非生物胁迫下黄瓜(Cucumis sativus)四个自噬基因的表达及生物信息学分析  

Expression and Bioinformatics Analysis of Four Autophagy Genes in Cucumber(Cucumis sativus) under Abiotic Stress

作　　者：杨悦 王瑀 丁国华[1] Yang Yue;Wang Yu;Ding Guohua(School of Life Science and Technology,Harbin Normal University,Harbin,150025)

机构地区：[1]哈尔滨师范大学生命科学与技术学院,哈尔滨150025

出　　处：《分子植物育种》2020年第14期4547-4554,共8页Molecular Plant Breeding

摘　　要：自噬是一种存在于真核生物中进化保守的分解代谢过程,在植物处于逆境胁迫以及生长发育受阻时起到至关重要的作用。通过水杨酸诱导,从黄瓜中分离出四个自噬基因:ATG8C1、ATG8C2、ATG8F、ATG8C,为揭示这些自噬基因在非生物胁迫下的作用,本研究选取黄瓜幼苗为试材,分别进行以下激素和胁迫处理,并用qRT-PCR检测基因的表达:(1)用10 mmol/L SA和0.2%MeJA滴加叶片进行激素诱导处理,于0 h、6 h、9 h和24 h在滴加的位置取材;(2)用200 mmol/L NaCl进行盐胁迫处理,用20%PEG6000进行干旱胁迫处理,于0 h、6 h和24 h取黄瓜幼苗的根、茎和叶;(3)用黑暗进行碳饥饿处理,于0 h、6 h和24 h取黄瓜幼苗的根、茎和叶。结果表明,在SA和Me JA处理下,黄瓜四个自噬基因都呈现上调表达;在盐和干旱处理下,四个自噬基因的表达有所不同,在黄瓜根和叶中基本呈现上调表达,在茎中多数呈现下调表达。在碳饥饿处理下,四个自噬基因呈现大幅度上调表达。该结果说明在不同的胁迫下,自噬的发生途径有所不同。本研究还利用ExPasy在线工具ProParam和ProtScale分析了四个自噬蛋白的氨基酸、理论分子量和等电点等理化参数,利用MEGA5软件建立了系统进化树,对四个自噬基因进行了初步的生物信息学分析。通过以上试验可以为进一步研究黄瓜自噬基因参与植物抗逆胁迫提供分子生物学的理论基础。Autophagy is an evolutionarily conserved catabolic process in eukaryotes, which plays a crucial role in plant growth and development under stress. Induced by salicylic acid, isolated from four autophagy genes in cucumber;ATG8 C1, ATG8 C2, ATG8 F, ATG8 C3, to reveal the autophagy genes under abiotic stress, this article selects the cucumber seedlings as test materials, processing, make the following hormone and stress respectively. qRT-PCR is used to test the expression of genes:(1) With 10 mmol/L SA and 0.2% MeJA plus a hormone treatment of blades, at0 h, 6 h, 9 h and 24 h in the drops and the location of the materials.(2) Salt stress was treated with 200 mmol/L NaCl, and drought stress was treated with 20% PEG6000. Roots, stems and leaves of cucumber seedlings were collected at 0 h, 6 h and 24 h.(3) Carbon starvation was conducted in the dark. Roots, stems and leaves of cucumber seedlings were taken at 0 h, 6 h and 24 h. The results showed that four autophagy genes were up-regulated in cucumber under SA and Me JA treatment. Under the treatment of salt and drought, the expressions of four autophagy genes were different, and the expressions were basically up-regulated in the root and leaf of cucumber, and downregulated in most of the stem. Under the carbon starvation treatment, the expression of four autophagy genes was significantly up-regulated. This also indicates that autophagy occurs in different ways under different stresses. In this paper, ExPasy online tools ProParam and ProtScale were used to analyze the amino acid, theoretical molecular weight, isoelectric point and other physical and chemical parameters of the four autophagy proteins, and MEGA5 software was used to establish the phylogenetic tree for the preliminary bioinformatics analysis of the four autophagy genes. The above experiments can provide a theoretical basis for the further study of cucumber autophagy genes involved in plant stress resistance.

关 键 词：黄瓜(Cucumis sativus) 自噬 非生物胁迫 基因表达 

分 类 号：S642.2[农业科学—蔬菜学]