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Heterology Expression of the Arabidopsis C-Repeat/Dehydration Response Element Binding Factor 1 Gene Confers Elevated Tolerance to Chilling and Oxidative Stresses in Transgenic Tomato1

机译:拟南芥C-重复/脱水反应元件结合因子1基因的异源表达赋予转基因番茄1寒冷和氧化胁迫较高的耐受性。

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摘要

In an attempt to improve stress tolerance of tomato (Lycopersicon esculentum) plants, an expression vector containing an Arabidopsis C-repeat/dehydration responsive element binding factor 1 (CBF1) cDNA driven by a cauliflower mosaic virus 35S promoter was transferred into tomato plants. Transgenic expression of CBF1 was proved by northern- and western-blot analyses. The degree of chilling tolerance of transgenic T1 and T2 plants was found to be significantly greater than that of wild-type tomato plants as measured by survival rate, chlorophyll fluorescence value, and radical elongation. The transgenic tomato plants exhibited patterns of growth retardation; however, they resumed normal growth after GA3 (gibberellic acid) treatment. More importantly, GA3-treated transgenic plants still exhibited a greater degree of chilling tolerance compared with wild-type plants. Subtractive hybridization was performed to isolate the responsive genes of heterologous Arabidopsis CBF1 in transgenic tomato plants. CATALASE1 (CAT1) was obtained and showed activation in transgenic tomato plants. The CAT1 gene and catalase activity were also highly induced in the transgenic tomato plants. The level of H2O2 in the transgenic plants was lower than that in the wild-type plants under either normal or cold conditions. The transgenic plants also exhibited considerable tolerance against oxidative damage induced by methyl viologen. Results from the current study suggest that heterologous CBF1 expression in transgenic tomato plants may induce several oxidative-stress responsive genes to protect from chilling stress.
机译:为了提高番茄(Lycopersicon esculentum)植物的胁迫耐受性,将包含由菜花花叶病毒35S启动子驱动的拟南芥C-重复/脱水反应元件结合因子1(CBF1)cDNA的表达载体转移到番茄植物中。 CBF1的转基因表达已通过Northern和Western印迹分析证实。通过存活率,叶绿素荧光值和自由基伸长率的测量,发现转基因T1和T2植物的耐寒性程度显着高于野生型番茄。转基因番茄植株表现出生长迟缓的模式。但是,经过GA3(赤霉素)处理后,它们恢复了正常生长。更重要的是,与野生型植物相比,经GA3处理的转基因植物仍表现出更高的耐寒性。进行减性杂交以分离转基因番茄植物中异源拟南芥CBF1的响应基因。获得了CATALASE1(CAT1),并在转基因番茄植株中显示出活化。在转基因番茄植株中也高度诱导了CAT1基因和过氧化氢酶活性。在正常或寒冷条件下,转基因植物中的H2O2水平均低于野生型植物。转基因植物还表现出对甲基紫精诱导的氧化损伤的相当大的耐受性。当前研究的结果表明,转基因番茄植株中异源CBF1的表达可能诱导了几种氧化应激反应基因,以保护其免受低温胁迫的影响。

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