| Glycometabolism was a complex metabolic pathway in plants, it was a result of comprehensive expression from various factors including inside and outside, however,little is known about its metabolic regulation mechanism of exogenous suagr effect on the grape plantlet in vitro glycometabolism. In this study, the ‘red globe’(Vitis vinifera L.) test-tube seedlings was used as raw materials, the experiment treatments were adopted by different glucose concentrations(0 g/L, 10 g/L, 20 g/L, 40 g/L) and same concentration of sugar(20 g/L), glucose(20 g/L) and fructose(20 g/L), we measured the activity size of different enzymes related glycometabolism, chlorophyll contents, the changes of morphological indicators such as plant height and internode length, in order to further clarify that the main factors effected on the the red globe tube seedlings sugar metabolic strengthusing. Meanwhile, using RNA-seq,transcriptome sequencing technology analyses were performed, we selected the related pathways genes of sugar metabolism or sugar signaling, and combined with the expression conditions of various amount of gene in different treatments, to study their molecular regulation mechanism in the sugar metabolic pathway, and understood the relationship between the autotrophic and heterotrophic, which lay a theoretical foundation for better regulating the growth and development of tube seedlings. The main results were as follows:1. Different treatment were determined, we observed the changes of glycometabolism related enzyme activity, such as SS, SPS, AI, NI, G6 PDH, PFK,RuBp various enzymes during the red globe tube seedlings growth and development process, and concluded that there were some significant differences, at the early stage and medium-term of the processed material development, the enzyme activity basic showed gradually rising trend, at the late stage of development, the enzyme activity showed a slight downward trend.2. Determination of different treatments, the conclusion could be drawn that the changes of endogenous hormones, ZT, IAA and ABA contents showed a certainsignificant difference among in different sugar treatments during the red globe tube seedlings in the entire development process, and reflected the differences in sugar metabolism, in terms of different concentrations of sugar, the each enzyme activity of G20 and G40 was slightly higher than the other two; For different kinds of sugar, PFK and G6 PDH enzyme activities of G20 were slightly higher than the other two processing, other was more significant in F20 and S20, this reflected the metabolic model was more close to both.3. By sequencing, about 31.65 Gb clean and high-quality reads were generated,and 68.31%-73.71% of the data were mapped to the reference genome from various treatment, 1. A total of 27131 genes were found from six different libraries, which has25898 genes were previously found on the reference genome; Compared with the Wayne figures of different kinds sugar, we found 1023, 1717, 1217 different expression genes in the F20 vs S20, G20 vs F20 and G20 vs S20, respectively, and compared to the Wayne figures of different sugar glucose concentrations, we found G0, G10, G20, G40 libraries contains 22539, 19258, 22367, 22730 expression genes,respectively. The sequencing of these genes, we used the GO software to genome functions annotation, and filtered according to their respective functions, and selected different glycometabolism processing, from the results, we concluded the ABA, JA,photosynthetic pathway was the most significant on red earth tube seedlings sugar metabolism, the gene numbers involved in these three pathways was the superior, 273,181 and 131, respectively; Meanwhile, we aslo found that the genes related to the sugar metabolism key enzyme SS, SPS, NI, G6 PDH, PFK, and plant endogenous hormones IAA and ZT, CTK, GA, ETH, and protein kinases, HXK, PP2 A, and so on.These genes involved in regulating the whole process of sugar signaling. |
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