Genes used for regulating and controlling synthesis and transport of nicotine in tobaccos and application thereof
A nicotine and gene technology, applied in the field of gene technology and botany, can solve problems affecting nicotine content, achieve good content traits, and improve content traits
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Embodiment 1
[0035] Based on high-throughput sequencing to obtain the small RNA sequences of tobacco roots and leaves after topping and leaf damage, as well as the genome sequences of two ancestral species of tobacco, a large number of tobacco gene sequences can be found, which can form a stable hairpin structure, and its stem can Production or expression of small RNA sequences (mature sequences). The hairpin structures formed by 6 gene sequences are shown in Figure 1-6 . These predicted hairpin structures were stable (all MEF values were less than −24.82). The expression sequences of these genes were detected, especially in the roots and leaves of topping and leaf damage tobacco, which indicated that the expression of these genes was probably induced by these treatments.
Embodiment 2
[0037] The small RNA sequences generated by the expression of the above six genes are predicted by the target genes, and all target the key protein-coding genes in the tobacco nicotine synthesis and transport pathway, including QPT (quinolinate phosphirobosyltransferase), PMT (putrescine N-methyltransferase) and MATE (multi antimicrobial extrusion family protein) genes (Table 1). A large number of studies have shown that these genes are the key functional genes of nicotine synthesis (QPT and PMT) and transport (MATE), respectively (Dwey and Xie, 2013, Phytochemistry). The predicted values of the 6 genes and their target genes found in the present invention are all between 0-2.0 (generally, the close value is 3.0, exceeding this value indicates that the binding result is unreliable), and all of them regulate the target genes by cleavage. At the same time, the analysis results of the degradome data obtained in the experiment showed that 6-105 cut fragments were detected (the c...
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