Division of Plant Biotechnology, ICAR-Indian Institute of Pulses Research, Kanpur, 208 024, India.
Division of Basic Sciences, ICAR-Indian Institute of Pulses Research, Kanpur, 208 024, India.
BMC Plant Biol. 2021 Jan 11;21(1):39. doi: 10.1186/s12870-020-02815-4.
Chickpea (Cicer arietinum L.) is the second most widely grown pulse and drought (limiting water) is one of the major constraints leading to about 40-50% yield losses annually. Dehydration responsive element binding proteins (DREBs) are important plant transcription factors that regulate the expression of many stress-inducible genes and play a critical role in improving the abiotic stress tolerance. Transgenic chickpea lines harbouring transcription factor, Dehydration Responsive Element-Binding protein 1A from Arabidopsis thaliana (AtDREB1a gene) driven by stress inducible promoter rd29a were developed, with the intent of enhancing drought tolerance in chickpea. Performance of the progenies of one transgenic event and control were assessed based on key physiological traits imparting drought tolerance such as plant water relation characteristics, chlorophyll retention, photosynthesis, membrane stability and water use efficiency under water stressed conditions.
Four transgenic chickpea lines harbouring stress inducible AtDREB1a were generated with transformation efficiency of 0.1%. The integration, transmission and regulated expression were confirmed by Polymerase Chain Reaction (PCR), Southern Blot hybridization and Reverse Transcriptase polymerase chain reaction (RT-PCR), respectively. Transgenic chickpea lines exhibited higher relative water content, longer chlorophyll retention capacity and higher osmotic adjustment under severe drought stress (stress level 4), as compared to control. The enhanced drought tolerance in transgenic chickpea lines were also manifested by undeterred photosynthesis involving enhanced quantum yield of PSII, electron transport rate at saturated irradiance levels and maintaining higher relative water content in leaves under relatively severe soil water deficit. Further, lower values of carbon isotope discrimination in some transgenic chickpea lines indicated higher water use efficiency. Transgenic chickpea lines exhibiting better OA resulted in higher seed yield, with progressive increase in water stress, as compared to control.
Based on precise phenotyping, involving non-invasive chlorophyll fluorescence imaging, carbon isotope discrimination, osmotic adjustment, higher chlorophyll retention and membrane stability index, it can be concluded that AtDREB1a transgenic chickpea lines were better adapted to water deficit by modifying important physiological traits. The selected transgenic chickpea event would be a valuable resource that can be used in pre-breeding or directly in varietal development programs for enhanced drought tolerance under parched conditions.
鹰嘴豆(Cicer arietinum L.)是第二大广泛种植的豆类,干旱(限制水分)是导致每年产量损失 40-50%的主要限制因素之一。脱水应答元件结合蛋白(DREBs)是一类重要的植物转录因子,可调节许多胁迫诱导基因的表达,在提高非生物胁迫耐受性方面发挥着关键作用。本研究通过 rd29a 启动子驱动的拟南芥脱水应答元件结合蛋白 1A(AtDREB1a 基因)转化鹰嘴豆,获得了一批转基因鹰嘴豆品系,旨在提高鹰嘴豆的耐旱性。对一个转基因事件及其对照品系的后代进行了评估,主要评估了赋予耐旱性的关键生理特性,如植物水分关系特性、叶绿素保持、光合作用、膜稳定性和水分利用效率在水分胁迫下的表现。
通过农杆菌介导法转化鹰嘴豆,共获得了 4 个稳定遗传的转基因鹰嘴豆株系,转化率为 0.1%。通过聚合酶链式反应(PCR)、Southern 杂交和反转录聚合酶链式反应(RT-PCR)分别对整合、转录和调控表达进行了验证。与对照相比,转基因鹰嘴豆品系在严重干旱胁迫(胁迫水平 4)下具有更高的相对含水量、更长的叶绿素保持能力和更高的渗透调节能力。在相对严重的土壤水分亏缺下,转基因鹰嘴豆品系的光合作用不受影响,涉及到 PSII 量子产量的提高、饱和辐照度下的电子传递速率以及叶片中相对较高的相对含水量,这也表明了其耐旱性的提高。在一些转基因鹰嘴豆品系中,较低的碳同位素分馏值表明其具有较高的水分利用效率。与对照相比,表现出更好的水分利用效率的转基因鹰嘴豆品系在水分胁迫下具有更高的种子产量。
通过精确的表型分析,包括非侵入性叶绿素荧光成像、碳同位素分馏、渗透调节、更高的叶绿素保持和膜稳定性指数,可以得出结论,通过修饰重要的生理特性,AtDREB1a 转基因鹰嘴豆品系能够更好地适应水分亏缺。选择的转基因鹰嘴豆事件将成为一种有价值的资源,可用于预繁殖或直接在品种发展计划中,提高在干旱条件下的耐旱性。