CDNA-AFLP Analysis Of The RN-type Cytoplasmic Male Sterile Line And Its Maintainer Line In Soybean

Cytoplasmic Male Sterility(CMS)is the basis of heterosis utilization.Research on CMS of Soybean in China began in 1980 s.After continuous efforts of soybean breeders,we have released 18 hybrid soybean varieties by using "three lines" methods,which were widely used in production and made China is the world leader in this field.But some problems in the process of hybrid soybean industrialization still existed and need to be solved.Lack of sterile cytoplasm and mechanism of cytoplasmic male sterility were still unclear.Some of the key problems in hybrid soybean production could not break through and restricted the development of industrialization of hybrid soybean.So it is very important to accelerate the male sterility genes clone and explain the mechanism of CMS.This study was analyzing the mechanism of Cytoplasmic Male Sterility with RN type cytoplasm soybean by cDNA-AFLP molecular markers at first.Materials used sterile line JLCMS9 A and its maintainer line JLCMS9 B which were more than 10 backcross generation with heterogeneous nucleus CMS.Using 64 pairs of primers analyses of differential expression of genes in bud of sterile line and its maintainer line.And combined bioinformatics methods for deferentially expressed genes were screened and validated.Got the deferentially expressed genes fragments.It established the theoretical foundation on the study of molecular mechanism of CMS in soybean.The conclusions were summed up as following:1.Improved template preparation,per-amplification,selective amplification and electrophoresis testing procedures with cDNA-AFLP molecular markers.Developed a set of suitable system for differential expression of genes for CMS in soybean.2.Using sterile line and its maintainer line buds cDNA as templates,64 pairs of primers for amplification,12 differential expression fragments were got.There were 8 fragments specific expression in sterile line and 4 fragments specific expression in maintainer line.By sequencing and homology,all these fragments with high homology can be founded in Genbank.Among of them 8 fragments were homologous with known genes and 4 with unknown functions.3.Sequence analysis of the 8 known fragments shows that the functions involve meiosis,ATP synthesis,protein modification and other related processes.The fragment A1 which was specific expression in sterile line which was sequence homology with encoding ATPC.The fragment B1 which was specific expression in maintainer line which was sequence homology with encoding Arabidopsis Asy1.Of these 2 genes are related to ATP synthesis and meiosis which can affect pollen growth and development.So we predicted these 2 genes might relevant to the soybean sterility genes or played an important role in fertility regulation.And the fragment A4 which was specific expression in sterile line and located in the mitochondria.We indicated this fragment might be associated with sterility too.