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Transgenic euramerican populus trichocarpa*populus deltoides for resisting apocheima cinerarius pests

A transgenic, poplar inchworm technology, applied in genetic engineering, plant genetic improvement, angiosperms/flowering plants, etc., to achieve the effects of reducing pesticide use, delaying pest resistance, and improving insect resistance

Inactive Publication Date: 2013-05-08
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] WIN4 is a protein induced by poplar leaves after trauma. It has only been described as a nutrient storage protein. There is no literature to report that it has resistance to one of the main leaf-eating pests of poplar - Lepidoptera poplar inchworm ability

Method used

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  • Transgenic euramerican populus trichocarpa*populus deltoides for resisting apocheima cinerarius pests
  • Transgenic euramerican populus trichocarpa*populus deltoides for resisting apocheima cinerarius pests
  • Transgenic euramerican populus trichocarpa*populus deltoides for resisting apocheima cinerarius pests

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1: the acquisition of transgenic hybrid poplar

[0023] 1. Cloning of wound response protein gene cDNA of hybrid poplar (Populus trichocarpa female parent×Populus del toides male parent)

[0024] According to the partial cDNA sequence (L20233.1) of the hybrid poplar (Populus trichocarpa×Populus deltoides) wound response protein registered in Genbank, primers were designed, using the total RNA of hybrid poplar (Populus trichocarpa×Populus deltoides) leaves as a template, with Forward:5' -atgtcgagcgttaacttggtag-3'; (SEQ ID No.2) and Reverse:5'-tcattcacaagccaaacgtg-3'; (SEQ ID No.3) are upstream and downstream primers, and a specific band of the expected size is obtained by RT-PCR ( The nucleotide sequence represented by SEQ IDNo.1), the result is as follows figure 1 shown.

[0025] The PCR product (nucleotide sequence represented by SEQ ID No.1) was recovered from the gel, and the recovered PCR product was ligated to the pJET1.2 vector, and the ligated produc...

Embodiment 2

[0041] Example 2: Insect resistance of a transgenic European and American hybrid poplar (transgenic hybrid poplar) resistant to poplar inchworm pests to poplar inchworm pests

[0042] Capability research

[0043] 1. Group breeding of poplar inchworms

[0044] The egg masses of Populus deltoides×Populus nigra (control plant) were used to hatch the egg masses of Populus deltoides×Populus nigra (control plants) and put the newly hatched 1st instar larvae of Populus deltoides in a clean and dry jar. Feed the larvae and tie the mouth of the bottle tightly with air-permeable gauze to ensure that the leaves are fresh and sufficient. There were 30 larvae per bottle, and 6 replicates were set. Place the culture bottle in a biochemical incubator at (25±2)°C and relative humidity (RH) 70%-80%. When feeding for 3 weeks, the body weight and body length of poplar inchworm larvae were counted for each treatment; the survival situation was counted before pupation, and the mortality rate wa...

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Abstract

The invention discloses transgenic euramerican populus trichocarpa*populus deltoides for resisting apocheima cinerarius pests. The transgenic euramerican populus trichocarpa*populus deltoides is obtained by the following methods of taking total RNA (ribonucleic acid) of leaves of populus trichocarpa*populus deltoides (populus trichocarpa*populus deltoides) as a template, taking SEQ ID No.2,3 as primers, obtaining SEQ ID No. 1 by RT-PCR (reverse transcription-polymerase chain reaction) and being bonded to pJET1.2 to obtain recombinant plasmid and leading E.coli strain TOP10, screening positive clones, and screening bacterial colony containing resistance plasmids; carrying out PCR validation and double enzyme validation on extracted plasmids, choosing correct recombinant, naming as pBI121-WIN4 and transferring into agrobacterium tumefaciens to carry out colonial PCR verification, converting the obtained bacterial strain into euramerican populus trichocarpa*populus deltoides to obtain the transgenic euramerican populus trichocarpa*populus deltoides for resisting the apocheima cinerarius pests. The transgenic euramerican populus trichocarpa*populus deltoides for resisting apocheima cinerarius pests disclosed by the invention has a strong ability for resisting the apocheima cinerarius pests.

Description

technical field [0001] The invention belongs to the biotechnology of plant variety improvement, in particular to a transgenic European-American hybrid poplar resistant to the poplar inchworm pest. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 王洁华杨少辉宋英今
Owner TIANJIN UNIV
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