja-Bonija D, et al. (2025). Climate-induced shifts in long-term tropical tree reproductive phenology: Insights from species dependent on and independent of biotic pollination. Funct Ecol 00:1-13. https://doi.org/10.1111/1365-2435.70090 Martins G, Yuliarto M, Melia T, et al. (2025) Pollination and mating dynamics unveiled by orchard-wide pedigree reconstruction in Acacia crassicarpa. Tree Genet Genomes 21:7. https://doi.org/10.1007/s11295-025-01692-y Martins AE, Camargo MGG, Morellato LPC. (2021) Flowering Phenology and the Influence of Seasonality in Flower Conspicuousness for Bees. Front Plant Sci 11:1-11. https://doi.org/10.3389/fpls.2020.594538 Nyabera LA, Nzuki IW, Runo SM, et al . (2021) Assessment of genetic diversity of pumpkins( Cucurbita spp.) from western Kenya using SSR molecular markers. Mol Biol Rep 48:2253-2260. https://doi.org/10.1007/s11033-021-06245-9 Primack RB. (2003) Relationships among flowers, fruits, and seeds. Annu Rev Ecol Syst 18:409-430. https://doi.org/10.1146/annurev.es.18.110187.002205 Paul JC, Amy MI, David WI. (2014) Shifts in flowering phenology reshape a subalpine plant community. Proc Natl Acad Sci USA 111: 4916-4921. https://doi.org/10.1073/pnas.1323073111 Pearse IS, Koenig WD, Funk KA, et al. (2015) Pollen limitation and flower abortion in a wind-pollinated, masting tree. Ecology 96:587-593. https://doi.org/10.1890/14-0297.1 Pederick AL, Brown GA. (2013) Seed Production in Radiata Pine Seed Orchards in Australia. Aust For 39:164-179. https://doi.org/10.1080/00049158.1976.10674149 Ruotsalainen S. (2014) Increased forest production through forest tree breeding. Scand J For Res 29:333-344. https://doi.org/10.1080/02827581.2014.926100 Song J, Ratcliffe B, Kess T, Lai BS, Korecký J, et al. (2018) Temporal quantification of mating system parameters in a coastal Douglasfir seed orchard under manipulated pollination environment. Sci Rep 8:11593. https://doi.org/10.1038/s41598-018-30041-4 Torimaru T, Wennström U, Lindgren D, et al. (2012) Effects of male fecundity, interindividual distance and anisotropic pollen dispersal on mating success in a Scots pine ( Pinus sylvestris ) seed orchard. Heredity 108:312-321. https://doi.org/10.1038/hdy.2011.76 Tania DF, Katharina BB, John EC, et al. (2024) Distance-dependent mating but considerable pollen immigration in an isolated Quercus rubra planting in Germany. Eur J Forest Res 143:1447-1460. https://doi.org/10.1007/s10342-024-01704-x Wang F, Zhang SY, Zhu P, et al. (2023) The effects of fertility and synchronization variation on seed production in two Chinese fir clonal seed orchards. Sci Rep 13:627. https://doi.org/10.1038/s41598-022-27151-5 Wang XY, Morin X, Zhang J, et al. (2023) Geographical patterns and determinants in plant reproductive phenology duration. Front Plant Sci 14:1-8. https://doi.org/10.3389/fpls.2023.1199316 Wang XY, Bjorkman AD, Li X, et al . (2025) Shifts in plant reproductive phenology induced by multiple global change factors depend on phenological niche andpollination mode. J Plant Ecol 18:rtaf048. https://doi.org/10.1093/jpe/rtaf048 Wang F, Zhang SY, Zhu P, et al . (2023) The effects of fertility and synchronization variation on seed production in two Chinese fir clonal seed orchards. Sci Rep 13:627. https://doi.org/10.1038/s41598-022-27151-5 Wang XR, Torimaru T, Lindgren D, et al . (2010) Marker-based parentage analysis facilitates low input ‘breeding without breeding’ strategies for forest trees. Tree Genet Genomes 6:227-235. https://doi.org/10.1007/s11295-009-0243-8 Wu YQ, Weng YH, Hennigar C, et al. (2015) Benefit–cost analysis of a white spruce clonal seed orchard in New Brunswick. Canada. New Forests 46, 141–156. https://doi.org/10.1007/s11056-014-9453-5. Yang BN, Su NH, Qi JD, et al. (2020) Improve dgenetic distance-based spatial deployment can effectively minimize inbreeding in seed orchard. For Ecosyst 7:1-11. https://doi.org/10.1186/s40663-020-0220-0 Yang T, Wang PC, Wang WY, et al. (2023) Early growth evaluation and biomass allocation difference between clones and families in Cupressus funebris . Eur J For Res 142:839-850. https://doi.org/10.1007/s10342-023-01563-y Yang ML, Liu JC, Gao MK, et al . (2025) Pollen limitation influences the divergences in mating system and floral traits between two sympatric Halenia species. J Plant Ecol 18:rtae098. https://doi.org/10.1093/jpe/rtae098 Zoe D, Anita C, Kathleen MK, et al. (2023) Reproductive strategies and their consequences for divergence, gene flow, and genetic diversity in three taxa of Clarkia. Heredity 131: 338-349. https://doi.org/10.1038/s41437-023-00649-y Tables Table 1 Paternity Analysis of Open-Pollinated Progeny Type PIR /% SR /% OR /% NUAIP PCFP /% DSO 70.89 27.81 72.19 8 5.80 USO 76.67 18.30 81.70 12 8.00 DSO represents the dwarf seed orchard, USO represents the undwarfed seed orchard, PIR represents the paternity identification rate, SR represents the selfing rate, OR represents the outcrossing rate, NUAIP represents the number of unique alleles in progeny, and PCFP represents the proportion of contaminated foreign pollen. Table 2 Genetic Diversity of Parental and Progeny Populations in the 1.5-Generation Cypress Clonal Seed Orchard Locus Na Ne I Ho He F Parent Progeny Parent Progeny Parent Progeny Parent Progeny Parent Progeny Parent Progeny 1-C95 6 8 2.608 3.356 1.164 1.352 0.111 0.046 0.617 0.702 0.820 0.935 C83 8 10 7.401 8.310 2.037 2.198 0.556 0.441 0.865 0.880 0.358 0.499 CYP52 3 3 2.527 2.920 0.989 1.085 0.852 0.739 0.604 0.657 -0.410 -0.125 CYP84 7 8 3.837 4.835 1.515 1.685 0.630 0.736 0.739 0.793 0.148 0.073 CYP174 11 12 7.290 8.451 2.164 2.279 0.667 0.847 0.863 0.882 0.227 0.040 CYP293 7 7 3.574 3.458 1.492 1.474 0.926 0.877 0.720 0.711 -0.286 -0.234 F015 6 6 3.170 2.158 1.407 0.973 0.667 0.096 0.684 0.537 0.026 0.822 F042 9 9 2.899 3.229 1.509 1.533 0.630 0.602 0.655 0.690 0.039 0.129 JT04 4 4 2.690 2.591 1.125 1.029 0.519 0.115 0.628 0.614 0.175 0.813 JT05 3 4 1.561 2.156 0.630 0.932 0.444 0.789 0.359 0.536 -0.237 -0.472 YW03 4 4 1.911 1.834 0.822 0.863 0.593 0.521 0.477 0.455 -0.243 -0.146 CF02 4 4 2.876 3.068 1.194 1.240 0.852 0.621 0.652 0.674 -0.306 0.079 Mean 6.000 6.583 3.529 3.864 1.337 1.387 0.620 0.536 0.655 0.678 0.026 0.201 Na denotes the number of alleles; Ne denotes the effective number of alleles; I is Shannon’s diversity index; Ho is the observed heterozygosity; He is the expected heterozygosity; F is the fixation index. Additional Declarations No competing interests reported. Supplementary Files SupplementaryFile.docx 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-7253675","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":506821048,"identity":"7423f656-c96d-4d28-af23-110c32427e8d","order_by":0,"name":"Hao-bo Zhao","email":"","orcid":"","institution":"Chinese Academy of Forestry","correspondingAuthor":false,"prefix":"","firstName":"Hao-bo","middleName":"","lastName":"Zhao","suffix":""},{"id":506821050,"identity":"d6b184d2-5bae-47fe-91fc-f11dd796186d","order_by":1,"name":"sen Cao","email":"","orcid":"","institution":"Chinese Academy of Forestry","correspondingAuthor":false,"prefix":"","firstName":"sen","middleName":"","lastName":"Cao","suffix":""},{"id":506821051,"identity":"4c038ea7-0cb5-4b48-80f0-248aed612008","order_by":2,"name":"zhen Zhang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBElEQVRIiWNgGAWjYDACCSBmbJCQ42dvbP+QUGHDw8/fQJQWC2PJnsPHGB6cSZORnHGAKC0ViRtuuKUxPmw5bGPQkIBfh/zs5mMPf+6QSJw5g8fsQWLDeR4DhgOMHz7m4NbCOOdYuoHkGQnjfukec4PEHbd5zJkbmCVnbsOthVkix0zCsE1CduacMwYSiWdu81g2HGBj5sWjhU0i/5tEYpsE44YbOUAtbed4DA4k4NfCI5HDJnGwTUJxw420NKCWA4S1SEikmUk2tkmAAvmwQcKZZB7JGQeb8fpFfkbyM8mfbXWgqGx8+KPCzp6fv/ngh494tGADjA2kqR8Fo2AUjIJRgAEAh99Xbf2tSnwAAAAASUVORK5CYII=","orcid":"","institution":"Chinese Academy of Forestry","correspondingAuthor":true,"prefix":"","firstName":"zhen","middleName":"","lastName":"Zhang","suffix":""},{"id":506821053,"identity":"e10eb187-c4c2-4bcc-9ecf-897271d7c4dc","order_by":3,"name":"zhi-chun Zhou","email":"","orcid":"","institution":"Chinese Academy of Forestry","correspondingAuthor":false,"prefix":"","firstName":"zhi-chun","middleName":"","lastName":"Zhou","suffix":""},{"id":506821055,"identity":"d3114e03-eb4c-46b5-9add-af710ee127fe","order_by":4,"name":"guo-qing Jin","email":"","orcid":"","institution":"Chinese Academy of Forestry","correspondingAuthor":false,"prefix":"","firstName":"guo-qing","middleName":"","lastName":"Jin","suffix":""},{"id":506821057,"identity":"f8e9ae26-87e1-4d59-8348-892b8697a2b9","order_by":5,"name":"zhong-ping Feng","email":"","orcid":"","institution":"Chun'an County Forestry Farm Company Limited","correspondingAuthor":false,"prefix":"","firstName":"zhong-ping","middleName":"","lastName":"Feng","suffix":""}],"badges":[],"createdAt":"2025-07-30 13:53:45","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7253675/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7253675/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90377993,"identity":"a3aa2dfe-37e8-4514-935b-e17a4dcc0c94","added_by":"auto","created_at":"2025-09-02 06:22:59","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":13083601,"visible":true,"origin":"","legend":"\u003cp\u003eThe seed orchard and its location sampled in this study. DSO represents a dwarfed seed orchard, USO represents an undwarfed seed orchard.\u003c/p\u003e","description":"","filename":"Figure1Theseedorchardanditslocationsampledinthisstudy.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/67608a2c55fdce0027ea616e.png"},{"id":90379292,"identity":"3c22ad74-0163-422b-b133-18f713d097ab","added_by":"auto","created_at":"2025-09-02 06:30:58","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":848493,"visible":true,"origin":"","legend":"\u003cp\u003eDistributions of Parental and Paternal Contributions in the Seed Orchards. (a) represents the contributions of parental clones to reproductive capacity in the dwarf seed orchard over three years; (b) illustrates the effective paternal contributions within both dwarf and undwarfed seed orchards. DSO represents a dwarfed seed orchard, USO represents an undwarfed seed orchard.