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以新疆陆地棉为实验材料,通过花粉管通道法向优质棉花高代材料中导入了灰绿藜耐盐基因CgNHX.经过三年连续转导的结果显示:①大田注射期间的天气状况对转导成功影响较大.当昼夜温差较大且有露水时,花和铃的状态较好,导致结铃率、卡那霉素检测阳性率增加;②棉叶基因组DNA的提取质量与棉叶采后冷藏的时间有一定相关性.采后一周内提取的棉叶DNA质量较好,PCR检测获得30%的阳性率,而两周及以后的棉叶DNA质量降低,PCR检测未获得阳性结果;③在mRNA水平上检测转基因植株的表达较为困难.本实验中PCR检测阳性的30份样品,在RT-PCR检测中均未获得目的基因的明显特异扩增条带,只有部分样品在目的条带的位置出现弥散带型.本研究初步显示外源基因CgNHX已导入棉花受体中,但其表达情况还需进一步检测验证.
Xinjiang Upland cotton was used as experimental material to introduce CgNHX, a salt-tolerant salt-resistant gene of CgNHX, into pollen tube passageway. After three years of continuous transduction, the results showed that: (1) The success of the larger .When the temperature difference between day and night is large and dew, flowers and bell state is better, leading to the bell rate, kanamycin positive rate increased; ② cotton genomic DNA extraction quality and cotton harvest There was a certain correlation between the time of refrigerated storage and the quality of DNA extracted from cotton leaves within one week after harvest. The positive rate of 30% was obtained by PCR, while the quality of DNA of two weeks and later decreased, but no positive result was detected by PCR. It is difficult to detect the expression of transgenic plants at the mRNA level.In this experiment, 30 samples tested positive by PCR showed no obvious specific amplification bands of RT-PCR, only some of the samples Which showed that the exogenous gene CgNHX has been introduced into the cotton receptor, but its expression needs further testing and verification.