Studies On The Activities Of ASA Metabolism Relative Enzymes In Fruit And Leaf Regenration System On Raspberry

Plant vitamin C(ASA).Ascorbic acid not only provides the minority animals(including humans)who cannot synthesis AsA normally with the source of vitamin C,but also the content of AsA has become an important indicator to measure the quality of agricultural products.Raspberry is rich in vitamin C,for the study of raspberry fruit ASA synthsis metabolic enzyme gene in ASA accumulation regulation.Taking four raspberry cultivars as tested material.Determine ASA content and metabolism relative enzymes activities for four kinds fruits of raspberry of various ages.Establishment of regeneration system used Fertod Zamatos and Heritage.Discuss the plant growth regulators and dark periods produce influence for raspberry leaf establishment of regeneration system.Use the technology of gene engineeing on raspberry biosynthesis of vitamin C metabolism enzyme genes in raspberry leaf regeneration system for genetic transformation.Provide a theoretical basis for the improvement of fruit in the ASA content.The results were as follows:1)The experiment determined the ASA content of raspberry fruits of five ages and metabolism relative enzymes activities.The result showed that the content of AsA accumulated continually but first increased in the initial stage of plant development and then decreased,and the AsA content of Fertod Zamatos was higher than that of other three varieties in different development stages.The changes in GalLDH activity coincided with AsA accumulation rate during fruit development.GalL DH activity in Fertod Zamatos was higher than that in other varieties.GalLDH activity was significantly positive correlated with AsA accumulation in four varieties.The changes in DHAR activity coincided with AsA accumulation rate during fruit development.AsA contents and DHAR activities were significantly positive correlated with in different development stages of Fertod Zamatos and Heritage fruits.AsA contents and DHAR activities were considerably positively correlated in Summit and Tulameen fruits for four species.AsA contents had no clear pertinence with MDHAR,AAO and APX activities,but DHA content was significantly associated with ascorbate peroxidase(APX)activity.It is proposed that AsA contents were strongly affected by the GalLDH and DHAR activities,but MDHAR,AAO and APX activities were not the main factors in influencing the AsA contents in different raspberry fruit varieties.2)The experiment for leaves of raspberry callus induction.The medium added 30g/L sugar.Agar 7g/L and PH4.9-5.2.The optimal medium for the callus induction from Fertod Zamatheos was MS+6-BA0.4mg/L+2,4-D0.8mg/L and MS+6-BA0.5mg/L+NAA0.8mg/L.The induction rate reached 100%.When the cytokinin was TDZ,the optimal medium was MS+TDZ1.5 mg/L+IAA0.2mg/L.The induction rate was up to 57.69%.The TDZ composed with other auxin,Most of the leaves were not induction callus.the induction became browning with the next few days.3)The optimal medium for the callus induction from Heritage was MS+6-BA1.5mg/L+IBA0.2 mg/L.The induction rate reached 92%.The callus appeared yellow green,close texture and faster growth.When the medium added TDZ,the callus didn't induction from leaves.4)The buds was induced from two varieties of callus.The Fertod Zamatos leaves of callus induction of the shoot regeneration rate was 15%with MS+TDZ 2.0 mg/L+NAA 0.5 mg/L.The higher rate of regeneration rate with MS+BA 1.0 mg/L+NAA 0.5 mg/L,the regeneration rate was 22.72%.The Heritage leaves of callus induction of the shoot regeneration rate was 15%with MS+BA2.0 mg/L+NAA0.5 mg/L.The shoot regeneration rate was 10%with MS+TDZ2.0 mg/L+NAA0.5 mg/L.5)The medium in dark culture 7-12 days,then in the light of the conventional culture.The medium could increase the amount of callus.And speed up the formation of callus.