\u003c/p\u003e","description":"","filename":"Figure2DistributionsofParentalandPaternalContributionsintheSeedOrchards.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/40049495cccb501204c010ef.png"},{"id":90377989,"identity":"c25fffd5-c3d2-4b05-811f-973c113d8dc5","added_by":"auto","created_at":"2025-09-02 06:22:58","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":433550,"visible":true,"origin":"","legend":"\u003cp\u003ePollen Dispersal Distance in Dwarf Seed Orchards (DSO) and Non-dwarf Seed Orchards (USO)\u003c/p\u003e","description":"","filename":"Figure3PollenDispersalDistanceinDwarfSeedOrchardsDSOandNondwarfSeedOrchardsUSO.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/13a0e079484ccb8c3c34933c.png"},{"id":90377994,"identity":"617d68e1-4bce-4011-94ff-7cc401381ab6","added_by":"auto","created_at":"2025-09-02 06:22:59","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":14679158,"visible":true,"origin":"","legend":"\u003cp\u003eDuration of flowering period for each clone over three years. (a) represents the flowering duration of each clone in 2021, with February 15th as Day 1; (b) represents the flowering duration of each clone in 2024, with February 12th as Day 1; (c) represents the flowering duration of each clone in 2025, with February 3rd as Day 1.\u003c/p\u003e","description":"","filename":"Figure4Durationoffloweringperiodforeachcloneoverthreeyears.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/fbc2c046b227e7c85f5d74b7.png"},{"id":90377986,"identity":"e70ef0dc-f856-4706-8a6b-ef14d8504861","added_by":"auto","created_at":"2025-09-02 06:22:58","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1016296,"visible":true,"origin":"","legend":"\u003cp\u003eA shows the flowering duration of \u003cem\u003eCupressus funebris\u003c/em\u003e 1.5 - generation clonal seed orchard over three years, and b shows the distribution frequency of flowering synchrony index ranges among clones. (a) the x-axis is based on February 2nd of each year as Day 1, with subsequent days calculated accordingly. (b) represents the distribution frequency of each clone within the range of the flowering phenological synchrony index. \u003cem\u003eW\u003c/em\u003e\u003csub\u003e\u003cem\u003eij\u003c/em\u003e\u003c/sub\u003e\u003cem\u003e \u003c/em\u003edenotes the pairwise flowering synchrony index for each parental clone when serving as both the male and female parent.\u003c/p\u003e","description":"","filename":"Figure5ashowsthefloweringdurationofCupressusfunebris1.5generationclonalseedorchardoverthreeyearsandbshowsthedistributionfrequencyoffloweringsynchronyindexrangesamongclones.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/5a5b535ac4a43872721083a6.png"},{"id":90379291,"identity":"3d0491dc-7783-49f5-ba76-92d80123b022","added_by":"auto","created_at":"2025-09-02 06:30:58","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":2086744,"visible":true,"origin":"","legend":"\u003cp\u003eFlowering Synchrony Index of Each Clone as Parental and Self-Pollinated Lines Over Three Years\u003c/p\u003e","description":"","filename":"Figure6FloweringSynchronyIndexofEachCloneasParentalandSelfPollinatedLinesOverThreeYears.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/92f659eae188efdf735ff37f.png"},{"id":90377990,"identity":"cd29fb71-be70-4a48-b8a9-32540535ab1d","added_by":"auto","created_at":"2025-09-02 06:22:58","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1012807,"visible":true,"origin":"","legend":"\u003cp\u003eFactorial Analysis of the Main Meteorological Factors and Phenological Phases of Flowering during the Flowering Period. FIF represents the initial flowering period of female cones, FFF represents the full flowering period of female cones, FEF represents the end flowering period of female cones, FTFP represents the total flowering period of female cones, MIF represents the initial flowering period of male cones, MFF represents the full flowering period of male cones, MEF represents the end flowering period of male cones, and MTFP represents the total flowering period of male cones.\u003c/p\u003e","description":"","filename":"Figure7FactorialAnalysisoftheMainMeteorologicalFactorsandPhenologicalPhasesofFloweringduringtheFloweringPeriod.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/de6b5af7e8c6b503eaf950e5.png"},{"id":90377991,"identity":"5d2cb8e4-8261-452d-933e-799c526af5ec","added_by":"auto","created_at":"2025-09-02 06:22:59","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":3045507,"visible":true,"origin":"","legend":"\u003cp\u003eAnnual variations in precipitation, mean daily temperature, and the ratio of male to female cone flowering during the flowering period. (a) , (c) , and (e) indicate the flowering ratio of female-to-male cones in relation to precipitation for the years 2021, 2024, and 2025, respectively; (b) , (d) , and (f) indicate the flowering ratio of female-to-male cones in relation to daily mean temperature for the same years.\u003c/p\u003e","description":"","filename":"Figure8Annualvariationsinprecipitationmeandailytemperatureandtheratioofmaletofemaleconefloweringduringthefloweringperiod.png","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/d431552a547983aeb1345cc1.png"},{"id":97366684,"identity":"4b7d9795-95ce-4816-9b55-1dde8b77ca6f","added_by":"auto","created_at":"2025-12-03 15:57:43","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":34071218,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/78f2dc9d-c2e0-4ca0-9c7c-29780585b40a.pdf"},{"id":90377988,"identity":"8938a22f-02fb-440b-920d-3d1da490374e","added_by":"auto","created_at":"2025-09-02 06:22:58","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":13496,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFile.docx","url":"https://assets-eu.researchsquare.com/files/rs-7253675/v1/fb9d4e5d54e391083b1b7a02.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003ePhenological Synchrony, Parental Contribution, and Pollen Dispersal Pattern affects the Genetic Composition of \u003cem\u003eCupressus funebris\u003c/em\u003e Dwarf Seed Orchard Progenies\u003c/p\u003e","fulltext":[{"header":"Key Message","content":"\u003cp\u003eThis study demonstrates that the low flowering synchrony in dwarf seed orchards is primarily attributable to inter-clonal variation and to temperature and precipitation conditions during the anthesis period. Although the progeny population of dwarf orchards retains the high genetic diversity of the parental clones, unbalanced gametic contributions within the orchard predispose the offspring to increased self- or inbreeding. Moreover, compared with non-dwarf orchards, pollen dispersal distances in dwarf orchards are significantly reduced. These findings underscore the necessity for improving clonal selection, spatial arrangement, and shoot growth management in dwarf seed orchards.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"},{"header":"1 Introduction","content":"\u003cp\u003eSeed orchards, special - purpose tree populations for seed production, are the \"output delivery system\" of breeding populations (Allender 2011; Li et al. 2012; Heuchel et al. 2022). For conifers lacking clonal propagation materials, seed orchards link genetic improvement with artificial forest establishment. They contain the best individuals (breeding values), and theoretically, their offspring should have high genetic gain and diversity to enhance major economic traits or adaptability. This is crucial for global seed production programs\u0026nbsp;(El-Kassaby and Reynolds 1990;\u0026nbsp;Ruotsalainen 2014).\u003c/p\u003e\n\u003cp\u003eTo produce seeds in a predictable manner, in open - pollinated seed orchards, balanced mating probabilities between all parents, low - level inbreeding, and random mating are expected. Each parent should contribute equally to the offspring, ensuring a broad genetic base and the expected genetic value (Lindgren and Prescher 2005;\u0026nbsp;Chen et al. 2018). This requires several conditions: (a) all parents provide equal numbers of male and female gametes for seed production; (b) female flower fertilization timing is synchronized with pollen dispersal; (c) the seed orchard parents have sufficient genetic diversity to pass on to the offspring; and (d) the seed orchard is isolated from natural stands of the same species to prevent external pollen contamination. As observed in many tree breeding programs, these expectations for seed orchards are rarely fully met (Funda et al. 2011; Torimaru et al. 2012;\u0026nbsp;Lisa and Keith 2017, Song et al. 2018; Bruna\u0026nbsp;et al. 2020). Parental phenology, pollen quantity, and seed yield in seed orchards can vary significantly among parent sources and genotypes. Parental contributions to the genetic pool of the offspring are often unbalanced, potentially leading to a loss of genetic diversity (Heike and Dagmar 2022;\u0026nbsp;Chatterjee et al. 2025). When selecting parents for seed orchards, it is essential to optimize a large number of parental materials and evaluate offspring genetic traits to maximize gains while maintaining high diversity levels (Yang et al. 2020; Chaloupková and Lstibůrek 2022; Lee\u0026nbsp;et al. 2022).\u003c/p\u003e\n\u003cp\u003ePhenology is a key part of a plant's life history and a major fitness factor that affects reproductive success (Primack 2003; Paul et al. 2014;\u0026nbsp;Du et al. 2024; Martins et al. 2025). Differences in phenological variation and flowering synchrony can impact the frequency of gene exchange between parents and the genetic composition of seeds in seed orchards. These differences can also reduce effective population size, cause year - to - year changes in mating systems, increase self - pollination probabilities, and raise the risk of external pollen contamination (Barrett et al. 2011;\u0026nbsp;Darlin\u0026nbsp;et al. 2024). These issues are particularly evident in conifer seed orchards where wind is the main pollen dispersal agent (Li et al. 2012; Heuchel et al. 2022;\u0026nbsp;Wang et al. 2023). In conifers, 34% - 52% of parents typically contribute to 80% of the offspring in seed orchards, showing a clear imbalance (Wang et al. 2023; Martins et al. 2025). Pollen dispersal and female cone flower pollination usually occur within two to three weeks. During this time, factors such as rainfall, wind direction, and wind speed can become critical, alter random mating in seed orchards, and increase pollen contamination (Pearse et al. 2015;\u0026nbsp;Tania\u0026nbsp;et al. 2024).