CAJ | 학술논문

大豆再生受基因型限制，高效稳定再生体系建立难、遗传转化效率低，影响生物技术的应用。为获得与大豆再生相关基因，解析大豆再生基因分子机制，从而建立高效、稳定再生体系，为生物技术在大豆分子育种中的应用奠定基础。文章利用抑制性消减杂交技术（Suppression subtractive hybridization，SSH），以东农50子叶节为材料构建消减杂交cDNA文库，从文库中随机挑取917个阳性克隆，PCR鉴定插入片段大部分集中在100~750 bp，利用BLAST在GenBank数据库进行序列相似性比对，经过同源性比对归并后得到411个高质量ESTs片段，功能分析表明，这些基因与信号转导、葡萄糖、蛋白质大分子生物合成代谢，光、叶形态发生相关；调控细胞凋亡、细胞自身防御、细胞壁分化；参与各种激素及细胞分裂素介导的信号通路。
대두재생수기인형한제，고효은정재생체계건립난、유전전화효솔저，영향생물기술적응용。위획득여대두재생상관기인，해석대두재생기인분자궤제，종이건립고효、은정재생체계，위생물기술재대두분자육충중적응용전정기출。문장이용억제성소감잡교기술（Suppression subtractive hybridization，SSH），이동농50자협절위재료구건소감잡교cDNA문고，종문고중수궤도취917개양성극륭，PCR감정삽입편단대부분집중재100~750 bp，이용BLAST재GenBank수거고진행서렬상사성비대，경과동원성비대귀병후득도411개고질량ESTs편단，공능분석표명，저사기인여신호전도、포도당、단백질대분자생물합성대사，광、협형태발생상관；조공세포조망、세포자신방어、세포벽분화；삼여각충격소급세포분렬소개도적신호통로。
It is the world recognized chal enge to set up genetic transformation of soybean regeneration system with the problem of low efficiency. In order to establish a highly efficient and stable regeneration system, in this study, cotyledonary node of DN50 were used to construct suppression subtractive cDNA library, and 917 positive clones were screened. The length of most of the insert fragments was focused on 100-750 bp. For further ESTs screening, 411 differential y expressed functional ESTs were found and predicted by BLAST (Basic Local Alignment Search Tool) in GenBank. The ESTs were annotated by similarity to orthologs and paralogs detected with Blastn and Blastx. Function analysis showed that genes were annotated by similarity to signal transduction, macromolecules glucose, and protein synthesis and metabolism, light and leaf morphogenesis, regulation of apoptosis, the celldefense, and cellwal differentiation, participate in a variety of hormones and cytokinin-mediated signaling pathways.