DELLA蛋白介导的海岛棉新型遗传转化体系的建立  

Establishment of a Novel DELLA Protein-mediated Genetic Transformation System in Gossypium barbadense

作　　者：姑扎丽努尔·吐鲁洪 张佳伟 李悦[1] 王晓云 张霞[1] Guzhalinuer•TULUHONG;ZHANG Jia-Wei;LI Yue;WANG Xiao-Yun;ZHANG Xia(College of Life Sciences,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学生命科学学院,乌鲁木齐830052

出　　处：《农业生物技术学报》2024年第11期2681-2690,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32060044)。

摘　　要：植物遗传转化技术的关键是植株再生体系的建立,而植物遗传转化体系中抗生素的使用不利于植物细胞分化,从而增加了植株再生体系建立的难度。DELLA蛋白GbGAI4(GAI:gibberellic acid insensitive)基因的过表达赋予植株矮化的特性是一种肉眼可见、十分容易辨认的植物形态变化,有望作为形态筛选标记用于植株遗传转化体系的建立。本研究首先通过构建pTA7002-GbGAI4载体,通过农杆菌(Agrobacterium)介导法侵染海岛棉(Gossypium barbadense)'新海16'棉花胚尖,借助地塞米松特异性诱导GbGAI4基因表达,初步筛选出具有植株矮化性状的转化植株,并进一步借助RT-PCR技术鉴定出转基因阳性植株,从而建立一种新型棉花遗传转化体系。在此基础上,选择生长调控因子(growth-regulating factors,GRF)GRF4作为基于GbGAI4介导的遗传转化系统的转基因示例基因,获得(4±0.6)%的基因转化率。本研究为棉花分子育种工作提供一种新的遗传转化技术平台。Establishment of plant regeneration system is essential for plant genetic transformation,and the use of antibiotics in plant genetic transformation systems is detrimental to cell differentiation,thus exacerbating the difficulty of establishing plant regeneration systems.Overexpression of the DELLA protein GbGAI4(GAI:gibberellic acid insensitive)gene confers plant dwarfism,which is a visible and easily recognizable morphological phenotype in plants,and is expected to be used as a morphological screening marker in plant genetic transformation systems.In this study,pTA7002-GbGAI4 vector was constructed,the embryo tips of island cotton(Gossypium barbadense)'Xinhai16'was infiltrated by Agrobacterium-mediated method,the expression of GbGAI4 gene was induced by dexamethasone,and then preliminarily screened out the transformed plants with dwarf characters,and further identified the transgenic plants with RT-PCR technology,so as to establish a noval genetic transformation system in cotton.Based on this,the growthregulating factors(GRF)GRF4 was selected as the example gene for transgenic transformation based on the GbGAI4-mediated genetic transformation system,and a gene transformation rate of(4±0.6)%was obtained.This study provides a new genetic transformation platform for molecular breeding of cotton.

关 键 词：新型遗传筛选标记 DELLA蛋白 海岛棉 农杆菌介导法 胚尖 

分 类 号：S5[农业科学—作物学]