Gene transfer method for promoting insect resistance of soybean

A technology of transgenic and insect resistance, applied in the field of transgenic, can solve the problem of difficulty in obtaining regenerated plants, and achieve the effect of improving breeding efficiency and inhibiting growth and development.

Inactive Publication Date: 2006-12-27
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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AI Technical Summary

Problems solved by technology

In the study of transferring the target gene on soybean, people mainly use Agrobacterium-mediated or gene gun method, but since the tissue culture of soybean has always been a difficult problem to break through, it is very difficult to use Agrobacterium-mediated or gene gun method to transgene in soybean. It is difficult to obtain regenerated plants, but the use of pollen tube passage method can directly transform fertilized eggs or zygotes of soybeans to obtain transgenic seeds, and it is easy to obtain regenerated plants from seeds, overcoming the difficulty of tissue culture
However, there are not many reports in the literature at home and abroad on the use of pollen tube passage method to transfer the target gene into soybean.

Method used

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  • Gene transfer method for promoting insect resistance of soybean
  • Gene transfer method for promoting insect resistance of soybean
  • Gene transfer method for promoting insect resistance of soybean

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] 1) Transform soybean varieties Bogao and Nannong 73-935 (soybean varieties Both Bogao and Nannong 73-935 are well-known public varieties, which are sold by the National Soybean Improvement Center (Nanjing).

[0028] 2) From the T1 generation, identify the kanamycin resistance of the transgenic plants at the seedling stage, and further identify the kanamycin-resistant plants by PCR. The PCR amplification system contains 0.25mM dNTP, MgCl2, 1U Taq enzyme, 0.5μM Primer , 100-200ng DNA, total volume 25μl, PCR amplification reaction conditions: pre-denaturation at 95°C for 5min, followed by 95°C for 15s, 52°C for 59s, 72°C for 2min, 30 cycles, 72°C for 10min, and the PCR identification result was selected as Positive material ( figure 1 ).

[0029] 3) In the T2 generation, the kanamycin resistance identification is still performed on the transgenic plants at the seedling stage, and the kanamycin-resistant plants are identified by PCR. The PCR amplification system is the sa...

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Abstract

The invention relates to a transgenic method to improve soybean insect-resistance, which belongs to soybean transgenic breeding method. The invention includes switching pyranone synthetase gene g2ps1 into soybean by pollen tube port method, performing chanamyn certification to transgenic plant seedling stage, screening plant which can resist Kana, further performing PCR and RT-PCR certification, and breeding transgenic insect-resistance soybean materials by co-operating insect-resistance determination of indoor tobacco cutworm by feeding. The consequence of breeding insect indicates that a distinctive difference existed ( distinctive level P>P0.05) between the larval weight and pupal weight fed by two transgenic materials and those fed for comparison, compared with comparison, the transgenic materials can distinctively reduce the growth of tobacco cutworm.

Description

1. Technical field [0001] The present invention---a transgenic method for improving soybean insect resistance, belongs to soybean transgenic breeding method. 2. Technical background [0002] Soybean is rich in protein and oil. It is an important source of protein and edible oil for people, and it is also an important industrial raw material. It has a long history of planting. Among them, soybean leaf-eating pests are the main pests in soybean production in my country. Soybean leaf-eating pests are harmful to soybeans in all soybean producing areas in my country, especially in the southern and Huanghuai soybean areas. In 1990, the bean hornworm broke out in Xinxiang, Henan (Liu Zhen, 1991); in 1992-1993, the cotton bollworm occurred in North China; in 1966, the large bridge-builder broke out in Peixian County; In 1993, the bean leaf roller broke out; in 1994 and 1998, the occurrence of Spodoptera litura was relatively large. In 2001 and 2002 alone, more than 800,000 mu of s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N15/52C12N15/82C12Q1/68A01H1/04
CPCY02A40/146
Inventor 喻德跃张曼孙海燕阚贵珍王慧
Owner NANJING AGRICULTURAL UNIVERSITY
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