CAJ | 학술논문

V-H+-ATPase在植物生长、发育和胁迫响应等生物学过程中发挥重要作用。本研究通过序列比对,从谷子中克隆到V-H+-ATPase E亚基基因SiVHA-E。进化树分析显示,SiVHA-E基因在进化上属于E1/E3亚族,与玉米V-H+-ATPase E亚基基因ZmVHA-EL亲缘关系较近。表达谱分析结果显示,SiVHA-E在高盐、茉莉酸甲酯(MeJA)、水杨酸(SA)和脱落酸(ABA)处理下表达上调,在低温和低氮处理下表达下调。亚细胞定位分析表明 SiVHA-E 定位于液泡膜上。遗传转化拟南芥的耐盐性鉴定表明,在盐处理条件下,转基因株系的种子萌发率、幼苗主根长、植株鲜重及存活率显著高于野生型的。与野生型拟南芥植株相比, SiVHA-E过表达植株体内Na+含量减少,体内相对含水量提高。此外, ABA萌发试验结果显示,在种子萌发后期,SiVHA-E过表达植株对ABA更加敏感。研究表明,谷子SiVHA-E可以显著提高拟南芥耐盐性,这可能与其正向调控ABA信号途径以及减少植株体内Na+积累和水分散失有关。
V-H+-ATPase재식물생장、발육화협박향응등생물학과정중발휘중요작용。본연구통과서렬비대,종곡자중극륭도V-H+-ATPase E아기기인SiVHA-E。진화수분석현시,SiVHA-E기인재진화상속우E1/E3아족,여옥미V-H+-ATPase E아기기인ZmVHA-EL친연관계교근。표체보분석결과현시,SiVHA-E재고염、말리산갑지(MeJA)、수양산(SA)화탈락산(ABA)처리하표체상조,재저온화저담처리하표체하조。아세포정위분석표명 SiVHA-E 정위우액포막상。유전전화의남개적내염성감정표명,재염처리조건하,전기인주계적충자맹발솔、유묘주근장、식주선중급존활솔현저고우야생형적。여야생형의남개식주상비, SiVHA-E과표체식주체내Na+함량감소,체내상대함수량제고。차외, ABA맹발시험결과현시,재충자맹발후기,SiVHA-E과표체식주대ABA경가민감。연구표명,곡자SiVHA-E가이현저제고의남개내염성,저가능여기정향조공ABA신호도경이급감소식주체내Na+적루화수분산실유관。
The V-H+-ATPase plays an important role in processes of plant growth, development and response to stresses. In this research,SiVHA-E,a V-H+-ATPase E subunit gene, was cloned from millet by the Blast analysis against GenBank database. Phy-logenetic tree showed that the gene belongs to E1/E3 subgroup and is close withZmVHA-EL, a V-H+-ATPase E subunit from maize. The quantitative Real-time PCR (qRT-PCR) analysis revealed that the expression levels ofSiVHA-Ewere up-regulated under treatments of high-salt, exogenous MeJA, SA, and ABA hormones, while down-regulated under stresses of cold and low nitrogen. Protein subcellular localization analysis using protoplast showed that SiVHA-E is located on tonoplast. The results of salt tolerance assay showed that the germination rate ofSiVHA-E transgenic lines was significantly higher than that of wild type plant under salt stress. During seedlings period, the root lengths were significantly longer as well as fresh weight and survival rate were significantly higher in transgenic lines than in wild type plant under salt treatment. Compared with wild type plant, trans-genic plant reduced the content of Na+and increased the relative water content inside cells. In addition, the results of germination experiment used ABA showed thatSiVHA-E transgenicArabidopsis was more sensitive to ABA than wild type plant during post-germination. In short, overexpressingSiVHA-Ein transgenicArabidopsis lines enhances salt tolerance, which might be relates to positive regulation of ABA signaling pathway or reduction of Na+ accumulation and water loss in transgenic plants.