【作者】 王翔；

【导师】 任正隆；

【作者基本信息】 四川农业大学 ， 作物遗传育种， 2005， 硕士

【摘要】 本实验以利用“单体附加系染色体作载体转移外源DNA”的染色体工程新方法创造的小麦-黑麦远缘杂交后代中选育出的一个品系R149为供试材料,在细胞生物学和分子生物学的基础上,利用染色体C-分带技术,基因组原位杂交技术,蛋白质分离技术,特异PCR技术等对R149品系作了较为系统的初步研究,试图发现该品系中特异的外源DNA片断,为“单体附加系染色体作载体转移外源DNA”育种方法提供一个可靠的证据,充分证实“小片段易位诱导理论”的可行性。结果如下: 1.用传统的细胞学技术检测R149品系结果表明:并无大片断的黑麦染色体片断,但其表现出的某些黑麦特性说明R149品系具有小片段易位。 2.利用基因组原位杂交对材料R149,川农号系列小麦进行分析,发现对照的原位杂交结果清楚地显示一对1RS/1BL易位染色体;而品系R149原位杂交结果显示在两对染色体的特定位置,具有较为规则的杂交位点,初步判断有可能是小片段易位。 3.根据黑麦的特异重复序列psc119.1,pscH20设计引物,对R149品系进行扩增,psc119.1、pscH20能特异的扩增出黑麦的特异条带,进一步确定R149确实为黑麦小麦小片段易位系。 4.对扩增出的条带进行克隆、测序,并与黑麦扩增出的条带进行序列进行比较,结果显示回收条带的DNA序列与数据库中公布的序列同源性可达95%,从而更为确切地证实R149含有黑麦的染色体小片断。 5.采用SDS-PAGE电泳对供试材料检测发现:并无黑麦的特征谷蛋白,R149具有与最初亲本绵阳11相同的非常优异的面包加工品质,品质得分10。 6.进一步证明利用八倍体小黑麦与普通小麦的后代中选择单体附加系,再利用附加单体遗传学上的不稳定性可以选育出比较理想的小片段易位系,与其它几种染色体工程诱发易位反应的途径相比,具有相对简单,高效,易位片断小的特点,其在育种学上具有深远的意义。更多还原

【Abstract】 Advanced wheat cultivar R149 whose parents are common wheat (Triticum aestivum) and rye (Secale cereals) is bred by new chromosome engineering ways "The use of monosomic rye addition lines for transferring rye chromatin into wheat" . Using the C-banding technique, genome in situ hybridization, and protein seperation technique and PCR studied R149 cultivar systematically. In order to verify the theory "induction of small-segment-translocation" . The studies substantiated feasibility of the theory.1. The study results suggested that the monosomic added rye chromosomes in wheat could induce translocation between wheat and rye chromosomes effectively and that the added monosomic could induce small segment translocation (SS translocation) with high frequency. The theory "induction of SS translocation" is significant in plant breeding compare with other ways of induction of translocation.2. The results of traditional cytology techniques showed that R149 has a normal cytology behavior and stable genetic system. Whereas, the plants exhibited phenotypic alteration compared with its wheat parent.3. The results of the GISH indicated that the control materials showed alien chromosome clearly and R149 had a relative regular hybridization site along the special chromosomes. Therefore, we could conclude that it is possible that R149 had some segments of the rye chromosome.4. Using the primers of specific repeated sequence from S. cereale amplified the R149, the results showed that pscll9.1、 pscH20 could amplified the products correctly.5. Based on SDS-PAGE analysis, R149 showed no rye special glutenin protein.However, it has a terrific quality of making bread wheat.更多还原