摘要
[目的]通过实时PCR(qRT-PCR)筛选海州常山叶片不同非生物胁迫条件下稳定的内参基因。[方法]利用已有的转录组数据,筛选出17个候选内参基因,在不同非生物胁迫(盐害、干旱和热害)下进行qRTPCR,通过利用GeNorm、NormFinder、BestKeeper及ReFinder软件分析筛选不同胁迫中最适的内参基因,选择CtNHX1基因,验证所筛选的内参基因的稳定性。[结果]RPL、AP-2盐害胁迫下最稳定,MDH、AP-2干旱胁迫下最稳定,UBCE2、ACT热害胁迫下最稳定;综合来看,非生物处理中,AP-2、UBCE2是最稳定的参考基因。[结论]AP-2、UBCE2可作为海州常山叶片非生物胁迫研究中基因定量使用的内参基因,为进一步开展海州常山分子机制研究、耐盐基因的开发利用奠定基础。
[Objective]To select the stable internal reference genes by real-time PCR(qRT-PCR)under dif-ferent abiotic stress conditions in Clerodendrum trichotomum leaves.[Method]Based on previous tran-scriptome data,17 reference genes were selected under different abiotic stresses(salt,drought and heat)for C.trichotomum.Four reference gene analysis software i.e.GeNorm,NormFinder,BestKeeper and Re-Finder were used to select the optimal internal reference gene,to select the genes for Na+/H+Exchanger1(ClNHX1)and to verify the stability of internal reference genes elected.[Result]Genes RPL and AP-2 were the most stable under salt stress;MDH and AP-2 were the most stable under drought stress;UBCE2 and ACT were the most stable under heat stress.For abiotic treatments,AP-2 and UBCE2 were the most stable reference genes.[Conclusion]AP-2 and UBCE2 can be used as internal reference genes for gene quantification in abiotic stress studies of C.trichotomum leaves.This study can provide reliable normaliza-tion for gene expression analysis and ensured more accurate data for further molecular mechanism re-search in C.trichotomum.
作者
华雅洁
岳远征
杨秀莲
何卿
HUA Ya-jie;YUE Yuan-zheng;YANG Xiu-lian;HE Qing(Co-Innovation Center of Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing210037,Jiangsu,China;Jiangsu Province Key Lab.of Environmental Engineering,Jiangsu Provincial Academy of Environmental Science,Nanjing210036,Jiangsu,China)
出处
《林业科学研究》
CSCD
北大核心
2022年第2期194-202,共9页
Forest Research
基金
江苏省“六大人才高峰”高层次人才项目(NJ-066)
国家林业局公益性行业科研专项项目(201404109)
江苏高校优势学科建设工程资助项目(PAPD)。
关键词
海州常山
胁迫
内参基因
实时荧光PCR
Clerodendrum trichotomum
abiotic stress
reference genes
qRT-PCR
