CAJ | 학술논문

直链淀粉由于其独特的理化性质，在工业上有着广泛的应用.郭志鸿[1]等利用RNA干扰技术抑制马铃薯中两类淀粉分支酶亚基基因Sbe1和Sbe2的表达，有效提高了马铃薯的直链淀粉含量.本研究利用RNA干扰技术，构建马铃薯SBEⅠ( the ndogenesis starch branching enzymeⅠ)和SBEⅡ( the endogenesis starch branching enzyme Ⅱ)单基因以及SBEⅠ和SBEⅡ双基因的RNA干扰双元表达质粒载体.采用农杆菌介导法将重组基因转化马铃薯品种Désirée后，得到3个转化系列.分别对转基因马铃薯植株进行PCR鉴定，对转基因淀粉表观直链淀粉含量和凝胶化特性等性质进行分析.结果表明，利用RNA干扰技术单独抑制SBEⅠ或SBEⅡ基因的表达，不能提高直链淀粉含量，甚至会导致其含量降低.而当SBEⅠ和SBEⅡ基因表达同时受到抑制时，超过50%的转基因株系直链淀粉含量高于对照.转化株系表观直链淀粉的含量最高可达到49.1%，与对照(28.5%)相比提高了20.6%.
직련정분유우기독특적이화성질，재공업상유착엄범적응용.곽지홍[1]등이용RNA간우기술억제마령서중량류정분분지매아기기인Sbe1화Sbe2적표체，유효제고료마령서적직련정분함량.본연구이용RNA간우기술，구건마령서SBEⅠ( the ndogenesis starch branching enzymeⅠ)화SBEⅡ( the endogenesis starch branching enzyme Ⅱ)단기인이급SBEⅠ화SBEⅡ쌍기인적RNA간우쌍원표체질립재체.채용농간균개도법장중조기인전화마령서품충Désirée후，득도3개전화계렬.분별대전기인마령서식주진행PCR감정，대전기인정분표관직련정분함량화응효화특성등성질진행분석.결과표명，이용RNA간우기술단독억제SBEⅠ혹SBEⅡ기인적표체，불능제고직련정분함량，심지회도치기함량강저.이당SBEⅠ화SBEⅡ기인표체동시수도억제시，초과50%적전기인주계직련정분함량고우대조.전화주계표관직련정분적함량최고가체도49.1%，여대조(28.5%)상비제고료20.6%.
Amylose is extensively used in industrial applications because of it’s unique physicochemical property. Zhihong Guo inhibted expression of two potato starch branching enzyme subunit genes Sbe1 and Sbe2 by using RNA interference technology,effectively improved the amylase content in potato. To develop transgenic potato plants with high-amylose starch in tubers, the endogenesis starch branching enzymeⅠ( SBEⅠ) and Ⅱ( SBEⅡ) genes were, single or simutaneously,targeted for RNA mediated gene silencing by expression of RNA interference genes in potato. In this study,RNA interference was used for gene inhibition of the starch branching enzymeⅠ( SBEⅠ) andⅡ( SBEⅡ) genes. Three SBE-RNA interference plasmids were constructed and transformed into potato cultivar Désirée. Three transformed series were obtained and analyzed by PCR. The level of silencing of SBEⅠ and SBEⅡ in transgenic plants was investigated by analysis of apparent amylose content. The results showed that silencing of SBEⅠ or SBEⅡ alone had yielded a very low frequency of high-amylose lines, and even a reduction in amylose content in most of transgenic starches. However,production of high-amylose potato lines were be achieved by simultaneous inhibition of SBEⅠ and SBEⅡ. In this transgenic series,more than 50% of the transgenic lines showed that amylose content was higher than thire control. The highest amylose content in one line was 49. 1%,increasing 20. 6% in comparison with untransformed starch ones(28. 5%).