Positional Clone And Genetic Analysis Of Ess Mutant Of Arabidopsis Thaliana

1.Positional Clone and Genetic Analysis of The Arabidopsis thaliana ess mutantAnther deveopment is involved in the most complex pathway in plant growth,the researching results of which could give insight into the other aspects of the plant development.The progress in Anther development research has been accelerated by the genetic analysis of those anther mutants.Though a few genes cloned from those anther mutants,does it mean that a new step of anther development be uncovered whenever any novel mutants were found related to anther development and the corresponding genes were cloned. The outline of anther development is just figured with those steps by this way. In recent, several restorer fertility genes were cloned from crops, which could restore the CMS male sterility and all belong to nuclear gene, these genes translate into protein containing PPR motifs locating into mitochondria and regulate the mitochondria RNA processing or post-translation involved in CMS. Because the gene ESS is just this kind of protein containing PPR motifs, the studying of ess mutant must bring into some clues about anther development and also make a progress in understanding the plant male sterility involved in mitochondria and PPR protein. Uncovering the function of ESS gene maybe give some helps in the artificial control on male fertility because the data of homologous comparing analysis indicate that ESS gene not be a protein taking part in metabolism or a block in cell structure, but a factor or cofactor in some kind of regulators. Male fertility is always focused attention upon only because some of the studying can be used in crops production. The findings earning from the model plant such as arabidopsis can be used in crops smoothly because of the evolution consensus. Positional clone is an important method for discovering novel genes. There is a prominent breakthrough in arabidopsis mapping, which make a standard protocol for positional clone in arabidopsis. Although several inland Labs have carry out Arabidopsis mapping research, to today there is not reported that mapping using CAPC information was published in the country. In our studying, we not only used the CAPC data in positional clone, but also developed about 30 novel makers based on CAPC and new protocol of heterogeneous double helix analysis for SNPs or small InDels. Our mapping work also gives a example for how to do positional clone in arabidopsis.The mutant ess selected from EMS mutant genesis population. The results of ess phenotype analysis and characterizing by light microscope are the following: Theearly siliques are shorter, silique numbers increase and the seeds products reduce.the flowering period is delayed, the secondary branch of main stem is just as high as main stem, and the secondary growing is weakened in ripeness. In the opening flower, the filaments are shorter, the anthers are brown, pollens reduce sharply, but the carpels are normal. Pollen germination is normal.the ess mutant is defect in male reproduction because the silique become natural when pollinating with normal pollen. Abnormal microspores are found in stage 11. In stage 14, anther wall is collapsed and crimpled, the cavities are found under epidermis and the sterility pollens are obvious. The character of short siliques and reduced seeds is caused by few of pollens.By positional clone, the mutant ess was localized on the right of BAC clone of F12K11 and the left of F4H5 between the maker UN201 (200054 lbp) and UN67(2067844bp) in the chromosome I . the region comes cross about 67kb. In the Sequence-based map of arabidopsis genes with mutant phenotypes, ESS is between KT-06-50(EMB1444, 1867128-73696bp) and AT 1(MOM, 2501739-1108lbp). All the data indicate that ess be a novel anther mutant, which show a new pathway in anther development. There are 24 candidate genes in this region, only the note of the ATlg06710.1 gene give a clue related to male sterility. The sequenced result proved that there is a mutant of G to A at 2683rd nt in the AT1G06710.1 (284 lnt), which induce that 895th Glu(E) changs to Lys(K) in the translation protein(895AA). Data from homogenous and structure analysis mean that ATIgO6710.1 be a protein containing 10 PPR motifs, and control the anther development by regulating the processing of some kind of unknown RNA like bicoid RNA or post-translation in the translation of unknown proteins.2 Molecular Checking on the offspring of the parents from anther culture of Hanfeng-xiangqing hybrid riceAFLP named from amplified restriction fragment length polymorphism is one of the most effective, stabilizing and credible methods in DNA fingerprinting. AFLP can be used in many aspect such as plasma checking, relative analysis, seed quality monitoring. Offspring of the parents from anther culture of Hanfeng-xiangqing hybrid...