The Arabidopsis CCCH Type Zinc Finger Protein OPW (only Pollen Wall) Gene Is A Key Gene For Microspore Development

Eighth stage of anther development is freed from microspore tetrad haploid microspores cell development,haploid microspores enters pollen sac,independent small spore pollen wall structure in dress,the cavitation occurrence in microspore,after microspore twice in the cell mitosis to form three nuclear pollen grains.This is a mature pollen development process,if there is little gametophyte dysplasia in this process,can cause male sterility of arabidopsis thaliana.This paper in T-DNA insertion mutants Salk₀59463 strain,screening to two male sterile mutant strains,with T-on the DNA sequence of a pair of primers for PCR identification showed no T-insert DNA in the genome.Through background purification and genetic analysis found that the two male sterile mutant strains is controlled by the same single recessive gene,is the main cause of infertility in anther development of 8,microspore cytoplasm content gradually decrease and eventually disappear,to anther development period,12 medicine indoor small spore pollen wall shell,only one reason the mutant named opw?only pollen wall?,the method of using map-based cloning of opw gene localization,the results show that opw genes located on chromosome 2 T28M21 molecular markers and 12 KB interval between T3G21,according to TAIR network data,there are 21 genes between the annotations,by cloning sequencing the genes and the area in between,found that opw 1 mutant exons the At2g40140 gene coding sequence of the genome between 289 th and 290 th base by inserting an A base,and opw-2 mutant exons the At2g40140 gene coding sequence of the genome between 412 th and 413 th base by inserting A T base,coding sequence moved code makes the 424 th to 426 th bases termination codon,therefore At2g40140 is coding opw candidate genes.Further according to the design of complementary carrier gene sequences,and into agrobacterium,via agrobacterium infect Ler came x opw 1 the F1 plants,T0 generation seeds are obtained.According to TAIR network data,At2g40140 coding is a zinc finger protein CCCH,located in the nucleus,the protein was proved to play a role in the way of salt stress,is mediated arabidopsis resist plant diseases and insect pests of CPK CPK3 downstream genes in the calcium signal channel.For OPW protein amino acid sequence analysis found that OPW gene encoding amino acid sequence is not across the membrane structure domain;The evolutionary tree analysis showed that OPW genes and At2g37200 have close kinship.