Study On Biotechnology Of "Mopan" Persimmon

Dormant buds formed in the same year of Mopan persimmon trees were taken as explants. Effects of explants form and basal media on primary culture were studied. Factors affecting subculture micropropagation were studied by using tube seedlings as materials- The suited ingredient of culture medium for subculture micropropagation of forepart and metaphase were established Some factors affecting adventitious shoot regeneration were studied. Adventitious shoot regeneration systems from in vitro leaves and stems without internode of tube seedlings were obtained. 2 ACC synthase gene fragments were successfully cloned from, mature Mopan persimmon fruit, Here all have been done is in order to improve the persimmon performance of storage and transportation and simplify the management of storage and transportation. The mean results were as follows:Dormant bud explants taken from Mopan persimmon trees after being peelled squamas and 3 leaf primordia off were inoculated on MS, MS(1/2N) and 1/2MS all supplemented with 7.5 mg/L BA. Primary cultures were all obtained on the 3 media. The growth state of explants inoculated on MS(1/2N)+ BA 7.5 mg/L medium was the best. They then were transferred on MS (1/2N) +ZT 1.0 mg/L +IAA.04 mg/L+BA1.0mg/L medium. The mean multiplication reached 2.37 and the average height of tube seedlings reached 11.8 mm after being subcultured 2 times.Tube seedlings having been cultured on MS(1/2N) + ZT 1.0 mg/L + LAA 0.1 mg/L + BA 1.0 mg/L medium for about 10-12 times then were transferred on MS + ZT 1.0 mg/L + IAA 0.1 mg/L + BA 1.0 mg/L medium. The multiplication increased 44.4%, at the same time the average height of shoots increased 5 mm. The results showed that the effect of the 4 factors on subculture micropropagation was in the order of basal medium, ZT, IAA and BA. Compared with MS(1/2N), MS basal medium was more beneficial to forepart subculture micropropagation.Tube seedlings having been cultured on MS + ZT 1.0 mg/L + IAA 0.1 mg/L + BA 1.0 mg/L medium about 10~12 times then were inoculated on DKW + ZT 1.0 mg/L + IAA 0.1 mg/L + BA 1.0 mg/L medium. The multiplication increased 43.1% , and the average height of shoots increased 6 mm. The results of R value showed that the DKW basal medium wasthe most suitable medium to the metaphase subculture micropropagation of "Mopan" tube seedlings.Effects of basal media, ZT and IAA on adventitious shoot regeneration were studied through orthogonal design trial L9(34). The results showed that the effect of ZT concentration was the greatest and the basal medium took the second place. But the concentration of IAA was more effective on the forming of callus than basal medium and ZT. The upper 2 expanding leaves of tube seedlings, cut into 4 mm X 4 mm and inoculated with abaxial layer downside may have the greatest regeneration efficiency. The differentiation percentage and the average adventitious shoot number per explants on MS(1/2N) + ZT 4.0 mg/L were 76.76% and 1.26 respectively.The upper 2 extending stems without internode on tube seedlings, cut about 4mm in length and lay on the MS(1/2N) + ZT 4.0 mg/L + IAA 0.1 mg/L medium may have the greatest regeneration efficiency. The differentiation percentage and the average adventitious shoot number per explants were 83.65% and 2.25 respectively. The effect of ZT concentration was the greatest and the basal medium was greater than IAA. But the concentration of IAA was more effective on the forming of callus than basal medium and ZT.2 ACC synthase gene fragments M-ACS1 and M-ACS2 were successfully cloned from mature Mopan persimmon fruit. The identities of nucleotide acid and amino acid were 98% and 99% respectively. The nucleotide acid identities of M-ACS1 and M-ACS2 with DK-ACS1 were 69% and 70% respectively. They both encoded 224 amino acid. They also have the conserved amino acid regions and invariant amino acid residues of ACC synthase existing in other plants, and with higher identity in polypeptide leveE The amino acid identities with DK-ACS1 were 98% and 99% respectively.