Bioinformatics Analysis Of BAHD Family In Pear And Preliminary Functional Verification Of PbCCoAOMT1

The content of stone cells in ’Dangshan Su’ pear(Pyrus bretschneideri cv.Dangshan Su)fruit is one of the key factors influencing its intrinsic quality.Transport and deposition of lignin are closely related to the development of stone cells.Acyl-CoA-dependent acyltransferases(BAHD),are responsible for acylation of lignin,anthocyanins,alkaloids and other secondary metabolites.Ferulic coenzyme-A transferase(FMT)is one of the members of BAHD family.It was found that transferring AsFMT encoding lignin monomer from Angelica sinensis into poplar could change the lignin structure.Caffeoy coenzyme A-O-methyltransferase(CCoAOMT)is an important factor in the biosynthesis pathway of lignin monomers.At present,there are few studies on the function of lignin metabolism in the pear BAHD acyltransferase family.Through the study on the regeneration of adventitious buds of ’Dangshan Su’ leaves and the exogenous gene AsFMT from Angelica sinensis transferring into ’Dangshan Su’ pear,effects of AsFMT on lignin formation and stone cell development were determined.The identification,sequence characteristics and expression analysis of BAHD family members of the the pear were carried out at the whole genome level,and the functional verification of cloned PbCCoAOMT1 was carried out.Following are major results of this study:1.95 of pear BAHD family members(PbACT1-95)were identified at the genome level which were divided into seven classes.The spatial and temporal expression patterns of PbACTs family members were analyzed.It was found that the expression changes of three genes PbACT3,PbACT6 and PbACT41 were consistent with the changes of stone cells and lignin content in fruit,suggesting that they could participate in the lignin biosynthesis and stone cell development.2.The results showed that all the PbACT has the HXXXD or DFGWG domain,mainly located in cytoplasm and chloroplast.Through the analysis of chromosome localization and duplication events,it was found that PbACTs,distributions was found on 17 chromosomes of pear,and the main type of PbACT gene duplications was segmental duplication.3.The pCAMBIA1301a-PbCCoAOMT1 recombinant plasmid was conducted,which was transferred into wild type Arabidopsis thaliana.Compared with the wild type,the inflorescence stem cell wall thickness,cross-section staining,the root length and lignin content in PbCCoAOMT1 transgenic plants,were significantly increased.4.qRT-PCR results show that PbCCoAOMT1 relative expression level present the rising first then decreasing during fruit development,and expression quantity of PbCCoAOMT1 reaches the peak at 47 days after flowering.