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园艺学报 ›› 2014, Vol. 41 ›› Issue (8): 1563-1572.

• 果树 • 上一篇    下一篇

超量表达欧李ChPSY基因促进转基因番茄类胡萝卜素合成的研究

张建成1,王鹏飞1,郝燕燕1,杨淑一1,郑 斌1,牛自勉2,杜俊杰1,*   

  1. ZHANG Jian-cheng1,WANG Peng-fei1,HAO Yan-yan1,YANG Shu-yi1,ZHENG Bin1,NIU Zi-mian2,and DU Jun-jie1,*
  • 收稿日期:2014-04-16 出版日期:2014-08-25 发布日期:2014-08-25

Overexpression of ChPSY Gene from Cerasus humilis Improved Carotenoids Synthesis in Transgenic Tomato

(1山西农业大学园艺学院,山西太谷 030801;2山西省农业科学院现代农业研究中心,太原 030031)   

  1. (1College of Horticulture,Shanxi Agricultural University,Taigu,Shanxi 030801,China;2Research Centre of Modern Agriculture,Shanxi Academy of Agricultural Sciences,Taiyuan 030031,China)
  • Received:2014-04-16 Online:2014-08-25 Published:2014-08-25

摘要: 将从欧李果实中分离的ChPSY cDNA经过序列分析、酶切之后,连接到植物表达载体PMV,构建了植物重组载体pBI-ChPSY,并通过根瘤农杆菌EHA105介导转化番茄,获得了12株抗性植株。PCR检测和Southern杂交结果显示,12株抗性植株均为阳性,说明外源基因ChPSY整合到转化植株的基因组中。RT-PCR分析表明,ChPSY基因在转化植株果实中得到表达。HPLC分析表明,转ChPSY基因番茄果实中总类胡萝卜素含量增加了1.62 ~ 3.04倍,八氢番茄红素、番茄红素、β–胡萝卜素和α–胡萝卜素含量分别增加了4.87、2.10、2.59和3.25倍。转化植株中内源类胡萝卜素合成基因的表达也受到广泛的影响。ChPSY基因超量表达有效促进了番茄果实中类胡萝卜素的合成和积累。

关键词: 欧李, 番茄, 八氢番茄红素合成酶基因, 遗传转化, 类胡萝卜素

Abstract: The ChPSY cDNA from the Cerasus humilis fruit was analyzed,digested and inserted into plant expression vector PMV,resulting in the plant recombinant vector pBI-ChPSY,which was introduced into tomato via Agrobacterium-mediated transformation. Twelve putatively independent transgenic plants were generated. PCR and Southern blot analysis showed 8 out of these putative transgenic lines were positive,which revealed that ChPSY gene was integrated into the transgenic lines genome. RT-PCR analysis indicated that ChPSY gene was expressed in the fruits of these transgenic lines. HPLC analysis showed that the total amount of carotenoids in the fruit of transgenic tomato was 1.62 to 3.04-fold higher than the non-transformed controls,at the same time,phytoene,lycopene,β-carotene and α-carotene levels were also increased by 4.87,2.10,2.59 and 3.25 folds,respectively. In addition,the expression of endogenous genes involved in carotenoid synthesis was affected in the transgenic lines. These results suggested that overexpression of ChPSY significantly enhanced the carotenoids biosynthesis and accumulation in transgenic tomato fruits.

Key words: Cerasus humilis, tomato, phytoene synthease gene, genetic transformation, carotenoids

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