\u003c/p\u003e\n\u003cp\u003eIn recent years, tree dwarfing has been used to address the problem of tall trees in seed orchards and difficulties in seed collection. It's an important part of modern intensive seed orchard management (Han et al. 2013; Wu et al. 2015; Wang et al. 2025). Methods like topping, pruning, girdling, and hormone induction are used to suppress apical dominance and control upward growth. This makes it easier to harvest cones and conduct artificial pollination. However, it also causes female and male cone flowers to shift downward (Kolpak et al. 2015). But tree - breeding programs rarely focus on flower - related management in dwarfed seed orchards. It's still unclear whether pollen and mating patterns change in these orchards. This information is crucial for making genetic management decisions in seed orchards, such as dwarfing, thinning, artificial pollination, and clone arrangement (Chen et al. 2022; Tania et al. 2024).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eCupressus funebris\u003c/em\u003e is a key afforestation and ecological tree species in southern China. It has excellent wood quality, strong adaptability, and can tolerate drought and poor soils, making it highly effective for afforestation and ecological restoration in barren mountains (Yang et al. 2023). However, significant clonal differences in reproductive capacity and flowering phenology have been observed in \u003cem\u003eCupressus funebris\u003c/em\u003e. Factors such as multiple branches, poor ventilation and light penetration in the inner canopy, and adverse conditions like temperature drops and rainfall during the flowering period may affect mating probabilities among parents in dwarfed seed orchards (Huntsman and Leslie 2023; Martins et al. 2021). To ensure seed quality for afforestation and develop rational genetic management strategies for dwarfed seed orchards, long - term monitoring of reproductive phenology and flowering synchrony within seed orchards is essential. It is also crucial to evaluate the impact of these factors on seed production, as well as pollen dispersal patterns and mating systems among parents. The objectives of our study are to: 1) Characterize differences in flowering synchrony and identify major climatic factors in dwarfed \u003cem\u003eCupressus funebris\u003c/em\u003e seed orchards. 2) Assess parental contributions to the genetic pool of offspring. 3) Examine changes in mating mechanisms and pollen dispersal patterns following tree management practices. Our findings offer new insights into the mating mechanisms of dwarfed seed orchards and provide valuable references for flower - related management practices.\u003c/p\u003e"},{"header":"2 Material and methods","content":"\u003cp\u003e\u003cstrong\u003e2.1 Study Area and Material Sources\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study materials were obtained from a \u003cem\u003eCupressus funebris\u003c/em\u003e seed orchard located in Chun'an County, Zhejiang Province (119°03'57″ E, 29°32'24″N). The orchard covers an area of 3.7 hm², with an altitude ranging from 250 to 300 m. It is situated on a gentle slope with a gradient of less than 15°, facing south - southeast. The soil conditions of the seed orchard were determined, with mountain red soil (pH 4.74) containing 20.08 g/kg organic matter, 1.12 g/kg total nitrogen, 0.49 g/kg total phosphorus, 95.87 mg/kg alkaline - hydrolyzed nitrogen, 283.92 mg/kg available phosphorus, 93.36 mg/kg available potassium, 212.90 mg/kg exchangeable calcium, and 108.55 mg/kg total boron.\u003c/p\u003e\n\u003cp\u003eThe improved seed orchard was established in 2015. After genetic testing of the parental materials from the first - generation seed orchard (established in 1990) with 45 parents, 27 genotypes were retained to form this improved clonal seed orchard. The seed orchard is divided into five blocks, with a total of 1,680 trees planted. The clones are arranged in a random spatial arrangement within the orchard. Each clone is repeated 10 - 15 times within a plot, and the distance between ramets of the same clone is over 20 m, with a tree spacing of 4 m × 4 m. A pollen - free zone of over 300 m surrounds the seed orchard. The seed orchard first flowered and produced cones in 2018. In 2020, the mother trees in blocks 2 - 5 underwent topping to facilitate cone harvest and pollination management, forming the Dwarfed Seed Orchard (DSO). Specifically, the top 1 - 2 whorls were cut, with the cutting height controlled at 2.5 m, with no further treatment (Fig. 1) . Block 1 was undwarfized seed orchards (USO). The management of both seed orchards remained consistent.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2 Phenological Observation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePhenological observations were conducted during the flowering stage in 2021, 2024, and 2025. For each clone, five ramets were selected. The following method was used: one first - order branch in the middle - upper part of the canopy was chosen in each of the four directions (east, south, west, and north). These branches were observed daily until the end of the flowering period (Li et al. 2012) .The flowering phases were determined as follows: (1) When 0% - 30% of female cone scales crack or 0% - 30% of male cones shed pollen, it's the onset of flowering. (2) When 30% - 75% of female cone scales crack or 30% - 75% of male cones shed pollen, it's full flowering. (3) The end of flowering for female cones is when 75% - 100% of female cone scales close, thicken, and turn dark purple - red. For male cones, it's when 75% - 100% of pollen is shed, and the flower branches turn yellowish - brown and wilt. The entire flowering period, which included the beginning, full, and end of flowering phases, was recorded. At the end of the flowering period, the number of female cone flowers and male cone flower spikes on each observed branch were recorded (Beren et al. 2020).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3 Sampl\u003c/strong\u003e\u003cstrong\u003ees of mating systems\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSample collection was carried out in two parts. In the first part, in May 2023, we collected the current year's needles from 27 parents in the dwarfed seed orchard and stored them in a -80 ℃ freezer for DNA extraction. During the cone maturation period, we collected cones from all clones and processed the seeds for sowing and seedling cultivation. When the seedlings reached a height of 20 cm, we randomly selected 20 seedlings from each clones, forming a total of 540 offspring individuals for genetic diversity analysis between parental and offspring generations in the seed orchard.\u003c/p\u003e\n\u003cp\u003eThe second part involved selecting five identical clones from both non - dwarfed and dwarfed plots. We marked the locations of the mother trees and collected current - year needles. Upon cone maturation, we collected cones from the middle - upper part of the tree, air - dried them in the laboratory, and gathered over 1,000 seeds per tree. After sowing and seedling cultivation, 30 offspring seedlings were randomly chosen from each mother tree, forming 150 offspring individuals. Young tissues were collected and stored in a -80 ℃ freezer for DNA extraction. These samples were used for paternal analysis and pollen dispersal pattern research in dwarfed and non - dwarfed seed orchards.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4 DNA Extraction, Primer Screening, and SSR-PCR Amplification\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDNA was extracted from parent trees and seedling progeny in the seed orchard using an improved CTAB plant genomic DNA extraction kit (Aidlab, Beijing, China), and DNA quality and concentration were measured using a NanoDrop 2000 spectrophotometer. For parent - offspring relationship analysis and genetic diversity assessment, 12 SSR markers were used, with primer information listed in Table S1. PCR amplification and detection followed previously described methods (Yang et al. 2016). Parent and offspring DNA samples were amplified with primers, and the amplified products were separated by electrophoresis on a Qsep100\u003csup\u003eTM\u003c/sup\u003e automatic nucleic acid protein analysis system (BIOptic, Taiwan).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5 Genetic Diversity Parameter Calculation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter exporting and converting the genotyping data from the Qsep100TM system, genetic diversity parameters were calculated using GenAlex 6.5. These parameters included the number of alleles (\u003cem\u003eNa\u003c/em\u003e), effective number of alleles (\u003cem\u003eNe\u003c/em\u003e), Shannon’s diversity index (\u003cem\u003eI\u003c/em\u003e), observed heterozygosity (\u003cem\u003eHo\u003c/em\u003e), expected heterozygosity (\u003cem\u003eHe\u003c/em\u003e), fixation index (\u003cem\u003eF).\u0026nbsp;\u003c/em\u003ePolymorphism information content (\u003cem\u003ePIC\u003c/em\u003e) for microsatellite loci were assessed using Cervus 3.0 (Zoe et al. 2023).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6 Paternity Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCERVUS 3.0 was used to conduct parentage analysis of free - pollinated offspring within a 70% - 80% confidence interval, determining paternal contributions. Paternity Identification Rate (\u003cem\u003ePIR\u003c/em\u003e), outcrossing rate (\u003cem\u003eOCR\u003c/em\u003e), selfing rate (\u003cem\u003eSR\u003c/em\u003e), Number of Unique Alleles in Progeny (\u003cem\u003eNUAIP\u003c/em\u003e) and pollen contamination rate by foreign pollen (\u003cem\u003ePCFP\u003c/em\u003e) were calculated for non - dwarfed and dwarfed plots. Pollen dispersal distance was also calculated based on the actual distances between parents in the seed orchard (Heike and Dagmar 2022).