冬瓜抗氧化基因的克隆及其在非生物胁迫下的表达分析

Analysis on Cloning and Expression of Oxidation Resistance Genes in Wax Gourd Under Abiotic Stress

【目的】鉴定、克隆冬瓜BhiOXR基因,明确其在非生物胁迫下的表达特征,为深入研究冬瓜BhiOXR基因的功能奠定基础。【方法】从冬瓜基因组鉴定BhiOXR基因家族成员,设计扩增BhiOXR基因CDS的全长引物,通过RT-PCR克隆BhiOXR基因。以低温(4℃)、高温(40℃)、盐胁迫(150 mmol/L NaCl溶液)和干旱胁迫(10%PEG6000溶液)处理冬瓜高代自交系B227幼苗,利用qRT-PCR技术分析冬瓜BhiOXR基因分别在上述4种非生物胁迫处理下(处理0、4、8、12、24 h)的表达特征。【结果】鉴定到6个冬瓜BhiOXR成员,并进行全长基因克隆和生物信息学分析。qRT-PCR分析结果表明,BhiOXR1响应低温、高温和干旱胁迫,不响应盐胁迫;BhiOXR2a、BhiOXR2b和BhiOXR4响应低温、盐和干旱胁迫,不响应高温胁迫;BhiOXR3和BhiOXR5响应低温、高温和盐胁迫,不响应干旱胁迫。【结论】明确了冬瓜BhiOXR基因受低温、高温、盐和干旱等非生物胁迫诱导表达,并且不同BhiOXR基因响应的非生物胁迫有差异,由此推测6个BhiOXR成员可能在抵御不同非生物胁迫过程中所表现的抗氧化作用不尽相同,为进一步探究冬瓜BhiOXR基因的生物学功能奠定基础。

【Objective】Identification and cloning of BhiOXR genes from wax gourd and clarification of its expression characteristics under abiotic stresses would lay a foundation for further studying the function of BhiOXR genes in wax gourd.【Method】BhiOXR gene family members from wax gourd reference genome were identified,full-length CDS primers were designed to clone the BhiOXR genes of the wax gourd by RT-PCR.Seedlings of wax gourd inbred line B227 were treated with low temperature(4℃),high temperature(40℃),salt stress(150 mmol/L NaCl solution) and drought(10% PEG6000 solution).The expression characteristics of BhiOXR genes under the above mentioned abiotic stresses treatment time:0,4,8,12,24 h were analyzed by using qRT-PCR.【Result】Six BhiOXR members were identified from wax gourd reference genome,and the fulllength genes were further cloned and bioinformatically analyzed.qRT-PCR analysis results showed that BhiOXR1 responded to low temperature,high temperature and drought stress,but did not respond to salt stress.BhiOXR2a,BhiOXR2b and BhiOXR4 responded to low temperature,salt and drought stresses but not to high temperature stress;BhiOXR3 and BhiOXR5responded to low temperature,high temperature and salt stresses but not to drought stress.【Conclusion】It is clarified that BhiOXR genes are induced by abiotic stresses as low temperature,high temperature,salt and drought,and different BhiOXR genes have differences in response to abiotic stresses.Therefore,it is speculated that the six BhiOXR members may have different antioxidant effects in the process of resisting different abiotic stresses,which lays a foundation for further exploring the biological functions of BhiOXR genes in wax gourd.