Method for improving insect resistance capability of hybrid poplar

A technology of ability and poplar, which is applied in the field of improving the insect resistance of hybrid poplar, can solve the problems of wound-induced proteins with unknown functions, and achieve the effects of improving insect resistance, delaying pest resistance, and reducing the use of pesticides

Inactive Publication Date: 2013-02-13
TIANJIN UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] POP3 is a wound-induced protein with unknown function, an

Method used

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  • Method for improving insect resistance capability of hybrid poplar
  • Method for improving insect resistance capability of hybrid poplar
  • Method for improving insect resistance capability of hybrid poplar

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: the acquisition of turning POP3 poplar

[0023] 1. Cloning of P. Trichocarpa wound response protein gene cDNA

[0024] According to the European aspen wound response protein cDNA sequence (AJ276517) registered in Genbank, a Blastn search was carried out in the draft genome of Populus trichocarpa (P. Trichocarpa), and the obtained sequence showed that it was a predicted protein with unknown function , its amino acid sequence is (XP 002314976), and its gene sequence is (XM 002314940).

[0025] Using the mRNA of Populus trichocarpa (P. Trichocarpa) leaves as a template, Forward:5'-atggcaaccagaactccaaa-3'; (SEQ ID No.2) and Reverse:5'-tagtagagaaagtagtctatcacaagacgc-3'; (SEQ ID No.3 ) as upstream and downstream primers, and a specific band of expected size (nucleotide sequence represented by SEQ ID No.1) was obtained by RT-PCR. The results are as follows figure 1 shown.

[0026] The PCR product (nucleotide sequence represented by SEQ ID No.1) was recovered f...

Embodiment 2

[0042] Embodiment 2: Research on the insect resistance of the POP3 poplar saplings

[0043] 1. Group rearing of American white moth

[0044] Hatch the eggs of white moth in room at room temperature, put the newly hatched 1st instar larvae of white moth in clean and dry jars, and feed the larvae with leaves of transgenic POP3 poplar saplings and non-transgenic poplar saplings respectively. Air-permeable gauze tightly binds the mouth of the bottle to ensure that the leaves are fresh and sufficient. 30 per bottle, 6 replicates. Place the culture bottle in a biochemical incubator at (25±2)°C and relative humidity (RH) 70%-80%. When feeding for 3 weeks, count the body weight and body length of each processing white moth larvae; count the survival situation before pupation, and calculate the mortality rate; record the leaf weight and remaining leaf weight for each feeding, and calculate the food intake compared with the control; After pupation, the pupation rate and average pupal...

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Abstract

The invention discloses a method for improving the insect resistance capability of hybrid poplar, which comprises the following steps: obtaining SEQ ID No.1 through RT-PCR with the mRNA of the leaves of poplar as a template and SEQ ID No.2 and No.3 as primers and connecting pMD18-T, leading the obtained recombinant plasmid into E.coli strain DH5alpha, screening positive clones, extracting plasmid and carrying out double enzyme digestion, connecting the SEQ ID No.1 and pBI121 subjected to double enzyme digestion, transforming E.coli strain DH5alpha, screening bacterial colony containing resistance plasmid, extracting plasmid for PCR verification and double enzyme digestion verification, choosing the correct recon, naming the correct recon as pBI121-POP3 and transferring the correct recon into agrobacterium tumefaciens for colony PCR identification; transforming the hybrid poplar by the obtained strain to obtain transgenic seedlings. The method provided by the invention can improve the insect resistance capability of the hybrid poplar.

Description

technical field [0001] The invention belongs to the biotechnology of plant variety improvement, in particular to a method for improving the insect resistance ability of hybrid poplar Background technique [0002] Poplar is the most widely distributed and most adaptable tree species in the world, and it is one of the important fast-growing timber tree species in my country. However, with the continuous deterioration of the environment and the frequent occurrence of natural disasters, the harm of leaf-eating pests and stem-boring pests has become the biggest obstacle to the development of high-yield poplar forests in large areas. Therefore, the use of genetic engineering methods to cultivate new varieties of poplar with insect resistance is an important way to control insect pests. Due to the long growth cycle of forest plants, they always face the invasion of various diseases and insect pests in the surrounding environment. Therefore, in the long-term evolution process, they...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 王洁华杨少辉宋英今
Owner TIANJIN UNIV
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