Preliminary Study On In Vitro Culture And Genetic Transformation Of Jasmine

Jasmine often refers to the double-petal jasmine[Jasminum sambac(L.)Ait]of the subtropics,which is also the city flower of Fuzhou.The jasmine flower is a natural perfume plant,and the root,the stem and the leaf of the jasmine flower have higher medicinal value,and can be used for clearing away heat and toxic materials,promoting blood circulation,removing blood stasis,regulating menstruation and inducing diuresis,and the like;and the jasmine is the commercial value of the flower cake,the cosmetic and the like due to the strong fragrance of the jasmine,the cellar-made scented tea,the refined essential oil,Therefore,the cultivation area of the jasmine in the tea-producing area of China is increasing.Because the jasmine is only flowering,it is rare and firm,so it is not capable of sexual reproduction,and the breeding method commonly used in the production has the advantages of high seedling rate,fast hair root,simple and convenient operation and the like.but the traditional breeding method is easy to cause the occurrence of the variety degradation of the jasmine,can not maintain the good character of the female parent,and the like,and is not beneficial to the cultivation of the new variety.In addition,the flower of jasmine with a single color also restricted the innovation of the ornamental shape.The modern biotechnology provides an efficient and practical new way for the innovation and breeding of the ornamental plants in the garden,but the application research in the jasmine is almost blank.In view of this,the most basic bottleneck in the application of the biological technology of the jasmine is the establishment of the body regeneration system and the genetic transformation system,and lays the foundation for further developing the excellent gene resource of the jasmine and the cultivation of new varieties.The main research contents and the results of the study are as follows:(1)The best disinfection time and phytohormone ratio and the best culture medium for callus proliferation and differentiation were selected as explants for callus induction.The results showed that the contamination rate and the browning rate were the lowest when the leaf was treated for 12 min and the stem section was treated for 15 min,and the WPM medium supplemented with 20 g/L sucrose,6.5 g/L agar,2 mg/L 6-BA,0.2mg/L NAA was the best in inducing the callus and the wound tissue to proliferate.and the induction rate of the stem segment is higher than that of the leaf,and the induction rate of the stem segment of the jasmine stem is 100 percent.The medium for callus induction and differentiation was WPM 2 mg/L 6-BA 0.2 mg/L NAA 20 g/L sucrose 6.5g agar,and the induced differentiation rate was 8%?10%.(2)In the course of the experiment,it is difficult to differentiate the phenomena of bud and root in the callus,and the 8 members(JsWOX4,c7735,c19299,c136885,c92402,c16725,JsWOX1,c76574)The expression pattern of the different period of the formation and differentiation of the healing tissue was studied,and the potential role of the WOX gene family in the dedifferentiation and redifferentiation of the plant tissues was studied.The results showed that the 8 gene family members expressed in the process of the formation of the healing tissue,indicating that these genes were more or less involved in the regulation of the related pathway in this process.The expression of JsWOX4,c7735,c19299,c136885,c92402 and c16725 in the family members is the first to decrease in this process,and the expression of the JsfWOX4 gene is the highest.The expression of JsWOX1,c76574 in the family members has continued to rise in this process.(3)According to the expression of the WOX gene family in the formation and differentiation of the jasmine-wound tissue,the JsWX4 and JsWOX1 genes with obvious change are selected,and the expression position and the distribution of the cell layer in the healing tissue of 2 months after induction and differentiation are observed through in-situ hybridization technology.The potential function of its product in the tissue of jasminum is studied.The results showed that both the JsWX4 and JsWOX1 genes were expressed in the outside tissue of the tissue of the jasmine,and the JsWOX4 marked the most wound tissue,indicating that the healing tissue cells mainly originated from the cambium-forming stem cells of the stem phloem,and also observed,The JsWOX1 gene is only expressed in the newly dedifferentiated part of the cell,which indicates that the two genes play a different role in the differentiation of the jasmine-healing tissue.(4)The genetic transformation of JsWOXI and JsWOX4 as the target gene was further studied.The plant expression vector of the two genes was constructed to transform the Jasmine tissue,and GUS staining was performed on the transformed healing tissue(the transformed plant of the control plasmid)and the GFP fluorescence detection and the fluorescence quantitative detection(fusion expression label).The results show that the expression of JsWOX4 and JsWOX1 gene can promote the expression of JsWOX4,but the expression of JsWOX4 and JsWOX1 can promote the expression of JsWOX4.In addition,there was a green root in the differentiation of the JsWOX1 gene.