Hsieh Tsai-Hung, Lee Jent-Turn, Yang Pei-Tzu, Chiu Li-Hui, Charng Yee-yung, Wang Yu-Chie, Chan Ming-Tsair
National Graduate Institute of Life Sciences, National Defense Medical Center, 115, Taiwan, Republic of China.
Plant Physiol. 2002 Jul;129(3):1086-94. doi: 10.1104/pp.003442.
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 T(1) and T(2) 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 GA(3) (gibberellic acid) treatment. More importantly, GA(3)-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 H(2)O(2) 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.
为了提高番茄(Lycopersicon esculentum)植株的胁迫耐受性,将一个含有由花椰菜花叶病毒35S启动子驱动的拟南芥C-重复/脱水响应元件结合因子1(CBF1)cDNA的表达载体转入番茄植株。通过Northern杂交和Western印迹分析证实了CBF1的转基因表达。通过存活率、叶绿素荧光值和根伸长测量发现,转基因T(1)和T(2)植株的耐寒程度明显高于野生型番茄植株。转基因番茄植株表现出生长迟缓的模式;然而,在赤霉素(GA(3))处理后它们恢复了正常生长。更重要的是,与野生型植株相比,经GA(3)处理的转基因植株仍表现出更高的耐寒程度。进行了消减杂交以分离转基因番茄植株中异源拟南芥CBF1的响应基因。获得了过氧化氢酶1(CAT1),并在转基因番茄植株中表现出激活状态。CAT1基因和过氧化氢酶活性在转基因番茄植株中也被高度诱导。在正常或寒冷条件下,转基因植株中的H(2)O(2)水平均低于野生型植株。转基因植株还表现出对甲基紫精诱导的氧化损伤的相当耐受性。当前研究结果表明,转基因番茄植株中异源CBF1的表达可能诱导多个氧化应激响应基因以抵御低温胁迫。