Related gene for regulating and controlling wood development and application thereof
A gene and exogenous gene technology, applied in the field of biotechnology and bioenergy, can solve the problems of large environmental pollution
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Embodiment 1
[0168] The cloning of the gene expression sequence of embodiment 1PtMAN6 and promoter
[0169] 1. Sequence information
[0170] The complete CDS sequence of PtMAN6 is as follows:
[0171]
[0172] or
[0173]
[0174]
[0175] The amino acid sequence of PtMAN6 protein is as follows:
[0176]
[0177] or
[0178]
[0179] The cloned PtMAN6 promoter sequence is as follows:
[0180]
[0181]
[0182] 2. Vector construction
[0183] 2.1 Overexpression vector
[0184] 35S:PtMAN6 vector map see figure 1 ; 35S:PtMAN6-GFP vector map figure 2 .
[0185] 2.2 Antisense expression vector
[0186] 35S:PtMAN6AS map see image 3 .
[0187] 2.3 Transient expression vector
[0188] MAN6G transient expression vector see Figure 4 , MAN6sG transient expression vector see Figure 5 .
Embodiment 2
[0189]Example 2 Overexpression of PtMAN6 reduces the content of wood lignin and crystalline cellulose
[0190] Western detection proves that the PtMAN6 overexpression (OE) vector and antisense (AS) expression vector are successfully transferred into poplar, and can be expressed in plants, and further play a role ( Figure 6 A).
[0191] Analysis of transgenic plants showed that compared with wild-type plants, the petioles and stems of OE plants became softer ( Figure 6 B, 6C and 6D), while AS plants were the opposite ( Figure 6 B, 6C and 6E), indicating that PtMAN6 is involved in the regulation of stem mechanical strength. Further slice analysis showed that the content of lignin in the petiole of OE plants decreased ( Figure 6 F-H). Similarly, lignin deposition in stems also decreased ( Figure 7 A-I). The determination of lignin and crystalline cellulose content of mature wood further proved the function of PtMAN6, and overexpression of this gene resulted in a decrea...
Embodiment 3
[0192] The biomass of embodiment 3 transgenic poplar does not change significantly
[0193] The present embodiment has measured the biomass of poplar, and the result shows, with respect to wild type, the height and girth of 1.5 year old transgenic poplar wood all have no significant change ( Figure 8 ).
[0194] Figure 8 Shows the height and girth analysis results of 1.5-year-old transgenic poplar wood, wherein, Figure 8 A shows the overexpression of PtMAN6 poplar (OE), the antisense expression of PtMAN6 poplar (AS) and the wild type (WT) poplar have no significant difference in trunk height; Figure 8 B shows that there is no significant difference in the circumference of the base of the trunk of OE, AS and WT poplar, and the statistical analysis uses T test.
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