Isolation And Function Identification Of Abscission Zone Related Gene GhBOP1 Of Gossypium Hirsutum L.

Cotton boll-settingnumber per plant is one of the important traits for determining yield. However, abscission rate of cotton buds and bolls is naturally high due to environment stresses and physiology changes. Thereby, improving cotton boll-setting trait is an important role in increasing cotton productivity. Here, a cotton homolog of Arabidopsis BOP gene relating differentiation of abscission zone is isolated by PCR, which encoding protein contains BTB and ANK domains revealed by bioinformatics analysis. T he phylogenetic tree analysis of Gh BOP1 as an entry showed that the protein sequences from variousplantshave high identity, exhibiting slow evolution in amino acid changes.1. We isolated the regulation of differentiation related gene GhBOP1 from the abscission zone of Gossypium hirsutum, bioinformatics analysis showed that GhBOP1 protein sequence contains BTB and ANK domain highly conserved; phylogenetic analysis showed that the protein amino acid sequence homology with many plants have a high degree of similarity, indicating that this kind of protein evolution. The use of qRT-PCR for tissue specific expression analysis showed that Gh BOP1 gene in root, stem, Ye Hehua had little expression in tissues and organs, but in the abscission zone of the dominant expression, expression of leaf expression level of 20 times.2. the cDNA sequence of sequence length of the 341 bp, using DNAMAN to design the primers, and use of Gossypium hirsutum Variety CCRI 35 proposed RNA reverse transcription cDNA as template, PCR amplified target fragments, and the virus vector pYL-156 to connect, and transferred into Escherichia coli DH5 a competent cells, the plasmid was extracted by electroporation into the Agrobacterium tumefaciens strain GV3101, yielding strain。 using virus induced gene silencing(VIGS) expression of cotton GhBOP1 gene interference technology, at the same time the low salt stress analysis, the emergence of a large number of control leaves off, while Gh BOP1 gene silencing plants grew normally. The results of VIGS interference in cotton petiole after processing, and paraffin sections of plant tissue showed that the abscission zone of gene expression was inhibited.3. GhBOP1 full-length coding sequence was amplified by PCR and GFP fusion gene was obtained pGhBOP1-GFP fusion sequence of the vector, and injected subcellular localization in tobacco leaves was observed and found GhBOP1-GFP fusion protein in the nucleus and cytoplasm are expression.(4) the cloning vector pGEM-T-GhBOP1 and the vector pBin438,gel extraction DNA fragment and vector fragment, eukaryotic expression vector pBin438-GhBOP1 was successfully constructed. The construction of pBin438-GhBOP1 expression vector into Agrobacterium strain LBA4404 by electroporation in the success of the pBin438-GhBOP1, will be transformed into tobacco by leaf disc transformation method, get the tobacco plants, the PCR and RT-PCR were detected.In a word, the advantages of Gh BOP1 gene expression in abscission zone, and have the function to regulate the formation of abscission zone.