Hormonal Effect Of Tissue Culture Cells Of Toona Sinensis And Analysis Of The Distribution Of Flavonoids In Adult Leaves

Toona sinensis was used as the experimental research material to explore the effects of different hormones on tissue culture,callus induction,active substance content and related isoenzyme activities and zymogram,while also affecting the trunk of red,black and green oil the content and distribution of flavonoids in the adult leaves of the parts were determined and analyzed.1.Through the selection of the medium and the ratio of different plant hormones,a high-efficiency rooting and buds proliferation medium and a hormone medium that was conducive to the robust cultivation of buds seedlings were selected.The results showed that WPM was more conducive to the growth of tissue culture seedlings of toon than MS and 1 / 2MS culture media.The best rooting hormone medium was MS + IBA0.5mg / L + NAA 0.25 mg / L,at which the rooting rate of sterile seedlings was 86.7 %;Proper increase of gibberellin concentration would be beneficial to the growth of cluster buds.The best combination and concentration of proliferation hormones was add 1.0mg / L6-BA,0.1mg / LNAA and 0.5mg / LGA3 to the MS medium.The multiple was up to 5.9 times,and the stem segments of tissue culture seedlings cultured in MS + 1.0mg / LKT + 0.1mg / LNAA + 0.5mg / LGA3 +0.5g /LAC hormone medium when the rooting tissue culture seedlings were cultivated stout,dark green leaves,vigorous growth,when further bud cultivation,it was found that the combination of 6-BA and IAA were better than the combination with NAA. When the concentration of 6-BA was 2.0mg / L and the concentration of IAA was0.05 mg / L,the buds were cluster the strong buds effect was the best.At this time,the buds height were 3.34 ± 0.13 cm,and the multiplication factor was 3.88.2.During callus induction,cell differentiation culture,and preliminary determination of metabolic substance content,it was found that WPM was not suitable as a basic medium for callus induction.When the addition amount of 2.4-D in MS medium was 1.5 mg / L,the tissue culture seedlings has the best callus induction effect,and the outdoor growth toon has the best induction effect when the concentration of 2.4-D is 2.0mg / L.Among the callus cell clusters with buds spots,there were good effect of budding buds formation after culturing in MS + 1.0mg /L6-BA + 0.1 mg / LNAA medium.In MS + 1.0mg / LKT + 0.1mg / LNAA medium,both leaves callus cell clusters and the bud-dot cell clusters could produce multiple white lateral roots,and the buds spots would differentiate and grow into a single robust new plant.When the leaves callus were rapidly differentiated and cultured,it grows rapidly in MS + 0.5mg / LKT + 1.0 mg / L6-BA + 0.1mg / LNAA + 0.05 mg / L IAA hormone medium.During the second subculture,the GA3 concentration was changed,there was a tendency to change the content of total flavonoids and total saponins in secondary metabolites,the content of these two substances measured in callus cells produced by MS + 1.0mg / LKT + 0.1mg / LNAA + 0.5g / L AC relatively high.3.After added 2.4-D alone and used 6-BA and NAA in combination,the zymogram and enzyme activity of isozymes POD and PPO in three growth periods(6days,12 days and 18 days)of callus induction it has the following effects: after hormone-induced callus,the POD and PPO enzyme activities and zymogram bands measured in the cells were significantly different due to incubation time and hormone type.The overall enzyme activity measured after 12 days of induction was higher.In MS + 1.5mg / L 2.4-D medium,the highest measured POD enzyme activity in callus cells,POD enzyme after gel electrophoresis,6-7 enzyme spectrum appeared in each group,the band Ⅱ,Ⅲ,Ⅴ And Ⅵ were relatively stable in different hormone-induced cells;PPO enzymes showed 9 enzyme profiles as a whole,among which bands Ⅰ,Ⅱ,Ⅲ and Ⅸ changed greatly due to different hormone types and induction time,while bands Ⅳ and Ⅵ,Ⅶ and Ⅷ were relatively stable in each group.4.Analyze and determine the flavonoids contained in different growth parts(upper part,middle part and lower part)of Toona sinensis of different strains(Black oil toona,Red oil toona,and Green oil toona)by different stages of high-performance liquid chromatography.It provides a reference for studying the dynamic changes of flavonoids in Toona sinensis.In the experiment,the main conclusions has shown: the flavonoids in the middle leaves of the Red oil toona > the upper leaves> the lower leaves.Both flavonoids were measured in the upper leaves of the tree trunk in Black oil toon and Green oil toona.In a single substance,rutin and quercetin were higher in Black oil toona and Red oil toona,and the highest part was in the upper leaves of Black oil toona,and isoquercetin was higher in Green oil toona,and the highest was in the upper leaves of Green oil toona.The results of comprehensive measurement of the three substances in the upper leaves of Black oil toona showed that the parts and varieties with higher flavonoid content were upper leaves of Black oil toona> middle leaves of Red oil toona > upper leaves of Green oil toona.