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Application of hda909 gene of Populus trichocarpa in improving antifungal ability of plants
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A technology of poplar and plant, applied in the field of genetic engineering, can solve the problems of long growth cycle, high genetic heterozygosity, and difficulty in meeting the requirements
Active Publication Date: 2022-05-13
NORTHEAST FORESTRY UNIVERSITY
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Due to the long growth cycle of forest trees and high genetic heterozygosity, it is often difficult to meet the requirements of directional cultivation of new tree varieties for different purposes by using conventional breeding methods
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Embodiment 1
[0034] Cloning of HDA909 Gene of Populus trichocarpa
[0035] 1. Experimental materials
[0036] The plant materials are sterile tissue culture seedlings of wild-type Populus trichocarpa. See Table 1 for strains and vectors, and Table 2 for main reagents.
[0037] Table 1 strains and carriers
[0038]
[0039]
[0040] Table 2 main reagents
[0041]
[0042] 2. Solution preparation
[0043] (1) 0.1% DEPC aqueous solution: fully mix ultrapure water and DEPC stock solution at a ratio of 9:1, stir for 2 hours, and sterilize for 20 minutes.
[0044](2) 5M NaCl solution: Dissolve 14.625g of NaCl in 45mL of 0.1% DEPC water, dilute to 50mL, incubate at 37°C for 12h, and sterilize for 20min.
[0045] (3) 75% ethanol solution: Mix 0.1% DEPC water and absolute ethanol in a ratio of 1:3 after high temperature sterilization and cooling to room temperature, and store at -20°C.
[0046] 3. Experimental method
[0047] 3.1. Extraction of Populus trichocarpa total RNA
[0048]...
Embodiment 2
[0086] Genetic transformation of Populus trichocarpa with HDA909 gene and its functional analysis
[0087] 1. Experimental materials
[0088] Tissue culture seedlings of wild-type Populus trichocarpa. Alternaria alternata belongs to the subdivision Deuteromycota, its conidiophores are brown or blue-brown inverted rod-shaped, mostly solitary or in bundles, and conidia are mostly in clusters of 3 to 6 There are 1 to 2 mediastinums and 3 to 4 diaphragms. purchased from the Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry. The pROKII vector was purchased commercially. The main reagents are shown in Table 10.
[0089] Table 10 main reagents
[0090]
[0091] Populus trichocarpa rooting medium: WPM medium 2.41g / L, sucrose 20g / L, agar powder 5.8g / L, pH=5.75, IBA0.1mg / L.
[0092] Populus trichocarpa differentiation medium: WPM medium 2.41g / L, sucrose 20g / L, agar powder 5.8g / L, pH=5.75, 6-BA 0.3mg / L, IBA 0.02mg / L and TDZ 0.0008mg / L .
[00...
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Abstract
The invention discloses the application of Populus trichocarpa HDA909 gene in improving the antifungal ability of plants, and relates to the technical field of genetic engineering. According to the Populus trichocarpa HDA909 gene sequence, the present invention constructs the Populus trichocarpa HDA909 overexpression vector, adopts the Agrobacterium-mediated method to introduce the recipient plant, so that the HDA909 gene in the transgenic plant is overexpressed, and antifungal plants are obtained. Using Alternaria to infect plants, it was found that the number of plaques and plaque area of transgenic plants were significantly reduced, and the degree of damage suffered by Alternaria infection was relatively light. The invention proves that the overexpression of Populus trichocarpa HDA909 gene can improve the antifungal ability of plants, and provides a new idea for further creating antifungal transgenic plants in the future.
Description
technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of Populus trichocarpa HDA909 gene in improving the antifungal ability of plants. Background technique [0002] With the development of society and economy, environmental pollution is becoming more and more serious, and the original forest tree species will encounter environmental stresses that are significantly different from those under natural selection. In order to ensure that the living environment and quality of life of human beings will not be degraded, human beings must learn to assist trees to better and faster adapt to various environmental stress factors that are intensified and emerging with the modernization of human society. Due to the long growth cycle of forest trees and high genetic heterozygosity, it is often difficult to meet the requirements of cultivating new tree varieties for different purposes by using conventional breeding meth...
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