‘粉红亚都蜜’葡萄NAC转录因子基因VvDRL1的功能初步分析

Subcellular Localization and Functional Analysis of a NAC Gene VvDRL1 from Vitis vinifera ‘Yatomi Rosa’

以欧洲葡萄‘粉红亚都蜜’为材料,利用同源克隆法获得1个NAC转录因子,命名为VvDRL1（Gen Bank：XP-002281816）。该基因全长840 bp,编码280个氨基酸,与‘黑比诺’葡萄中该基因的同源性为100%,但是基因组DNA序列中存在6处单核苷酸突变。瞬时转化洋葱表皮细胞进行亚细胞定位分析,VvDRL1基因主要在细胞核内表达,属于核蛋白;通过农杆菌介导的叶盘法获得稳定整合的转基因烟草,T2代转基因植株生长迟缓,株高、叶面积和叶片细胞面积约为野生型烟草的60%、34%和31%,转基因植株叶片出现向内卷曲现象,利用石蜡切片进行显微结构观察,转基因植株主叶脉上表皮下缺失2~3层厚壁细胞,且海绵组织内的空隙明显多于野生型植株。研究结果表明,VvDRL1在调控植株的形态发育方面起着重要作用。

In this study, a NAC gene, VvDRL1, was isolated from Vitis vinifera ‘Yatomi Rosa＇. The length of full VvDRL1 was 840 bp, encoding 280 amino acids, which shared a 100% similarity in cRNA and 6 single nucleotide mutation in DNA with ‘Pinot Noir＇ . Subcellular localization showed that VvDRL1 is a nuclear protein. Transgenic tobacco with VvDRL1 was obtained by Agrobacterium-mediated method. Compared with the wild type, the growth of T2-generation transgenic tobacco presented delayed with the plant height, leaf area and mesophyll cells area of about 60%, 34% and 31% in comparison with wild tobacco plants. In addition, the transgenic tobacco showed curly leaves. Through paraffin sections observation, it is found that the upper epidermis in main leaf veins of transgenic tobacco lacked 2 - 3 layers sclerenchyma ceils and had significantly more space in the spongy tissue compared with the wild type. Based on the above study, it is inferred that VvDRL1 gene play an important role inthe regulation of plant morphological development.