河南农业科学 ›› 2023, Vol. 52 ›› Issue (1): 125-133.DOI: 10.15933/j.cnki.1004-3268.2023.01.013

• 园艺 • 上一篇    下一篇

青花菜花蕾发育LncRNAs 内参基因筛选

裴徐梨1,冯鹏宇1,唐征2,李亚梅1,焦鹏1,荆赞革1,2   

  1. (1.昆明学院农学 与生命科学学院,云南 昆明 650214;2.温州科技职业学院 农业与生物技术学院,浙江 温州 325006)
  • 收稿日期:2022-09-21 出版日期:2023-01-15 发布日期:2023-03-09
  • 通讯作者: 荆赞革(1983-),男,河南灵宝人,副研究员,博士,主要从事蔬菜分子生物学研究。E-mail:jingzange@aliyun.com
  • 作者简介:裴徐梨(1990-),女,云南昭通人,讲师,博士,主要从事蔬菜分子生物学研究。E-mail:xuliP1990@163.com
  • 基金资助:
    云南省地方本科高校(部分)基础研究联合专项(2018FH001-044);浙江省自然科学基金项目(LY18C150006);浙江省农业
    (蔬菜新品种选育)新品种选育重大科技专项子项目(2021C02065-4-2);温州市科技项目(2019ZX007-2);浙江省教育厅一
    般项目(Y202045786)

Selection of LncRNAs Reference Genes in Development of Flower Bud in Broccoli

PEI Xuli1,FENG Pengyu1,TANG Zheng2,LI Yamei1,JIAO Peng1,JING Zange1,2   

  1. (1.College of Agriculture and Life Sciences,Kunming University,Kunming 650214,China;2.College of Agriculture and Biotechnology,Wenzhou Vocational College of Science and Technology,Wenzhou 325006,China)
  • Received:2022-09-21 Published:2023-01-15 Online:2023-03-09

摘要: 在分析青花菜花蕾发育过程长链非编码RNA(LncRNAs)的表达中,筛选稳定表达的内参基因至关重要。通过前期高通量测序筛选出16个候选内参基因,利用qRT-PCR技术检测其表达水平,并结合geNorm、NormFinder和BestKeeper 3个软件对16个LncRNAs在青花菜花蕾不同部位和不同发育时期的表达稳定性进行分析。结果表明,在花蕾不同部位中,geNorm算法认为XLOC_000400、XLOC_008536、XLOC_034059为最适内参基因;NormFinder计算XLOC_021473、XLOC_000400、XLOC_034059为最佳内参基因;BestKeeper显示XLOC_025578表达最稳定。在花蕾不同发育时期中,geNorm算法认为XLOC_000108、XLOC_039609、XLOC_021473为最适内参基因;NormFinder计算XLOC_021473、XLOC_039609、XLOC_000108为最佳内参基因;BestKeeper显示XLOC_034059表达最稳定。综合评估后,在花蕾不同部位和不同发育时期中适宜的内参基因对分别为XLOC_000400/XLOC_008536XLOC_039609/XLOC_021473

关键词: 青花菜, 内参基因, 长链非编码RNA, 花蕾

Abstract: In order to preliminarily analyze the expression of LncRNAs during the flower bud development of broccoli,it is very important to screen the stably expressed internal reference genes. In this study,16 candidate internal reference genes were screened by high‑throughput sequencing.The expression levels were detected by qRT‑PCR. And the expression stability of 16 LncRNAs in different parts and different developmental stages of broccoli flower bud was analyzed by geNorm,NormFinder and BestKeeper softwares.The results showed that in different parts of bud,geNorm demonstrated that XLOC_000400,XLOC_008536and XLOC_034059 were suitable reference genes;NormFinder recommended XLOC_021473,XLOC_000400 and XLOC_034059 as reference genes;BestKeeper showed that the expression of XLOC_025578 was the most stable.In different bud development stages,geNorm demonstrated that XLOC_000108,XLOC_039609 and XLOC_021473 were suitable reference genes;NormFinderrecommended XLOC_021473,XLOC_039609 and XLOC_000108 as reference genes; BestKeeper showed that the expression of XLOC_034059 was the most stable. After a comprehensive evaluation,XLOC_000400/XLOC_008536 and XLOC_039609/XLOC_021473 were the most stable reference genes for different parts and different developmental stages,respectively.

Key words: Broccoli, Reference gene, Long non?coding RNA, Flower bud

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