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Method for identifying resistance function of aphid-resisting genes through transforming hairy roots of soybean

A technology of aphid-resistant gene and hairy root, which is applied in the field of identifying the resistance function of the aphid-resistant gene by transforming soybean hairy root, can solve problems such as difficult control of aphids, and achieve simple and convenient implementation, accurate identification, convenient and reliable identification Effect

Inactive Publication Date: 2013-10-09
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

According to the literature, soybean aphids are specialized and overwinter in the roots of several Rhamna plants; the identification of soybean aphid resistance and the reproduction of aphids are all on intact plants, and the propagation of aphids in intact plants has the disadvantages of being difficult to control.

Method used

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Examples

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Embodiment 1

[0024] The soybean receptor is selected from the aphid-susceptible variety Williams82, and the method for identifying the function of the Rag1 aphid-resistant gene using soybean hairy roots, the steps are as follows:

[0025] 1) Obtain soybean cotyledons: select healthy and mature soybean grains without disease spots, rinse them with clean water, disinfect with 75% alcohol for 30 seconds, and use 0.1% Ca(ClO) 2 Disinfect for 20 minutes, shake about 5 times during the period, wash with sterile water to remove Ca(ClO) 2 Residue, washed 5 times, inoculated in the germination medium, light 16h / day, 25±3℃; the composition of the germination medium is: MSB+7g / L agar+30g / L sucrose, the pH value is 5.8; The cotyledon is the cotyledon formed after 5 days of aseptic germination of mature soybean seeds.

[0026] 2) Agrobacterium rhizogenes infection and transformation to induce hairy roots: cut the cotyledon from the hypocotyl with a sterile scalpel blade, and cut a wound with a radius ...

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Abstract

The invention discloses a method for identifying the resistance function of aphid-resisting genes by transforming hairy roots of soybean and relates to the field of methods for crop tissue culture and transgenosis. According to the method, the hairy roots are cultured by infecting mature seeds of the soybean by agrobacterium rhizogenes, and the effect of the aphid-resisting genes to resist against soybean aphids is verified through the transgenic hairy roots. The method mainly includes a cotyledon obtaining and culturing technique for the mature seeds of the soybean, an infection transformation technique for the agrobacterium rhizogenes, a degerming and continuous culturing technique for the transgenic hairy roots, a transgenic hairy root aphid inoculation technique, and an identification technique for the aphid-resisting functional genes of the hairy roots. The method is simple, easy to operate, free of limitation of seasons and capable of effectively and quickly identifying the effect of the target genes to resist against the soybean aphids through the transgenic hairy roots and providing reference for resistance identification of other crops.

Description

technical field [0001] The invention relates to plant tissue culture and transgenic methods, in particular to a method for quickly identifying whether a target gene is resistant to soybean aphids through soybean hairy roots. Background technique [0002] Soybean has been cultivated in my country for a long time and has always been an important economic crop. However, soybeans are infested by many diseases and insect pests during the growth process, resulting in a decline in its yield and quality. Among them, soybean aphid is a kind of pest that is more harmful to soybean growth, resulting in reduced photosynthetic rate of soybean leaves, curly shrinkage, plant dwarfing, and reduced number of pods. Other viruses carried by it (soybean mosaic virus, dwarf disease etc.) exacerbated the loss of soybean yield, so it is urgent to excavate aphid-resistant genes from existing soybean germplasm resources. Kim et al. locked Rag1 in an interval of about 115kb through fine mapping, and...

Claims

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Application Information

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IPC IPC(8): A01K67/033A01H4/00
Inventor 钟云鹏程琳静闫军辉王彪武天龙
Owner SHANGHAI JIAO TONG UNIV
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