Physio-biochemical responses and crop performance analysis in chickpea upon botanical priming

Physio-biochemical responses and crop performance analysis in chickpea upon botanical priming | 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 Article Physio-biochemical responses and crop performance analysis in chickpea upon botanical priming Kamini Kaushal, Kumari Rajani, Ravi Ranjan Kumar, Anand Kumar, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3914755/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Apr, 2024 Read the published version in Scientific Reports → Version 1 posted 8 You are reading this latest preprint version Abstract Chickpea is a highly nutritious protein-rich source and one of the major crops to alleviate global malnutrition. It is the third-largest legume cultivated globally in arid and semi-arid environments. Chickpea productivity is affected by several factors, among which poor seed quality is one of the significant factors contributing to its decrease in productivity. Seed quality is essential for better crop establishment and higher yields, particularly in the uncertain climate change. The present study investigated the role of botanical priming in enhancing seed quality by investigating physio-biochemical responses and crop performance in chickpeas. The study compared the effects of botanical priming with hydropriming, seed treatment with bavastin and control on medium-vigour chickpea seeds. A detailed physiological (germination percentage, root and shoot length, vigour index) and biochemical (amylase, protease, dehydrogenase, phytase, and lipid peroxidation) analysis was carried out in the laboratory to assess the effect of priming treatments. SDS-PAGE was used to compare seed storage protein expression in primed and control seeds. Seeds treated with 1% turmeric leaf extract displayed a higher germination rate (94.5%) than the control and other priming treatments. Turmeric-primed seeds showed higher seedling length, vigour index, enzyme activity, and lower MDA content. SDS-PAGE analysis revealed the expression of two minor polypeptides of the albumin and three minor polypeptides of the globulin subunit. Moreover, field experiments indicated increased crop growth, vigour, and days to 50% flowering, yield and its attributing traits in turmeric-primed seeds. The study demonstrates that botanical priming can increase the yield of chickpeas by up to 16% over the control group. This study proves that implementing low-cost and eco-friendly seed priming techniques can significantly enhance the genetic potential of chickpeas by improving their physiological and biochemical activities during seed germination and crop growth. Therefore, chickpea growers must adopt botanical priming techniques to improve seed quality and crop performance. This study unequivocally establishes the efficacy of botanical priming as a powerful tool for augmenting chickpea growth. Moreover, this approach is environmentally sustainable and can help conserve natural resources long-term. Therefore, this new approach must be widely adopted across the agricultural industry to ensure sustainable and profitable farming practices. Biological sciences/Biotechnology Biological sciences/Plant sciences botanical priming turmeric dehydrogenase globulin vigour index SDS PAGE Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction Global climate change and water scarcity generally affect crop performance from germination and eventually reduce grain yield and quality with losses of up to 40–50% in crop productivity. Chickpea, a cool-season legume, are an excellent source of protein (18–20%) and the minerals phosphorus, calcium, magnesium, iron, and zinc that play a significant role in achieving global food security 1 . Over 90% of the world's chickpeas are grown in arid and semi-arid regions, making them vulnerable to various stresses 2 , 3 . Even though it is considered the candidate crop for rainfed conditions, the uncertain climatic condition adversely affects the crop's performance and make it vulnerable to several biotic and abiotic stresses 4 . Successful seedling and crop establishment are critical to higher crop production 5 . Poor seedling establishment is one of the most significant barriers primarily caused by subpar seed quality 6 . The improper storage is the primary factor for quick loss of seed vigour, which ultimately affects germination and crop performance. Moreover, crop germination due to adverse climatic conditions and delayed sowing affects production and productivity. Seed quality enhancement techniques provide a comprehensive solution to unlock the full genetic potential of seeds. Seed quality enhancement, a pre-sowing treatment, enhances germination and crop vigour during early planting and contributes to a higher crop yield 7 . Seed priming, a technique in seed enhancement, involves the initial absorption of water by seeds to initiate the early stages of germination. However, this absorption is insufficient for radical emergence, and the seeds return to their original moisture content 8 . During the priming process, biochemical changes occur, including activating enzymes, producing compounds that promote growth, metabolizing molecules that inhibit germination, and repairing damaged cells 9 . The imbibition of seeds triggers a cascade of metabolic processes, such as the activation of hydrolytic enzymes (such as amylase, protease, lipase, dehydrogenase, and phytase), which lead to the hydrolysis of stored starch, lipids, proteins, polyphosphates, and other storage materials, converting them into simpler forms that readily absorbed by the embryo and ultimately influencing seed vigour 10 . Alpha-amylase breaks down starch into sugars for the developing embryo 11 . Proteolytic enzymes use seed storage proteins 12 , while dehydrogenase catalyzes the stored products during the anaerobic phase of seed germination 13 . Phytase is an enzyme that unequivocally converts phytate into inositol/phosphoric acid, making a remarkable contribution to seed germination and growth 14 . On the other hand, lipid peroxidation is an unequivocally detrimental process that destabilizes the membrane and degrades proteins, leading to inevitable cell death, which hinders the capacity for ionic transport 15 . Similarly, the hydrolysis of seed storage proteins such as globulin and albumin unequivocally generates free amino acids, essential for germination and seed vigour 16 . Over the past few decades, there has been a significant increase in the utilization of non-hazardous chemicals and fertilizers as alternatives to enhance crop productivity in the agriculture system. Among these alternatives, priming has emerged as a highly effective method to tackle this issue. Numerous efficacious priming agents, such as salts, polyamines, hormones, compatible solutes, and aqueous plant extracts, have been documented by various researchers 17 . Botanical priming, a priming technique that employs plant extracts as its agents, has proven particularly advantageous. This botanical priming method can stimulate metabolic processes, is non-toxic and environmentally friendly, and has excellent potential to control pathogenic microorganisms. There are few reports on using turmeric rhizome and neem seed extracts on aphid control, plant growth and crop yield. However, the effect of leaf extracts on seed physiology and plant growth in pulse crops has yet to be studied. No studies have investigated globulin and albumin expression in chickpeas during priming and germination at different durations. In the present investigation, we hypothesized that, first, the effect of turmeric and neem leaf extracts as seed priming agents positively influences physiological parameters and enhances seed vigour in chickpeas. Second, the improved enzymatic activities during botanical priming lead to increased seed germination and growth. Third, priming also increases the expression of different seed storage proteins compared to non-primed conditions. Our objective was to investigate the effect of botanical priming on physiological, biochemical and yield in chickpeas, for which we performed comprehensive laboratory and field experiments. Results Effect of botanical priming on seed germination and seedling characteristics. The present study was carried out to analyze the effects of seed priming with botanicals (turmeric, neem) on the physiological parameters of chickpeas and compare it with conventional hydropriming and fungicide seed treatment. In the present investigation, physiological parameters, viz germination percentage and seedling characteristics, were significantly increased by priming the seeds with botanicals, as shown in Table 1 . The botanical primed seed had significantly higher seed germination and seedling growth than the control. Seeds primed with turmeric and neem leaves aqueous extract had higher germination and seedling growth followed by hydro priming. The results demonstrate that turmeric had a higher germination percentage (94.5%) than control (82.5%), indicating that botanical priming successfully enhanced the germination rate by 15% over control. Seedling characteristics, including root length, shoot length, seedling length, seedling dry weight and vigour indices, were significantly similarly affected by botanical priming. Table 1 shows marked enhancement in seedling length by 29%, vigour index Ι by 42% and vigour index II by 63% were observed in turmeric primed seed over control. Table 1 Impact of different priming agents on germination percentage, root length, shoot length, seedling length, dry weight and vigor indices was estimated on 8th day of germination. Data presented in a column are means of four replicates, where each replicate of the treatment contains 100 seeds. Means in each column, followed by different lower-case alphabets are significantly different at the 0.05% probability level using Post Hoc test. Where T 1 = No priming (control), T 2 = Conventional practice (Bavistin@2g/kg), T 3 = Turmeric leaf priming, T 4 = Neem leaf priming, T 5 = On-farm priming (Hydropriming). Treatments Germination (%) Root length (cm) Shoot length (cm) Seedling length (cm) Dry weight (g) VI Ι VI ΙΙ T 1 82.5 b 17.5 a 10.73 c 24.77 c 0.41 c 2050.14 d 33.24 c T 2 83.5 b 15.07 c 10.97 c 26.99 c 0.43 c 2226.8 c 35.52 c T 3 94.5 a 18.16 a 13.77 a 31.93 a 0.6 a 2905.71 a 54.13 a T 4 86.75 b 16.02 b 13.38 a 29.4 b 0.51 b 2226.8 c 44.44 b T 5 86.75 b 14.04 d 12.02 b 26.06 c 0.49 b 2549.21 b 42.68 b S.E.M. 2.02 0.27 0.32 0.8 0.01 57.97 1.24 C.D. 5% 6.1 0.8 0.96 2.41 0.04 174.75 3.73 C.V. 4.66 3.28 5.22 5.74 5.28 4.85 5.89 Biochemical changes upon botanical priming. The dynamic changes in hydrolytic enzymes and MDA content are shown in Fig. 1 – 5 . Figure 1 shows that botanical priming and hydropriming significantly affected amylase activity, which increased with the increase in priming duration. In contrast, the amylase activity was decreased as the germination proceeded. Conversely, the amylase activity was constant at different priming hours in non-primed and bavistin-treated seeds; however, they showed a similar decreasing trend during germination as botanicals primed seed. Compared to non-primed seeds, maximum amylase activity was observed in turmeric-primed seeds, and it was increased to 4.8-fold at 12 hours and 5.2-fold at 18 hours of priming. The amylase activity was also assessed during germination, and it showed a decreasing trend with the increasing days of germination, and it was highest in turmeric primed seed. The highest amylase activity, 0.71 mg maltose/min, was observed on the first day of germination on the first day, and its activity decreased to 1.7-fold on the third day and 2.0-fold on the fifth day of germination. Botanical priming significantly affected the protease activity (P < 0.05; Fig. 2 ). The protease activity significantly increased with increasing priming duration and till the third day of germination; after that, on the fifth day of germination, irrespective of treatments, the protease activity decreased. The protease activity was maximum during priming durations and germination days in seeds primed with 1% aqueous extract of turmeric leaves. Its activity increased to 1.3-fold in 6 hr, 1.8-fold in 12 hr, 2-fold in 18 hr, 1.2-fold on the first day and 3.2-fold on the third day compared to control in turmeric primed seed. On the fifth day, there was a drastic reduction in the protease activity, which decreased by 1.8-fold compared to the third day of the same treatment. A similar pattern was observed in 1% aqueous extract of neem leaves and hydro-priming, whereas activity was constant throughout the priming duration in control and bavistin-treated seed. In the current study, botanical priming significantly affected dehydrogenase activity (P < 0.05; Fig. 3 ) and differed among treatments. The dehydrogenase activity was significantly increased with priming duration and days of germination. The maximum dehydrogenase activity of 0.98 mg formazan/min was observed on the fifth day of germination in seeds primed with 1% aqueous extract of turmeric leaves. Its activity increased to 1.84-fold