红苞凤梨实时荧光定量pcr分析中内参基因的筛选-热带亚热带植物.pdfVIP

热带亚热带植物学报 2017, 25(3): 250 ~ 256 Journal of Tropical and Subtropical Botany 红苞凤梨实时荧光定量PCR 分析中内参基因的筛选 李瑞雪, 余三淼, 李夏, 熊颖媛, 蔺珍, 唐琰琪, 马均* ( 四川农业大学风景园林学院，成都 611130) 摘要：为筛选表达稳定的内参基因，以红苞凤梨(Ananas comosus var. bracteatus)不同发育时期的全绿、全白苗为材料，对10 个组成型表达基因EF1 、UBQ、ACT 、GADPH、Histone 、TUA、TUB、18S、elf-5A、α-tubulin 进行筛选，并分析PetF 基因 的表达模式。结果表明，10 个候选内参基因在红苞凤梨全绿、全白苗不同生长阶段中的表达稳定性不同。红苞凤梨不同生 长时期以Histone 和α-tubulin 为最适内参基因，而全绿苗和全白苗的对比分析以18S、EF1 和α-tubulin 为最理想的内参组合。 PetF 基因在红苞凤梨发育过程及绿、白苗对比分析中的表达水平变化趋势一致，因此，所选的内参基因是合适的。 关键词：红苞凤梨；实时荧光定量PCR ；内参基因 doi: 10.11926/jtsb.3659 Screening of Reference Genes in Ananas comosus var. bracteatus for qRT-PCR * LI Rui-xue, YU San-miao, LI Xia, XIONG Ying-yuan, LIN Zhen, TANG Yan-qi, MA Jun (College of Landscape Architecture, Sichuan Agricultural Uniersity, Chengdu 611130, China) Abstract: In order to screen appropriate reference gene for real-time qPCR (qRT-PCR) in Ananas comosus var. bracteatus at different development stages, the expression stability of 10 housekeeping genes, such as EF1, UBQ, ACT , GADPH, Histone, TUA, TUB, 18S, elf-5a, and α-tubulin, were detected, and the expression of PetF in seedling of A. comosus var. bracteatus was studied. The results showed that the expression stability of these genes in albino and green seedlings was different at development stages. Histone and α-tubulin were optimum reference genes at different development stages. Compared between green and albino seedlings, the ideal reference gene combination were 18S, EF1 and α-tubulin. The expression of PetF gene had the same variation trends in green and albino seedlings at three development stages by using screening reference genes. So, it was confirmed that the reference genes screened were suitable for qRT-PCR of A. comosus var. bracteatus