作物学报 ›› 2022, Vol. 48 ›› Issue (12): 3004-3017.doi: 10.3724/SP.J.1006.2022.13060
悦曼芳1,2(), 张春2(), 郑登俞2, 邹华文1(), 吴忠义2()
YUE Man-Fang1,2(), ZHANG Chun2(), ZHENG Deng-Yu2, ZOU Hua-Wen1(), WU Zhong-Yi2()
摘要:
bHLH (basic helix-loop-helix)是植物中一个重要的转录因子家族, 在调控植物生长发育、逆境胁迫及信号转导过程中发挥着重要作用。目前, 动物中大部分bHLH转录因子功能已明确, 但是在植物中, 尤其是玉米中的研究报道较少。在前期工作中, 我们对玉米生长发育过程中的4个关键时期进行了根系表型鉴定和转录组测序分析, 发现转录因子ZmbHLH91在六叶期(V6)、十二叶期(V12)和抽雄期(VT)的相邻时期间的表达均差异显著, 且ZmbHLH91在V6, V12和VT这些根系生长发育活跃期的表达量较高, 推测该基因可能调控玉米根系的生长发育。为探究ZmbHLH91基因在根系生长和抵抗逆境胁迫方面的作用, 本研究克隆了ZmbHLH91 (AC: NC_AQL05369)基因, 该基因全长2112 bp, 具有bHLH转录因子家族特有的保守结构域。实时荧光定量PCR (RT-qPCR)分析表明, ZmbHLH91在玉米根中的表达量最高, 其在三叶期幼根中的表达量高于抽雄期成熟根。在高盐、渗透、低温以及脱水胁迫处理下, 玉米幼苗中ZmbHLH91的表达均上调。在无胁迫处理的1/2 MS培养基上, ZmbHLH91异源表达拟南芥与野生型拟南芥的根长无明显差别, 而在梯度浓度NaCl和甘露醇处理的培养基上, ZmbHLH91异源表达拟南芥的根均长于野生型, 且差异显著; 在土壤中进行干旱和高盐处理后, 转基因拟南芥比野生型拟南芥表现出更好的生长状态、更高的过氧化物酶(POD)活性和更高的绿叶率。推测ZmbHLH91基因可能参与响应高盐、干旱以及渗透胁迫等逆境条件。ZmbHLH91基因在茉莉酸(jasmonic acid, JA)、脱落酸(abscisic acid, ABA)等激素处理下均上调表达, 推测ZmbHLH91基因可能参与响应JA和ABA激素信号。在梯度浓度JA处理的培养基上, 转基因拟南芥的根长均长于野生型, 且差异显著。酵母双杂交实验表明ZmbHLH91与ZmMYC2相互作用, ZmMYC2是JA信号通路中重要的蛋白, 由此推测ZmbHLH91蛋白可能参与JA信号通路。综上所述, ZmbHLH91可能参与高盐、干旱和渗透胁迫应答及JA信号转导途径。本研究为进一步解析ZmbHLH91在玉米中的生物学功能提供了重要的参考依据。
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