1 Department of Biology, College of Science, Sultan Qaboos University , Muscat, Oman .
2 Department of Crop Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University , Muscat, Oman .
DNA Cell Biol. 2018 Jun;37(6):524-534. doi: 10.1089/dna.2018.4159. Epub 2018 Mar 29.
Although date palm is a relatively salt-tolerant plant, the molecular basis of this tolerance is complex and poorly understood. Therefore, this study aimed to identify the genes involved in salinity tolerance using a basic yeast functional bioassay. To achieve this, a date palm cDNA library was overexpressed in Saccharomyces cerevisiae cells. The expression levels of selected genes that make yeast cells tolerant to salt were subsequently validated in the leaf and root tissues of date palm seedlings using a quantitative PCR method. About 6000 yeast transformant cells were replica printed and screened on a synthetic minimal medium containing 1.0 M of NaCl. The screening results showed the presence of 62 salt-tolerant transformant colonies. Sequence analysis of the recombinant yeast plasmids revealed the presence of a group of genes with potential salt-tolerance functions, such as aquaporins (PIP), serine/threonine protein kinases (STKs), ethylene-responsive transcription factor 1 (ERF1), and peroxidases (PRX). The expression pattern of the selected genes endorsed the hypothesis that these genes may be involved in salinity tolerance, as they showed a significant (p < 0.05) overexpression trend in both the leaf and root tissues in response to salinity. The genes identified in this project are suitable candidates for the further functional characterization of date palms.
尽管椰枣树是一种相对耐盐的植物,但这种耐受性的分子基础很复杂,目前还不太清楚。因此,本研究旨在使用基本的酵母功能生物测定来鉴定与耐盐性相关的基因。为此,在酿酒酵母细胞中过表达椰枣 cDNA 文库。随后,使用定量 PCR 方法在椰枣幼苗的叶片和根组织中验证了使酵母细胞耐受盐的选定基因的表达水平。大约 6000 个酵母转化细胞被复制并在含有 1.0 M NaCl 的合成最小培养基上筛选。筛选结果显示存在 62 个耐盐转化菌落。对重组酵母质粒的序列分析表明存在一组具有潜在耐盐功能的基因,如水通道蛋白 (PIP)、丝氨酸/苏氨酸蛋白激酶 (STKs)、乙烯响应转录因子 1 (ERF1) 和过氧化物酶 (PRX)。选定基因的表达模式支持了这些基因可能参与耐盐性的假设,因为它们在叶片和根组织中均表现出显著(p < 0.05)的盐胁迫诱导过度表达趋势。本研究中鉴定的基因是进一步对椰枣进行功能表征的合适候选基因。