Application of miR156 and related biomaterial thereof in regulating disease resistance of plants
A biomaterial and disease resistance technology, applied in the application field of regulating plant disease resistance, can solve problems such as unreported research, and achieve the effect of enriching forest tree breeding resources
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[0066] Example 1, the application of miR156 in regulating the disease resistance of birch
[0067] 1. Construction and identification of transmiR156 birch
[0068] The Gateway technology was used to construct the overexpression vector pGWB2-miR156, and the genetic transformation of birch was carried out by using the zygotic embryo method mediated by Agrobacterium tumefaciens. The miR156-transformed birch was identified by PCR and quantitative PCR. Specific steps are as follows:
[0069] 1. The construction of miR156 birch
[0070] (1) Construction of overexpression vector pGWB2-miR156
[0071] The 192bp DNA fragment containing the mature sequence of miR156 (the DNA fragment shown in the 221-412 position of sequence 3) was constructed into the entry vector pENTR (purchased from Invitrogen Company) by Gateway technology, and then used GatewayLR Clonase™ II Enzyme Mix (purchased from Invitrogen Co., Ltd.) carried out homologous recombination of a DNA fragment with a size of 19...
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