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March 1, 2023
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Pepper (potatoL.) is a vegetable of great economic importance and high value in the medicinal industry, chemical additives and other industries. However, heat stress during seed germination and loss of seed vigor during storage affect seed germination, development, yield and economic value. To meet the demands of industries, it is essential to explore the potential molecular mechanism of thermotolerance and vigor of pepper seeds to develop good quality pepper seeds. In the present study, we found that pepperCaHsfA1dThe gene is highly expressed during seed development and its expression decreases as it enters the germination stage. Heat induced a decrease in the expression ofCaHsfA1dgene during seed germination. However, the overexpression ofCaHsfA1dnoArabidopsissignificantly improved seed thermotolerance and improved thermotolerant transcription levels (AtHsfA2yAtHsp18.2) and antioxidant (EnglishSTU5) during seed germination after heat stress. Also,CaHsfA1dtransgenicArabidopsisthe seeds showed greater resistance to accelerated aging and what is its possible role in the germination vigor of the seeds. Research on seed germination under environmental stress and seed vigor are new topics in pepper seed science, and our study provides a reference for further study of the defense mechanism of pepper seeds and plant breeding.
The seed is the reproductive systemspermatophyteand acts as a prominent character in the continuation of the species. For agricultural production, good seed quality is a fundamental guarantee for the successful reproduction of high-quality crops and agricultural products (Finch-Savage and Bassel, 2016). Seed quality assessment indicators include seed vigor, seed germination, seed dormancy, water content, etc. (Hampton, 2002; Rashidetal., 2018). Seed germination is a vital step in the life cycle of horticultural crops, as it determines the right time for crops to enter the agroecosystem. Once the seeds find a favorable environment for growth, they sprout immediately after imbibition (Chenetal., 2019). Adverse environmental factors in nature generally weaken the germination capacity of seeds, mainly due to the uncertainty of climate change (Lamichaneyetal., 2021; Rashidetal., 2018; Zhaoetal., 2021). With the intensification of global warming, high temperatures have become a notable challenge for the thermotolerance of seeds in food production (Moroetal., 2021; Sakaietal., 2022). Seed germination is closely related to seed vigor, and high vigor seeds increase the germination rate of seeds kept for a long time (Zhouetal., 2012a). However, seed vigor gradually decreases and seed germination capacity decreases significantly during germplasm conservation (Waterworthetal., 2011; Zhouetal., 2012b). When seed vigor decreases, it shows more sensitivity to adverse environmental conditions during germination, and even loses its germination capacity (Chenetal., 2012; Sarkaretal., 2019; Zhouetal., 2012a, 2012b), which seriously affects the ability reproductive. crop growth and yield.
Seed thermotolerance and germination vigor have been the subject of many studies and the molecular mechanisms underlying the transition from seed to seedling development have also been investigated (Chenetal., 2020b, 2019; Zhouetal., 2012a).Arabidopsis AtPER1reduces seed germination by inhibiting abscisic acid (ABA) catabolism and gibberellin (GA) biosynthesis in seeds (Chenetal., 2020a). Some studies have also emphasized the importance ofMONTEgenes in seed development and germination (Yuanetal., 2008; Zhouetal., 2012b). In addition, heat shock proteins (Hsps) have been related to improving seed thermotolerance (Guoetal., 2015a; Haqetal., 2019; Huangetal., 2019; Kimetal., 2013) and have shown their participation in vigor. of seed germination ( Hsuetal., 2010; Sarkaretal., 2019; Zhaoetal., 2021; Zhouetal., 2012a; Zhuetal., 2016). For example,Arabidopsisseeds with overexpression ofNelumbo nucifera NnHsp17.5showed higher germination vigor and seed thermotolerance (Zhouetal., 2012a). The germination process was significantly reduced with the silencing ofCI-sHsps(cytosolic class I Hsp) in rice (Oryza sativaL.), while a decrease in thermotolerance was also observed (Sarkaretal., 2019). Furthermore, three cytosolichsp70sit has also been indicated that they function in the germination process under high salt conditions (Zhaoe et al., 2021). These identified genes provide some reference value for the molecular mechanism of seed germination in response to abiotic stress. Recent studies show that reactive oxygen species (ROS) may be the main reason for seed deterioration, which is related to loss of seed vigor (Baillye et al., 2008; Chenetal., 2012; El-Maarouf-Bouteau et al., 2011). ROS are ubiquitous oxidizing agents and oxidize most macromolecules, including DNA, proteins and lipids, leading to cell damage and disruption (Chenetal., 2012; Wenjingetal., 2020). Elimination of ROS with excess ROS scavengers can increase the longevity of transgenic seeds (Chenetal., 2012; Leeetal., 2010; Zhouetal., 2012b). Furthermore, ROS in the plant are generated under exposure to abiotic stresses such as heat, salt and drought (Gaietal., 2020). Heat stress can also promote the accumulation of ROS in plant seeds, mainly H2O2EU2−attenuate the germination capacity of seeds (Zhangetal., 2020a). Meanwhile, seed germination after heat stress is enhanced by application of gamma-aminobutyric acid to suppress heat-induced ROS overaccumulation (Zhangetal., 2020a). The vital functions of the heat shock transcription factor (Hsf) in response to abiotic stress in plants have been explored (Gaietal., 2020; Guoetal., 2015b; Scharfetal., 2012; Yueetal., 2021). Hsfs improve plant heat tolerance by regulating the expression of downstream genes (e.g.,Hsps, Hsfs, youGSTU) and maintain it in the steady state of the antioxidant system (Gaietal., 2020; Zhangetal., 2019). As previously described, regulatory pathways related to Hsps and ROS play an important role in increasing seed thermotolerance and germination vigor; however, the role of Hsfs in seed vigor is still unclear.