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.7 Statistical Analysis of Flowering Synchrony\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe flowering synchrony among parents as male, female, and self - pollination was analyzed using the flowering synchrony index (\u003cem\u003eW\u003csub\u003eij\u003c/sub\u003e\u003c/em\u003e) proposed by Li et al. (Li et al.2012).\u003c/p\u003e\n\u003cp\u003e\u003cimg width=\"624\" height=\"52\" src=\"data:image/png;base64,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\" alt=\"image\"\u003e\u003c/p\u003e\n\u003cp\u003eIn the formula,\u0026nbsp;\u003cem\u003eW\u003csub\u003eij\u0026nbsp;\u003c/sub\u003e\u003c/em\u003eis the flowering synchrony index between the male cones of the \u003cem\u003ei\u003c/em\u003e-th clone and the female cones of the \u003cem\u003ej\u003c/em\u003e-th clone;\u0026nbsp;\u003cem\u003eM\u003csub\u003eki\u003c/sub\u003e\u003c/em\u003e is the flowering frequency of the male cones of the \u003cem\u003ei\u003c/em\u003e-th clone on day \u003cem\u003ek\u003c/em\u003e;\u0026nbsp;\u003cem\u003eP\u003csub\u003ekj\u003c/sub\u003e\u003c/em\u003e is the flowering frequency of the female cones of the \u003cem\u003ej\u003c/em\u003e-th clone on day \u003cem\u003ek\u003c/em\u003e; and \u003cem\u003en\u003c/em\u003e is the number of days from the earliest flowering start to the latest flowering end between clones \u003cem\u003ei\u003c/em\u003e and \u003cem\u003ej\u003c/em\u003e. When \u003cem\u003ei\u0026nbsp;\u003c/em\u003e= \u003cem\u003ej\u003c/em\u003e, the index represents the self - pollination flowering synchrony index\u0026nbsp;\u003cem\u003eW\u003csub\u003ei\u003c/sub\u003e\u003c/em\u003e. A\u0026nbsp;\u003cem\u003eW\u003csub\u003eij\u003c/sub\u003e\u003c/em\u003e value of 1 means complete overlap of male and female cone flowering periods, while a value of 0 means no overlap.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.8 Collection and Analysis of Climatic Data\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA micro - automatic weather station was used to record weather factors during the flowering period. The average high temperature (AHT) was the mean of the highest daily temperatures. The average low temperature (ALT) was the mean of the lowest daily temperatures. DMA stands for daily average temperature. Rainfall amount (RA) was the sum of daily rainfall. The number of rainy days (NRD) was counted. Average wind speed (AWS) was the mean of wind speeds, and average humidity (AH) was the mean of humidity levels during the flowering period.\u003c/p\u003e"},{"header":"3 Results","content":"\u003cp\u003e\u003cstrong\u003e3.1 Reproductive Capacity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDuring the three observation years, in the dwarfed seed orchard, 51.85% to 66.67% of clones contributed over 85% of female cones, and 44.44% to 51.85% of clones contributed over 85% of male cones. The parental contribution curves for female and male cones deviated from the ideal (Fig. 2-a), showing unbalanced gamete contributions. Paternal contribution analysis (Fig. 2-b) showed that in the dwarfed plot, paternal contribution ranged from 0.99% to 14.85%, with 23.81% of fathers producing over 50% of the offspring. In the non - dwarfed plot, paternal contribution was between 0.87% and 13.04%, with 24% of fathers producing over 50% of the offspring. Compared to the non - dwarfed plot, the dwarfed plot had fewer effective pollen - producing fathers, reducing gene exchange. Overall, a small number of clones contributed more pollen during reproduction.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.2 Mating Pattern\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe dwarfed seed orchard is mainly characterized by outcrossing, with an outcrossing rate of 72.19%. Among the offspring, 101 individuals (70.89% identification rate) matched with parental clones, and 8 offspring-specific alleles (\u003cem\u003eNa\u003c/em\u003e) were found. The pollen contamination rate was 5.80%. In contrast, the non-dwarfed seed orchard had a higher outcrossing rate of 81.70%, with 115 offspring (76.76% identification rate) matching the parentals and 12 offspring-specific alleles detected. Its pollen contamination rate was 8.00% (Table 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.3 Pollen Dispersal Patterns\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePollen propagation among seed orchard clones is random without a specific pollination direction. The highest proportion of offspring occurs when effective pollen dispersal is 30 - 40 m. In non - dwarfed plots, the average distance between mother trees and pollen donors was 35.03 m, with a maximum pollination distance of 68.82 m and 64.86% of effective pollination within 40 m. In dwarfed plots, the maximum pollination distance was 60.00 m, the average pollination distance was 33.35 m, and 75.76% of effective pollination occurred within 40 m (Fig. 3). Dwarfing reduces the effective pollen dissemination distance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.4 Parental Contribution to the Offspring Gene Pool\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn the parental population of the dwarfed seed orchard, the number of observed alleles (\u003cem\u003eNa\u003c/em\u003e) ranged from 3 to 11, with an average of 6.000, and the average number of effective alleles (\u003cem\u003eNe\u003c/em\u003e) was 3.529. Among 540 offspring, Na ranged from 4 to 11, averaging 6.583, which is 9.717% higher than the parental average. Effective allele numbers were 9.493% higher. Genetic diversity was comparable to the parental population, indicating no reduction in offspring genetic diversity (Table 2). Offspring largely retained parental genetic diversity. However, offspring heterozygosity (\u003cem\u003eHo\u003c/em\u003e) was lower (0.536) than parental Ho (0.620), showing a decrease in offspring heterozygosity. Both parental and offspring populations had positive fixation indices (\u003cem\u003eF\u003c/em\u003e), 0.026 and 0.201, respectively. This suggests potential excess homozygosity and inbreeding, but with relatively low heterozygote deficiency.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.5 Flowering Synchrony Index\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDue to clonal differences, male and female cone flowering durations vary. Over three observation years, pollen shedding lasted 5 - 11, 9 - 14, and 10 - 12 days, respectively. The female cone pollination period was 5 - 12, 9 - 13, and 9 - 11 days (Fig. 3a). The full flowering period accounted for the largest proportion, with male cone full flowering comprising 36.36% - 66.67% of the total flowering period and female cone full flowering comprising 37.50% - 71.42% (Fig. 4).\u003c/p\u003e\n\u003cp\u003eIn 2021, 2024, and 2025, the flowering synchrony index (\u003cem\u003eW\u003csub\u003ei\u003c/sub\u003e\u003c/em\u003e\u003csub\u003ej\u003c/sub\u003e) for clone combinations ranged from 0 to 0.89, 0 to 0.72, and 0 to 0.69, respectively, with averages of 0.26, 0.39, and 0.42. Overall, the flowering synchrony index was low. In 2021, 41.54% of clone combinations had a flowering synchrony index above 0.3, while 58.46% had an index below 0.3, and only 23.84% exceeded 0.4. In 2024, 74.36% of clone combinations had an index above 0.3, 25.64% below 0.3, and 48.72% exceeded 0.4. In 2025, 83.76% of clone combinations had an index above 0.3, 16.24% below 0.3, and 62.11% exceeded 0.4. (Figure 5 - b).\u003c/p\u003e\n\u003cp\u003eWithin the same year, the flowering synchrony indices between clones as male and female parents were similar, with variations within 5%. The self - pollination flowering synchrony indices for each clone were 0.341, 0.472, and 0.492 over the three years, higher than when they served as male or female parents. This indicates increasing self - pollination risk with orchard age (Fig. 6).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.6 Climatic Impact Factors\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFactorial analysis of major meteorological factors during the flowering period and flowering phenology shows that rainfall and temperature are key drivers of the flowering synchrony index (Fig. 7). Interestingly, in years with good weather, the shorter full - flowering period leads to a lower synchrony index. Conversely, in years with rainy and cold conditions, the synchrony index rises. Among the three observed years, two with low temperatures and rainfall showed similar increases in the flowering synchrony index (Fig. 8). During the flowering period, the frequency of rainfall events is independent of floral duration; however, cumulative precipitation can prolong anthesis. Daily mean temperature inversely (DMT) modulates floral longevity, with elevated thermal regimes shortening the flowering window in clonal genotypes. Genetically mediated clonal effects consequently diminish phenological synchrony.\u003c/p\u003e"},{"header":"4 Discussion ","content":"\u003cp\u003e\u003cstrong\u003e4.1 Imbalanced Parental Contribution to the Offspring Gene Pool\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCompared to other conifer seed orchards, the genetic diversity of \u003cem\u003eCupressus funebris\u003c/em\u003e parental populations is slightly higher or similar, indicating a proper selection intensity of high - gain parents from the first - generation seed orchard for the improved one. Despite fewer parents than the breeding population (\u003cem\u003eHo\u003c/em\u003e, 0.567) (Yang \u003cem\u003eet al.