Study On Establishment Of Phalaenopsis Regeneration System And Transformation Of Blue Forming Genes

Phalaenopsis aphrodite,due to the limitation of genetic resources,lacks blue varieties,and uses molecular methods to introduce foreign genes into Phalaenopsis so that it can obtain new anthocyanin synthesis capabilities to overcome the limitations of gene types in the plant gene pool.And due to the fact that apartheid cannot directly obtain blue plant forming genes of other species,it provides a theoretical basis for the directed improvement of plant color breeding.In this study,the Phalaenopsis ’big chil’ was used as the material,and the plant expression vectors of blue forming genes from different sources were used to form different combinations for transient transformation of Phalaenopsis.The genetic regulation of blue forming genes on the color of butterfly orchids was initially explored and suitable for stability Combination of transformed vectors to construct a stable expression vector for plant transformation;at the same time,the effects of material selection,disinfection method,basic medium,6-BA and NAA of Phalaenopsis explants on the induction,proliferation and rooting of adventitious buds were studied.Explore the establishment of a reproductive reproduction system of Phalaenopsis,set different hygromycin concentrations,and select the optimal selection pressure for protocorm-like species.The results obtained are as follows:1.After transient transformation of Ma DFR and Ma F3’5’H vector combination from grape hyacinth,petal color becomes darker,and further anthocyanin content measurement results show that the transformed anthocyanin content is 1.32 times of wild type,thus The combination suitable for stable transformation is selected,namely Ma DFR + Ma F3’5’H.2.Constructed a plant binary expression vector p CAMBIA1301-Ma DFR-Ma F3’5’H for subsequent stable transformation of Phalaenopsis.3.(1)The pedicel is suitable as an adventitious bud-inducing explant,and its bud emergence rate is better than that of buds,scapes,and calyxes;(2)First treated with 2% hydrogen peroxide solution for 15 minutes,then treated with 10% sodium hypochlorite solution for 10 minutes,disinfect The effect is good,and the pedicel pollution rate is low,26.6%;(3)The optimal medium for pedicel induction is: Huabao No.1 + Huabao No.2 + 6-BA 3 mg / L + NAA 0.05 mg / L,the germination rate 78.9%;(4)The optimal medium for adventitious bud proliferation is: Huabao No.1 + 6-BA 5 mg / L + NAA 0.05 mg / L,and the proliferation coefficient is 3.60;(5)The optimal medium for seedling rooting is: 1 / 2MS + NAA 0.2 mg / L,the rooting rate is 73.7%.Established a regeneration system for Phalaenopsis ’big chil’.