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园艺学报 ›› 2024, Vol. 51 ›› Issue (4): 748-760.doi: 10.16420/j.issn.0513-353x.2023-0343

• 遗传育种·种质资源·分子生物学 • 上一篇    下一篇

冬瓜实时荧光定量PCR内参基因的筛选与评价

何长霞1,2,罗 陈2,闫晋强2,刘文睿2,王 敏2,谢大森2,吴智明1,*,江 彪2,*   

  1. 1仲恺农业工程学院园艺园林学院,广州 510225;2广东省农业科学院蔬菜研究所,广东省蔬菜新技术研究重点实验室,广州 510640
  • 出版日期:2024-04-25 发布日期:2024-04-26
  • 基金资助:
    广东省自然科学基金青年提升项目(2023A1515030049);广东省基础与应用基础研究基金-区域联合基金青年基金项目(2021A1515111214);2022年广东省级乡村振兴战略专项资金种业振兴项目(2022-NPY-00-028);广东省农业科学院中青年学科带头人项目(R2020PY-JG003);广东省农业科学院优秀博士项目(R2021YJ-YB2004)

Screening and Evaluation of Reference Genes for Real-time Quantitative PCR in Wax Gourd

HE Changxia1,2,LUO Chen2,YAN Jinqiang2,LIU Wenrui2,WANG Min2,XIE Dasen2,WU Zhiming1,*,and JIANG Biao2,*   

  1. 1College of Horticulture and Landscape Architecture,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China;2Guangdong Key Laboratory for New Technology Research on Vegetables,Vegetable Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China
  • Published:2024-04-25 Online:2024-04-26

摘要: 运用3种统计学软件GeNorm、NormFinder和BestKeeper分析冬瓜9个内参基因(18SrRNA、ACT、CYP、EF-1α、F-box、GAPDH、TUA、TUB和UBQ)在冬瓜不同组织(根、茎、叶、雄花、雌花和子房)、不同发育时期果实(授粉后0、10和20 d)、生物胁迫(疫霉侵染)和非生物胁迫(盐、高温和低温处理)植物叶片样品中的表达稳定性。结果表明,在不同组织中,EF-1α的表达稳定性最好;在不同发育时期果实和疫霉处理植物叶片中,F-box的表达稳定性最好;在盐胁迫处理植物叶片中,18SrRNA表现出较强的稳定性;在高温胁迫处理植物叶片中,ACT的表达稳定性最好;在低温胁迫处理植物叶片中,TUA的稳定性最好。

关键词: 冬瓜, 实时荧光定量PCR, 内参基因

Abstract: Three statistical software including GeNorm,NormFinder,and BestKeeper were applied to analyze the expression stability of nine internal reference genes(18SrRNA,ACT,CYP,EF-1α,F-box,GAPDH,TUA,TUB,and UBQ)in wax gourd. The plant samples of plants were taken in different tissues (root,stem,leaf,male flower,female flower,ovary),fruits at different developmental stages including (0 d,10 d,and 20 d after pollination)of wax gourd,and leaves under biotic stress(Phytophthora melonis infection),and abiotic stress(salt,high temperature,and low temperature treatment). The obtained results showed that the expression stability of EF-1α was the best in different tissues. The expression stability of F-box was the best in the fruits at different developmental stages and the leaves under P. melonis infection. By comparison,18SrRNA showed strong stability in the leaves under salt stress,while the expression stability of ACT was the best in the leaves under high temperature stress and TUA had the best stability in the leaves under low temperature stress.

Key words: wax gourd, real-time quantitative PCR, reference genes

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