\u003c/em\u003e 2016), the seed orchard maintains comparable genetic diversity (\u003cem\u003eHo\u003c/em\u003e, 0.620), showing a broad genetic base. However, significant gamete contribution imbalance exists: 51.85% - 66.67% of clones contribute over 85% of female cones, and 44.44% - 51.85% contribute over 85% of male cones (Gömöry et al. 2003). This may lead to a few parents dominating the offspring gene pool, reducing genetic diversity. Similar cases have been reported in other conifer seed orchards (Chen et al. 2022; Wang et al. 2023). This study also reveals that the offspring's observed heterozygosity (\u003cem\u003eHo\u003c/em\u003e = 0.536) is lower than the parents' (\u003cem\u003eHo\u003c/em\u003e = 0.620), and the inbreeding coefficient (\u003cem\u003eF\u003c/em\u003e) is positive for both, suggesting excess homozygosity and inbreeding risk. The decrease in offspring heterozygosity mainly stems from the imbalanced parental contribution to the offspring gene pool (Nyabera et al. 2021;\u0026nbsp;Heuchel et al. 2022).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4.2 Pollen Dispersal Patterns and Paternal Reproductive Success\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mating system affects the reproductive level and genetic structure of parent trees in seed orchards. The extent of effective pollen dispersal within seed orchards is limited, and pollen from external unknown sources has a wide dispersion range, leading to imbalanced paternal contributions (Lisa and Keith 2017). Funda and El-Kassaby indicated that 20% of parental clones in seed orchards can produce up to 80% of the cones (Funda et al. 2012). In this study, over three years of investigation in the seed orchard, a smaller number of clones were found to contribute a larger amount of pollen during reproduction. The parental balance curves for female and male cone quantities deviated significantly from the ideal state (Fig. 1). The offspring parentage analysis results showed that in the dwarfed plots, the pollen source could be determined for 70.89% of the offspring, and in the non-dwarfed plots, the pollen source could be determined for 76.67% of the offspring. The range of the number of offspring produced by the mother tree and the candidate father tree was 1 to 11, with the majority producing only one offspring. The parental reproductive contribution rate ranged from 3.57% to 42.31%, indicating an imbalance in paternal contributions within the seed orchard, especially in the dwarfed plots, where the frequency of gene exchange was reduced. The contribution of the paternal candidate trees decreased with the increase of the distance between the mother tree and the candidate tree. The wind direction between the mother tree and the pollen donor was also an important factor determining the paternal contribution. This is because in wind-pollinated tree species, pollen wind direction is affected by the prevailing wind direction (Heuchel et al. 2022). This phenomenon also occurs in the western larch experimental forest (Ben\u0026nbsp;et al. 2008), but the influencing factors are different. The low pollination efficiency and asynchronous flowering among clones are the main reasons for the imbalance in paternal reproductive output within the seed orchard. Gene flow through the dynamic dispersal of pollen and seeds helps the genetic variation of plant populations (Huang et al. 2018;\u0026nbsp;Chen et al. 2018; Bruna et al. 2020). The pollen dispersal range in \u003cem\u003eCupressus funebris\u003c/em\u003e seed orchards is relatively wide, with a pollination pattern similar to that of other wind-pollinated tree species, such as \u003cem\u003ePinus massoniana, Pinus tabuliformis\u003c/em\u003e, and \u003cem\u003eAbies\u003c/em\u003e. This is different from the traditional pollination pattern of tall trees (Armstrong et al. 2024; Yang et al. 2025). In dwarfed seed orchards, the widespread pattern of pollen dispersal is likely due to the significantly higher plant density in the seed orchard compared to natural populations, and the lower position of male and female cones on dwarfed trees compared to non-dwarfed trees.\u003c/p\u003e\n\u003cp\u003eStudies show that the outcrossing rate in \u003cem\u003eCupressus funebris\u0026nbsp;\u003c/em\u003eseed orchards is lower than in other conifers. This suggests that the genetic diversity of parental clones in the seed orchard isn’t being effectively and stably transmitted. Even with a fixed - clone design, there's a risk of inbreeding and pollen contamination. The flowering asynchrony among parents exacerbates this (Li et al. 2012;\u0026nbsp;Luc 2019). Also, the low density of flowering plants in the seed orchard is a significant factor increasing the inbreeding rate (Tania et al. 2024). Parental flowering synchrony, flower quantity, and selective fertilization may cause non - random mating and inbreeding. Importantly, the seed orchard has been subjected to topping and pruning. The maximum and average pollen dispersal distances in non - dwarfed plots are greater than those in dwarfed plots. Thus, dwarfing reduces effective pollen dispersal and increases self - pollination risk. This implies that more attention should be paid to parental clone arrangement and spacing in dwarfed seed orchards. Pollen dispersal and paternal reproductive success are key for seed orchard management (Wang et al. 2023; Martins et al. 2025). To boost seed yield and quality, a \"wide - row, close - plant\" spacing is more suitable for conifers (El-Kassaby et al. 2010). After dwarfing, the fruit - bearing layer shifts downward, and male/female cones enter a \"relatively dense zone.\" So, more pollination space is needed to reduce inbreeding risk.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4.3 Effects of Flowering Synchrony and Climatic Factors\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDuring flowering, synchrony among individuals impacts gene flow and may be influenced by changing climatic conditions (Pearse et al. 2015;\u0026nbsp;Tania\u0026nbsp;et al. 2024). Low synchrony may reduce resource competition among clones but decreases pollination efficiency and affects offspring genetic gain (Wang et al. 2023). High synchrony can enhance pollination efficiency through resource sharing (Li et al. 2012). However, over the three - year observation period, overall synchrony in the seed orchard was low. Differences in flowering synchrony among clones and between years were observed, indicating that varying flowering times among clones is common in forest tree seed orchards. This asynchrony can reduce effective parental mating combinations, increase self - pollination and external pollen contamination, and thus lower the genetic gain of seeds (Wang et al. 2010).\u003c/p\u003e\n\u003cp\u003eThe flowering synchrony index is closely related to environmental factors such as temperature, rainfall, and altitude. In years with low temperatures and high rainfall (e.g., 2024 and 2025), the flowering synchrony index was higher. This is because adverse weather conditions slowed the flowering process of the clones. Those clones that were ready or already in the flowering stage experienced a longer flowering period due to the impact of the weather. The cold and rainy weather typically lasted for 5 - 8 days. Once the weather improved, the male and female strobili of the clones opened in a more concentrated manner, leading to a relatively higher flowering synchrony index. In contrast, in sunny years (e.g., 2021), the flowering synchrony was lower. This is attributed to the fact that under favorable weather conditions, clones capable of pollen shedding or female flower pollination completed their flowering process in a shorter time frame, making clonal differences more pronounced. This finding differs from studies on trees like \u003cem\u003ePinus radiata\u003c/em\u003e, which generally suggest that high temperatures and humidity reduce synchrony (Pederick and Brown\u0026nbsp;2013;\u0026nbsp;Beren et al. 2020). Therefore, the flowering management of \u003cem\u003eCupressus funebris\u003c/em\u003e seed orchards should be tailored to the local climate. Monitoring meteorological data during the flowering period, forecasting changes in synchrony, and implementing measures such as assisted pollination or flowering induction can enhance pollination efficiency (Codesido and Merlo 2005).\u003c/p\u003e"},{"header":"5 Conclusions","content":"\u003cp\u003eIn \u003cem\u003eCupressus funebris\u003c/em\u003e dwarfed seed orchards, gamete contributions are unbalanced. The genetic composition of offspring is jointly influenced by flowering synchrony, parental contributions, and pollen dispersal patterns, with clonal differences playing a leading role. Climatic factors also have complex effects on flowering synchrony. Although dwarfing does not significantly alter pollen dispersal distances, it lowers the fruit - bearing layer, causing male and female cones to enter a \"relatively dense zone.\" This requires more extensive pollen cloud dispersal to reduce inbreeding risks. Consequently, in \u003cem\u003eCupressus funebris\u003c/em\u003e dwarfed seed orchards, clone selection should be optimized based on cone quantities, phenology, and parent origins. Additionally, greater attention should be paid to branch management and plant spacing to enhance ventilation and light penetration. Combined with flowering - period management, such as assisted pollination and phenological regulation, these measures can improve the genetic gain and stability of seed orchards.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are deeply grateful to the Research Group of Forest Tree Genetic Breeding and Cultivation of the Research Institute of Subtropical Forestry, Chinese Academy of Forestry for their great assistance in laboratory aspects.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe roles and contributions of each author must be described in the subsequent manner: The authors confirm contribution to the paper as follows: study conception and design: Z Z; data collection: H-B Z, S C, G-Q J and Z-P F; analysis and interpretation of results: Z Z, H-B Z, S C, G-Q J and Z-C Z; draft manuscript preparation: Z Z, H-B Z, S C. All authors reviewed the results and approved the final version of the manuscript. An author name can appear multiple times, and each author name must appear at least once.