摘要
【目的】ethylene insensitive 3/ethylene insensitive-like 1(EIN3/EIL1)是乙烯信号通路的重要成员,克隆并分析其在木薯块根采后生理性变质(post-harvest physiological deterioration,PPD)过程中的表达情况,可以为深入研究乙烯信号在木薯块根PPD过程中的功能提供参考。【方法】通过RT-PCR技术从木薯栽培品种SC8中克隆得到了木薯MeEIN3.1基因,然后对MeEIN3.1基因的遗传进化关系、结构域、蛋白质结构、理化性质等进行分析。对MeEIN3.1蛋白进行亚细胞定位,并通过荧光定量PCR和酵母双杂交技术对MeEIN3.1基因在木薯块根PPD过程中的表达水平以及下游互作转录因子进行分析。【结果】MeEIN3.1基因的长度为1452 bp,编码483个氨基酸残基,等电点和分子质量分别为5.08和55.12 ku,氨基酸序列包含7个EIN3/EIL1结构域,和橡胶HbEIN3-like基因的亲缘关系最近,序列一致性为69.35%。MeEIN3.1蛋白的亚细胞定位结果显示该基因位于细胞核。荧光定量PCR结果显示,在木薯块根的PPD过程中MeEIN3.1基因的表达量和对照0 h相比,表现为显著上升趋势,即MeEIN3.1基因的表达受到PPD过程的诱导。另外,酵母双杂交结果显示MeEIN3.1能够与MeERF1.2和MeERF1.3产生互作。【结论】MeEIN3.1基因的表达在采后过程中受到诱导,推测其通过下游转录因子MeERF1.2和MeERF1.3进行乙烯信号转导来参与木薯块根的PPD过程。
【Objective】Ethylene insensitive 3/ethylene insensitive-like 1(EIN3/EIL1)is a pivotal member of the ethylene signaling pathway.Cloning and analyzing its expression in post-harvest physiological deterioration(PPD)of cassava storage roots could serve as a reference for in-depth exploration of the functional role of ethylene signaling in the PPD process of cassava storage roots.【Meth-od】MeEIN3.1 gene was cloned from Manihot esculenta cv.SC8 by RT-PCR for its genetic evolutionary relationship analysis and pre-diction of physicochemical properties,conserved domain,structure,and subcellular localization of MeEIN3.1 protein by bioinformat-ics tools.Furthermore,its subcellular localization was verified by Agrobacterium tumefaciens infection method,and the expression of MeEIN3.1 protein in PPD process and its downstream interacting transcription factors were analyzed via qRT-PCR and yeast 2-hybrid assay.【Result】The results showed that the open reading frame(ORF)of MeERF3.1 was 1452 bp in length and encoded 483 amino acid residues.The molecular weight and isoelectric point of MeEIN3.1 protein was 55.12 ku and 5.08,respectively.The MeEIN3.1 protein contained 7 EIN3/EIL1 domain,presenting high sequence similarity with HbEIN3-like(69.35%).Subcellular localization showed that MeEIN3.1 protein was localized in nucleus.The expression of MeEIN3.1 gene presented a significant uptrend during the PPD process.The results of yeast 2-hybrid showed that MeEIN3.1 protein interacted with the downstream transcription factor MeERF1.2 and MeERF1.3.【Conclusion】MeEIN3.1 gene might be involved in the PPD process of cassava roots through ethylene signal transduction by downstream transcription factors MeERF1.2 and MeERF1.3.
作者
赖锦涛
杨静琳
罗佳科
陈志晟
叶晓雪
颜彦
曾坚
胡伟
LAI Jintao;YANG Jinglin;LUO Jiake;CHEN Zhisheng;YE Xiaoxue;YAN Yan;ZENG Jian;HU Wei(Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region,Shaoguan University,Shaoguan,Guangdong 512005,China;School of Biology andAgriculture,Shaoguan University,Shaoguan,Guangdong 512005,China;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou,Hainan 571101,China)
出处
《福建农林大学学报（自然科学版）》
CAS
CSCD
北大核心
2024年第1期7-14,共8页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
广东省基础与应用基础研究基金(2021A1515011236,2023A1515010336)
海南省自然科学基金面上项目(320MS098,322MS128)
中央级公益性科研院所基本科研业务费专项(1630052022027)
广东省普通高校重点领域专项(2022ZDZX4047)
韶关学院重点项目(SZ2022KJ05)
韶关学院博士启动项目(99000615)
国家自然科学基金(31901537)
校级大学生创新创业训练计划项目(Sycxcy2022089)。
关键词
木薯
乙烯
EIN
活性氧
采后生理性变质
cassava
ethylene
ethylene insensitive
reactive oxygen species
post-harvest physiological deterioration
