灰绿藜耐盐基因CgNHX转导新疆陆地棉的初步研究

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凶CgNHX已导入棉花受体中，但其表达情况还需进一步检测验证．

Preliminary Studies on Cotton (Gossypium hirsutum)Transformation with Salt Tolerance Gene CgNHX from

By Using Xinjiang Upland cotton as experimental materials, the salt-tolerant gene CgNHX from Chenopodium glaucum was introduced into the high-quality cotton lines through the pollen-tube pathway. After 3 years＇ DNA injection, results showed that： ①The weather condition within the injection period in field had great effect on the success of transduction. With the larger temperature difference between day and night, and with some dew, tile flower and boll would be in a good developmental condition, and which resulted in a higher boll-setting and kanamycin-resistant rate ; ②The quality of genomic DNA of cotton leaf was co-related with the time duration of low-temperature storage of leaf after sampling. DNA extraction within one week of post-harvest had higher quality and the PCR-positive rate was 30 %, while the DNA quality was much poor and with no PCR-positive sample when extraction in more than one week; ③It was difficult to detect the expression of t ransgenic plants at mRNA level. With 30 PCR-positive samples we got in this study, there had no sample with obviously specific amplification band of target gene in RT-PCR test, only some samples showed diffusion bands at the right position. This study preliminarily showed that the foreign gene CgNHX have been introduced into cotton material, but the expression of CgNHX still need to be verified with further test.