辣椒MADS-box转录因子CaRIN基因克隆、表达与功能分析
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1.重庆市农业科学院蔬菜花卉研究所;2.重庆市农业科学院生物技术研究所

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重庆市农业科学院绩效激励引导专项项目(cqaas2022jxjl002);国家现代农业产业技术体系项目(CARS-24-A-04);


Cloning, Expression, and Function analysis of MADS-box Transcription Factor CaRIN Gene from Capsicum annuum
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    摘要:

    【目的】为探究MADS-box家族基因RIN的表达特征和功能,解析其对辣椒类胡萝卜素代谢的影响。【方法】基于辣椒果实发育转录组,通过RT-PCR克隆辣椒MADS-box转录因子CaRIN基因CDS全长,对其进行生物信息学、表达模式、亚细胞定位、转录活性等分析,并探讨VIGS诱导CaRIN基因沉默对类胡萝卜素代谢的影响。【结果】(1)CaRIN基因CDS全长732 bp,编码243个氨基酸,蛋白分子量为27.95 kD,等电点(pI)为7.06,其编码蛋白具有典型的MEF2_like MADS结构域,属MICK-type型转录因子;(2)CaRIN基因主要在花和果实中表达,具有组织特异性;CaRIN定位在细胞核,并且具有转录激活活性。(3)CaRIN基因启动子具有ABRE等多个激素应答元件,外源脱落酸和乙烯利均能加速果实转红,诱导CaRIN及相关基因高表达;(4)VIGS诱导沉默CaRIN基因后,类胡萝卜素代谢途径基因PSY1、CCS、PDS、CRTZ、LCYB和NCED1表达水平降低为对照组的0.27-0.59倍,且果实总类胡萝卜素含量(0.379 mg/g)较对照(0.650 mg/g)显著降低。【结论】CaRIN可能是辣椒果实类胡萝卜素代谢过程的重要调控因子。

    Abstract:

    【Objective】 The study aims to explore the expression characteristics and function of RIN gene of MADS-box, and analyze its impact on the metabolism of carotenoids in pepper. 【Methods】 Based on the transcriptome of pepper fruit development, the full-length CDS of MADS-box transcription factor CaRIN gene related to chili ripening was cloned. Then the bioinformatics, expression patterns, subcellular localization, and transcriptional activation activity were conducted, and the effects of VIGS-induced CaRIN gene silencing on carotenoid metabolism were investigated. 【Results】 (1) The CDS region of CaRIN gene was 732 bp in length. The CaRIN gene encoded a protein containing 243 amino acids with a protein molecular weight of 27.95 kD and an isoelectric point (pI) of 7.06. The CaRIN protein has typical MEF2_ Like MADS domain, belonging to the MICK type transcription factor. (2) CaRIN gene is mainly expressed in flowers and fruits with tissue specificity. CaRIN was located in the nucleus and had transcriptional activation activity. (3) The promoter of CaRIN gene had several hormone response elements such as ABRE, and exogenous ABA and ethephon could accelerate the red turning of fruit and induce the high expression of CaRIN and related genes. (4) After VIGS-induced gene silencing of CaRIN, the expression levels of PSY1, CCS, PDS, CRTZ, LCYB and NCED1 of the carotenoid metabolic pathway genes were decreased by 0.27-0.59 times and the total carotenoid content in fruits (0.379 mg/g) was also significantly decreased compared with the control group (0.650 mg/g). 【Conclusion】 CaRIN may be an important regulatory factor of carotenoid metabolism in capsicum fruit.

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  • 收稿日期:2024-02-22
  • 最后修改日期:2024-04-03
  • 录用日期:2024-04-08
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