作物学报 ›› 2017, Vol. 43 ›› Issue (06): 839-848.doi: 10.3724/SP.J.1006.2017.00839
余建,刘长英,赵爱春,王传宏,蔡雨翔,余茂德*
YU Jian,LIU Chang-Ying,ZHAO Ai-Chun,WANG Chuan-Hong,CAI Yu-Xiang,YU Mao-De*
摘要:
1-氨基环丙烷-1-羧酸氧化酶(ACO)作为关键酶,能够催化1-氨基环丙烷-1-羧酸(ACC)形成乙烯。为探究桑树MnACO基因在桑树生长发育和抵御外界胁迫中的功能,本研究构建了pMnACO::GUS的植物表达载体并转化拟南芥。采用GUS组织染色法鉴定转基因拟南芥不同生长阶段及胁迫处理后的GUS活性。通过PCR克隆得到MnACO1和MnACO2启动子片段,它们分别为1518 bp和1429 bp,启动子区域有大量的TATA-box、CAAT-box和其他响应外界刺激的顺式作用元件。GUS活性分析显示MnACO启动子能驱动GUS在拟南芥中表达;MnACO1启动子在拟南芥的根、叶片、花瓣、花药、花丝、柱头以及果荚中均有表达,且活性较MnACO2强;MnACO2启动子在果荚中无表达。转MnACO1和MnACO2植株经不同逆境处理后GUS表达活性不同,转MnACO1植株的GUS活性随处理延长时间而减弱,转MnACO2植株GUS活性随处理时间延长而增强。qRT-PCR检测2周苗龄的桑幼苗在经过胁迫处理后MaACO1和MaACO2的基因表达量,发现MaACO基因的表达模式与MnACO启动子GUS活性变化趋势一致。本研究结果表明,MnACO为诱导型启动子,MnACO1兼具组成型启动子特性,MnACO2兼具组织特异型启动子特性。MnACO1在转基因植株中对胁迫响应能力更强,预示着可将其用来调控改良桑树品种抗逆性靶基因;MaACO2可能与果实成熟有关,可将其启动子作为果实特异性启动子对桑椹品质进行合理改良。
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