作物学报2018，Vol.44Issue(2)：218-226,9.DOI:10.3724/SP.J.1006.2018.00218
棉花半乳糖基转移酶基因GhGalT1启动子的克隆及表达分析
Cloning and Expression Analysis of Galactosyltransferase Gene GhGalT1 Promoter in Cotton
摘要
Abstract
Glycosytransferases (GTs) transfer an activated sugar donor to a specific acceptor to form glucosidic bond, which are regulated by various abiotic and biotic stresses, and may play a role in plant responses to changes in living conditions. In this study, a 539 bp fragment of GhGalT1 5′-flanking sequence was isolated from upland cotton Coker 312 by PCR, designated pGhGalT1. Analysis of pGhGalT1 sequence by PlantCARE revealed it contained not only putative CAAT box, TATA box se-quence, but also MBS, HSE, TC-rich repeats, MYCCONSE and CGTCA-motif cis-acting element which involved in drought, heat, dehydration, defense and stress responsiveness. Thus, we constructed it into pBI101-GUS vector and formed pGhGalT1::GUS fusion expression vector (pBI101-pGhGalT1-GUS), then transferred the vector into Arabidopsis by the Agro-bacterium-mediated floral dip method, and successfully obtained positive transgenic plants by screening test of resistance to kanamycin and PCR detection.Histochemical assay of T3generation of transgenic Arabidopsis revealed that GUS activities were mainly accumulated in root tips of primary and lateral roots in 5- to 15-day-old seedlings, and less strongly in cotyledons and rosette leaves. The histochemical staining results and the assay of quantitative GUS activity and GUS gene expression under abiotic stresses and hormone treatments revealed that the GhGalT1 promoter was salt-/osmotic-/6-BA-/MeJA-/BL-inducible. These findings contribute to the selection of a suitable promoter for crop molecular improvement.关键词
棉花/糖基转移酶/启动子/顺式作用元件/GUS组织化学染色/GUS定量分析Key words
cotton(Gossypium hirsutum L.)/glycosytransferase/promoter/cis-acting element/GUS histochemical staining/quan-titative GUS assay引用本文复制引用
秦丽霞,李静,张换样,李盛,竹梦婕,焦改丽,吴慎杰..棉花半乳糖基转移酶基因GhGalT1启动子的克隆及表达分析[J].作物学报,2018,44(2):218-226,9.基金项目
本研究由国家自然科学基金项目(31601350),山西省基础研究项目(2015021152)和国家转基因生物新品种培育重大专项(2016ZX08005)资助.This study was supported by the National Natural Science Foundation of China(31601350),the Shanxi Province Fundamental Research Foundation(2015021152),and the National Major Project for Developing New GM Crops(2016ZX08005). （31601350）