Pepper (potatoL.) is a vegetable of great economic importance, being also used in the production of natural capsaicinoids, widely used in medicine, food, condiment and insect repellent. However, being a sessile plant, pepper is constantly subject to various environmental stresses during its germination and growth phases, mainly for summer cultivation in tropical or even temperate regions of the world. Previously, we evaluated the function of pepperCaHsfA1don the thermotolerance of seedlings (Gaietal., 2020). Although the current study is exploring the role of pepperCaHsfA1din improving seed germination tolerance at high temperatures and resistance to accelerated aging (AA) treatment. We characterize the expression pattern ofCaHsfA1dgene in the development and germination phase of pepper seeds. Analyzes of thermotolerance and treatment with AA showed thatCaHsfA1dThe gene was involved in improving seed vigor after AA treatment and conferring resistance to heat stress during seed germination. These findings present a new perspective on the biological function ofhsfgene and provides a new molecular mechanism for seed thermotolerance and germination vigor.
Plant materials and growing conditions.
wild typeArabidopsis thaliana(ecotype Columbia, WT) andCaHsfA1d- overexpressionArabidopsisseedlings were routinely cultivated in plant growth chambers at 60 to 70% relative humidity at 22/18°C light/dark. so allArabidopsisthe seeds were produced under the same conditions and were collected from the dried silicas at the same time. HimArabidopsisthe seeds used in this study germinated in a sterile environment. For aseptic treatment, seeds were superficially sterilized in 75% ethanol for 30 seconds, then soaked.
expression profiles ofCaHsfA1din pepper
CaHsfA1dTranscription levels were analyzed during pericarp, placenta and seed development at 20, 25, 30, 35, 40, 45, 50, 55 and 60 DAF (Fig. 1A and Fig. S1). the transcription ofCaHsfA1dwas predominantly detected in developing seeds and absent in the pericarp.CaHsfA1dwas expressed in the prepubertal placenta, however, transcription was not observed in the placenta at 40 DAF. TimeCaHsfA1dwas ubiquitously expressed throughout the pepper seed development process. HimCaHsfA1dthe transcript was
Seed germination represents the beginning of plant growth and development, while it is the most vulnerable stage of the plant's life. The seed is a unique reproductive and in vitro organ of plants, seed germination is related to the reproduction and evolution of the plant population, related to the quality and yield of crops related to people's lives. Pepper is a widely cultivated crop all over the world, but the high temperature environment seriously affects the germination of its seeds and causes severe cutbacks in pepper production.
In summary, we analyzed the expression pattern ofCaHsfA1dgene in pepper and identified its biological function in the seed.CaHsfA1dit could be used as a candidate gene to improve the thermotolerance of pepper seeds during the germination phase and could participate in increasing the longevity of pepper seeds. The potential molecular mechanism of thermotolerance and vigor of pepper seeds has not been previously characterized and, therefore, our analyzes provide a reference for further studies.
This work was funded by theChina National Key R&D Program(Not.2016YFD0101900) it's himNational Natural Science Foundation of China(Not.31772309, Not.31860556).
Contribution Statement Authored CRediT
Wen Xian Gai:Conceptualization, Formal analysis, Research, Software, Visualization, Writing – original draft.Abânico Yang:Formal analysis, Methodology, Software, Visualization.Mohamed Ali:Data curation, research, validation, visualization, writing: proofreading and editing.Aqel Ahmad:Validation, visualization, writing: proofreading and editing.Zhenhui Gong:Conceptualization, fundraising, validation, writing: proofreading and editing.
Declaration of conflicting interests
The authors declare that they have no known competing financial interests or personal relationships that could appear to influence the work reported in this article.
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