棉花新陆早17号愈伤组织中盐响应相关基因的表达分析

摘要

[Objective]Callus is an ideal experimental material for studying plant development and gene regulation, so it will be helpful to elucidate the gene expressive characteristics by exploring the effects of salt stress on gene expression of cotton callus. [Method] The calli of cotton Xinluzaol7 variety was treated with different NaCl concentrations and then the expression levels of GhBADH, GhATPase - b, GhNHX1 and GhPSCS of non - embryogenic and embryogenic calli of the cotton were investigated by using semi -quantitative RT - PCR method. [ Result] Medium added with 100 mmoloL - 1 NaCl can improve the growth of Non - embryogenic calli and embryogenic calli; In new induced non - embryogenic calli, the expression level of GhNHX1 first decreased and then increased slightly, while GhATPase -b, GhBADH and GhPSCS expression level were up - regulated with the different NaCI treatment times; In repeatedly subcultured non - embryogenic calli, the expression level of GhNHX1 and GhBADH had the same trend as that of GhNHX1 in new non -embryogenic calli, however GhATPase - b and GhP5CS were also up - regulated; In embryogenic calli, the expression level of GhATPase - b was rising with the extending NaCl stress time, and other salt response related genes first reduced and then increased. [Conclusion] Non -embryogenic calli and embryogenic calli grew better and differentiated faster in medium added with low concentration of NaCl than the control. There are different gene expressive characteristics at developmental stages of cotton calli, which indicates that these calli may have the specific responses to NaCl stress.%[目的]愈伤组织是一种研究植物生长发育及基因调控的理想实验材料,开展盐胁迫对棉花愈伤组织基因表达影响的研究,有助于阐明棉花愈伤组织基因的表达特性.[方法]对棉花新陆早17号品种非胚性愈伤和胚性愈伤进行盐胁迫处理,采用半定量RT-PCR技术,对棉花不同基因GhATPase-b、GhBADH、GhNHX1和GhP5 CS盐响应相关基因在不同NaC1浓度胁迫0、14和28 d时的表达情况进行分析.[结果]100 mmol/LNaC1促进棉花非胚性愈伤和胚性愈伤的生长分化；在新诱导的非胚性愈伤中,GhNHX1的表达水平先降低后小幅升高,而GhA TP ase-b、GhBADH及GhP5CS相对表达量随处理时间延长均呈上调表达；在继代多次的非胚性愈伤中,GhNHX1与GhBADH在处理期间相对表达量先降低后升高,GhATPase-b与GhP5CS相对表达量随处理时间延长均呈上调表达；胚性愈伤中,仅GhA TPase-b的表达量随处理时间增加持续升高,其余基因在处理期间相对表达量先降低后升高.[结论]适量NaC1可促进棉花非胚性愈伤和胚性愈伤的生长分化；不同阶段的棉花愈伤组织基因表达模式的不同,表明这些愈伤组织可能采用不同的机制响应NaCl的胁迫.