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克隆和表达小拟南芥种子中半乳糖醇合成酶基因（AnGolS1）

Molecular cloning and expression of an encoding galactinol synthase gene (AnGolS1) in seedling of Ammopiptanthus nanus.

机构信息

Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District 110866, P.R. China.

Key Laboratory of Protected Horticulture of Ministry of Education, No. 120 Dongling Road, Shenhe District 110866, P.R. China.

出版信息

Sci Rep. 2016 Oct 27;6:36113. doi: 10.1038/srep36113.

DOI:10.1038/srep36113
PMID:27786294
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5081558/
Abstract

Based on the galactinol synthase (AnGolS1) fragment sequence from a cold-induced Suppression Subtractive Hybridization (SSH) library derived from Ammopiptanthus nanus (A. nanus) seedlings, AnGolS1 mRNA (including the 5' UTR and 3' UTR) (GenBank accession number: GU942748) was isolated and characterized by rapid amplification of cDNA ends polymerase chain reaction (RACE-PCR). A substrate reaction test revealed that AnGolS1 possessed galactinol synthase activity in vitro and could potentially be an early-responsive gene. Furthermore, quantitative real-time PCR (qRT-PCR) indicated that AnGolS1 was responded to cold, salts and drought stresses, however, significantly up-regulated in all origans by low temperatures, especially in plant stems. In addition, the hybridization signals in the fascicular cambium were strongest in all cells under low temperature. Thus, we propose that AnGolS1 plays critical roles in A. nanus low-temperature stress resistance and that fascicular cambium cells could be involved in AnGolS1 mRNA transcription, galactinol transportation and coordination under low-temperature stress.

摘要

基于从小叶白蜡（Ammopiptanthus nanus）幼苗低温诱导抑制差减杂交文库中克隆的半乳糖醇合酶（AnGolS1）片段序列，采用快速扩增 cDNA 末端聚合酶链反应（RACE-PCR）法分离并鉴定了 AnGolS1 mRNA（包含 5'UTR 和 3'UTR）（GenBank 登录号：GU942748）。体外酶活性测定表明 AnGolS1 具有半乳糖醇合酶活性，可能是一个早期响应基因。实时荧光定量 PCR（qRT-PCR）分析表明，AnGolS1 对低温、盐和干旱胁迫有响应，但低温能显著诱导其在各器官中表达上调，尤其在植物茎中。此外，低温下各组织束间形成层细胞中杂交信号最强。因此，我们推测 AnGolS1 在小腊抵抗低温胁迫中发挥重要作用，束间形成层细胞可能参与低温胁迫下 AnGolS1 mRNA 的转录、半乳糖醇的运输和协调。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/548c68c72cf5/srep36113-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/d48714046065/srep36113-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/b298c4da790b/srep36113-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/18d7e153ac7f/srep36113-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/e548f3029722/srep36113-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/24f4f431dd28/srep36113-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/548c68c72cf5/srep36113-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/d48714046065/srep36113-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/b298c4da790b/srep36113-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/18d7e153ac7f/srep36113-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/e548f3029722/srep36113-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/24f4f431dd28/srep36113-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2c9e/5081558/548c68c72cf5/srep36113-f6.jpg

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