in 18 hr and 1.3-fold on the fifth day of germination compared to the control. To confirm the activation of dormant phytase zymogens induced by botanical priming, we investigated the dynamic changes in phytase activity during priming durations and days of germination. As shown in Fig. 4 , the phytase activity significantly increased with increasing priming duration and days of germination. The maximum activity was observed in turmeric primed seed during the fifth day of germination, and it was 0.74 µM trypsin/min (1.9-fold increased over control at a particular time). Compared to the control, its activity in turmeric primed seed increased to 1.7-fold in 6 hours, 2.4-fold in 12 hours, and 2.9-fold in 18 hours of priming. The phytase remained active during germination and increased by 2-fold on the first day, 2.1-fold on the third day and 1.9-fold on the fifth day of germination. Botanically primed seed showed a significant reduction in lipid peroxidation activity, measured in terms of MDA content (µmol MDA/gr F-W). In the present study, as shown in Fig. 5 , on increasing the priming duration, the MDA content was not found to change up to 12 hours in turmeric primed seed; after that, it decreased sharply at 18 hours of priming and during days of germination. Compared to the control, the MDA content decreased to 2.49-fold in 18 hours of priming, 3.4-fold on the first day, 4-fold on the third day, and 3.6-fold on the fifth day of germination and the maximum MDA content was observed in the case of the control seed (T 1 ). Effects of botanical priming on total protein and the expression of seed storage protein. The impact of botanical priming on total protein content was significant in our present work; as illustrated in Fig. 6 , a significant upsurge (p < 0.05) in protein content was evident during priming hours and days of germination with both turmeric and neem leaf aqueous extract. In this, the total protein content increased with the priming duration up to the first day of germination. However, upon increasing the days of germination, irrespective of the priming material, the total protein decreases significantly from the third to the fifth days. Meanwhile, in the control and bavistin-treated seeds, protein content did not vary much throughout the priming duration and during germination. The total protein content increased 1.4-fold on the first day after that, decreased to 1.1-fold on the third day and 1-fold on the fifth day of germination compared to the control. Seed storage proteins (SSP) of chickpeas, i.e., globulin and albumin, were fractionalized by the Osborne method 18 and the molecular weight of the protein was determined using SDS PAGE. Subunits separated from chickpea proteins are shown in Table 2 . Figure 7 shows that the expression of proteins was higher in primed seeds than in the control. In the present work, the estimated molecular weights of subunits of globulin, i.e., legumin, were (40 kDa, 39 kDa, 26 kDa, 23 kDa), vicillin Mw (50 kDa, 39 kDa, 19 kDa, 15 kDa), and glutelin (58 kDa, 55 kDa, 54 kDa). Three minor bands of 36 kDa, 42 kDa, and 56 kDa, the globulin subunits, were detected in the electrophoregram. Table 2 also reveals that the significant globulin subunits, i.e., 42 kDa and 56 kDa, were present in all primed seeds, whereas control and bavistin polypeptides of 42 kDa proteins were not observed. Our work in SDS-PAGE showed a progressive accumulation of the 11-S globulin during priming. In contrast, a slower accumulation of 11S occurred in control seeds, which resulted in an intense polypeptide of 56 kDa and 42 kDa in primed seeds. Table 2 Effect of botanical priming on the expression of seed storage proteins (globulin and albumin) detected through SDS-PAGE. Where (+) and ( – ) sign in each column indicates presence and absence of polypeptide of globulin and albumin sub unit during 6, 12 and 18 h duration of priming. Seed treatments includes T 1 = No priming (control), T 2 = Conventional practice (Bavistin@2g/kg), T 3 = Turmeric leaf priming, T 4 = Neem leaf priming, T 5 = On-farm priming (Hydropriming). Seed Storage protein Molecular Weight (kDa) T 1 T 2 Duration of priming (h) T 3 T 4 T 5 6 12 18 6 12 18 6 12 18 Globulin 36 + + + + + + + + + + + 42 – – + + + + + + + + + 56 + + + + + + + + + + + Albumin 12 + + + + + + + + + + + 14 + + + + + + + + + + + Effects of botanical priming on plant establishment, growth and yield of chickpea. An experiment with turmeric leaves extract priming in chickpea was compared with bavistin seed conventional seed treatment (+ control) and non-priming (control) under field conditions. Table 3 shows that turmeric priming significantly improved field emergence, plant height at different growth stages days to 50% flowering and yield attributing characters (Tables 3 & 4 ). Turmeric priming significantly increased the field emergence by 27%, plant heights at 15 DAS, 45 DAS, and maturity by 5%, 11%, and 4.7%, respectively, compared to the non-priming. About the days to 50% flowering, it was significantly (p < 0.05) affected by priming and obtained three days earlier than in the control plant (Table 3 ). The effect of turmeric leaf priming on yield-attributing characteristics in chickpeas is shown in Table 4 , which presents the data on-field performance, where turmeric priming considerably increased the number of pods per plant, weight of 100 seeds, biological yield, number of seeds per plant, economic yield, and harvest index compared to the control. In comparison to the control, turmeric priming significantly raised the 100 seed weight by 9.1%, the number of pods per plant by 23.4%, the number of seeds per plant by 15.5%, the seed weight per plant by 20.0%, and the harvest index by 18.1%. Table 3 Effect of turmeric priming on plant establishment and development. Field emergence %, plant height at 15, 30, 45 days and maturity, days to 50% flowering were observed. The field experiment was laid in RBD, and data presented in each column are means of 8 replicates (n = 8). Means in each column, followed by different lower-case alphabets, are significantly different at the 0.05% probability level using the Post Hoc test. Where, T 1 = No priming (control), T 2 = Conventional practice (Bavistin@2g/kg), T 3 = Turmeric leaf priming. Treatments Field Emergence (%) Plant height at 15 days (cm) Plant height at 30 days (cm) Plant height at 45 days (cm) Plant height at maturity (cm) Days to 50% Flowering T 1 70.16 c 12.77 b 20.34 24.7 b 56.1 ab 85.00 a T 2 77.85 b 12.84 b 21.31 25.75 ab 54.88 b 86.75 a T 3 88.86 a 13.37 a 21.4 27.4 a 58.75 a 82.5 b S.E.M. 2.43 0.16 0.59 0.70 1.00 0.77 C.D. 5% 7.36 0.48 NS 2.11 3.03 2.33 C.V. 8.70 3.41 7.90 7.58 5.00 2.57 Table 4 Effect of priming on yield attributing characters of chickpea variety PG-186 on number of pods/plant, seed weight, biological yield, yield kg/ha, economic yield and harvest index. Data in each column are means of 8 replicates, different lower-case alphabet shows significant level of difference at 0.05% probability level. Where T 1 = No priming (control), T 2 = Conventional practice (Bavistin@2g/kg), T 3 = Turmeric leaf priming. Treatments No. of Pods/plant Seed weight (g) Biological Yield (g) Yield (kg/ha) Harvest index T 1 84.38 b 17.01 43.28 b 2007.14 b 53.6 b T 2 85.00 b 16.96 48.39 ab 2039.62 b 51.51 b T 3 110.25 a 18.73 52.73 a 2324.87 a 65.47 a S.E.M. 4.20 0.85 1.95 68.61 2.69 C.D. 5% 12.73 NS 5.93 208.11 8.15 C.V. 12.73 13.72 11.48 9.14 13.36 Discussion Seed germination is an intricate process involving several physiological and biochemical changes modulated by several biochemical enzymes, and seed priming has been proven to alleviate the adverse effects of any stress during early seedling establishment. The present study suggested that seeds primed with botanicals accelerated seed germination rate and significantly enhanced seed vigour, as indicated by longer root lengths, shoot lengths, seedling length and seedling dry weight compared with the control (Table 1 ). Our findings are similar to previous results of botanical priming on black gram using neem and prosopus, demonstrating a significant increase in standard germination, shoot length, root length, seedling length and vigour 19 . Various physiological, biochemical, and molecular changes occurred during seed priming, contributing to improved germination rate and seedling vigour under various environmental conditions 20 . The increase in seedling characteristics was due to enhanced enzyme activity from bioactive substances like curcumin and phenols in the turmeric leaf extract. Similar results were reported in greengram 21 , clusterbean 22 , Vigna sinensis 23 , maize 23 and wheat 24 . The increase in dry weight with botanical treatment may be due to the faster growth and development of seedlings and the hike in vigour index 25 . The high vigour index in botanically primed seeds may be due to growth-promoting compounds, and secondary metabolites translocated during seedling growth. Priming improves chickpea starch metabolism by increasing amylase content during germination (Fig. 1 ). Similar results were obtained by Mukasa and coworkers in sugar beets 26 , where the level of amylase activity in primed sugar beets was 1.9 to 11.5 times higher than in the control group. In similar work on rice, seed priming increases the α-amylase activity and total soluble sugar content, resulting in a higher starch degradation process under chilling stress27. Seed priming induced primary memory, which activates pre-germinative metabolism in seed that triggers gibberellin biosynthesis, antioxidants 28 , 29 , protein synthesis 28 , amylase and protease activation (Fig. 2 ), which helps in radical protrusion and enhances the antioxidant defense system against DNA damage 30 . Proteins stored in the seed are utilized during germination to provide amino acids and amides for the embryo's development. Proteases play a crucial role in protein degradation during the maturation process. Protease activity was highest in primed seeds due to botanical priming (Fig. 2 ), which showed improved nitrogen metabolism in primed seeds, as reported in the pearl millet 31 . In our investigation, increasing priming duration led to enhanced protease activity (Fig. 2 ); similar results were obtained in beans where the proteolytic enzyme activity increased during the first seven days of seed germination 32 . Dehydrogenase enzymes are essential components of the electron transport chain, facilitating the transport of electrons and ATP production 33 , and their activity is considered a positive biomarker for testing seed viability and vigour 34 . The present study observed a relatively high amount of dehydrogenase after priming. Similar trends were reported in cucumber 35 and cowpea 36 , suggesting the role of priming in accelerating dehydrogenase activity. Seeds store phytic acid (phytate), the phosphorous storage form in the plants 37 , which can bind with essential cations like calcium, magnesium, and zinc, reducing their availability for digestion 38 . However, phytic acid is broken down by the enzyme phytase during germination, releasing cations, phosphates, and inositol utilized by the seedlings 39 . Priming significantly increased the activity of the phytase enzyme (Fig. 4 ); this may result from the de novo synthesis of phytase during germination 40 . A similar trend was reported for germinating rice 41 , lupin 42 , barley 40 and soybean 43 . A maximum of 7-fold increase in phytase activity was observed on the 10th day of germination in rice 41 . Significant differences in the phytase activity of wheat, rye, barley, and oats grains were observed, with rye grains showing the highest activity and oats being the lowest. After four days, wheat, barley, and oat activities increased approximately 4.5, 6, and 9-fold, respectively, and rye activities increased approximately 2.5-fold after three days of germination 44 . Seed priming significantly decreased the rate of lipid peroxidation in terms of MDA content (Fig. 5 ). MDA content was reduced after priming due to increased antioxidant enzyme activity 45 . In similar work, malondialdehyde content was 9% lower in primed pea seeds at 42 h of germination against unprimed seeds 46 . This decreased level of MDA indicated decreased lipid peroxidation, which helps maintain the integrity of the membrane in primed seeds 47 . In our finding, the protein content was significantly affected upon botanical priming, where the protein content was increased during priming and subsequently decreased during germination (Fig. 6 ). This was in accordance with the reports documented from black chickpea primed with MgO nanoparticles 28 . After priming, two minor bands of 12 kDa and 14 kDa subunits of albumin were identified (Fig. 7 ); our findings are similar to previous research 48 , where it was reported that chickpea 2S albumin (∼20 kDa) is composed of two polypeptides of 10 and 12 kDa. In similar findings, it was hypothesized that the extra peptide could be a peptide of 2S albumin with Mw 4–10 kDa 49 . Similar trends were reported in cucumber 35 and black beans 50 . The probable reason for the extra one band in all