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以广亲和水稻品种02428为材料,建立悬浮细胞系并分离出原生质体。以PEG为介导,将水稻联合固氮细菌粪产碱菌(Alcaligenes faecalis)A1501导入水稻原生质体中,经分裂、培养后再生成植株。实验结果表明,固氮细菌导入的水稻原生质体的分化频率可提高50%左右;电镜观察表明,A1501进入了水稻原生质体并存在于再生幼苗的根、叶细胞中;免疫检测及微生物学鉴定证明,由再生幼苗中分离出的细菌确实是原导入的A1501;~(15)N示踪试验确证了在再生幼苗内的A1501菌具有固氮活性,再生幼苗的内源激素含量比对照组明显提高;双向电泳谱结果表明,导入A1501后再生的水稻幼苗中产生了至少一种特异性蛋白。
The hybrid rice variety 02428 was used as a material to establish a suspension cell line and protoplast isolation. With PEG as the mediator, Alcaligenes faecalis A1501, a nitrogen-fixing bacteria of rice, was introduced into protoplasts of rice and the plants were regenerated after being splitted and cultured. The results showed that the frequency of protoplasts differentiation by rice-nitrogen-fixing bacteria could be increased by about 50%. Electron microscopy showed that A1501 entered the rice protoplasts and existed in the root and leaf cells of regenerated seedlings. The results of immunoassay and microbiological identification showed that A1501 The bacteria isolated from the regenerated seedlings were indeed A1501 introduced into the original seedlings. The ~ (15) N tracer confirms that the A1501 bacteria in the regenerated seedlings have nitrogen fixation activity, and the content of endogenous hormones in regenerated seedlings is significantly higher than that in the control group. The spectral results showed that at least one specific protein was produced in the rice seedlings regenerated after being introduced into A1501.