Transformation and expression of DGAT1-2 gene and its effect on the oil content of super-high oil corn
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摘要:
【目的】三酰甘油是油料作物油脂的主要成分,二酰基甘油酰基转移酶(DGAT)是催化三酰甘油合成唯一的关键酶和限速酶.探讨DGAT 基因对超高油玉米Zea mays L. 种子油分含量的影响,以进一步提高超高油玉米的含油率.【方法】采用花粉管通道法将DGAT1-2基因导入超高油玉米自交系,在盆栽条件下5~6叶期对T1代植株进行PCR检测筛选,Southern杂交进一步验证.成熟时收获10株自交授粉的转基因植株(T2代)籽粒,测籽粒平均含油率,通过t检验进行统计分析.【结果和结论】初步筛选获得假定转化植株163株,Southern杂交获29株转基因植株,平均转化率为1.44%.t检验结果表明,10株自交授粉的转基因植株(T2)籽粒平均含油率比非转基因植株籽粒平均含油率提高了6.19%,差异极显著(P<0.01).利用花粉管通道法导入DGAT1-2基因,可显著提高超高油玉米籽粒含油率,是一种切实有效提高超高油玉米含油量的新途径.
Abstract:
【Objective】Triacylgycerol (TAG) is the main oil ingredient of oil-bearing crops. Diacylgycerol acyltransferase (DGAT) is the sole crucial rate-limiting enzyme in the synthesis process of triacylgycerol. The effect of DGAT gene on the oil content of super-high oil corn was investigated to improve the oil content of super-high oil corn.【Method】DGAT1-2 gene was transferred into super-high oil corn inbred lines by pollen-tube-pathway. Transgenic plants were obtained through screening with PCR among T1 generation potted plants at the 5-6-leaf stage. Transgenic plants were verified by Southern-blotting. The average oil content of transgenic T2 seeds was analyzed by the t-test.【Result and conclusion】One hundred and sixty-three putative transgenic plants were obtained through screening with PCR among a large number of T1 generation potted plants at the 5-6-leaf stage. Twenty-nine transgenic plants were verified by Southern-blotting. The average transformation rate was 1.44%. The seeds of 10 transgenic T2 plants were harvested by self-pollination. The average oil content of transgenic T2seeds was significantly higher than that of the non-transgenic plants by the t-test (P<0.01), with the increase rate being 6.19%. The results provide a new practical and effective approach to improve the oil content of super high oil corn.