论文部分内容阅读
二酰基甘油酰基转移酶(DGAT)是生物体内三酰甘油(TAG)合成过程中的关键酶。本研究克隆陆地棉GhDGAT1基因308bp片段,构建了该基因含内含子结构的hpRNA干涉载体;应用花粉管通道法转化棉花,研究内源基因GhDGAT1沉默对棉花油分含量的影响。结果表明:1)经PCR及Southern杂交鉴定,转基因种子中GhDGAT1基因表达量受到显著抑制,种仁含油量下降,最多下降3.13%。2)种子油脂量显著减少的株系中,总蛋白含量及可溶性糖分含量,分别相对提高4.31%～9.77%和11.67%～23.01%。3)与野生型相比,转基因株系幼胚在发育后期鲜重降低,而可溶性蛋白含量升高。4)与对照相比,转基因植株的株高、第一果枝高度、果枝数均显著降低,但其他农艺性状与主要的经济性状均未受到显著影响。研究表明,通过调控GhDGAT1基因的表达可影响陆地棉的油脂合成,调节棉籽油分含量。
Diacylglycerol acyltransferase (DGAT) is a key enzyme in the synthesis of triacylglycerol (TAG) in vivo. In this study, we cloned the 308 bp fragment of GhDGAT1 gene of upland cotton (Gossypium hirsutum L.), and constructed the hpRNA interference vector containing this intron structure. The pollen tube pathway was used to transform cotton and the effect of endogenous GhDGAT1 silencing on oil content of cotton was studied. The results showed that: 1) The expression of GhDGAT1 gene in transgenic seeds was significantly inhibited by PCR and Southern blotting, and the oil content of seed kernel decreased by up to 3.13%. 2) The total protein content and soluble sugar content increased by 4.31% ~ 9.77% and 11.67% ~ 23.01% in the lines with significantly reduced oil content. 3) Compared with the wild type, the immature embryos of the transgenic lines decreased in fresh weight and the content of soluble protein increased in later development. 4) Compared with the control, the plant height, the height of the first fruit branch and the number of fruit branches of the transgenic plants decreased significantly, but the other agronomic characters and the main economic characters were not significantly affected. Studies have shown that regulation of GhDGAT1 gene expression can affect the uptake of upland cotton oil synthesis, regulating cottonseed oil content.