treated seeds is priming, resulting in the synthesis of lost proteins and some new ones. Chickpea seeds primed with aqueous leaf extract of turmeric had higher field emergence and plant heights during early establishment and at maturity than the control (Table 3 ). In the present study, turmeric leaf extract priming improved the seedling growth attributes by triggering the biosynthesis of nucleic acid, proteins, and hydrolytic enzymes and consequentially enhanced the cell division, cell enlargement, and metabolic activity and increased the photosynthetic process of the plant, resulting in increased uptake of more nutrients by efficient and more robust roots. In similar works on nanoparticle priming, AgNPs accumulated in the seeds might activate the metabolic events vital for seed germination and seedling growth 51 . Significant differences were observed in the number of pods, seed yield, and harvest index in turmeric primed seed over control (Table 4 ). The results are in accordance with the studies on maize, where seeds primed with prosopis and moringa leaf extracts led to higher seed yield and yield-related parameters 52 . Physiologically active substances in the turmeric leaf may have activated the embryo growth, resulting in early seedling emergence from the soil. The early growth of roots is vital for establishing a robust and efficient root system, which contributes to the development of higher seedling vigour. The elasticity of the cell walls plays a significant role in ensuring effective water absorption, which is essential for the healthy growth of plants. As a result, the seedlings are better equipped to cope with the challenges of their environment, leading to stronger and healthier plant growth53. A similar observation was made in blackgram 25 , greengram 54 and okra 55 . Conclusions This study conclusively demonstrates the effectiveness of botanical priming as a low-cost and eco-friendly technique to improve seedling attributes, hydrolytic enzymes, and yield significantly. Multiple lab and field studies suggest that botanicals can internalize seed coats and support water uptake inside seeds, accelerating various enzymatic activities and promoting seed germination. The study also indicated that priming with botanicals increases the root length due to the presence of phenol, which indirectly influences the germination % and increases the vigour index Ι and ΙΙ. Furthermore, as evidenced by biochemical activities, it can be hypothesized that the active ingredients in the botanicals, such as phenol and curcumin, accelerate the enzymes' activities, total proteins, and seed storage proteins. Since the low concentrations of botanicals help support early seedling establishment and prevent the attack of fusarium wilt disease during plant establishment, this leads to a healthy plant population and growth, ultimately resulting in higher crop yield. Therefore, botanical priming could be a cost-effective means of increasing the production and productivity of rainfed chickpea crops, which can further support the sustainable development of agriculture and improve the farmers' socio-economic condition. The study can serve as a boon for botanical priming applications for sustainable agricultural practices and the Agri-seed industry in the future. Material and methods Experimental materials. The present research used a medium-vigour seed lot of the chickpea cv. PG-186. The seed lot that exhibited less than 85% germination was considered medium-vigour and the standard germination test was conducted as per the ISTA standard (2020) 56 . Further, all the experiments and field study complies with local and national guidelines and regulations. Priming materials and techniques. Fresh turmeric and neem leaves were shaded for 5–6 days and then dried in a hot air oven for 2–3 hours at 60ºC. Dried leaves were ground into powder, dissolved in 100 mL distilled water, and left overnight at room temperature. Further, the filtrate was used as the priming agent. Hydropriming was carried out by soaking the seeds in distilled water. The seeds without priming and farmers' standard practice of 2g/kg bavistin treatment were taken as control. Seeds were placed in petri dishes between moist filter paper for 18 hours at (20 ± 1°C) and (80–85%) relative humidity. They were then air-dried for 48 hours to their original moisture content. Physiological analysis of chickpea upon priming. Physiological analysis was carried out of all the treatments, namely T 1 (without priming), T 2 (bavistin treatment), T 3 (1% aqueous turmeric leaf extract), T 4 (1% aqueous neem leaf extract), and T 5 (hydropriming) by following completely randomized design. All the experiments were conducted in four replicates. Seeds of different treatments were placed in rolled paper towels and kept in a germination chamber at 20 ± 1°C and 85% RH for a standard period of 8 days by following International Seed Testing Association (ISTA) protocols. All the physiological evaluations were carried out on the 8th day of germination. Seedling Characteristic Determination. The seeds were assessed for different morphological indexes of seedlings, such as germination percentage, root length (RL), shoot length (SL), seedling length (RL + SL), seedling dry weight, and seedling vigour indices I and II. Once the dry weight of the seedlings was determined, ten of them were carefully wrapped in wax paper and kept in a hot air oven maintained at a precise temperature of 80 ± 2°C for 17 hours. After this, the seedlings were allowed to cool for 45 minutes in desiccators before being weighed using an electronic scale. The dry mass of each seedling was calculated and expressed in grams. Additionally, the germination rate (the percentage of average germinated seed out of all tested seeds at the end of the entire test) was calculated, and seedling vigour indices (seedling vigour index I and seedling vigour index II) were estimated using the standard formulae 57 . Assays of Biochemical Activities Amylase activity was measured using Bernfeld's method 58 with some modifications. 1g of seed sample was ground in 10 ml of 10 mM CaCl 2 , and the supernatant was used for enzymatic activities. Starch and enzyme solutions were incubated at 27ºC for 30 min, and DNS reagent was added. After heating and adjusting the solution, optical density was measured at 560 nm, and a maltose standard curve was used. The enzymes were extracted from a 1g seed sample in acetone and centrifuged to assess protease activity 59 . Add casein solution to each tube to assay enzyme activity and incubate at 35°C. The enzyme solution was added and incubated again. TCA and sodium carbonate with FCR were added and incubated. The solution was filtered, and optical density was measured using a UV-Vis spectrophotometer at 600 nm. The dehydrogenase activity of primed and non-primed seed was quantified 60 , where imbibed seed samples of 200 mg were soaked in a freshly made, pH-7.0, 0.2% TTC solution in 10 ml. The solution was then incubated in the dark for 3 hours at 30°C. After draining the TTC solution, acetone was added to each tube for crushing, and the sample was incubated overnight before being centrifuged for 10 minutes at 10,000 rpm. Using a UV-Vis spectrophotometer, collect the supernatant and measure the absorbance at 480 nm. The phytase activity was measured using a modified method 61 . One gram of seed sample was homogenized in 0.1M sodium acetate buffer (pH 5.0) and centrifuged. Phytase activity was determined by adding buffer and sodium phytate solution, incubating, adding a crude enzyme, and incubating and measuring phosphate liberation with the ammonium molybdate method. MDA content was determined using 20% TCA and 0.5% TBA solution to quantify lipid peroxidation 62 . The seed sample was ground in 4 ml of 1% TCA solution. After centrifuging, the supernatant was collected. 1 ml of (20% TCA and 0.5% TBA) was added to each sample. The absorbance was accurately measured at 532 nm and 600 nm for specific and non-specific samples, following a rigorous incubation and centrifugation process. MDA content was measured in moles/ml. Calculation: $$\text{M}\text{D}\text{A} \left(\text{m}\text{M}\right)= \frac{{A}_{532}-{A}_{600}}{155} \times 100$$ The extinction coefficient of this MDA-TBA abduct at 532 nm is 155 mM-1cm-1. Characterization of seed storage proteins. The total protein was estimated from chickpea seed, and a standard curve was produced using Bradford's standard solution 63 . A sample of dried chickpea seeds was ground using a hammer mill; fine powder was obtained, passed through a 0.185 mm mesh grid, and kept in air-tight plastic containers at room temperature to prevent spoilage. Chickpea flours were defatted overnight using a horizontal shaker with hexane in a ratio of 1:10 and then washed twice with ethyl ether, followed by drying for 1 hour at -20ºC. Protein fractions were obtained using the Osborne (1907) fractionation method 64 . Seed flours were extracted by stirring in borate buffer at pH-7.6 with NaCl and sodium azide for 2 hours, followed by centrifugation at 30,000 rpm for 30 minutes. Polyacrylamide Gel Electrophoresis analysis. SDS-PAGE was carried out using the 5% stacking gel and 12% resolution gel 65 . Sample solutions were prepared from 10 mg of freeze-dried protein extract or precipitates dissolved in 1 ml sample buffer (distilled water, 0.5 M Tri-HCl pH 6.8, glycerol, 10% SDS, 1% bromophenol blue and β-mercaptoethanol heated at 98°C for 10 min, then applied to the sample wells. Electrophoretic migration was monitored at a constant current (12 mA/gel) for 1.5 to 2 h. SDS gels were stored for two hours, and distaining was done for 12 hours. Field experiment. The field research trial used a randomized complete block design with eight replications and three priming treatments: T 1 (dry seed as control), T 2 (bavistin treated as positive control), and T 3 (turmeric leaf extract primed). The seed was sown in 5 rows per plot, with a plot size of 2.8×1.8 m 2 . Field emergence percentage (no. of seedlings emerged/total no. of seed sown) was calculated 15 days and 30 days after sowing. Five randomly selected plants from each replicated plot of all treatments were tagged to take all the observations, such as plant height (at 15 DAS, 30 DAS, 45 DAS and maturity) and yield parameters. The biological yield was calculated before threshing by taking the total weight of harvested crop plants from the net plot area. The biological yield was given in kg ha − 1 . After sun drying for a few days, the harvested crop from the respective net plot was threshed with a thresher. Seed yield was recorded and expressed as kg/ha. The Harvest Index (HI) was calculated using formulae- $$\varvec{H}\varvec{I}=\frac{\varvec{B}\varvec{i}\varvec{o}\varvec{l}\varvec{o}\varvec{g}\varvec{i}\varvec{c}\varvec{a}\varvec{l} \varvec{Y}\varvec{i}\varvec{e}\varvec{l}\varvec{d}}{\varvec{E}\varvec{c}\varvec{o}\varvec{n}\varvec{o}\varvec{m}\varvec{i}\varvec{c} \varvec{Y}\varvec{i}\varvec{e}\varvec{l}\varvec{d}} \times 100$$ Statistical analysis. The collected data were analyzed using the statistical software SPSS and the analysis of variance technique. The treatment means were compared using a post-hoc test at a 5% significance level to determine whether there were any significant differences between them. Furthermore, a graphical representation of the data was created using Microsoft Excel to provide a clear and visual presentation of the results. Declarations Acknowledgements The authors acknowledge Bihar Agricultural University, Sabour, for providing the infrastructural facilities to carry out the research work and the BAU communication number of the manuscript. The author also acknowledges the fund support by the Science and Engineering Research Board, Government of India. Data Availability All data generated or analysed during this study are included in this published article and its supplementary information files. Author’s contributions Kamini Kaushal: Conducted the experiments, collected the data, played a crucial role in data interpreting and compiling the results. Kumari Rajani: Conceptualized the research idea and designed the experiments, significantly contributing to drafting and finalizing the manuscript. Ravi Ranjan Kumar: Designed the experiments and prepared and refined the manuscript draft. Anand Kumar: Designed field experiments and data analysis of the field data recorded. Tushar Ranjan: He contributed to preparing protein samples, executing experiments, and proofreading the manuscript. Vinod Kumar: Actively participated in collecting the research material for this project. Feza Ahmad: Provided proofreading of the manuscript and intellectual input. Vikas Kumar: Contributed to the standardization of biochemical assays and Aman Kumar: Conducted field experiments and recorded high-quality data accurately. Competing interests The authors declare no conflict of interest. References Agrawal, T., Kumar, A., Kumar, S., Kumar, A., Kumar, R.R., Kumar, S. & Singh, P.K. Correlation and path coefficient analysis for grain yield and yield components in chickpea ( Cicer arietinum L. ) under normal and late sown conditions of Bihar. International Journal of Current Microbiology and Applied Science , 7 (2), 1633–1642, https://doi.org/10.20546/ijcmas.2018.702.197 (2018). Krishnamurthy, L., Johansen, C. & Sethi, S.C. 