CAJ | 학술논문

为了解外源DNA导入对受体水稻基因组DNA序列的影响,探索受体基因组发生碱基变异的规律,为外源DNA 诱导水稻变异的研究提供数据。利用改良的花粉管通道法将玉米基因组DNA导入水稻受体日本晴中,获得的2个水稻变异株B1和B2,经AFLP分析,得到14条特异条带,对其序列分析发现：14条特异序列测序共得到3791个碱基,其中有144个碱基发生了突变,碱基总突变率为3.80%。特异带的产生是由于限制性内切酶识别序列的碱基或选择性碱基发生突变,或DNA片段的插入或缺失所致。发生单碱基突变的频次显著高于双碱基突变、连续三碱基突变和连续多碱基插入或缺失。发生碱基置换的位点频率(87.04%)显著高于碱基的插入(3.70%)和缺失(9.26%)。在碱基置换中,转换的频率(76.47%)显著高于颠换(23.53%)。上述结果表明,外源DNA导入水稻细胞可诱导受体碱基发生突变,且发生碱基置换的位点频率最高,在碱基置换中以转换为主。
위료해외원DNA도입대수체수도기인조DNA서렬적영향,탐색수체기인조발생감기변이적규률,위외원DNA 유도수도변이적연구제공수거。이용개량적화분관통도법장옥미기인조DNA도입수도수체일본청중,획득적2개수도변이주B1화B2,경AFLP분석,득도14조특이조대,대기서렬분석발현：14조특이서렬측서공득도3791개감기,기중유144개감기발생료돌변,감기총돌변솔위3.80%。특이대적산생시유우한제성내절매식별서렬적감기혹선택성감기발생돌변,혹DNA편단적삽입혹결실소치。발생단감기돌변적빈차현저고우쌍감기돌변、련속삼감기돌변화련속다감기삽입혹결실。발생감기치환적위점빈솔(87.04%)현저고우감기적삽입(3.70%)화결실(9.26%)。재감기치환중,전환적빈솔(76.47%)현저고우전환(23.53%)。상술결과표명,외원DNA도입수도세포가유도수체감기발생돌변,차발생감기치환적위점빈솔최고,재감기치환중이전환위주。
In order to understand the changes of the genomic DNA of rice caused by the introduction of exogenous DNA,and explore the variation regularity of the recipient genome nucleotide, provide data for mutation by exogenous DNA introduction,two rice mutative materials B1 and B2 were obtained by using a modified “pollen-tube pathway” to transfer maize genomic DNA into rice receptor Nipponbare. Fourteen specific bands were got in the two mutant materials by AFLP analysis. And its sequence analysis revealed that 14 specific sequences included 3 791 bp, including 144 base mutation, the base mutation rate was 3. 80%. specific bands resulted from base mutation of selective bases or restriction endonuclease recognition sequences,insertion or deletion of DNA fragments. The frequency of single base mutation was significantly higher than that of double bases mutation,three sequential bases mutation and multiple sequential bases mutation. The site frequency of base substitution ( 87. 04%) was significantly higher than that of base insertion(3. 70%) or deletion (9. 26%). In all of the base substitutions, the frequency of transition (76. 47%) was significantly higher than that of transversion(23. 53%). The above results indicated that transferring alien species DNA into rice cell could induce base mutation of the receptor, and the site frequency of base substitution was the highest,the dominant type of base substitution was transition.