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the project “Research on intelligent and high-yield management technology for seed orchards of southern coniferous tree species” of the National Key R\u0026amp;D Program of China during the 14th Five-year Plan period (2023YFD2200601-4), and the project “Breeding of new high-carbon-sequestration and high-quality timber tree varieties” of the Zhejiang Provincial Forest Tree New Variety Breeding Program during the 14th Five-year Plan period(2021C02070-8).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data and materials used in this study are available upon reasonable request from the corresponding authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors gave their informed consent to this publication and its content.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAllender C (2011) The Second Report on the State of the World's Plant Genetic Resources for Food and Agriculture. Exp Agr 47:574. https://doi.org/10.1017/s0014479711000275\u003c/li\u003e\n\u003cli\u003eArmstrong C, Ganasamurthy S, Wigley K, et al. (2024) The microorganisms and metabolome of \u003cem\u003ePinus radiata\u003c/em\u003e Pollen. Environ microbiome 19: 103.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1186/s40793-024-00656-4\u003c/li\u003e\n\u003cli\u003eBruna IA, Miguel LMF, Antonio SZ, et al. (2020) The effects of pollen dispersal and mating pattern on inbreeding depression and hybrid vigor in \u003cem\u003eBalfourodendron riedelianum\u003c/em\u003e (Engl.) Engl. (Rutaceae). Conserv Genet 21:305\u0026ndash;317. https://doi.org/10.1007/s10592-020-01250-0\u003c/li\u003e\n\u003cli\u003eBarrett JW, Knowles P, Cheliak WM. (2011) The mating system in a black spruce clonal seed orchard. Can J Forest Res 17:379-382. https://doi.org/10.1139/x87-065\u003c/li\u003e\n\u003cli\u003eBeren S, Richard M, Amir A, et al\u003cem\u003e.\u003c/em\u003e (2020) Flowering phenology in a Eucalyptus loxophleba seed orchard, heritability and genetic correlation with biomass production and cineole: breeding strategy implications. Sci Rep-UK 10: 15303.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1038/s41598-020-72346-3\u003c/li\u003e\n\u003cli\u003eBen SK, Funda T, Chen C, et al. (2008) Pollination dynamics in a Douglas-fir seed orchard as revealed by pedigree reconstruction. Ann Forest Sci 65: 808.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1051/forest/2010044\u003c/li\u003e\n\u003cli\u003eChatterjee T, Chakrabarty S, Mitra B, et al. (2025) Unveiling the association of mangrove flowering phenology on the dynamics of insect abundance. Acta Oecologica 126:104052. https://doi.org/10.1016/j.actao.2024.104052\u003c/li\u003e\n\u003cli\u003eChen XB, Sun X, Dong L, et al. (2018) Mating patterns and pollen dispersal in a Japanese larch (Larix kaempferi) clonal seed orchard: a case study. Sci China Life Sci 61:1011-1023. https://doi.org/10.1007/s11427-018-9305-7\u003c/li\u003e\n\u003cli\u003eChen SF, Wataru I, Yamato U, et al. (2022) Pollen dispersal patterns and male reproductive success based on pedigree reconstruction in a hybrid larch (\u003cem\u003eLarix gmelinii var. Japonica \u0026times; L. kaempferi\u003c/em\u003e) open‑pollinated seed orchard. Tree Genet Genomes 18:33. https://doi.org/10.1007/s11295-022-01566-7\u003c/li\u003e\n\u003cli\u003eChaloupkov\u0026aacute; K, Lstibůrek M. (2022) Spatial optimization of genetic thinning in seed orchards. Ann Forest Sci 79:37. https://doi.org/10.1186/s13595-022-01158-4\u003c/li\u003e\n\u003cli\u003eCodesido V, Merlo EFJ. (2005) Variation in Reproductive Phenology in a Pinus radiata D. Don Seed Orchard in Northern Spain. Silvae Genet 54:246-256.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1515/sg-2005-0035\u003c/li\u003e\n\u003cli\u003eDu YJ, Zhang RC, Tang XR et al. (2024) The mid-domain effect in flowering phenology. Plant Diversity 46:502-509. https://doi.org/10.1016/j.pld.2024.05.005\u003c/li\u003e\n\u003cli\u003eDarlin UGZ, Alexandre MS, Bruno CR. (2024) Mating system and inbreeding depression in \u003cem\u003eHymenaea stigonocarpa\u003c/em\u003e. Tree Genet Genomes 20: 6.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1007/s11295-024-01639-9\u003c/li\u003e\n\u003cli\u003eEl-Kassaby YA, Funda T, Lai BSK (2010) Female reproductive success variation in a Pseudotsuga menziesii seed orchard as revealed by pedigree reconstruction from a bulk seed collection. J Hered 101:164-168. https://doi.org/10.1093/jhered/esp126\u003c/li\u003e\n\u003cli\u003eFunda FT, Liewlaksaneeyanawin L C, Fundova F I, et al. (2011) Congruence between parental reproductive investment and success determined by DNA-based pedigree reconstruction in conifer seed orchards. Can J Forest Res 41:380-389.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1139/x10-190\u003c/li\u003e\n\u003cli\u003eFunda T. (2012) Seed orchard genetics. Forest Sci 59: 1-12.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1079/pavsnnr20127013\u003c/li\u003e\n\u003cli\u003eG\u0026ouml;m\u0026ouml;ry D, Bruch\u0026aacute;nik R, Longauer R. (2003) Fertility variation and flowering asynchrony in Pinus sylvestris: consequences for the genetic structure of progeny in seed orchards.\u003cem\u003e For \u003c/em\u003eEcol Manage 174:117-126. https:// doi. org/ 10. 1016/ S0378- 1127(02) 00031-2\u003c/li\u003e\n\u003cli\u003eHeuchel A, Hall D, Zhao W, et al. (2022) Genetic diversity and background pollen contamination in Norway spruce and Scots pine seed orchard crops. Forestry research 2: 67-78. https://doi.org/10.48130/fr-2022-0008.\u003c/li\u003e\n\u003cli\u003eHeike L, Dagmar S. (2022) Flowering behavior of clones in a Norway maple (\u003cem\u003eAcer platanoides\u003c/em\u003e) seed orchard and mating system analysis using nuclear SSR markers. Eur J Forest Res 141:561-569. https://doi.org/10.1007/s10342-022-01459-3\u003c/li\u003e\n\u003cli\u003eHan S, Kang K, Kim C, et al. (2013) Effect of top-pruning in a clonal seed orchard of \u003cem\u003ePinus koraiensis\u003c/em\u003e. Ann Forest Sci 51:155-156. https://doi.org/10.15287/afr.2008.157\u003c/li\u003e\n\u003cli\u003eHuntsman SV, Leslie AB .The ontogeny of disparity in Cupressaceae seed cones. New Phytol 242:2301-2311. https://doi.org/10.1111/nph.19482\u003c/li\u003e\n\u003cli\u003eHuang L, Song J, Sun Y, et al. (2018) Pollination dynamics in a Platycladus orientalis seed orchard as revealed by partial pedigree reconstruction. Can J Forest Res 48:952-957. https://doi.org/10.1186/s40793-024-00656-4\u003c/li\u003e\n\u003cli\u003eKolpak ES, Smith J, Albrecht JM, et al. (2015) High-density miniaturized seed orchards of Douglas-fir. New For 46:121-140. https://doi.org/10.1007/s11056-014-9452-6\u003c/li\u003e\n\u003cli\u003eLi W, Wang X R, Li Y. (2012) Variation in Floral Phenological Synchronization in a Clonal Seed Orchard of \u003cem\u003ePinus tabuliformis \u003c/em\u003ein Northeast of China. Silvae Genet 61:133-142. https://doi.org/10.1515/sg-2012-0017\u003c/li\u003e\n\u003cli\u003eLindgren D, Prescher F. (2005) Optimal clone number for seed orchards with tested clones. Silvae Genet 54:80-92. https://doi.org/10.1515/sg-2005-0013\u003c/li\u003e\n\u003cli\u003eLisa A, Keith W. (2017) Pollen gene flow, male reproductive success, and genetic correlations among offspring in a northern red oak (\u003cem\u003eQuercus rubra\u003c/em\u003e L.) seed orchard. PLoS ONE 12:e0171598. https://doi.org/10.1371/journal.pone.0171598\u003c/li\u003e\n\u003cli\u003eLee K, Kim IS, Kang KS (2022) Pedigree reconstruction and spatial analysis for genetic testing and selection in a \u003cem\u003eLarix kaempferi \u003c/em\u003e(Lamb.) Carri\u0026egrave;re plantation. BMC Plant Biol 22:152. https://doi.org/10.1186/s12870-022-03530-y\u003c/li\u003e\n\u003cli\u003eLuc EP. (2019) Hybrid breakdown and inbreeding depression for seed yield and early growth in 2nd-generation interspecific hybrids of larch (\u003cem\u003eLarix. \u003c/em\u003e\u0026times;\u003cem\u003e eurolepis\u003c/em\u003e \u0026times; \u003cem\u003eL. \u003c/em\u003e\u0026times;\u003cem\u003e eurolepis\u003c/em\u003e). Ann Forest Sci 76:1-10. https://doi.org/10.1007/s13595-019-0853-8\u003c/li\u003e\n\u003cli\u003eMartins AE, Loiola PdP, Pareja-Bonija D, et al. (2025). Climate-induced shifts in long-term tropical tree reproductive phenology: Insights from species dependent on and independent of biotic pollination. Funct Ecol 00:1-13. https://doi.org/10.1111/1365-2435.70090\u003c/li\u003e\n\u003cli\u003eMartins G, Yuliarto M, Melia T, et al. (2025) Pollination and mating dynamics unveiled by orchard-wide pedigree reconstruction in Acacia crassicarpa. Tree Genet Genomes 21:7. https://doi.org/10.1007/s11295-025-01692-y\u003c/li\u003e\n\u003cli\u003eMartins AE, Camargo MGG, Morellato LPC. (2021) Flowering Phenology and the Influence of Seasonality in Flower Conspicuousness for Bees. Front Plant Sci 11:1-11. https://doi.org/10.3389/fpls.2020.594538\u003c/li\u003e\n\u003cli\u003eNyabera LA, Nzuki IW, Runo SM, et al\u003cem\u003e.\u003c/em\u003e (2021) Assessment of genetic diversity of pumpkins(\u003cem\u003eCucurbita\u003c/em\u003e spp.) from western Kenya using SSR molecular markers. Mol Biol Rep 48:2253-2260. https://doi.org/10.1007/s11033-021-06245-9\u003c/li\u003e\n\u003cli\u003ePrimack RB. (2003) Relationships among flowers, fruits, and seeds. Annu Rev Ecol Syst 18:409-430. https://doi.org/10.1146/annurev.es.18.110187.002205\u003c/li\u003e\n\u003cli\u003ePaul JC, Amy MI, David WI. (2014) Shifts in flowering phenology reshape a subalpine plant community. Proc Natl Acad Sci USA 111: 4916-4921.