摘要
为了筛选适用于雪茄烟叶(BESNO H382)基因定量研究的内参基因,以温室栽培条件下雪茄烟叶的不同组织器官(旺长期根、茎、叶,盛花期花萼、花冠、柱头、子房、雄蕊、雌蕊及种子)及不同盐处理条件下(300 mmol/L NaCl处理不同时间)的样品为材料,采用实时荧光定量PCR技术对从雪茄烟叶转录组数据中筛选到的10个候选内参基因进行分析,同时使用geNorm、NormFinder和BestKeeper等3种软件综合评价这些候选基因的表达稳定性。结果表明,RPL14A和UBC27最适合作为雪茄烟叶不同组织的内参基因;UBC28最适合作为雪茄烟叶不同盐处理条件下的内参基因。分别以UBC27、UBC28和RPL14A为内参基因考察了胁迫相关基因脯氨酸合成酶基因(P5CS)在盐胁迫下的表达情况,发现P5CS上调表达且相对3个内参基因的表达量和变化趋势均较为一致。
In order to screen stable reference genes in cigar tobacco(BESNO H382)for quantitative real-time PCR(qRT-PCR),different tissues(such as roots,stalks and leaves at fast growing stage,sepals,corollas,stigmas,ovaries,stamens and pistils at full-bloom stage and seeds)of cigar tobacco cultured in greenhouse and leaves under different salt stresses(300 mmol/L NaCl treated for different durations)were used as study materials.Ten candidate reference genes screened from transcriptome data of cigar tobacco were analyzed by qRT-PCR,and the expression stabilities of those candidate genes were comprehensively evaluated by geNorm,NormFinder and BestKeeper.The results showed that RPL14A and UBC27 were the best reference genes for the different organs of cigar tobacco,and UBC28 was the best reference gene for cigar tobacco under the different salt stresses.Furthermore,the expression of stress related proline synthase gene P5CS under the salt stress was analyzed by using UBC27,UBC28 or RPL14A as a reference gene separately.The expression of P5CS was upregulated,and the expression levels of P5CS relative to the three reference genes were similar with comparable variation tendencies.
作者
张慧
金静静
徐国云
翟妞
郑庆霞
刘萍萍
陈千思
金立锋
周会娜
ZHANG Hui;JIN Jingjing;XU Guoyun;ZHAI Niu;ZHENG Qingxia;LIU Pingping;CHEN Qiansi;JIN Lifeng;ZHOU Huina(Zhengzhou Tobacco Research Institute of CNTC,Zhengzhou 450001,China)
出处
《烟草科技》
CAS
CSCD
北大核心
2022年第5期17-24,共8页
Tobacco Science & Technology
基金
国家烟草专卖局科技重大专项项目“烟草氯离子调控功能基因研究及低氯离子含量的育种材料创制”[110202001018(JY-01)]。
