【作者】 朱飞舟；

【导师】 陈立云； 肖国樱； 罗峰；

【作者基本信息】 湖南农业大学 ， 作物遗传育种， 2004， 硕士

【摘要】 以转入了多个抗病基因—rch10、rac22、b-rip、β-glu的超级稻恢复系9311、E32和母本培矮64s及所配制的杂种为试验材料，进行分子检测及农艺性状考察，得出主要结论如下： 1 PCR和Dot-Southern杂交证明9311、E32及其杂种的基因组中整合有β-glu基因。根据连锁遗传的原理，转基因9311、E32及杂种的基因组中同时含有rch10、rac22、b-rip基因。 2 转基因9311和E32的T₂代基因组中整合有β-glu基因，证明外源基因可以稳定遗传。培矮64s／转基因E32中检测有β-glu基因，证明外源基因可以通过杂交转移。 3 通过PCR，还将转基因9311和E32的T₂代群体中的β-glu基因阳性株筛选出来。 4 农艺性状考察的结果表明：转基因E32、培矮64s／转基因E32、培矮64s／转基因9311的生育期极显著地短于对照；转基因E32、培矮64s／转基因E32、培矮64s／转基因9311的株高显著或极显著地矮于对照；转基因9311、E32及杂种的某些经济性状的数值也低于对照。但是，可以筛选到农艺性状较好的植株。 5 对生物素标记探针的制备方法进行了探讨，认为用PCR法制备的探针浓度高、信号强，且无需与待测DNA同源的DNA片段。因此，PCR法是制备生物素标记探针的好方法。更多还原

【Abstract】 Taking super rice restoring lines-9311 and E32, into which 4 disease-resistant genes, rch10, rac22, b-rip and B -glu, have been transformed, sterile line- Pei’ai 64s and their hybrids as experimental materials, molecular assay has been done and agronomic traits have been assessed. The conclusions are drawn out as follows:1 PCR and Dot-southern blotting analysis showed that B -glu gene had integrated into the genomes of transgenic 9311 E32 and their hybrids. On the basis of theory of genetic linkage, rchl0 rac22 b-rip genes had also integrated into the genomes of transgenic 9311 E32 and their hybrids.2 The genomes of T2 generation of transgenic 9311 and E32 were integrated with B -glu gene suggested that foreign genes could be inherited stably. B -glu gene could be detected in Pei’ai64s/transgenic E32 showed that foreign gene could be transferred by ordinary crossing.3 By PCR , the B -glu gene positive plants of T2 generation of transgenic 9311 and E32 were screened out.4 The assessment of agronomic traits showed that the growth duration of transgenic E32. Pei’ai64s/ transgenic E32, Pei’ai64s/transgenic 9311 was extremely significantly shorter than their controls, and the plant height of transgenic E32, Pei’ai64s/ transgenic E32, Pei’ai64s/ transgenic 9311 was significantly or extremely significantly shorter than their controls, and the value of some economic characters of transgenic 9311, E32 and their hybrids was also lower than their controls. However, the plants whose agronomic traits were good can be gained.5 The method of making biotin-label probe was discussed. It was concluded that the probe made by PCR had some advantages that density was high, sign was intensive and it’s no need of making fragments having the same sequence of detected DNA.更多还原