番茄qRT-PCR内参基因的筛选

发布时间：2018-04-18 04:07

本文选题：番茄 + 实时荧光定量PCR　；参考：《江苏农业学报》2017年02期

【摘要】：以高温、低温、盐胁迫、TYLCV接种处理和对照的番茄幼苗为植物材料,通过实时荧光定量RT-PCR(Real-time fluorescence quantitative PCR,qRT-PCR)方法分析了常用的8个内参基因肌动蛋白基因(ACT)、甘油醛-3-磷酸脱氢酶基因(GAPDH)、泛素基因(UBI)、18S核糖体RNA基因(18S)、转录延伸因子基因(EF-1a)、β-微管蛋白基因(TUB)、表达蛋白基因(EXP)和接合素蛋白复合物基因(CAC)在不同样本中的表达情况。利用ge Norm和Norm Finder软件分析筛选表达稳定性较高的内参基因。结果表明,在不同胁迫处理条件下,UBI和ACT的稳定性较高;在同样的胁迫处理中,8个内参基因的表达稳定性不同。综合而言,虽然UBI和ACT的稳定性同样较高,但在TYLCV接种处理试验中ACT的稳定性相对较差,而TUB和GAPDH表达较稳定,故TUB和GAPDH可在TYLCV接种处理试验中作为番茄qRT-PCR的内参基因。
[Abstract]:Tomato seedlings inoculated with TYLCV at high temperature, low temperature and salt stress were used as plant materials.By using real-time RT-PCR(Real-time fluorescence quantitative PCRQRT-PCRR, eight common domestic reference gene actin genes, Actr, glyceraldehyde 3-phosphate dehydrogenase, UBIN 18s ribosomal RNA gene, transcription extension factor gene EF-1a, 尾 -tubulin protein were analyzed by using real-time fluorescence quantitation method. The results were as follows: GAPDHN, UBIN 18s ribosomal RNA gene, transcriptional extension factor gene EF-1aa1, 尾 -tubulin protein, 尾 -tubulin.The expression of TUBN, express protein gene (exp) and conjugate protein complex gene (CAC) in different samples.GE Norm and Norm Finder software were used to analyze and screen stable internal reference genes.The results showed that the stability of UBI and ACT was higher under different stress conditions, and the stability of expression of 8 internal reference genes was different in the same stress treatment.In general, although the stability of UBI and ACT was also high, the stability of ACT was relatively poor in TYLCV inoculation test, while the expression of TUB and GAPDH was stable. Therefore, TUB and GAPDH could be used as internal reference genes of qRT-PCR in TYLCV inoculation test.
【作者单位】：南京农业大学园艺学院;江苏省农业科学院蔬菜研究所;
【基金】：国家自然科学青年基金项目(31401884) 江苏省自然科学青年基金项目(BK20140739) 江苏省自主创新基金项目[CX(14)5012] 江苏省农业科学院基本科研业务专项[ZX(15)2003]
【分类号】：S641.2

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