Establishment Of Rice Cell Suspension Line And CDNA Of OsPLD Gene Clone And Expression

Rice is most important crop in world,suspension cell is a necessary tools to modern molecule biology.PLD is very important for higher plants in growth and development.Today,it wasn't reported that study on Nipponbare(Japonica)cell suspension line and OsPLD gene functional.The article study on establishment of cell suspension line,cryopreservation and OsPLD gene clone,organs expression analysis and function recognization in Yeast mutant,material is Nipponbare(Japonica).Result: (1)Explant is rice mature embryo,embryogenic calls was induced on culture mediums of NB +2mg/L 2,4-D+3%Suc+7%Agar and pH is 5.8,after subcultured 2 mouthes in the conditions of 25℃and dark ,a lot of calli were got.Calli of light-yellow,structure crisp is necessary for cells suspension culture in the conditions of 26℃,dark and 110～130rpm/min.After 3 monthes,ideal cells suspension line was got.Suspension cell was cryopreservation by one-step procedure contain pre-culture of pre-culture solution containing Abiscisic acid (0.26μg/L)for 5 day with treatment of chemical cryoprotectants(DMSO(10%)+PEG(10%)+Suc(10%)),rehydration at -30℃for two hours before quenching in LN for long-storage.Suspension cell is ideal material for transgenic and offering a population of near-identical cells for studies of the cell cycle. (2)OsPLD genes is super family genes,at least contains 16 members,PLD is very important for higher in growth and development.Material is seeding, OsPLD V₁,α₁,λ,P₁ was cloned by RT-PCR,every cDNA lengh of genes is 2550bp,2439 bp,2451 bp,3606 bp;Organs expression analysis were experimented in root,stem,leaf,flower,cell of mature embryo initiation by semi-qualititative,results is OsPLDV1 was found in flower,another three genes were found in all above organs;Gene function was identity with Yeast mutant,results is 4 genes cann't express,this showed PLD genes has indifferention.The experimemt results has benefit for OsPLD family genes members.