CAJ | 학술논문

为研究尾穗苋凝集素(ACA)在植物中可能的抗虫作用,通过RH-PCR克隆了ACA cDNA并通过RACE分析证实了cDNA序列的正确性.构建了ACA基因的韧皮部特异表达载体pBCACAc并通过根癌杜菌介导转化了烟草(Nicotiana tabacum L.).PCR和Southern blot分析结果证明,ACA基因已经整合到转化再生植物的基因组中,其插入插贝数1~4个不等.对转基因烟草叶片蛋白时行行免疫反应的结果表明,ACA基因已被转录和翻译.用桃蚜(Myzuspersicae Sulzer)对转基因烟草离体叶片进行了的接虫试验结果表明,测试过的78%的烟草对桃蚜口密度增长的平均抑制率在75%以上,在抗性植株上观察到有桃蚜若虫死亡的现象.以上结果表明,ACA基因是一个有效的抗蚜基因,在作物抗蚜分子育种具有应具应用价值.
위연구미수현응집소(ACA)재식물중가능적항충작용,통과RH-PCR극륭료ACA cDNA병통과RACE분석증실료cDNA서렬적정학성.구건료ACA기인적인피부특이표체재체pBCACAc병통과근암두균개도전화료연초(Nicotiana tabacum L.).PCR화Southern blot분석결과증명,ACA기인이경정합도전화재생식물적기인조중,기삽입삽패수1~4개불등.대전기인연초협편단백시행행면역반응적결과표명,ACA기인이피전록화번역.용도아(Myzuspersicae Sulzer)대전기인연초리체협편진행료적접충시험결과표명,측시과적78%적연초대도아구밀도증장적평균억제솔재75%이상,재항성식주상관찰도유도아약충사망적현상.이상결과표명,ACA기인시일개유효적항아기인,재작물항아분자육충구유응구응용개치.
To investigate the possible function of the agglutinin from Amaranthus caudatus L. (ACA) in plant defending against insect pests,ACA cDNA was cloned by RT-PCR and the 5′ and 3′ sequences were confirmed by rapid amplification of cDNA ends (RACE). The phloem-specific expression vector of ACA gene, pBCACAc, was constructed based on the plant binary vector pBC438 and transfered into tobacco plants via Agrobacterium-mediated transformation method. Results from PCR and Southern blotting analysis showed that ACA gene was integrated into the genomes of transformed plants and the transgene integration varied from one to four estimated copies per genome. Western blotting analysis indicated that ACA gene was transcribed and translated in the transgenic plants. The bioassay of Myzus persicae Sulzer on detached leaves demonstrated that the 78% transgenic tobacco plants displayed an average aphid-resistant rate of more than 75%. Some apterous progeny of M. persicae were found dead on the resistant plants.These results indicate that ACA gene should be an effective aphid-resistant gene and could be valuable for application in crop breeding for aphid resistance.