CAJ | 학술논문

研究利用芒属植物(芒02381和获04005)叶绿体全基因组测序的结果,开发12对InDel标记.采用正交设计法优化cpInDel-PCR体系,得到模板DNA、dNTPs、Mg2+、引物、TaqDNA聚合酶5个关键因素在体系中的最佳组合.12对InDel标记对43份芒属和2份甘蔗材料进行扩增,共扩增出538条条带,其中3对引物扩增出13条多态性条带,占总数的2.4％.将所有位点读带转化为数值矩阵并用UPGMA法聚类,在遗传相似系数1.4的水平上,和南获聚为一类,而芒和与五节芒、双药芒、红山茅、尼泊尔芒、甘蔗聚为一类,4个具有杂交种特征的材料单独聚为一类.试验开发的芒属植物cpInDel标记可用于芒与获的区分,指导芒属植物的杂交育种.
연구이용망속식물(망02381화획04005)협록체전기인조측서적결과,개발12대InDel표기.채용정교설계법우화cpInDel-PCR체계,득도모판DNA、dNTPs、Mg2+、인물、TaqDNA취합매5개관건인소재체계중적최가조합.12대InDel표기대43빈망속화2빈감자재료진행확증,공확증출538조조대,기중3대인물확증출13조다태성조대,점총수적2.4％.장소유위점독대전화위수치구진병용UPGMA법취류,재유전상사계수1.4적수평상,화남획취위일류,이망화여오절망、쌍약망、홍산모、니박이망、감자취위일류,4개구유잡교충특정적재료단독취위일류.시험개발적망속식물cpInDel표기가용우망여획적구분,지도망속식물적잡교육충.
In this paper,we have sequenced the whole-genoine of the Miscanthus(M.sinensis No.02381、M.sacchariflorus No.04005)chloroplast and developed 12 pairs of InDel markers.The orthogonal design was employed to optimize the cpInDel-PCR system,the best combination of the five key factors in this system,such as template DNA,dNTPs,Mg2+,primers,TaqDNA polymerase were obtained.A list of 43 Miscanthus and 2 sugarcane materials were used to amplify by 12 pairs of InDel markers and 538 bands were amplified totally.13 polymorphic bands were amplified using three pairs of primers,which account for 2.4％ of the total.All sites and bands were converted to number matrix and clustered using UPGMA,at the level of genetic similarity coefficient of 1.4.M.sacchariflorus and M.lutarioriparius were clustered together.M.sinensis was clustered as one class with Andersson,M.floridulus,M.nudipes,M.paniculatus,M.nepalensis and sugarcane.The four materials with hybrid features were clustered separately together in the level of genetic similarity coefficient of 1.4.The developed Miscanthus cpInDel markers can be used to identify Miscanthus and guide Miscanthus crossbreeding.