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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-3914755","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":273222980,"identity":"c49fda6e-0fcc-4526-b8a6-82e5815cad29","order_by":0,"name":"Kamini Kaushal","email":"","orcid":"","institution":"Bihar Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Kamini","middleName":"","lastName":"Kaushal","suffix":""},{"id":273222981,"identity":"f15b159a-2ae4-4d70-a531-7db91d969586","order_by":1,"name":"Kumari Rajani","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEElEQVRIiWNgGAWjYHCCBAYGAwbGBiDrAAODhBxI6MAD4rQwg7UYg7UkEGEVWAsIJDZAjcEJzNsPPPxcUMAgO7///MEDP/dYpM8PO/wQaIudnG4Ddi0yZxKSpWcYMBhvuJHMcLDnmUTuxttpBkAtycZmB7BrkWBISJDmMWBI3CAB9AvPAaCW2QkgLQcSt+HSwv8g+TdIy/z+wwwH/xyQSDecnf4BvxaJhDSwLQ0HkhkOA21JkJfOIWCLxIM0ax4DCZBfDA7LHJAw3CCdU3AgwQCPX/hzkm/z/LEBhtjBxx/fHKiTl5+dvvnDhwo7OVxaGBh4EsChAAcGYJUGuJSDADuaYfIN+FSPglEwCkbBSAQAkqdh8omMYvIAAAAASUVORK5CYII=","orcid":"","institution":"Bihar Agricultural University","correspondingAuthor":true,"prefix":"","firstName":"Kumari","middleName":"","lastName":"Rajani","suffix":""},{"id":273222982,"identity":"a64c1037-2a86-44d9-b50a-db3e549cc9ae","order_by":2,"name":"Ravi Ranjan Kumar","email":"","orcid":"","institution":"Bihar Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Ravi","middleName":"Ranjan","lastName":"Kumar","suffix":""},{"id":273222983,"identity":"8fe37a63-fac4-4f1a-a944-2e70f1eed335","order_by":3,"name":"Anand Kumar","email":"","orcid":"","institution":"Bihar Agricultural University Sabour","correspondingAuthor":false,"prefix":"","firstName":"Anand","middleName":"","lastName":"Kumar","suffix":""},{"id":273222984,"identity":"04ea005c-43c3-445c-8858-48f7bd41199b","order_by":4,"name":"Tushar Ranjan","email":"","orcid":"","institution":"Bihar Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Tushar","middleName":"","lastName":"Ranjan","suffix":""},{"id":273222985,"identity":"8d0fd2e2-536d-4f70-945c-a31602ed4e37","order_by":5,"name":"Vinod Kumar","email":"","orcid":"","institution":"Bihar Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Vinod","middleName":"","lastName":"Kumar","suffix":""},{"id":273222986,"identity":"85499a9b-3492-408d-94ae-876a8f515b6f","order_by":6,"name":"Feza Ahmad","email":"","orcid":"","institution":"Bihar Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Feza","middleName":"","lastName":"Ahmad","suffix":""},{"id":273222987,"identity":"f8a30e1f-e86b-4977-96b3-23d84f17c4e5","order_by":7,"name":"Vikash Kumar","email":"","orcid":"","institution":"Bihar Agricultural University Sabour","correspondingAuthor":false,"prefix":"","firstName":"Vikash","middleName":"","lastName":"Kumar","suffix":""},{"id":273222988,"identity":"8d6d9085-a261-45ad-8a60-d8d22f3505d8","order_by":8,"name":"Aman Kumar","email":"","orcid":"","institution":"Bihar Agricultural University Sabour","correspondingAuthor":false,"prefix":"","firstName":"Aman","middleName":"","lastName":"Kumar","suffix":""}],"badges":[],"createdAt":"2024-01-31 18:16:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3914755/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3914755/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-024-59878-8","type":"published","date":"2024-04-23T18:05:11+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":51304215,"identity":"40ca7577-9dc5-4de5-bda4-07f747f8f7a8","added_by":"auto","created_at":"2024-02-19 08:12:08","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":30748,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of different priming agents on amylase activity during 6 h, 12 h and 18 h duration of priming and 1\u003csup\u003est\u003c/sup\u003e, 3\u003csup\u003erd\u003c/sup\u003e, 5\u003csup\u003eth\u003c/sup\u003e days of germination in primed and non-primed seed of chickpea variety PG-186. Data presented are means of four replicates with standard deviation. Within each treatment, different letters indicate significant differences by Post Hoc test at P=0.05 levels. Where T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/5602b2059421b108fb7aab54.png"},{"id":51304214,"identity":"06b826ce-6c3a-4d26-b614-f23d3a4c8db9","added_by":"auto","created_at":"2024-02-19 08:12:08","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":29322,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of different priming agent on protease activity during 6 hr, 12 hr and 18 hr duration of priming and 1\u003csup\u003est\u003c/sup\u003e, 3\u003csup\u003erd\u003c/sup\u003e, 5\u003csup\u003eth\u003c/sup\u003e days of germination in primed and non -primed seed of chickpea variety PG-186. \u0026nbsp;Error bars are the representative of standard deviation of four replicates. Within treatment, different letters indicate significant differences by Post Hoc test at P =0.05 levels. Where T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/d6b71540b8d162a88c06d6c7.png"},{"id":51304219,"identity":"6c0e9780-d877-4f69-8203-b721369d79aa","added_by":"auto","created_at":"2024-02-19 08:12:09","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":28716,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of botanical priming on dehydrogenase activity during 6 h, 12 h and 18 h duration of priming and 1\u003csup\u003est\u003c/sup\u003e, 3\u003csup\u003erd\u003c/sup\u003e, 5\u003csup\u003eth\u003c/sup\u003e days of germination in primed and non -primed seed of chickpea variety PG-186.vertical bar presented are means of four replicates with standard deviation. Within each treatment, different letters indicate significant differences by Post Hoc test at P =0.05 levels. Where T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/3d345e12e9df1e18d4800956.png"},{"id":51304222,"identity":"f17d7074-9b9a-46df-a134-42c912269382","added_by":"auto","created_at":"2024-02-19 08:12:09","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":21574,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of different priming agent on phytase activity during 6 h, 12 h and 18 h duration of priming and 1\u003csup\u003est\u003c/sup\u003e, 3\u003csup\u003erd\u003c/sup\u003e, 5\u003csup\u003eth\u003c/sup\u003e days of germination in primed and non -primed seed of chickpea variety PG-186. \u0026nbsp;Error bars are the representative of standard deviation of four replicates. Within each treatment, different letters indicate significant differences by Post Hoc test at P=0.05 levels. Where T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/6def09e1e7b6299d41d8ed8d.png"},{"id":51304216,"identity":"87ef0633-124f-47ac-bedf-1af6ebd9825d","added_by":"auto","created_at":"2024-02-19 08:12:08","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":30192,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of different priming agent on MDA content during 6 h, 12 h and 18 h duration of priming and 1\u003csup\u003est\u003c/sup\u003e, 3\u003csup\u003erd\u003c/sup\u003e, 5\u003csup\u003eth\u003c/sup\u003e days of germination in primed and non -primed seed of chickpea variety PG-186. Data presented are means of four replicates with standard deviation. Within each treatment, different letters indicate significant differences by Post Hoc test at P =0.05 levels. Where T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/a1169bc059a34ce9092b0eaf.png"},{"id":51304221,"identity":"7fd92e09-7429-4b12-9c12-45447a63aa90","added_by":"auto","created_at":"2024-02-19 08:12:09","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":29367,"visible":true,"origin":"","legend":"\u003cp\u003eImpact of different priming treatment on total protein content in chickpea during 6 h, 12 h and 18 h duration of priming and 1\u003csup\u003est\u003c/sup\u003e, 3\u003csup\u003erd\u003c/sup\u003e, 5\u003csup\u003eth\u003c/sup\u003e days of germination in primed and non -primed seed of chickpea variety PG-186. Data presented in each column are means of four replicates with standard deviation, within treatments different letter indicates significant difference by post-Hoc test at P=0.05% level. Where T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/68b5146b722fc6ccd41722a0.png"},{"id":51304223,"identity":"eb7cf5f7-2855-43d1-b6a7-166086f48a6d","added_by":"auto","created_at":"2024-02-19 08:12:09","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1288473,"visible":true,"origin":"","legend":"\u003cp\u003e12 % of SDS- PAGE of chickpea seed storage protein extracted and characterized by Osbornes methods using the borate buffer. Shows the effect of different priming treatment on the expression of globulin and albumin sub unit during 6,12,18 hour of priming.12kDa and 14kDa are the sub units of albumin and 36kDa, 42kDa and 56kDa are the globulin sub units. Seed treatments includes T\u003csub\u003e1\u003c/sub\u003e = No priming (control), T\u003csub\u003e2\u003c/sub\u003e = Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e = Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e = Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e = On-farm priming (Hydropriming).\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/b5f729e91486cf93af3be4f7.png"},{"id":51304224,"identity":"7d6fd37b-758b-47f8-b88e-2552021a3cf5","added_by":"auto","created_at":"2024-02-19 08:12:09","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":186499,"visible":true,"origin":"","legend":"\u003cp\u003ePossible mechanism upon botanical seed priming in chickpea\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/db3a8302f6873eaa02bae50f.png"},{"id":55398952,"identity":"9d586663-7780-4109-b154-9dc269c2e17f","added_by":"auto","created_at":"2024-04-26 18:05:22","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2111043,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/2a7c4203-180a-48e9-9029-9b41a790fe54.pdf"},{"id":51304217,"identity":"e4452f8f-ba46-4eb9-8549-dd1adf95b27c","added_by":"auto","created_at":"2024-02-19 08:12:08","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":16906,"visible":true,"origin":"","legend":"","description":"","filename":"1.PhysiologicalData.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/9010cf2d9a2bf96351a2e907.xlsx"},{"id":51304220,"identity":"9a54e573-d80b-4ca7-b924-68d3233a9a16","added_by":"auto","created_at":"2024-02-19 08:12:09","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":89786,"visible":true,"origin":"","legend":"","description":"","filename":"2.BiochemicalData.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/0df5c2c461b53214efb320f3.xlsx"},{"id":51304218,"identity":"33c2ccf9-67a0-46dc-9057-651fb4114c5f","added_by":"auto","created_at":"2024-02-19 08:12:08","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":22428,"visible":true,"origin":"","legend":"","description":"","filename":"3.FieldData.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-3914755/v1/612c6750f9139664a485eb48.xlsx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Physio-biochemical responses and crop performance analysis in chickpea upon botanical priming","fulltext":[{"header":"Introduction","content":"\u003cp\u003eGlobal climate change and water scarcity generally affect crop performance from germination and eventually reduce grain yield and quality with losses of up to 40\u0026ndash;50% in crop productivity. Chickpea, a cool-season legume, are an excellent source of protein (18\u0026ndash;20%) and the minerals phosphorus, calcium, magnesium, iron, and zinc that play a significant role in achieving global food security\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Over 90% of the world's chickpeas are grown in arid and semi-arid regions, making them vulnerable to various stresses\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e,\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Even though it is considered the candidate crop for rainfed conditions, the uncertain climatic condition adversely affects the crop's performance and make it vulnerable to several biotic and abiotic stresses\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eSuccessful seedling and crop establishment are critical to higher crop production\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Poor seedling establishment is one of the most significant barriers primarily caused by subpar seed quality\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The improper storage is the primary factor for quick loss of seed vigour, which ultimately affects germination and crop performance. Moreover, crop germination due to adverse climatic conditions and delayed sowing affects production and productivity. Seed quality enhancement techniques provide a comprehensive solution to unlock the full genetic potential of seeds. Seed quality enhancement, a pre-sowing treatment, enhances germination and crop vigour during early planting and contributes to a higher crop yield\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eSeed priming, a technique in seed enhancement, involves the initial absorption of water by seeds to initiate the early stages of germination. However, this