Studies On The Function Of Rice Floral Developmental Gene PSD1 Using RNAi Technique

1kb(包括正反向目的片断和内含子)的片断,表明目的片断已经以正向和反向连接到载休中。将该载体分别命名为pTCK303-ARI-SRⅠ和pTCK303-ARⅡ-SR质粒,用引物P3与P4或者P5与P6进行pcr检测,能得到目的片断R11或R12。这表明载体pTCK303-ARI-SRⅠ和pTCK303-ARⅡ-SRⅡ已经成功转化到农杆菌细胞中去。二；用携带载体pTCK303-ARI-SR1|和pTCK303-ARⅡ-SRⅡ的农杆菌浸染转化水稻植株日本晴和psd1成熟胚诱导的愈伤组织,并对其进行低浓度潮霉素(30mg/L)筛选后再高浓度(50mg/L)筛选,共培养种子500粒左右,获得了抗性愈伤78粒,有待进一步生根,移栽,检测和表型观察。ABSTRACTThe specification of each type of floral organ is a key process in flower development.During the last few years,knowledge of the molecular and genetic mechanisms that underlie floral inductiong,floral patterning and floral organ indentity in dicot species has been exploded.The "ABC" model for the determination of floral organ identities has been established by working in two dicot species Arabidopsis thaliana and Antirrhinum majus,and developed into "A-E"model.However,molecular developmental studies are incomplete in monocots. Rice is not only one of the most important food crops in the word,but also a model plant for the study of molecular developmental biology in monocots. So the acquisition and research on the rice floral reproductive mechanism play an imoortant role in the plant developmental biology and molecular biology.psd1 was found in a double haploid (DH)population from the cross between Taiwanjing and GUI630．The phenotype of psdl mutant plants seemed to be normal . However, the spikelets of the psdl were larger than the normal ones,and its stamens and pistils were totally degenerated,with many lays of glumes instead.It si obvious that PSD1is the key gene controlling floral organ primordial differentiation and development in rice ,and probably takes its effects in the first stage of the floral primordium .Our research team has acquired the candidate gene by fine mapping.Rhese studies will analyze the expression and function of PSD1．The article has conformed the function of PSD1 by RNA interference (RNAi). The gene fragments RI1 with length of 267bp and RI2 with length 337bp were cloned from cDNA of wild-type PSD1,using two primers each designed according to the require of RNAi. Meanwhile ,sence and antisence of RI were linked to the vector of Ptck303 individually,RNAi expression vector pTCK303-AR-SR was constructed. Then using an Agrobacterium-mediated transformation system,we have introduced the expression vector into the mature embryo calli of japonica rice variey and mature embryo calli of wild-type PSD1 .The calli were selected on the NB media plus with light concentration hygromycin(30ml/L)and then with high concenteation hygromycin (50ml/L).There are 78 resistant calli abtained and they need to be development into rice plantlet and removed into rice paddy for further study.