Cloning and Expression Analysis of bZIP Transcription Factor from Finger Millet (Eleusine coracana L.) Under Abiotic Stress Conditions

J. Lolita C.
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Abstract

Basic leucine zipper (bZIP) transcription factors comprise one of the largest gene families in plants. They play a key role in almost every aspect of plant growth and development and also in biotic and abiotic stress tolerance. In this study, we were attempted to study characterization of bZIP, a transcription factor from a climate smart cereal finger millet (Eleusine coracana L.). Seeds of Eleusine coracana (finger millet) was purchase from local market and were grown under field conditions drought and salt stress conditions. In this study, EcbZIP gene was isolated from finger millet, cloned into DH5α cells, screened by using colony PCR and expression analysis in response to two abiotic stresses was carried out by using qRT PCR. EcbZIP coding DNA sequence and protein sequence were retrieved from NCBI Nucleotide Database and Genpept of Accession number KP033192.1 and AJP67539.1 and validated by using SMART (simple modular architecture tool) Domain Tool. Cloning and expression studies were carried out using standardized molecular biology protocol. Results depicted that EcbZIP transcription factor showed significant upregulation under both salt and drought stress conditions, indicating that it plays an important role in tolerance towards these stresses. In conclusion, expression analysis of bZIP gene from finger millet seed cultivar ML-365 showed 5-fold upregulation to salt stress to drought stress and 8-fold upregulation to salt stress. Hence, it can serve as a candidate gene for improving abiotic stress tolerance and can be helpful in enhancing the crop productivity under stress conditions.
小谷子bZIP转录因子的克隆及表达分析在非生物胁迫条件下
碱性亮氨酸拉链(bZIP)转录因子是植物中最大的基因家族之一。它们几乎在植物生长发育的各个方面发挥着关键作用,也在生物和非生物胁迫耐受中发挥着关键作用。在本研究中,我们试图研究气候智能型谷物小谷子(Eleusine coracana L.)的转录因子bZIP的特征。从当地市场购买小谷子种子,在干旱和盐胁迫条件下进行田间种植。本研究从谷子中分离出EcbZIP基因,克隆到DH5α细胞中,利用集落PCR进行筛选,并利用qRT PCR分析其在两种非生物胁迫下的表达情况。从NCBI核苷酸数据库和Genpept中检索EcbZIP编码DNA序列和蛋白序列,登录号为KP033192.1和AJP67539.1,使用SMART (simple modular architecture tool) Domain tool进行验证。采用标准化的分子生物学方案进行克隆和表达研究。结果表明,EcbZIP转录因子在盐胁迫和干旱胁迫条件下均表现出显著上调,表明其在耐盐胁迫和干旱胁迫中发挥重要作用。综上所述,小谷子种子品种ML-365 bZIP基因的表达分析显示,bZIP基因对盐胁迫对干旱胁迫的表达上调5倍,对盐胁迫的表达上调8倍。因此,它可以作为提高非生物胁迫耐受性的候选基因,有助于提高作物在逆境条件下的生产力。
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