absorption is insufficient for radical emergence, and the seeds return to their original moisture content\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. During the priming process, biochemical changes occur, including activating enzymes, producing compounds that promote growth, metabolizing molecules that inhibit germination, and repairing damaged cells\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The imbibition of seeds triggers a cascade of metabolic processes, such as the activation of hydrolytic enzymes (such as amylase, protease, lipase, dehydrogenase, and phytase), which lead to the hydrolysis of stored starch, lipids, proteins, polyphosphates, and other storage materials, converting them into simpler forms that readily absorbed by the embryo and ultimately influencing seed vigour\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Alpha-amylase breaks down starch into sugars for the developing embryo\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Proteolytic enzymes use seed storage proteins\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, while dehydrogenase catalyzes the stored products during the anaerobic phase of seed germination\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003ePhytase is an enzyme that unequivocally converts phytate into inositol/phosphoric acid, making a remarkable contribution to seed germination and growth\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. On the other hand, lipid peroxidation is an unequivocally detrimental process that destabilizes the membrane and degrades proteins, leading to inevitable cell death, which hinders the capacity for ionic transport\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Similarly, the hydrolysis of seed storage proteins such as globulin and albumin unequivocally generates free amino acids, essential for germination and seed vigour\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eOver the past few decades, there has been a significant increase in the utilization of non-hazardous chemicals and fertilizers as alternatives to enhance crop productivity in the agriculture system. Among these alternatives, priming has emerged as a highly effective method to tackle this issue. Numerous efficacious priming agents, such as salts, polyamines, hormones, compatible solutes, and aqueous plant extracts, have been documented by various researchers\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Botanical priming, a priming technique that employs plant extracts as its agents, has proven particularly advantageous. This botanical priming method can stimulate metabolic processes, is non-toxic and environmentally friendly, and has excellent potential to control pathogenic microorganisms.\u003c/p\u003e \u003cp\u003eThere are few reports on using turmeric rhizome and neem seed extracts on aphid control, plant growth and crop yield. However, the effect of leaf extracts on seed physiology and plant growth in pulse crops has yet to be studied. No studies have investigated globulin and albumin expression in chickpeas during priming and germination at different durations. In the present investigation, we hypothesized that, first, the effect of turmeric and neem leaf extracts as seed priming agents positively influences physiological parameters and enhances seed vigour in chickpeas. Second, the improved enzymatic activities during botanical priming lead to increased seed germination and growth. Third, priming also increases the expression of different seed storage proteins compared to non-primed conditions. Our objective was to investigate the effect of botanical priming on physiological, biochemical and yield in chickpeas, for which we performed comprehensive laboratory and field experiments.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e \u003cb\u003eEffect of botanical priming on seed germination and seedling characteristics.\u003c/b\u003e The present study was carried out to analyze the effects of seed priming with botanicals (turmeric, neem) on the physiological parameters of chickpeas and compare it with conventional hydropriming and fungicide seed treatment. In the present investigation, physiological parameters, viz germination percentage and seedling characteristics, were significantly increased by priming the seeds with botanicals, as shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The botanical primed seed had significantly higher seed germination and seedling growth than the control. Seeds primed with turmeric and neem leaves aqueous extract had higher germination and seedling growth followed by hydro priming. The results demonstrate that turmeric had a higher germination percentage (94.5%) than control (82.5%), indicating that botanical priming successfully enhanced the germination rate by 15% over control. Seedling characteristics, including root length, shoot length, seedling length, seedling dry weight and vigour indices, were significantly similarly affected by botanical priming. Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows marked enhancement in seedling length by 29%, vigour index Ι by 42% and vigour index II by 63% were observed in turmeric primed seed over control.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eImpact of different priming agents on germination percentage, root length, shoot length, seedling length, dry weight and vigor indices was estimated on 8th day of germination. Data presented in a column are means of four replicates, where each replicate of the treatment contains 100 seeds. Means in each column, followed by different lower-case alphabets are significantly different at the 0.05% probability level using Post Hoc test. Where T\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;No priming (control), T\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;On-farm priming (Hydropriming).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGermination\u003c/p\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRoot length\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eShoot length\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSeedling length\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eDry weight\u003c/p\u003e \u003cp\u003e(g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eVI Ι\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eVI ΙΙ\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT\u003c/b\u003e\u003csub\u003e\u003cb\u003e1\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e82.5\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.5\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.73\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.77\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.41\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2050.14\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e33.24\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT\u003c/b\u003e\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e83.5\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.07\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.97\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26.99\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.43\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2226.8\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e35.52\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT\u003c/b\u003e\u003csub\u003e\u003cb\u003e3\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e94.5\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.16\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.77\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e31.93\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.6\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2905.71\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e54.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT\u003c/b\u003e\u003csub\u003e\u003cb\u003e4\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e86.75\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.38\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e29.4\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.51\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2226.8\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e44.44\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT\u003c/b\u003e\u003csub\u003e\u003cb\u003e5\u003c/b\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e86.75\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.04\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26.06\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.49\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2549.21\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e42.68\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS.E.M.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e57.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.D. 5%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e174.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.V.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.89\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eBiochemical changes upon botanical priming.\u003c/b\u003e The dynamic changes in hydrolytic enzymes and MDA content are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows that botanical priming and hydropriming significantly affected amylase activity, which increased with the increase in priming duration. In contrast, the amylase activity was decreased as the germination proceeded. Conversely, the amylase activity was constant at different priming hours in non-primed and bavistin-treated seeds; however, they showed a similar decreasing trend during germination as botanicals primed seed. Compared to non-primed seeds, maximum amylase activity was observed in turmeric-primed seeds, and it was increased to 4.8-fold at 12 hours and 5.2-fold at 18 hours of priming. The amylase activity was also assessed during germination, and it showed a decreasing trend with the increasing days of germination, and it was highest in turmeric primed seed. The highest amylase activity, 0.71 mg maltose/min, was observed on the first day of germination on the first day, and its activity decreased to 1.7-fold on the third day and 2.0-fold on the fifth day of germination.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBotanical priming significantly affected the protease activity (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05; Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The protease activity significantly increased with increasing priming duration and till the third day of germination; after that, on the fifth day of germination, irrespective of treatments, the protease activity decreased. The protease activity was maximum during priming durations and germination days in seeds primed with 1% aqueous extract of turmeric leaves. Its activity increased to 1.3-fold in 6 hr, 1.8-fold in 12 hr, 2-fold in 18 hr, 1.2-fold on the first day and 3.2-fold on the third day compared to control in turmeric primed seed. On the fifth day, there was a drastic reduction in the protease activity, which decreased by 1.8-fold compared to the third day of the same treatment. A similar pattern was observed in 1% aqueous extract of neem leaves and hydro-priming, whereas activity was constant throughout the priming duration in control and bavistin-treated seed.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the current study, botanical priming significantly affected dehydrogenase activity (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05; Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) and differed among treatments. The dehydrogenase activity was significantly increased with priming duration and days of germination. The maximum dehydrogenase activity of 0.98 mg formazan/min was observed on the fifth day of germination in seeds primed with 1% aqueous extract of turmeric leaves. Its activity increased to 1.84-fold in 18 hr and 1.3-fold on the fifth day of germination compared to the control.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo confirm the activation of dormant phytase zymogens induced by botanical priming, we investigated the dynamic changes in phytase activity during priming durations and days of germination. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, the phytase activity significantly increased with increasing priming duration and days of germination. The maximum activity was observed in turmeric primed seed during the fifth day of germination, and it was 0.74 \u0026micro;M trypsin/min (1.9-fold increased over control at a particular time). Compared to the control, its activity in turmeric primed seed increased to 1.7-fold in 6 hours, 2.4-fold in 12 hours, and 2.9-fold in 18 hours of priming. The phytase remained active during germination and increased by 2-fold on the first day, 2.1-fold on the third day and 1.9-fold on the fifth day of germination.