The Prediction And Prelimilary Identification Of Fruit Dehiscence-related Genes In Oilseed Rape (Brassica Napus)

Fruit dehiscence of oilseed rape bring many disadvantages for breeding, therefore,to ravel the dehiscence process and genetic mechanisms underlying the regulation of fruit dehiscence in oilseed rape could suggest new approach to produce new dehiscence tolerant varieties of oilseed rape.Recent studies on Arabidopsis have identified several transcriptional factors involved in fruit dehiscence by regulating the proper differentiation of cell types in dehiscence zone.These transcriptional factors are FRUITFULL(FUL),SHATTERPROOF1(SHP1),SHATTERPROOF2(SHP2), INDEHISCENT(IND),ALCATRAZ(ALC).To obtain these genes list above in oilseed rape,BLAST procedure was carried out in similarity search in oilseed rape database using FUL,SHP1,SHP2,IND and ALC sequeces as queries respectively,thus some ESTs and cDNAs sequences sharing hingh similarity to queries respectively were obtained.BnSHP1 has been reported and the other candidate genes in Brassica napus containing BnFUL-a,BnFUL-b,BnFUL-c,BnFUL-d,BnSHP2,BnIND and BnALC were finally deduced by sequence assembly,subsequently cloning and RACE method.The results of RT-PCR showed that BnFUL-b,BnALC,BnSHP1 and BnSHP2 were all expressed in flower and fruit,moreover,BnFUL-b,BnALC and BnSHP2 were also expressed in other vegetative tissues.The southern bloting result showed that there are two duplication of BnSHP2 at least in genome of Brassica napus.The recombinant plasmid for overexpression of BnFUL-b and the recombinant plasmid for suppressed expression of BnALC were successfully transformed into Brassica napus by Agrobacterium Tumefaciens respectively for further identifying their functions.The ORF sequences without termination codons of BnFUL-b,BnIND and BnALC were inserted in the position between 35S promoter and sGFP(S65T)gene of the sGFP(S65T)vector respectively,so three transient-expression vectors were constructed and subsequently transferred into onion epidermal cell with particle bombardment.The expressiont of sGFP(S65T)were mainly restricted into the nucleus observed under fluorescence microscope,suggesting that BnFUL-b,BnIND and BnALC proteins were mainly located in the cell nucleus.