Research For Phenotype And Related Genes Of Ovp1 Mutant In Arabidopsis Thaliana

ovp1 mutant,which is kanamycin resistanct and has a segregation ration of 1:1,was screened from Ds insertion mutant library which contains genes of kanamycin resistance in previous work of our laboratory.Half of the pollen is normal,while the other half is sterility in the mutant.Genetic analysis and experiments showed that ovp1 is a mutant whose male gametophytes are totally lethal,while its female gametes are normal.Molecular experiments illustrated that Ds of the mutant is inserted into the second chromosome,with its 5' terminal is located at 23 bp downstream of start codon in at2g01560 and its 3' terminal at 1284 bp downstream of TAA in at2g01913.Result of RFLP showed that Ds insertion in ovp1 mutant leads to a long deletion of chromosome and this deletion is 271538 bp in length which contains 40 genes in total.However,identification and phenotypic observation of the corresponding T-DNA insertion mutants of these deleted genes revealed no sterile phenomenon in their male gametophytes.At the same time,SAIL?681?B03 the allelic mutant of at2g01913 which is located near the 3' terminal of Ds insertion was identified and observed.The result showed that this muant has the phenotype of partial pollen granule sterility.Result of qRT-PCR and semi-quantitative PCR showed that at2g01913 was down-regulated in ovp1 and SAIL?681?B03.It was known that SAIL?681?B03 is a homozygote of T-DNA insertion and has a phenotype of pollen granule sterility.Heterozygote SAIL?681?B03+/-also has a phenotype of pollen granule sterility(sterility rate 23.87%).Using GFP fusion protein,the protein encoded by at2g01913 was located in nucleus and cytomembrane.Meanwhile,full length of at2g01913 was cloned and transformed into SAIL?681?B03 for complementation test.