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBotanically primed seed showed a significant reduction in lipid peroxidation activity, measured in terms of MDA content (\u0026micro;mol MDA/gr F-W). In the present study, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, on increasing the priming duration, the MDA content was not found to change up to 12 hours in turmeric primed seed; after that, it decreased sharply at 18 hours of priming and during days of germination. Compared to the control, the MDA content decreased to 2.49-fold in 18 hours of priming, 3.4-fold on the first day, 4-fold on the third day, and 3.6-fold on the fifth day of germination and the maximum MDA content was observed in the case of the control seed (T\u003csub\u003e1\u003c/sub\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eEffects of botanical priming on total protein and the expression of seed storage protein.\u003c/b\u003e The impact of botanical priming on total protein content was significant in our present work; as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e, a significant upsurge (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in protein content was evident during priming hours and days of germination with both turmeric and neem leaf aqueous extract. In this, the total protein content increased with the priming duration up to the first day of germination. However, upon increasing the days of germination, irrespective of the priming material, the total protein decreases significantly from the third to the fifth days. Meanwhile, in the control and bavistin-treated seeds, protein content did not vary much throughout the priming duration and during germination. The total protein content increased 1.4-fold on the first day after that, decreased to 1.1-fold on the third day and 1-fold on the fifth day of germination compared to the control. Seed storage proteins (SSP) of chickpeas, i.e., globulin and albumin, were fractionalized by the Osborne method\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e and the molecular weight of the protein was determined using SDS PAGE. Subunits separated from chickpea proteins are shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Figure\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e shows that the expression of proteins was higher in primed seeds than in the control. In the present work, the estimated molecular weights of subunits of globulin, i.e., legumin, were (40 kDa, 39 kDa, 26 kDa, 23 kDa), vicillin Mw (50 kDa, 39 kDa, 19 kDa, 15 kDa), and glutelin (58 kDa, 55 kDa, 54 kDa). Three minor bands of 36 kDa, 42 kDa, and 56 kDa, the globulin subunits, were detected in the electrophoregram. Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e also reveals that the significant globulin subunits, i.e., 42 kDa and 56 kDa, were present in all primed seeds, whereas control and bavistin polypeptides of 42 kDa proteins were not observed. Our work in SDS-PAGE showed a progressive accumulation of the 11-S globulin during priming. In contrast, a slower accumulation of 11S occurred in control seeds, which resulted in an intense polypeptide of 56 kDa and 42 kDa in primed seeds.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of botanical priming on the expression of seed storage proteins (globulin and albumin) detected through SDS-PAGE. Where (+) and (\u003cb\u003e\u0026ndash;\u003c/b\u003e) sign in each column indicates presence and absence of polypeptide of globulin and albumin sub unit during 6, 12 and 18 h duration of priming. Seed treatments includes T\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;No priming (control), T\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Turmeric leaf priming, T\u003csub\u003e4\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Neem leaf priming, T\u003csub\u003e5\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;On-farm priming (Hydropriming).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eSeed Storage protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eMolecular Weight\u003c/p\u003e \u003cp\u003e(kDa)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"9\" nameend=\"c13\" namest=\"c5\"\u003e \u003cp\u003eDuration of priming (h)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c10\" namest=\"c8\"\u003e \u003cp\u003eT\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c13\" namest=\"c11\"\u003e \u003cp\u003eT\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003e\u003cb\u003eGlobulin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e\u0026ndash;\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eAlbumin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e+\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eEffects of botanical priming on plant establishment, growth and yield of chickpea.\u003c/b\u003e An experiment with turmeric leaves extract priming in chickpea was compared with bavistin seed conventional seed treatment (+\u0026thinsp;control) and non-priming (control) under field conditions. Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e shows that turmeric priming significantly improved field emergence, plant height at different growth stages days to 50% flowering and yield attributing characters (Tables\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e \u0026amp; \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Turmeric priming significantly increased the field emergence by 27%, plant heights at 15 DAS, 45 DAS, and maturity by 5%, 11%, and 4.7%, respectively, compared to the non-priming. About the days to 50% flowering, it was significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) affected by priming and obtained three days earlier than in the control plant (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The effect of turmeric leaf priming on yield-attributing characteristics in chickpeas is shown in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, which presents the data on-field performance, where turmeric priming considerably increased the number of pods per plant, weight of 100 seeds, biological yield, number of seeds per plant, economic yield, and harvest index compared to the control. In comparison to the control, turmeric priming significantly raised the 100 seed weight by 9.1%, the number of pods per plant by 23.4%, the number of seeds per plant by 15.5%, the seed weight per plant by 20.0%, and the harvest index by 18.1%.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of turmeric priming on plant establishment and development. Field emergence %, plant height at 15, 30, 45 days and maturity, days to 50% flowering were observed. The field experiment was laid in RBD, and data presented in each column are means of 8 replicates (n\u0026thinsp;=\u0026thinsp;8). Means in each column, followed by different lower-case alphabets, are significantly different at the 0.05% probability level using the Post Hoc test. Where, T\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;No priming (control), T\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Turmeric leaf priming.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eField Emergence\u003c/p\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePlant height at 15 days\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePlant height at 30 days\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePlant height at 45 days\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePlant height at maturity\u003c/p\u003e \u003cp\u003e(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eDays to 50% Flowering\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT 1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e70.16\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.77\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.7\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e56.1\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e85.00\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT 2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e77.85\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.84\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.75\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.88\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e86.75\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT 3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e88.86\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.37\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27.4\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e58.75\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e82.5\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS.E.M.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.D. 5%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.V.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of priming on yield attributing characters of chickpea variety PG-186 on number of pods/plant, seed weight, biological yield, yield kg/ha, economic yield and harvest index. Data in each column are means of 8 replicates, different lower-case alphabet shows significant level of difference at 0.05% probability level. Where T\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;No priming (control), T\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Conventional practice (Bavistin@2g/kg), T\u003csub\u003e3\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Turmeric leaf priming.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. of Pods/plant\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSeed weight\u003c/p\u003e \u003cp\u003e(g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBiological Yield\u003c/p\u003e \u003cp\u003e(g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eYield\u003c/p\u003e \u003cp\u003e(kg/ha)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHarvest index\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT 1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e84.38\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.28\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2007.14\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e53.6\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT 2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e85.00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e48.39\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2039.62\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e51.51\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eT 3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e110.25\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e52.73\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2324.87\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e65.47\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eS.E.M.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e68.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.D. 5%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e208.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eC.V.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eSeed germination is an intricate process involving several physiological and biochemical changes modulated by several biochemical enzymes, and seed priming has been proven to alleviate the adverse effects of any stress during early seedling establishment. The present study suggested that seeds primed with botanicals accelerated seed germination rate and significantly enhanced seed vigour, as indicated by longer root lengths, shoot lengths, seedling length and seedling dry weight compared with the control (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Our findings are similar to previous results of botanical priming on black gram using neem and prosopus, demonstrating a significant increase in standard germination, shoot length, root length, seedling length and vigour\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Various physiological, biochemical, and molecular changes occurred during seed priming, contributing to improved germination rate and seedling vigour under various environmental conditions\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The increase in seedling characteristics was due to enhanced enzyme activity from bioactive substances like curcumin and phenols in the turmeric leaf extract. Similar results were reported in greengram\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, clusterbean\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, \u003cem\u003eVigna sinensis\u003c/em\u003e\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, maize\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e and wheat\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The increase in dry weight with botanical treatment may be due to the faster growth and development of seedlings and the hike in vigour index\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The high vigour index in botanically primed seeds may be due to growth-promoting compounds, and secondary metabolites translocated during seedling growth. Priming improves chickpea starch metabolism by increasing amylase content during germination (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Similar results were obtained by Mukasa and coworkers in sugar beets\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, where the level of amylase activity in primed sugar beets was 1.9 to 11.5 times higher than in the control group. In similar work on rice, seed priming increases the α-amylase activity and total soluble sugar content, resulting in a higher starch degradation process under chilling stress27.