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为了获得转植酸酶基因(phyA)棉花高代纯合株系并验证其利用土壤中有机态磷的能力,对实验室获得的转phyA基因T4代材料进行了PCR筛选,在水培条件下研究了纯合株系植株根际分泌的植酸酶活性,并在田间进行了植株磷含量和棉花产量性状分析。结果表明,在PCR筛选的4个株系中,有2个株系为纯合株系,其后代均为PCR阳性植株。在以植酸盐为唯一磷源条件下,高表达phyA的转基因株系的植酸酶活性较野生型增加了1.5倍,不同生育时期转基因株系植株叶片磷含量中L6株系增加最多,苗期、现蕾期、花铃期和吐絮期分别增加了4.5%,5.95%,5.45%和8.73%。
In order to obtain the homozygous cotton line of phyA cotton and verify its ability to utilize organic phosphorus in the soil, the PCR-based screening of the T4 generation from the phyA gene was carried out in a hydroponic culture The phytase activity of rhizosphere secreted by homozygous plants was analyzed. Phosphorus content and cotton yield were analyzed in the field. The results showed that two of the four lines screened by PCR were homozygous lines, and all the progeny of the four lines were PCR positive plants. The phytase activity of transgenic plants with high expression of phyA increased by 1.5-fold compared with wild-type under the condition of phytate as the sole source of phosphorus. The L6 strain increased most of the leaf phosphorus content in the transgenic plants at different growth stages. Period, budding stage, boll stage and boll stage increased by 4.5%, 5.95%, 5.45% and 8.73% respectively.