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.1073/pnas.1323073111\u003c/li\u003e\n\u003cli\u003ePearse IS, Koenig WD, Funk KA, et al. (2015) Pollen limitation and flower abortion in a wind-pollinated, masting tree. Ecology 96:587-593. https://doi.org/10.1890/14-0297.1\u003c/li\u003e\n\u003cli\u003ePederick AL, Brown GA. (2013) Seed Production in Radiata Pine Seed Orchards in Australia. Aust For 39:164-179. https://doi.org/10.1080/00049158.1976.10674149\u003c/li\u003e\n\u003cli\u003eRuotsalainen S. (2014) Increased forest production through forest tree breeding. Scand J For Res 29:333-344. https://doi.org/10.1080/02827581.2014.926100\u003c/li\u003e\n\u003cli\u003eSong J, Ratcliffe B, Kess T, Lai BS, Koreck\u0026yacute; J, et al. (2018) Temporal quantification of mating system parameters in a coastal Douglasfir seed orchard under manipulated pollination environment. Sci Rep 8:11593. https://doi.org/10.1038/s41598-018-30041-4\u003c/li\u003e\n\u003cli\u003eTorimaru T, Wennstr\u0026ouml;m U, Lindgren D, et al. (2012) Effects of male fecundity, interindividual distance and anisotropic pollen dispersal on mating success in a Scots pine (\u003cem\u003ePinus sylvestris\u003c/em\u003e) seed orchard. Heredity 108:312-321. https://doi.org/10.1038/hdy.2011.76\u003c/li\u003e\n\u003cli\u003eTania DF, Katharina BB, John EC, et al. (2024) Distance-dependent mating but considerable pollen immigration in an isolated Quercus rubra planting in Germany. Eur J Forest Res 143:1447-1460. https://doi.org/10.1007/s10342-024-01704-x\u003c/li\u003e\n\u003cli\u003eWang F, Zhang SY, Zhu P, \u003cem\u003eet al.\u003c/em\u003e (2023) The effects of fertility and synchronization variation on seed production in two Chinese fir clonal seed orchards. Sci Rep 13:627. https://doi.org/10.1038/s41598-022-27151-5\u003c/li\u003e\n\u003cli\u003eWang XY, Morin X, Zhang J, \u003cem\u003eet al.\u003c/em\u003e (2023) Geographical patterns and determinants in plant reproductive phenology duration. Front Plant Sci 14:1-8.\u003c/li\u003e\n\u003cli\u003ehttps://doi.org/10.3389/fpls.2023.1199316\u003c/li\u003e\n\u003cli\u003eWang XY, Bjorkman AD, Li X, et al\u003cem\u003e.\u003c/em\u003e (2025) Shifts in plant reproductive phenology induced by multiple global change factors depend on phenological niche andpollination mode. J Plant Ecol 18:rtaf048. https://doi.org/10.1093/jpe/rtaf048\u003c/li\u003e\n\u003cli\u003eWang F, Zhang SY, Zhu P, et al\u003cem\u003e.\u003c/em\u003e (2023) The effects of fertility and synchronization variation on seed production in two Chinese fir clonal seed orchards. Sci Rep 13:627. https://doi.org/10.1038/s41598-022-27151-5\u003c/li\u003e\n\u003cli\u003eWang XR, Torimaru T, Lindgren D, et al\u003cem\u003e.\u003c/em\u003e (2010) Marker-based parentage analysis facilitates low input \u0026lsquo;breeding without breeding\u0026rsquo; strategies for forest trees. Tree Genet Genomes 6:227-235. https://doi.org/10.1007/s11295-009-0243-8\u003c/li\u003e\n\u003cli\u003eWu YQ, Weng YH, Hennigar C, \u003cem\u003eet al.\u003c/em\u003e (2015) Benefit\u0026ndash;cost analysis of a white spruce clonal seed orchard in New Brunswick. Canada. New Forests 46, 141\u0026ndash;156. https://doi.org/10.1007/s11056-014-9453-5.\u003c/li\u003e\n\u003cli\u003eYang BN, Su NH, Qi JD, et al. (2020) Improve dgenetic distance-based spatial deployment can effectively minimize inbreeding in seed orchard. For Ecosyst 7:1-11. https://doi.org/10.1186/s40663-020-0220-0\u003c/li\u003e\n\u003cli\u003eYang T, Wang PC, Wang WY, et al. (2023) Early growth evaluation and biomass allocation difference between clones and families in \u003cem\u003eCupressus funebris\u003c/em\u003e. Eur J For Res 142:839-850. https://doi.org/10.1007/s10342-023-01563-y\u003c/li\u003e\n\u003cli\u003eYang ML, Liu JC, Gao MK, et al\u003cem\u003e.\u003c/em\u003e (2025) Pollen limitation influences the divergences in mating system and floral traits between two sympatric Halenia species. J Plant Ecol 18:rtae098. https://doi.org/10.1093/jpe/rtae098\u003c/li\u003e\n\u003cli\u003eZoe D, Anita C, Kathleen MK, et al. (2023) Reproductive strategies and their consequences for divergence, gene flow, and genetic diversity in three taxa of Clarkia. Heredity 131: 338-349. https://doi.org/10.1038/s41437-023-00649-y\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Paternity Analysis of Open-Pollinated Progeny\u003c/p\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"547\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 18.7956%;\"\u003e\n \u003cp\u003eType\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17.1533%;\"\u003e\n \u003cp\u003e\u003cem\u003ePIR\u003c/em\u003e/%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10.9489%;\"\u003e\n \u003cp\u003e\u003cem\u003eSR\u003c/em\u003e/%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.8686%;\"\u003e\n \u003cp\u003e\u003cem\u003eOR\u003c/em\u003e/%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17.8832%;\"\u003e\n \u003cp\u003e\u003cem\u003eNUAIP\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 21.3504%;\"\u003e\n \u003cp\u003e\u003cem\u003ePCFP\u003c/em\u003e/%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 18.7956%;\"\u003e\n \u003cp\u003eDSO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17.1533%;\"\u003e\n \u003cp\u003e70.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10.9489%;\"\u003e\n \u003cp\u003e27.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.8686%;\"\u003e\n \u003cp\u003e72.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17.8832%;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 21.3504%;\"\u003e\n \u003cp\u003e5.80\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 18.7956%;\"\u003e\n \u003cp\u003eUSO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17.1533%;\"\u003e\n \u003cp\u003e76.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10.9489%;\"\u003e\n \u003cp\u003e18.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 13.8686%;\"\u003e\n \u003cp\u003e81.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17.8832%;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 21.3504%;\"\u003e\n \u003cp\u003e8.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eDSO represents the dwarf seed orchard, USO represents the undwarfed seed orchard, \u003cem\u003ePIR\u003c/em\u003e represents the paternity identification rate, \u003cem\u003eSR\u003c/em\u003e represents the selfing rate, \u003cem\u003eOR\u003c/em\u003e represents the outcrossing rate, \u003cem\u003eNUAIP\u003c/em\u003e represents the number of unique alleles in progeny, and\u0026nbsp;\u003cem\u003ePCFP\u003c/em\u003e represents the proportion of contaminated foreign pollen.\u003cbr\u003e\u0026nbsp;\u003cstrong\u003eTable 2\u003c/strong\u003e Genetic Diversity of Parental and Progeny Populations in the 1.5-Generation \u003cem\u003eCypress\u003c/em\u003e Clonal Seed Orchard\u003c/p\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"670\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 66px;\"\u003e\n \u003cp\u003eLocus\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cem\u003eNa\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 98px;\"\u003e\n \u003cp\u003e\u003cem\u003eNe\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 97px;\"\u003e\n \u003cp\u003e\u003cem\u003eI\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u003cem\u003eHo\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u003cem\u003eHe\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" style=\"width: 117px;\"\u003e\n \u003cp\u003e\u003cem\u003eF\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003eParent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003eProgeny\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003eParent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003eProgeny\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003eParent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003eProgeny\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003eParent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003eProgeny\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003eParent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003eProgeny\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003eParent\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003eProgeny\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003e1-C95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.608\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e3.356\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.164\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.352\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.111\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.046\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.617\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.702\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.820\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.935\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eC83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e7.401\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e8.310\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.037\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e2.198\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.556\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.441\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.865\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.880\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.358\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.499\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eCYP52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.527\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e2.920\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.085\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.852\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.739\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.604\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.657\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.410\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.125\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eCYP84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3.837\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e4.835\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.515\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.685\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.630\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.736\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.739\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.793\