\u003c/p\u003e \u003cp\u003eSeed priming induced primary memory, which activates pre-germinative metabolism in seed that triggers gibberellin biosynthesis, antioxidants\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e,\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, protein synthesis\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, amylase and protease activation (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), which helps in radical protrusion and enhances the antioxidant defense system against DNA damage\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Proteins stored in the seed are utilized during germination to provide amino acids and amides for the embryo's development. Proteases play a crucial role in protein degradation during the maturation process. Protease activity was highest in primed seeds due to botanical priming (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), which showed improved nitrogen metabolism in primed seeds, as reported in the pearl millet\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. In our investigation, increasing priming duration led to enhanced protease activity (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e); similar results were obtained in beans where the proteolytic enzyme activity increased during the first seven days of seed germination\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Dehydrogenase enzymes are essential components of the electron transport chain, facilitating the transport of electrons and ATP production\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, and their activity is considered a positive biomarker for testing seed viability and vigour\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The present study observed a relatively high amount of dehydrogenase after priming. Similar trends were reported in cucumber\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e and cowpea\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, suggesting the role of priming in accelerating dehydrogenase activity.\u003c/p\u003e \u003cp\u003eSeeds store phytic acid (phytate), the phosphorous storage form in the plants\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, which can bind with essential cations like calcium, magnesium, and zinc, reducing their availability for digestion\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. However, phytic acid is broken down by the enzyme phytase during germination, releasing cations, phosphates, and inositol utilized by the seedlings\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Priming significantly increased the activity of the phytase enzyme (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e); this may result from the de novo synthesis of phytase during germination\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. A similar trend was reported for germinating rice\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, lupin\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, barley\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e and soybean\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. A maximum of 7-fold increase in phytase activity was observed on the 10th day of germination in rice\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Significant differences in the phytase activity of wheat, rye, barley, and oats grains were observed, with rye grains showing the highest activity and oats being the lowest. After four days, wheat, barley, and oat activities increased approximately 4.5, 6, and 9-fold, respectively, and rye activities increased approximately 2.5-fold after three days of germination\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eSeed priming significantly decreased the rate of lipid peroxidation in terms of MDA content (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). MDA content was reduced after priming due to increased antioxidant enzyme activity\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. In similar work, malondialdehyde content was 9% lower in primed pea seeds at 42 h of germination against unprimed seeds\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. This decreased level of MDA indicated decreased lipid peroxidation, which helps maintain the integrity of the membrane in primed seeds \u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. In our finding, the protein content was significantly affected upon botanical priming, where the protein content was increased during priming and subsequently decreased during germination (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). This was in accordance with the reports documented from black chickpea primed with MgO nanoparticles\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. After priming, two minor bands of 12 kDa and 14 kDa subunits of albumin were identified (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e); our findings are similar to previous research\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, where it was reported that chickpea 2S albumin (\u0026sim;20 kDa) is composed of two polypeptides of 10 and 12 kDa. In similar findings, it was hypothesized that the extra peptide could be a peptide of 2S albumin with Mw 4\u0026ndash;10 kDa\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Similar trends were reported in cucumber\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e and black beans\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The probable reason for the extra one band in all treated seeds is priming, resulting in the synthesis of lost proteins and some new ones.\u003c/p\u003e \u003cp\u003eChickpea seeds primed with aqueous leaf extract of turmeric had higher field emergence and plant heights during early establishment and at maturity than the control (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). In the present study, turmeric leaf extract priming improved the seedling growth attributes by triggering the biosynthesis of nucleic acid, proteins, and hydrolytic enzymes and consequentially enhanced the cell division, cell enlargement, and metabolic activity and increased the photosynthetic process of the plant, resulting in increased uptake of more nutrients by efficient and more robust roots. In similar works on nanoparticle priming, AgNPs accumulated in the seeds might activate the metabolic events vital for seed germination and seedling growth\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Significant differences were observed in the number of pods, seed yield, and harvest index in turmeric primed seed over control (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The results are in accordance with the studies on maize, where seeds primed with prosopis and moringa leaf extracts led to higher seed yield and yield-related parameters\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Physiologically active substances in the turmeric leaf may have activated the embryo growth, resulting in early seedling emergence from the soil. The early growth of roots is vital for establishing a robust and efficient root system, which contributes to the development of higher seedling vigour. The elasticity of the cell walls plays a significant role in ensuring effective water absorption, which is essential for the healthy growth of plants. As a result, the seedlings are better equipped to cope with the challenges of their environment, leading to stronger and healthier plant growth53. A similar observation was made in blackgram\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, greengram\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e and okra\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThis study conclusively demonstrates the effectiveness of botanical priming as a low-cost and eco-friendly technique to improve seedling attributes, hydrolytic enzymes, and yield significantly. Multiple lab and field studies suggest that botanicals can internalize seed coats and support water uptake inside seeds, accelerating various enzymatic activities and promoting seed germination. The study also indicated that priming with botanicals increases the root length due to the presence of phenol, which indirectly influences the germination % and increases the vigour index Ι and ΙΙ. Furthermore, as evidenced by biochemical activities, it can be hypothesized that the active ingredients in the botanicals, such as phenol and curcumin, accelerate the enzymes' activities, total proteins, and seed storage proteins. Since the low concentrations of botanicals help support early seedling establishment and prevent the attack of fusarium wilt disease during plant establishment, this leads to a healthy plant population and growth, ultimately resulting in higher crop yield. Therefore, botanical priming could be a cost-effective means of increasing the production and productivity of rainfed chickpea crops, which can further support the sustainable development of agriculture and improve the farmers' socio-economic condition. The study can serve as a boon for botanical priming applications for sustainable agricultural practices and the Agri-seed industry in the future.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cp\u003e \u003cb\u003eExperimental materials.\u003c/b\u003e The present research used a medium-vigour seed lot of the chickpea cv. PG-186. The seed lot that exhibited less than 85% germination was considered medium-vigour and the standard germination test was conducted as per the ISTA standard (2020)\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Further, all the experiments and field study complies with local and national guidelines and regulations.\u003c/p\u003e \u003cp\u003e \u003cb\u003ePriming materials and techniques.\u003c/b\u003e Fresh turmeric and neem leaves were shaded for 5\u0026ndash;6 days and then dried in a hot air oven for 2\u0026ndash;3 hours at 60\u0026ordm;C. Dried leaves were ground into powder, dissolved in 100 mL distilled water, and left overnight at room temperature. Further, the filtrate was used as the priming agent. Hydropriming was carried out by soaking the seeds in distilled water. The seeds without priming and farmers' standard practice of 2g/kg bavistin treatment were taken as control. Seeds were placed in petri dishes between moist filter paper for 18 hours at (20\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C) and (80\u0026ndash;85%) relative humidity. They were then air-dried for 48 hours to their original moisture content.\u003c/p\u003e \u003cp\u003e \u003cb\u003ePhysiological analysis of chickpea upon priming.\u003c/b\u003e Physiological analysis was carried out of all the treatments, namely T\u003csub\u003e1\u003c/sub\u003e (without priming), T\u003csub\u003e2\u003c/sub\u003e (bavistin treatment), T\u003csub\u003e3\u003c/sub\u003e (1% aqueous turmeric leaf extract), T\u003csub\u003e4\u003c/sub\u003e (1% aqueous neem leaf extract), and T\u003csub\u003e5\u003c/sub\u003e (hydropriming) by following completely randomized design. All the experiments were conducted in four replicates. Seeds of different treatments were placed in rolled paper towels and kept in a germination chamber at 20\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C and 85% RH for a standard period of 8 days by following International Seed Testing Association (ISTA) protocols. All the physiological evaluations were carried out on the 8th day of germination.\u003c/p\u003e \u003cp\u003e \u003cb\u003eSeedling Characteristic Determination.\u003c/b\u003e The seeds were assessed for different morphological indexes of seedlings, such as germination percentage, root length (RL), shoot length (SL), seedling length (RL\u0026thinsp;+\u0026thinsp;SL), seedling dry weight, and seedling vigour indices I and II. Once the dry weight of the seedlings was determined, ten of them were carefully wrapped in wax paper and kept in a hot air oven maintained at a precise temperature of 80\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C for 17 hours. After this, the seedlings were allowed to cool for 45 minutes in desiccators before being weighed using an electronic scale. The dry mass of each seedling was calculated and expressed in grams. Additionally, the germination rate (the percentage of average germinated seed out of all tested seeds at the end of the entire test) was calculated, and seedling vigour indices (seedling vigour index I and seedling vigour index II) were estimated using the standard formulae\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eAssays of Biochemical Activities\u003c/h2\u003e \u003cp\u003eAmylase activity was measured using Bernfeld's method\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e with some modifications. 