Study On Establishment And Mechanism Of Tissue Culture And Rapid Propagation Of Acidosasa Edulis T.H.Wen.

Acidosasa edulis is one of Fujian’s unique bamboo species.It has a high level of protein and phosphorus and calcium,which is one of the most abundant nutrients in the bamboo shoots.It has high nutritional value and medicinal value.At the same time,Acidosasa edulis as a green plant has a high ornamental value and economic value.Plant tissue culture is an efficient breeding technology,which can shorten the breeding cycle of nursery stock,reduce the cost of nursery,accelerate the batching of plants,scale production,realize the industrialization and industrialization.In this experiment,different parts of Acidosasa edulis were used as explant materials.The main influencing factors and the best culture formula of the tissue culture process were studied.The mechanism of the determination of endogenous hormones was studied,and the aim of this study is to establish a complete tissue culture system for the regeneration of Acidosasa edulis.The second purpose is to provide an efficient production path for its scale.At the same time,it lay the foundation for further research and provide the basis and reference for the future research on tissue culture of other bamboo species,especially scattered bamboo.The main results of this study are as follows:(1)The optimal sampling time and explant site for bud initiation were In May selected Acidosasa edulis buds who are in the growing season of the year borning.At this point the pollution rate reached a minimum of 5%.The induction rate is the highest 93.37%and buds grow well.Inoculation of the best way to disinfect was 75%alcohol was used to soak for 25 s,and 0.1%HgCl2 solution was used for disinfection for 5.3 min.At this point the lowest pollution rate of 5.56%,the survival rate reached a maximum of 93.33%.The best basic medium was MS medium,the pollution rate reached the lowest 4%,the germination rate reached 80%and the bud growth was good.(2)The best preparation condition of callus culture of Acidosasa edulis was Take sterile stem segments as explants;0.05 mg·L-1 riboflavin(VB2)was added as an anti-browning agent to the MS medium,and the dark culture was carried out at 20℃.The callus rate was as high as 53.26%,and the callus strcture was dense and the browning rate was the lowest.(3)The best scheme for callus induction of Acidosasa edulis was MS + 2,4-D 2 mg·L-1 + 6-BA 0.5 mg·L-1 + KT 1 mg·L-1 + sucrose 30 g·L-1 + agar 7 g·L-1.The callus induction rate reached 23.33%.The effects of three kinds of hormones on callus induction were:2,4-D>6-BA>KT and 2,4-D were significantly affected by callus induction.It can be seen that 2,4-D is the most important factor of callus induction.(4)The best culture scheme for congestive bud Induction of Acidosasa edulis was MS + 6-BA 3 mg·L-1 + KT 1 mg·L-1 + TDZ 0.05 mg·L-1 + NAA 0.5 mg·L-1 + sucrose 30g·L-1 + agar 7 g·L-1。In this program,the induction rate of tufts was 93.33%,and the survival rate was higher.The effects of four kinds of hormones on the induction of shoots were:6-BA>KT>TDZ>NAA,6-BA,KT and TDZ had significant effects on the induction of shoots,and NAA was significantly affected.This indicated that cytokinin was the dominant factor in the induction stage of Acidosasa edulis buds.In this program,the contents of ZT,GA3 and IAA were the highest in the explants,and the content of endogenous hormone ABA was the lowest.In the process of bud induction,the contents of ZT,GA3 and IAA increased,and the content of ABA decreased.The ratio of GA3/ABA and GA3/IAA was higher than that of ABA/IAA,which indicated that the low level of abscisic acid and gibberellin were required at the stage of bud induction.(5)The best way to proliferate buds of Acidosasa edulis was MS+ 6-BA 2 mg·L-1 +KT 0.1 mg·L-1 + NAA 0.1 mg·L-1 + sucrose 30 g·L-1 +agar 7 g·L-1.Under this program,the multiplication factor reached a maximum of 4.0 times.Its survival rate of 90%.At this stage,the contents of ZT,GA3 and IAA in the explants showed a gradual increase trend,while the content of ABA did not change significantly,which was stable and did not show obvious inhibition to bud proliferation.The ratio of ZT/IAA was stable,while ABA/IAA showed a decreasing trend,indicating that the explants had good growth at this stage,and the explants were in the stage of bud differentiation.(6)The best scheme for regenerated planting rooting of Acidosasa edulis was 1/2MS + 6-BA 3 mg·L-1 + NAA 0.5 mg·L-1 + sucrose 30 g·L-1+ agar 7 g·L-1.At this time,the rooting rate of tissue culture seedlings was 93.33%,survival rate of 75%and the root growth was good.At this stage,the contents of ZT,GA3 and IAA in the explants showed a decreasing trend,while the content of ABA showed a trend of continuous growth.The ratio of GA3/ABA is decreasing,while the ratio of ABA/IAA is increasing,which indicates that the aboveground part of Acidosasa edulis is slowly growing or stopping the growth stage,but the ratio of GA3/ABA is decreasing.The underground root part is in the fast growing stage.(7)In regeneration plant transplanting process,the first time at room temperature does not open the bottle refining seedlings 5-8d,and then open the bottle refining 5-8 d,transplanted to the sterilized perlite:vermiculite:loam(1:1:1)matrix,placed in a cool and humid place culture,this time the survival rate of up to 63.2%.(8)In the process of tissue culture,the contents of ZT,GA3 and IAA in the bud induction and proliferation stage showed an increasing trend.Their range of change were 0.15-1.4 μg·gFW-1,0.24-1.2 μg·gFW-1,0.29-1.11 μg·gFW-1;ABA content is lower,0.21-0.49 μg·gFW-1.This indicated that the high concentration of ZT,GA3 and IAA and the lower level of ABA were needed in the bud induction and proliferation culture stages.And the contents of ZT,GA3 and IAA decreased to the rooting stage 0.57 μg·gFW-1,0.62 μg·gFW-1 and 0.45 μg·gFW-1,while the ABA content increased to 1.21 μg·gFW-1.This suggests that low con－centrations of ZT,GA3 and IAA and high concentrations of ABA are more suitable for rooting of yellow sweet bamboo tissue culture seedlings.The significant changes in GA3/ABA and ABA/IAA in the process of tissue culture could clarify the changes in the growth of aerial parts and underground parts of various stages.