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Method for simultaneously detecting emergence rate and insect resistance purity of transgenic Bt insect-resistant cottonseeds

A technology of germination rate and insect resistance, which is applied in the field of transgenic plant detection to achieve the effects of low cost, simple operation and simple equipment

Inactive Publication Date: 2013-05-22
COTTON RES INST HEBEI ACAD OF AGRI & FOREST SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No matter which of the above-mentioned detection methods can only detect the insect resistance of cotton seeds, but for cotton seed producers and sellers, it is often necessary to use visual inspection and sand bed culture method in addition to the detection of seed insect resistance. , towel culture method, incubator culture method, etc. to detect the germination rate of seeds

Method used

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  • Method for simultaneously detecting emergence rate and insect resistance purity of transgenic Bt insect-resistant cottonseeds
  • Method for simultaneously detecting emergence rate and insect resistance purity of transgenic Bt insect-resistant cottonseeds
  • Method for simultaneously detecting emergence rate and insect resistance purity of transgenic Bt insect-resistant cottonseeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Spray 0.4% kanamycin sulfate aqueous solution to measure the germination rate of transgenic Bt gene insect-resistant cotton seeds while measuring insect-resistant purity

[0019] 1. Preparation of 0.4% kanamycin sulfate aqueous solution: weigh 4 g of kanamycin sulfate powder reagent and 1 L of distilled water, and prepare 0.4% kanamycin sulfate aqueous solution;

[0020] 2. Preparation of wet sand: choose clean river sand and water and mix well at a ratio of 7:1 for later use. Wet sand can form a ball by hand, and it is better to loosen it and then loosen;

[0021] 3. Sowing: Spread the wet sand prepared in step 2 evenly on the bottom of a plastic basin with a diameter of 50 cm and a thickness of 3 cm; evenly place the seeds of transgenic Bt gene insect-resistant cotton Ji 1316 and non-insect-resistant cotton on the wet sand in different pots Variety Jimian 25 has 100 seeds each, with a seed spacing of 1.5 cm; then cover the seeds with wet sand prepared in st...

Embodiment 2

[0024] Example 2: Spray 0.45% kanamycin sulfate aqueous solution to measure the germination rate of transgenic Bt gene insect-resistant cotton seeds while measuring insect-resistant purity

[0025] Prepare 0.45% kanamycin sulfate aqueous solution according to the method in step 1 of embodiment 1, evenly spread 2cm thick wet sand on the bottom of the plastic basin in step 3, in step 4, the cultivation temperature is 30 ℃ and observe the insect resistance on the 15th day Property, all the other steps are identical with embodiment 1.

[0026] The germination rate of the seed of statistical treatment group and control group. Results In the treatment group and the control group, 88 seeds of the Bt-transgenic cotton Ji 1316 seeds germinated and grew into plants, and 86 seeds germinated and grew into plants in the non-Bt gene-resistant cotton Ji-mian 25 seeds, indicating that the Bt-transformed cotton The seed germination rate of Bt-resistant cotton Ji 1316 was 88%, and that of non-...

Embodiment 3

[0027] Example 3: Spraying 0.5% kanamycin sulfate aqueous solution to measure the germination rate of transgenic Bt gene insect-resistant cotton seeds while measuring insect-resistant purity

[0028] Prepare 0.5% kanamycin sulfate aqueous solution by the method in embodiment 1 step 1, select clean river sand and water in step 2 and mix well by 8:1 ratio for subsequent use, in step 3, spread 4cm thick evenly on the bottom of the plastic basin Wet sand, the culture temperature in step 4 is 20°C and the insect resistance is observed on the 15th day, and the rest of the steps are the same as in Example 1.

[0029] The germination rate of the seed of statistical treatment group and control group. Results In the treatment group and the control group, 89 seeds germinated and grew into plants in the Bt gene-transgenic cotton Ji 1316 seeds, and 85 seeds germinated and grew into plants in the non-Bt gene-transgenic cotton Ji Mian 25 seeds, indicating that the Bt-transformed cotton The ...

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Abstract

The invention discloses a method for simultaneously detecting the emergence rate and insect resistance purity of transgenic Bt insect-resistant cottonseeds. The method is characterized by comprising the following steps of: (1) preparing a kanamycin sulfate aqueous solution with concentration between 0.4 and 0.6 percent; (2) preparing wet sand; (3) sowing; and (4) culturing. Under a selective pressure applicable to antibiotics, the method adopts a mechanism that a product which is encoded with a neomycin phosphotransferase (npt II) gene carried by a transgenic Bt plant as a label can inactivate kanamycin sulfate and other aminoglycoside antibiotics to grow normally, but a non-transgenic Bt plant does not contain an npt II gene serving as a label and cannot grow normally, so that the insect resistance purity can be detected while the emergence rate of the transgenic Bt insect-resistant cottonseeds is detected. The method is easy and practical and is low in cost, and the detection accuracy of the method can be 100 percent.

Description

technical field [0001] The invention relates to the detection of transgenic plants, and relates to a method for simultaneously measuring the germination rate and insect resistance purity of transgenic insect-resistant cotton seeds. A method for testing the purity of insect-resistant antibiotic solutions. Background technique [0002] Bt transgenic insect-resistant cotton has been widely used in production and has effectively controlled the harm of cotton bollworm. However, the purity of insect resistance of insect-resistant cotton directly affects the yield and quality of cotton, so the detection of insect resistance of cotton seeds is an indispensable link in cotton production and management. [0003] As early as 1998, Wang Baomin and others from China Agricultural University had begun to use the double-antibody sandwich ELISA detection method to detect the insecticidal crystal protein of Bt cotton, and achieved preliminary results. Wang Kunbo et al. (2000) studied the in...

Claims

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Application Information

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IPC IPC(8): A01C1/02A01G7/00
Inventor 王凯辉郭宝生耿军义刘素恩赵存鹏马玉柱耿香利刘敏彦
Owner COTTON RES INST HEBEI ACAD OF AGRI & FOREST SCI
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