1g of seed sample was ground in 10 ml of 10 mM CaCl\u003csub\u003e2\u003c/sub\u003e, and the supernatant was used for enzymatic activities. Starch and enzyme solutions were incubated at 27\u0026ordm;C for 30 min, and DNS reagent was added. After heating and adjusting the solution, optical density was measured at 560 nm, and a maltose standard curve was used. The enzymes were extracted from a 1g seed sample in acetone and centrifuged to assess protease activity\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Add casein solution to each tube to assay enzyme activity and incubate at 35\u0026deg;C. The enzyme solution was added and incubated again. TCA and sodium carbonate with FCR were added and incubated. The solution was filtered, and optical density was measured using a UV-Vis spectrophotometer at 600 nm. The dehydrogenase activity of primed and non-primed seed was quantified\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e, where imbibed seed samples of 200 mg were soaked in a freshly made, pH-7.0, 0.2% TTC solution in 10 ml. The solution was then incubated in the dark for 3 hours at 30\u0026deg;C. After draining the TTC solution, acetone was added to each tube for crushing, and the sample was incubated overnight before being centrifuged for 10 minutes at 10,000 rpm. Using a UV-Vis spectrophotometer, collect the supernatant and measure the absorbance at 480 nm.\u003c/p\u003e \u003cp\u003eThe phytase activity was measured using a modified method\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. One gram of seed sample was homogenized in 0.1M sodium acetate buffer (pH 5.0) and centrifuged. Phytase activity was determined by adding buffer and sodium phytate solution, incubating, adding a crude enzyme, and incubating and measuring phosphate liberation with the ammonium molybdate method. MDA content was determined using 20% TCA and 0.5% TBA solution to quantify lipid peroxidation\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. The seed sample was ground in 4 ml of 1% TCA solution. After centrifuging, the supernatant was collected. 1 ml of (20% TCA and 0.5% TBA) was added to each sample. The absorbance was accurately measured at 532 nm and 600 nm for specific and non-specific samples, following a rigorous incubation and centrifugation process. MDA content was measured in moles/ml.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eCalculation:\u003c/h2\u003e \u003cp\u003e \u003cdiv id=\"Equa\" class=\"Equation\"\u003e \u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\text{M}\\text{D}\\text{A} \\left(\\text{m}\\text{M}\\right)= \\frac{{A}_{532}-{A}_{600}}{155} \\times 100$$\u003c/div\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe extinction coefficient of this MDA-TBA abduct at 532 nm is 155 mM-1cm-1.\u003c/p\u003e \u003cp\u003e \u003cb\u003eCharacterization of seed storage proteins.\u003c/b\u003e The total protein was estimated from chickpea seed, and a standard curve was produced using Bradford's standard solution\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. A sample of dried chickpea seeds was ground using a hammer mill; fine powder was obtained, passed through a 0.185 mm mesh grid, and kept in air-tight plastic containers at room temperature to prevent spoilage. Chickpea flours were defatted overnight using a horizontal shaker with hexane in a ratio of 1:10 and then washed twice with ethyl ether, followed by drying for 1 hour at -20\u0026ordm;C. Protein fractions were obtained using the Osborne (1907) fractionation method\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Seed flours were extracted by stirring in borate buffer at pH-7.6 with NaCl and sodium azide for 2 hours, followed by centrifugation at 30,000 rpm for 30 minutes.\u003c/p\u003e \u003cp\u003e \u003cb\u003ePolyacrylamide Gel Electrophoresis analysis.\u003c/b\u003e SDS-PAGE was carried out using the 5% stacking gel and 12% resolution gel\u003csup\u003e\u003cb\u003e\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e\u003c/b\u003e\u003c/sup\u003e. Sample solutions were prepared from 10 mg of freeze-dried protein extract or precipitates dissolved in 1 ml sample buffer (distilled water, 0.5 M Tri-HCl pH 6.8, glycerol, 10% SDS, 1% bromophenol blue and β-mercaptoethanol heated at 98\u0026deg;C for 10 min, then applied to the sample wells. Electrophoretic migration was monitored at a constant current (12 mA/gel) for 1.5 to 2 h. SDS gels were stored for two hours, and distaining was done for 12 hours.\u003c/p\u003e \u003cp\u003e \u003cb\u003eField experiment.\u003c/b\u003e The field research trial used a randomized complete block design with eight replications and three priming treatments: T\u003csub\u003e1\u003c/sub\u003e (dry seed as control), T\u003csub\u003e2\u003c/sub\u003e (bavistin treated as positive control), and T\u003csub\u003e3\u003c/sub\u003e (turmeric leaf extract primed). The seed was sown in 5 rows per plot, with a plot size of 2.8\u0026times;1.8 m\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Field emergence percentage (no. of seedlings emerged/total no. of seed sown) was calculated 15 days and 30 days after sowing. Five randomly selected plants from each replicated plot of all treatments were tagged to take all the observations, such as plant height (at 15 DAS, 30 DAS, 45 DAS and maturity) and yield parameters. The biological yield was calculated before threshing by taking the total weight of harvested crop plants from the net plot area. The biological yield was given in kg ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. After sun drying for a few days, the harvested crop from the respective net plot was threshed with a thresher. Seed yield was recorded and expressed as kg/ha. The Harvest Index (HI) was calculated using formulae-\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\varvec{H}\\varvec{I}=\\frac{\\varvec{B}\\varvec{i}\\varvec{o}\\varvec{l}\\varvec{o}\\varvec{g}\\varvec{i}\\varvec{c}\\varvec{a}\\varvec{l} \\varvec{Y}\\varvec{i}\\varvec{e}\\varvec{l}\\varvec{d}}{\\varvec{E}\\varvec{c}\\varvec{o}\\varvec{n}\\varvec{o}\\varvec{m}\\varvec{i}\\varvec{c} \\varvec{Y}\\varvec{i}\\varvec{e}\\varvec{l}\\varvec{d}} \\times 100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eStatistical analysis.\u003c/b\u003e The collected data were analyzed using the statistical software SPSS and the analysis of variance technique. The treatment means were compared using a post-hoc test at a 5% significance level to determine whether there were any significant differences between them. Furthermore, a graphical representation of the data was created using Microsoft Excel to provide a clear and visual presentation of the results.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors acknowledge Bihar Agricultural University, Sabour, for providing the infrastructural facilities to carry out the research work and the BAU communication number of the manuscript. The author also acknowledges the fund support by the Science and Engineering Research Board, Government of India.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analysed during this study are included in this published article and its supplementary information files.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor\u0026rsquo;s contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eKamini Kaushal: Conducted the experiments, collected the data, played a crucial role in data interpreting and compiling the results.\u003c/p\u003e\n\u003cp\u003eKumari Rajani: Conceptualized the research idea and designed the experiments, significantly contributing to drafting and finalizing the manuscript.\u003c/p\u003e\n\u003cp\u003eRavi Ranjan Kumar: Designed the experiments and prepared and refined the manuscript draft.\u003c/p\u003e\n\u003cp\u003eAnand Kumar: Designed field experiments and data analysis of the field data recorded.\u003c/p\u003e\n\u003cp\u003eTushar Ranjan: He contributed to preparing protein samples, executing experiments, and proofreading the manuscript.\u003c/p\u003e\n\u003cp\u003eVinod Kumar: Actively participated in collecting the research material for this project.\u003c/p\u003e\n\u003cp\u003eFeza Ahmad: Provided proofreading of the manuscript and intellectual input.\u003c/p\u003e\n\u003cp\u003eVikas Kumar: Contributed to the standardization of biochemical assays and\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAman Kumar: Conducted field experiments and recorded high-quality data accurately.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAgrawal, T., Kumar, A., Kumar, S., Kumar, A., Kumar, R.R., Kumar, S. \u0026amp; Singh, P.K. 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(1924). \u003c/li\u003e\n\u003cli\u003eLaemmli, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. \u003cem\u003eNature,\u003c/em\u003e\u003cstrong\u003e227\u003c/strong\u003e, 680-685, https://doi.org/10.1038/227680a0 (1970).\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"botanical priming, turmeric, dehydrogenase, globulin, vigour index, SDS PAGE","lastPublishedDoi":"10.21203/rs.3.rs-3914755/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3914755/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eChickpea is a highly nutritious protein-rich source and one of the major crops to alleviate global malnutrition. It is the third-largest legume cultivated globally in arid and semi-arid environments. Chickpea productivity is affected by several factors, among which poor seed quality is one of the significant factors contributing to its decrease in productivity. Seed quality is essential for better crop establishment and higher yields, particularly in the uncertain climate change. The present study investigated the role of botanical priming in enhancing seed quality by investigating physio-biochemical responses and crop performance in chickpeas. The study compared the effects of botanical priming with hydropriming, seed treatment with bavastin and control on medium-vigour chickpea seeds. A detailed physiological (germination percentage, root and shoot length, vigour index) and biochemical (amylase, protease, dehydrogenase, phytase, and lipid peroxidation) analysis was carried out in the laboratory to assess the effect of priming treatments. SDS-PAGE was used to compare seed storage protein expression in primed and control seeds. Seeds treated with 1% turmeric leaf extract displayed a higher germination rate (94.5%) than the control and other priming treatments. Turmeric-primed seeds showed higher seedling length, vigour index, enzyme activity, and lower MDA content. SDS-PAGE analysis revealed the expression of two minor polypeptides of the albumin and three minor polypeptides of the globulin subunit.\u003c/p\u003e \u003cp\u003eMoreover, field experiments indicated increased crop growth, vigour, and days to 50% flowering, yield and its attributing traits in turmeric-primed seeds. The study demonstrates that botanical priming can increase the yield of chickpeas by up to 16% over the control group. This study proves that implementing low-cost and eco-friendly seed priming techniques can significantly enhance the genetic potential of chickpeas by improving their physiological and biochemical activities during seed germination and crop growth. Therefore, chickpea growers must adopt botanical priming techniques to improve seed quality and crop performance. This study unequivocally establishes the efficacy of botanical priming as a powerful tool for augmenting chickpea growth. Moreover, this approach is environmentally sustainable and can help conserve natural resources long-term. Therefore, this new approach must be widely adopted across the agricultural industry to ensure sustainable and profitable farming practices.\u003c/p\u003e","manuscriptTitle":"Physio-biochemical responses and crop performance analysis in chickpea upon botanical priming","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-02-19 08:12:04","doi":"10.21203/rs.3.rs-3914755/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-03-07T05:50:20+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-02-25T06:39:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"d2543cec-ae14-46b0-8aa0-b39b9abd4fb7","date":"2024-02-22T08:37:22+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-02-22T08:03:51+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-02-22T07:46:31+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-02-16T07:07:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-02-16T07:04:59+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-01-31T18:06:11+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5650b8c5-155a-4aad-bd3c-aee88727c00d","owner":[],"postedDate":"February 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":28803260,"name":"Biological sciences/Biotechnology"},{"id":28803261,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2024-04-26T18:05:11+00:00","versionOfRecord":{"articleIdentity":"rs-3914755","link":"https://doi.org/10.1038/s41598-024-59878-8","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2024-04-23 18:05:11","publishedOnDateReadable":"April 23rd, 2024"},"versionCreatedAt":"2024-02-19 08:12:04","video":"","vorDoi":"10.1038/s41598-024-59878-8","vorDoiUrl":"https://doi.org/10.1038/s41598-024-59878-8","workflowStages":[]},"version":"v1","identity":"rs-3914755","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3914755","identity":"rs-3914755","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}