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Method for breeding aphid-resistant transgenic wheat and special vector

A technology of transgenic wheat and aphid resistance, which is applied in the direction of introducing foreign genetic material using vectors, botany equipment and methods, applications, etc., and can solve the problems of fecundity viability and lifespan reduction

Inactive Publication Date: 2011-08-17
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But according to Birch et al (Birch A N E, Geoghegan I E, Majerus M E N, Mcnicol J W, Hackett C, Gatehouse A M R, Gatehouse J A. Tri-trophic interaction involving pest aphids, predatory 2-spot ladybirds and transgenic potatoes expressing snowdrop lectin for aphid resistance. Molecular Breeding, 1999, 5: 75-83.) report, after the two-star ladybug eats the aphids on the transgenic potato, its fecundity, viability and lifespan of the ovum obviously reduce, thereby, transgenic plant is harmful to the ecological environment The impact of the

Method used

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  • Method for breeding aphid-resistant transgenic wheat and special vector
  • Method for breeding aphid-resistant transgenic wheat and special vector
  • Method for breeding aphid-resistant transgenic wheat and special vector

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Experimental program
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Effect test

Embodiment 1

[0044] Example 1. Preparation of cDNA clone of MhβFS gene

[0045] Mentha asiatica was purchased from Beijing Tanghe Herb Art Manor. The blunt-end vector pEASY-Blunt was purchased from Beijing Quanshijin Biotechnology Co., Ltd., and the product catalog number is CB101.

[0046] The total RNA of Asian peppermint was extracted and reverse transcribed into cDNA. Using the cDNA as a template, PCR amplification was performed on MhβFSF1 / MhβFS R1 with primers, and the PCR product obtained was the MhβFS gene.

[0047] MhβFS F1: 5’-ATGGCTACAAACGGCGTC-3’

[0048] MhβFS R1: 5'-TCAAAAGACTATGGCATCAACA-3'.

[0049] The amplification system is as follows: ddH 2 O 31.5μl, 5×PrimeSTAR Buffer(Mg 2+ plus) 10.0μl, dNTPMixture (2.5mM each) 4.0μl, MhβFS F1 (10μM) 1.5μl, MhβFS R1 (10μM) 1.5μl, reverse transcription product 1.0μl, PrimeSTAR HS DNA Polymerase (2.5U / μl) 0.5μl, Total 50.0μl.

[0050] PrimeSTAR high-fidelity enzyme amplification results in blunt-end fragments, so the recovered PCR product can be ...

Embodiment 2

[0051] Example 2. Transgenic wheat by gene bombardment

[0052] 1. The expression vector construction of gene bombardment transgenic wheat

[0053] The construction process of vector MhβFS 1-pG4AB and vector MhβFS 1+CTP-pG4AB is as follows: figure 1 Shown.

[0054] (1) Preparation of vector MhβFS 1-pG4AB

[0055] The rice variety is Nipponbare, purchased from the National Crop Germplasm Preservation Center of the Institute of Crop Science, Chinese Academy of Agricultural Sciences.

[0056] A. Preparation of rice Rubisco small subunit promoter (rbcS Promoter)

[0057] The rice leaf DNA was extracted, and PCR amplification was performed on rbcSP F1 / rbcSP R1 with primers to obtain the Rubisco small subunit promoter (rbcS Promoter).

[0058] rbcSP F1: 5’-TCGGGTCGAGGTGAACTTTA-3’;

[0059] rbcSP R1: 5’-AGATGCACTGCTCTGCACAC-3’;

[0060] Connect the rbcS Promoter to the T-easy vector, and perform sequencing verification on the recombinant vector. As a result, the correct recombinant vector is obtai...

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Abstract

The invention discloses a method for breeding aphid-resistant transgenic wheat and a special expression vector. A recombinant vector provided by the invention is obtained by inserting gene expression cassettes shown as nucleotides at sites 1 to 3,828 from the tail end 5' in SEQ ID NO (Sequence Identity Number): 1 into multiple cloning sites of a vector pG4AB. Experiments prove that the aphid resistance of the transgenic wheat into which the vector is transferred is obviously higher than that of wild wheat, so that the vector has great significance in the field of genetic breeding of plants.

Description

Technical field [0001] The invention relates to a method for cultivating aphid-resistant transgenic wheat and a special carrier. Background technique [0002] As one of the most important food crops, wheat is the main source of human protein intake; it is also rich in vitamin B, vitamin E, fiber, magnesium, phosphorus and other substances (Vasil IK, Anderson O D. Genetic engineering of wheat gluten. Trends in Plant Science, 1997, 2(8):292-297.). According to the "World Food and Sustainable Agriculture" conference held in 1997, the world wheat demand will increase at a rate of 1.6% per year, and will reach 775 million tons in 2020, an increase of 40% from 552 million tons in 1993 (Rosengrant MW, Agcaoili-Sombilla M, Perez N D. Global food projections to 2020: implications for investment. 1995, Washington, DC: IFPRI.). By then, my country's wheat demand will be about 140-150 million tons, an increase of 46-56 million tons over the current output, an increase of 49-60% (Gan Jishen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00
Inventor 夏兰琴喻修道马有志张保明
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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