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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。
A NAC transcription factor named VvDRL1 (Gen Bank: XP-002281816) was obtained by homologous cloning method using the European grape “Pink Yadu honey” as material. This gene is 840 bp in length and encodes a protein of 280 amino acids with 100% homology with that of ’Pinot Noir’ grape, but there are 6 single nucleotide mutations in the genomic DNA sequence. Transient transformation of onion epidermal cells for subcellular localization analysis, VvDRL1 gene is mainly expressed in the nucleus, is a nuclear protein; Agrobacterium-mediated leaf disk method to obtain stable and integrated transgenic tobacco, T2 generation transgenic plants growth retardation, plant height, leaf The area and area of leaf cells were about 60%, 34% and 31% of that of wild type tobacco. The curly leaves of transgenic plants were curled inwards. The microstructure of paraffin sections was observed. Thick-walled cells, and spongy tissues significantly more than the gaps in the wild-type plants. The results show that VvDRL1 plays an important role in the regulation of plant morphological development.