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.148\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.073\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eCYP174\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e7.290\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e8.451\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.164\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e2.279\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.667\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.847\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.863\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.882\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.227\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.040\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eCYP293\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3.574\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e3.458\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.492\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.474\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.926\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.877\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.720\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.711\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.286\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.234\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eF015\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3.170\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e2.158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.407\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.973\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.667\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.096\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.684\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.537\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.026\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.822\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eF042\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e3.229\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.509\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.533\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.630\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.602\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.655\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.690\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.039\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.129\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eJT04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.690\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e2.591\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.029\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.519\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.115\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.628\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.614\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.175\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.813\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eJT05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.561\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e2.156\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.630\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.932\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.444\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.789\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.359\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.536\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.237\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.472\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eYW03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.911\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e1.834\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.822\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.863\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.593\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.521\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.477\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.455\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.243\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.146\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eCF02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e2.876\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e3.068\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.194\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.852\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.621\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.652\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.674\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e-0.306\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.079\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e6.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e6.583\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e3.529\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e3.864\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e1.337\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1.387\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.620\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.536\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 50px;\"\u003e\n \u003cp\u003e0.655\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 48px;\"\u003e\n \u003cp\u003e0.678\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.026\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 56px;\"\u003e\n \u003cp\u003e0.201\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cem\u003eNa\u003c/em\u003e denotes the number of alleles; \u003cem\u003eNe\u0026nbsp;\u003c/em\u003edenotes the effective number of alleles; \u003cem\u003eI\u003c/em\u003e is Shannon\u0026rsquo;s diversity index; \u003cem\u003eHo\u003c/em\u003e is the observed heterozygosity; \u003cem\u003eHe\u003c/em\u003e is the expected heterozygosity;\u0026nbsp;\u003cem\u003eF\u003c/em\u003e is the fixation index.\u003cbr\u003e\u0026nbsp;\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"Cupressus funebris, dwarfed seed orchards, flowering synchrony, mating system, paternity analysis, pollen spread","lastPublishedDoi":"10.21203/rs.3.rs-7253675/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7253675/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eContext\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003e\u003cem\u003eCupressus funebris\u003c/em\u003e is a high - value conifer, is important for timber and ecological protection. In its seed production, optimizing parental composition in seed orchards and using “dwarfed” trees after topping are key to achieving high - yield and high - gain seeds. However, compared to traditional seed orchards, the parental contributions and pollen dispersal patterns in dwarfed seed orchards remain unclear.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eAims\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003eBy analyzing flowering synchrony, parental contributions, and pollen dispersal patterns in a dwarfed \u003cem\u003eCupressus funebris\u003c/em\u003e seed orchard, we investigated their effects on offspring genetics.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e \u003c/strong\u003eUsing three years of flowering-phenology observations, we quantified flowering synchrony and identified the principal climatic drivers governing its variation. We further characterised the mating system within the dwarf seed orchard, estimated the gametic contribution of each parent to the offspring cohort, and finally compared pollen dispersal patterns between dwarf and non-dwarf orchards.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eResults \u003c/strong\u003e\u003c/em\u003eThree years of monitoring revealed significant interclonal variation among parental ortets in the seed orchard with respect to both strobilus production and flowering phenology, indicating asymmetrical gametic contribution within the orchard. The low within-year synchrony index of male and female strobili is predominantly governed by clonal variation, as well as by significant effects of mean daily maximum temperature, mean daily minimum temperature, and cumulative precipitation during the flowering period. The offspring population in the dwarfed seed orchard maintained high genetic diversity but had inbreeding risk. Compared to the non - dwarfed orchard, spatial distance was a key factor affecting pollination success in the dwarfed one. After dwarfing, the spatial distribution of female and male cone flowers shifted downward, shortening the average pollen dispersal distance, reducing outcrossing rates, and increasing self - pollination probability.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eConclusion \u003c/strong\u003e\u003c/em\u003eBranch management and plant spacing in \u003cem\u003eCupressus funebris\u003c/em\u003e dwarfed seed orchards should be emphasized. Improving ventilation and light penetration, combined with flowering management, are essential management strategies.\u003c/p\u003e","manuscriptTitle":"Phenological Synchrony, Parental Contribution, and Pollen Dispersal Pattern affects the Genetic Composition of Cupressus funebris Dwarf Seed Orchard Progenies","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-02 06:22:54","doi":"10.21203/rs.3.rs-7253675/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":"46ec6a0e-94c4-4ef5-8239-6db1353a1a59","owner":[],"postedDate":"September 2nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-01T13:08:41+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-02 06:22:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7253675","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7253675","identity":"rs-7253675","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}