水杨酸引发提高低温下水稻种子萌发活力的生理与分子效应OA北大核心CSTPCD
Physiological and Molecular Effects of Salicylic Acid on Rice Seed Germination at Low Temperature
[目的]研究水杨酸(SA)引发对低温下水稻种子萌发活力的影响及生理响应,揭示SA引发对脱落酸(ABA)和赤霉素(GA)代谢途径相关基因以及细胞壁松弛基因的诱导模式,为水稻种子低温萌发研究提供理论依据.[方法]以籼型三系杂交水稻泰丰优 208 种子为材料,通过种子引发处理,分析SA对种子低温萌发活力及生理的影响,并通过qRT-PCR技术分析ABA、GA和扩展蛋白基因响应SA引发的表达模式.[结果]低温(15℃)显著推迟水稻种子萌发进程.在低温下萌发1 d种子中,其内源SA浓度是常温(28℃)下的1.7倍;但对于5 d的幼苗而言,低温下的SA浓度仅为常温下浓度的0.6%.SA引发可有效提高种子在低温下的萌发活力,尤其以 2 000 μmol·L-1 SA效果最为显著,该浓度显著提高了低温下种子的发芽指数、活力指数、芽长、根长、鲜重和干重,其中活力指数分别为未引发种子(CK1)和水引发种子(CK2)的3和2倍.在生理指标方面,SA 引发提高了低温萌发过程中种子的可溶性糖、脯氨酸以及活性氧含量,增加了总淀粉酶、β-淀粉酶、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,降低了丙二醛(MDA)含量.与CK1 相比,2 000 μmol·L-1 SA引发将种子ABA含量降低了79%,同时将IAA和GA1 含量增加了32.2%和2.66倍.在基因表达方面,对于2 000 μmol·L-1 SA引发的种子,ABA合成基因OsNCED2和OsNCED3的表达量分别比CK1 降低了94.26%和90.24%;而ABA分解基因OsABA8'ox2和OsABA8'ox3的表达量分别为CK1 的 5.9 和 3.9 倍.与CK1 相比,SA引发显著上调了GA合成基因OsCPS1、OsKAO和OsGA20ox1的表达量,并显著下调GA分解基因OsGA2ox2和OsGA2ox6的表达量.在几个候选的细胞壁松弛因子扩展蛋白基因中,除了OsEXPB11外,其余几个同源基因均在一定程度上被引发而上调表达.与CK1 相比,2 000 μmol·L-1 SA引发分别使OsEXPA2、OsEXPB4和OsEXPB6的表达量上调12.2、5.9和6.1 倍.[结论]SA引发可显著缓解低温对于水稻种子萌发和幼苗生长的影响.可能是由于SA提高SOD、CAT等抗氧化酶活性,降低MDA的产生,增加可溶性糖和脯氨酸的含量,进而增强种子和幼苗对于低温的耐受能力.另一方面,SA引发通过降低种子内源ABA含量,增加GA1 含量,增强总淀粉酶和β-淀粉酶活性,促进细胞壁松弛相关基因的表达,从而促进低温下的种子萌发和幼苗生长.
[Objective]The study investigated the impact of salicylic acid(SA)priming on the germination vigor and physiological response of rice seeds under low temperatures.It aimed to reveal the expression patterns of genes related to abscisic acid(ABA)and gibberellin(GA)metabolic pathways as well as cell wall relaxation genes by SA priming.This research provided a theoretical basis for the study of rice seed germination at low temperatures.[Method]Using indica three-line hybrid rice Taifengyou 208 seeds as materials,the effects of SA on seed germination vigor and physiology responses under low temperature were analyzed through seed priming treatment,and the expression patterns of genes related to ABA,GA and expansin in response to SA were analyzed by qRT-PCR.[Result]Low temperature(15℃)significantly delayed the germination process of rice seeds.In seeds germinated at low temperatures for one day,the endogenous SA concentration was 1.7 times higher than that at normal temperatures(28℃).However,for five-day-old seedlings,the SA concentration under low temperature was only 0.6%of that at normal temperatures.SA could effectively enhanced germination vigor of seeds at low temperature,with the most significant effects observed at 2 000 μmol·L-1 SA.This concentration significantly increased the germination index,vigor index,shoot length,root length,fresh weight,and dry weight of seeds under low temperature conditions.Notably,the vigor index was three times that of non-primed seeds(CK1)and two times that of water-primed seeds(CK2).In terms of physiological indexes,SA priming increased the contents of soluble sugar,proline and active oxygen,enhanced the activities of total amylase,β-amylase,superoxide dismutase(SOD)and catalase(CAT),and decreased the content of malondialdehyde(MDA).Compared with CK1,2 000 μmol·L-1 SA decreased the ABA content by 79%,and increased the IAA and GA1 contents by 32.2%and 2.66 times,respectively.In terms of gene expression,the expression levels of ABA synthesizing genes OsNCED2 and OsNCED3 were decreased by 94.26%and 90.24%compared with CK1 in seeds primed by 2 000 μmol·L-1 SA,respectively,whereas the expression levels of ABA decomposing genes OsABA8'ox2 and OsABA8'ox3 were 5.9 and 3.9 times higher than that of CK1,respectively.Compared with CK1,SA priming significantly upregulated the expression of GA synthesizing genes OsCPS1,OsKAO and OsGA20ox1,while it significantly downregulated the expression of GA decomposing genes OsGA2ox2 and OsGA2ox6.In several candidate genes encoding cell wall relaxation protein,e.t.expansin,all but OsEXPB11 were significantly upregulated to some extent by priming.Compared with CK1,2 000 μmol·L-1 SA increased the expression levels of OsEXPA2,OsEXPB4 and OsEXPB6 to 12.2,5.9 and 6.1 times,respectively.[Conclusion]SA priming can significantly alleviate the impact of low temperatures on rice seed germination and seedling growth,which is likely due to SA enhancing the activity of antioxidant enzymes such as SOD and CAT,reducing the production of MDA,and increasing the content of soluble sugars and proline,thereby strengthening the tolerance of seeds and seedlings to low temperatures.On the other hand,SA priming decreases endogenous ABA content,increases GA1 content,enhances the activities of total amylase and β-amylase,and promotes the expression of genes related to cell wall relaxation,thus facilitating seed germination and seedling growth at low temperature.
陈兵先;张琪;戴彰言;周旭;刘军
广东省农业科学院农业生物基因研究中心/广东省农作物种质资源保护与利用重点实验室,广州 510640
水稻低温胁迫种子引发水杨酸生理指标基因表达
ricelow temperature stressseed primingsalicylic acid(SA)physiological indexgene expression
《中国农业科学》 2024 (007)
1220-1236 / 17
广东省重点领域研发计划(2020B0202090003,2022B0202110003)、广东省自然科学基金(2022A1515012302)、广东省农业科学院农业优势产业学科团队建设项目(202132TD)
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