Study On Microspore Abortion Of Cytoplasmatic Male Sterility Line Yamian A

[Objective] The purpose of this test is to explore the causes of cytoplasmic male sterility (CMS) cotton from cell death and epigenetic point of view. Changes of cell autophagy genes were detected from the molecular level, and changes of DNA methylation in the bud in different developmental stages. On the one hand, This research was carried out in order to clarify the mechanism of CMS, and to provide experimental evidence for further study of floral organ development process PCD; On the other hand, to lay the foundation for better use of CMS in agriculture.[Material and Method] Yamian A, A new CMS cotton, produced from [G. hirsutum X (G arboreum X G. bickii)] tri-species hybrids, will be used as material in this experiment. The cotton has high sterility and rich genetic basis with G. arboreun and G. bickii background. The homology cloning method will be utilized to get atg6from Yamian A, which is related with cell atuophage, in addition, the expression profile of the atg6will be know by qRTf-PCR. The DNA methylation will be tested through MSAP.[Results](1)The2687bp atg6fragment is amplified by homology cloning, then BLAST with Gossyium in WGS database of NCBI. The results showed that the similarity with chromosome4in G. raimondii reached87%. atg6is very much closed to G. raimondii compared with Arabidopsis and O. sativa by DNAman analysis.(2)The anther abortion process is divided into three stages in this test, quantitative PCR was used to detect the atg6expression levels in each stage. It was found that atg6gene expression was first increased and then decreased in the bud during Abortion, and differences of atg6expression was extremely significant between abortion and the period before abortion.(3)the DNA methylation solution is testified by MSAP technology in flower. The results show that, in the Yamian A anther development process, the degree of methylation in gradually increased, and the difference reached a significant level.[conclusions] During the Anther Abortion of Yaminan A, atg6gene expression showed an upward trend, suggesting autophagy may play a role; the cell genome methylation level was significantly increased which mean the Anther Abortion may be affected by DNA methylation.