Cloning and physicochemical characterization of MePPD3gene from cassava(Manihot esculenta)
The subcellular localization and expression analysis of Psbp protein MePPD3(Manihot esculenta PsbP domain-containing protein 3,Phytozome database number:Manes.05G127800)were performed to explore whether MePPD3 is involved in disease resistance of cassava.MePPD3 gene was amplified by RT-PCR.Sequence analysis showed that MePPD3 gene was 807bp in length,encoding 268 amino acids,with PsbP domain located at position 106-266 aa.Bioinformatics analysis of MePPD3 protein was conducted by NetPhos 3.1 Server,SignalP 5.0 Server,TMHMM Servervr,PSIPRED and PHYRE2 online,respectively.The results indicated that MePPD3 protein contained 32 phosphorylation sites,5 glycosylation sites,and 1 transmembrane domain.The secondary structure of the protein was composed of 21.3%Helix(Helix),26.5%fold(strand)and 52.2%random curl(loop).The multiple sequence alignment,phylogenetic tree analysis and conserved domain analysis indicated that PPD3 protein had a high genetic relationship among different plants.Subcellular localization showed that MePPD3 protein was localized in chloroplast.QRT-PCR results revealed that the expression level of MePPD3 gene in different cassava tissues was significantly different,and was the highest in mature leaves.In addition,the expression of this gene was induced by Xanthomonas phaseoli pv.manihotis(Xpm),indicating MePPD3 is involved in the resistance of cassava to Xpm.After MePPD3 gene was silenced by VIGS technique,the leaf lesion area of silent plants was significantly smaller than that of the control plants,which implies that MePPD3negatively regulates cassava resistance to bacterial fusarium wilt caused by Xpm.
NETL
NSTL
万方数据
维普
