| Asparagus cochinchinensis,as a traditional Chinese medicinal material in my country,has the functions of nourishing yin and moisturizing dryness,clearing lungs and reducing fire,anti-oxidation,anti-bacterial and anti-inflammatory,anti-tumor,etc.It integrates medicinal,edible,feeding and ornamental use.Huge development potential.At present,the propagation of A.cochinchinensis is dominated by seed propagation and ramet propagation,but its seedling production is limited due to the long cycle and low propagation coefficient.Although some researchers have explored the rapid propagation of A.cochinchinensis in tissue culture,they have not yet established an effective rapid propagation technology system.This research system explored the appropriate conditions for aseptic germination and explant culture of A.cochinchinensis seeds,callus induction and adventitious bud formation,cluster bud induction and proliferation,test-tube seedling rooting and transplanting,etc.,in order to effectively construct the scale of this species The rapid propagation technology of seedlings required for chemical artificial planting provides the basis.The main results are as follows:1.Seed sterilization and germination culture conditions:Use mature and plump A.cochinchinensis seed culture materials for disinfection treatment.The appropriate disinfection method is:75%ethanol treatment for 30 s,2%sodium hypochlorite treatment for 20 min or 0.1%mercury liter treatment 10 min;Suitable seed germination medium is:MS+KT 0.1 mg·L-1+GA3 0.5 mg·L-1+6-BA 1.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1,p H 5.8?6.0,15 days of culture,the seeds begin to show white,about 36 days of culture,the sprouts grow to more than 3 cm,and grow well;the effect of three plant growth regulators on the germination of Asparagus seeds is in order of GA3>KT>6-BA,the effect of GA3reached a significant level by analysis of variance..2.Callus induction and adventitious bud formation conditions:take the stem segment without buds as explants for callus induction and adventitious bud induction.The callus induction medium is:1/2MS+NAA1.0 mg·L-1+6-BA 0.1 mg·L-1 sucrose 30 g·L-1+agar 7 g·L-1,p H 5.8-6.0;explants appear callus at 9 days,and the stems are fully healed at 21 days The size of the impact on the wound tissue is:the content of macro elements in the NAA?6-BA?medium,and the impact of NAA on the callus reached a significant level by analysis of variance(P<0.05).Differentiation medium:MS+NAA 0.3 mg·L-1+6-BA 0.5 mg·L-1+KT 0.5mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1,p H 5.8?6.0,the differentiation rate reaches 98.56%;the effect of three growth regulators on callus differentiation is 6-BA?KT?NAA,among which 6-BA and KT have significant effects on callus differentiation(P<0.05),it is an important factor for the differentiation of Asparagus callus.The proliferation medium is:NAA 0.3 mg·L-1+6-BA 0.5 mg·L-1+KT 0.5 mg·L-1+KH2PO4 30 mg·L-1+sucrose30 g·L-1+agar 7 g·L-1,p H 5.8?6.0,multiplication factor of 6.64.3.Bud-bearing stems to induce cluster buds and their proliferation conditions:take the bud-bearing stems as explants to induce and proliferate cluster buds.The axillary bud induction medium is:MS+NAA 0.1 mg·L-1+6-BA 1.0 mg·L-1+sucrose 30 mg·L-1+agar 7 mg·L-1,p H 5.8?6.0.The induction rate is 85.05%,the average number of buds is up to 2.87,the bud color is emerald green,and the growth is robust;the proliferation medium for cluster buds is:MS+6-BA 0.5 mg·L-1+IAA 1.5 mg·L-1+KT 1.5 mg·L-1+sucrose 30 g·L-1+agar 7g·L-1,p H 5.8?6.0,after 21 days of culture,its proliferation multiple reached 3.65,the effect of three growth regulators is:KT>6-BA>IAA,the effect of KT reached a significant level by analysis of variance(P<0.05).Adding 100 g·L-1 of banana mash,the multiplication factor reached 4.47,which was better than apple mash,coconut milk,and potato mash.When browning occurs during proliferation,adding PVP 1.0 g·L-1 can reduce the browning rate to 14.13%.4.Rooting culture conditions:Take rootless sprouts as explants for rooting culture.The highest rooting rate in 1/2MS is 78.59%,which is significantly different from that in White,MS,and 1/4MS medium(P<0.05);the appropriate rooting medium is:1/2MS+IAA 1.0 mg·L-1+IBA 1.5 mg·L-1+PUT 1.0 mg·L-1Sucrose 30 g·L-1+agar 7 g·L-1,p H 5.8?6.0,rooting rate 78.89%,three plant growth regulators for rooting The size of the induced effect is:IBA?PUT?IAA,all of which have a significant impact on the rooting of A.cochinchinensis(P<0.05);the addition of 0.5 g·L-1 activated carbon is conducive to thickening of the root system and luxuriant stems.5.Suitable transplanting substrate:use coir,peat,vermiculite,perlite and garden soil as substrate materials to mix and mix asparagus test-tube seedlings respectively.Through the analysis of the physical and chemical properties of each combination substrate,the growth status of test-tube plantlets is observed Based on the determination of physiological indicators,the substrate bulk density was significantly negatively correlated with the transplanted survival rate of test-tube plantlets(P<0.01),and the porosity of the substrate was positively correlated with the growth indicators of test-tube plantlets.In the combined substrate of coir-perlite-vermiculite(bulk density of 0.23 g·cm-3,total porosity of 65.07%,p H of 6.30, conductivity of 0.51 m S·cm-1),the survival rate of test-tube plantlets reached 87.10%,which was stronger than other substrates;the comprehensive evaluation of the membership function shows that the combination of coir:perlite:vermiculite=1:1:1 has the highest score(0.93),which is more conducive to the growth of test tube plantlets.Suitable for cultivation substrate. |
PDF Full Text Request