中国农业科学

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最新录用: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

陈兵先,张琪,戴彰言,周旭,刘军   

  1. 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

Physiological and Molecular Effects of Salicylic Acid on Rice Seed Germination at Low Temperature

CHEN BingXian, ZHANG Qi, DAI ZhangYan, ZHOU Xu, LIU Jun   

  1. Agro-biological Gene Research Center, Guangdong Academy of Agricultural Sciences/Guangdong Provincial Key Laboratory for Crop Germplasm Resources Preservation and Utilization, Guangzhou 510640

摘要: 【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74.7%。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OsNCED2OsNCED3的表达量分别比CK1降低了94.26%和90.24%;而ABA分解基因OsABA8’ox2OsABA8’ox3的表达量分别是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OsCPS1OsKAOOsGA20ox1的表达量,并显著下调GA分解基因OsGA2ox2OsGA2ox6的表达量。在几个候选的细胞壁松弛蛋白-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。与CK1相比,2 000 μmol·L-1 SA引发分别使OsEXPA2OsEXPB4OsEXPB6的表达量上调至原来的12.25.9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。


关键词: 水稻, 低温胁迫, 种子引发, 水杨酸, 生理指标, 基因表达

Abstract: 【ObjectiveTo study the effects of salicylic acid (SA) priming on the vigor of seed germination and physiological response of rice seeds at low temperature, and to reveal the induction mode of SA priming on genes related to abscisic acid (ABA) and gibberellin acid (GA) metabolic pathways and cell wall relaxation, which can provide a theoretical basis for the study of low temperature germination of rice seeds.MethodUsing indica hybrid rice Taifengyou 208 seeds as materials, the effects of SA on seed germination vigor and physiology were analyzed by seed priming, and the expression patterns of ABA, GA and expansion protein genes in response to SA were analyzed by qRT-PCR.ResultLow temperature (15 ℃) significantly delayed the germination process of rice seeds. The concentration of endogenous SA in seeds germinated at low temperature for 1 day was 1.7 times that at room temperature (28 ℃). However, the SA concentration at low temperature was only 0.6% of that at normal temperature for 5 d seedlings. SA could effectively improve germination vigor of seeds at low temperature, especially 2 000 μmol·L-1 SA, which significantly increased germination index, vigor index, bud length, root length, fresh weight and dry weight of seeds at low temperature, and vigor index of seeds at low temperature was 3 and 2 times of that of non-induced seeds (CK1) and water-induced seeds (CK2), respectively. 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 74.7%. In terms of gene expression, the expressions 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. The expression levels of ABA decomposing genes OsABA8 'ox2 and OsABA8' ox3 were 5.9 and 3.9 times that of CK1, respectively. Compared with CK1, SA significantly up-regulated the expression levels of GA synthesizing genes OsCPS1, OsKAO and OsGA20ox1, and significantly down-regulated the expression levels of GA decomposing genes OsGA2ox2 and OsGA2ox6. In several candidate genes encoding cell wall relaxation protein, e.t. expansin, except OsEXPB11, the other homologous genes were triggered and upregulated to some extent. 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.ConclusionSA induction could significantly alleviate the effects of low temperature on seed germination and seedling growth of rice. It was probably because SA increased the activity of antioxidant enzymes such as SOD and CAT, decreased the production of MDA, increased the contents of soluble sugar and proline, and thus enhanced the tolerance of seeds and seedlings to low temperature. On the other hand, SA prime decreased endogenous ABA content, increased GA1 content, enhanced the activities of total amylase and β-amylase, and promoted the expression of genes related to cell wall relaxation, thus facilitating seed germination and seedling growth at low temperature.


Key words: rice, low temperature stress, seed priming, salicylic acid (SA), physiological index, genes expression