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将豌豆DNA解旋酶45导入甘蔗(甘蔗属杂交种)可增强细胞膜热稳定性,应激反应基因的上调导致对非生物胁迫的耐受性。

Introduction of Pea DNA Helicase 45 Into Sugarcane (Saccharum spp. Hybrid) Enhances Cell Membrane Thermostability And Upregulation Of Stress-responsive Genes Leads To Abiotic Stress Tolerance.

作者信息

Augustine Sruthy Maria, Ashwin Narayan J, Syamaladevi Divya P, Appunu C, Chakravarthi M, Ravichandran V, Tuteja Narendra, Subramonian N

机构信息

Sugarcane Breeding Institute (ICAR), Coimbatore, Tamil Nadu, India.

出版信息

Mol Biotechnol. 2015 May;57(5):475-88. doi: 10.1007/s12033-015-9841-x.

Abstract

DNA helicases are motor proteins that play an essential role in nucleic acid metabolism, by providing a duplex-unwinding function. To improve the drought and salinity tolerance of sugarcane, a DEAD-box helicase gene isolated from pea with a constitutive promoter, Port Ubi 2.3 was transformed into the commercial sugarcane variety Co 86032 through Agrobacterium-mediated transformation, and the transgenics were screened for tolerance to soil moisture stress and salinity. The transgene integration was confirmed through polymerase chain reaction, and the V 0 transgenic events showed significantly higher cell membrane thermostability under normal irrigated conditions. The V 1 transgenic events were screened for tolerance to soil moisture stress and exhibited significantly higher cell membrane thermostability, transgene expression, relative water content, gas exchange parameters, chlorophyll content, and photosynthetic efficiency under soil moisture stress compared to wild-type (WT). The overexpression of PDH45 transgenic sugarcane also led to the upregulation of DREB2-induced downstream stress-related genes. The transgenic events demonstrated higher germination ability and better chlorophyll retention than WT under salinity stress. Our results suggest the possibility for development of increased abiotic stress tolerant sugarcane cultivars through overexpression of PDH45 gene. Perhaps this is the first report, which provides evidence for increased drought and salinity tolerance in sugarcane through overexpression of PDH45.

摘要

DNA解旋酶是一类驱动蛋白,通过提供双链解旋功能在核酸代谢中发挥重要作用。为提高甘蔗的耐旱性和耐盐性,将一个从豌豆中分离得到的带有组成型启动子Port Ubi 2.3的DEAD-box解旋酶基因,通过农杆菌介导转化法导入商业甘蔗品种Co 86032中,并对转基因植株进行土壤水分胁迫和盐胁迫耐受性筛选。通过聚合酶链反应确认了转基因整合情况,V0代转基因事件在正常灌溉条件下显示出显著更高的细胞膜热稳定性。对V1代转基因事件进行土壤水分胁迫耐受性筛选,结果显示与野生型(WT)相比,在土壤水分胁迫下,它们具有显著更高的细胞膜热稳定性、转基因表达、相对含水量、气体交换参数、叶绿素含量和光合效率。PDH45转基因甘蔗的过表达还导致DREB2诱导的下游胁迫相关基因上调。在盐胁迫下,转基因事件的发芽能力高于WT,叶绿素保留情况也更好。我们的结果表明通过过表达PDH45基因培育提高非生物胁迫耐受性甘蔗品种具有可能性。这可能是第一份通过过表达PDH45证明甘蔗耐旱性和耐盐性增强的报告。

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