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Method for collecting to-be-screened transgenic oilseed rape pollen in large scale by utilizing bees

A rapeseed pollen, large-scale technology, applied in application, honey harvesting, beekeeping, etc., can solve the problem that it is difficult to achieve full coverage, and achieve the effect of wide coverage, easy operation, and good collection randomness

Active Publication Date: 2021-12-31
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, screening for genetically modified ingredients in rapeseed planted in large areas generally uses multi-point single-plant sampling and mixed DNA extraction for detection, but it is difficult to achieve full coverage

Method used

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  • Method for collecting to-be-screened transgenic oilseed rape pollen in large scale by utilizing bees
  • Method for collecting to-be-screened transgenic oilseed rape pollen in large scale by utilizing bees
  • Method for collecting to-be-screened transgenic oilseed rape pollen in large scale by utilizing bees

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Experimental program
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Effect test

Embodiment 1

[0031] The pollen collection test site is the Hanchuan Base of the Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, located in Xujia Village, Chachahu District, Hanchuan City, Hubei Province. The test planting scale of transgenic rapeseed is 3 mu, and the GPS positioning is N30°40'41", E113°43 '39".

[0032] The pollen collection time was March 31, 2019.

[0033] Step 1: Prepare the hive. The bee variety is Apis mellifera, and the honeycomb with the queen bee is placed in the beehive. The beehive has at least 6 boxes of honeycombs, and the honeycombs are commensurate. Before taking the pollen for detection, remove the pollen in the honeycomb, and install a pollinator at the bee entrance and exit of the beehive.

[0034] Step 2: Select the rapeseed field to be screened, and place beehives at a certain distance the night before taking powder to stabilize the bee colony. See the location of beehives figure 1 .

[0035] Step 3: Before 9:00 a.m. in ...

Embodiment 2

[0041] Testing for genetically modified ingredients in canola pollen collected by bees

[0042] Step 1: Pollen DNA extraction. Genomic DNA was isolated and purified according to the instructions of the New Plant Genomic DNA Extraction Kit (CW0531M, Kangwei Century). Dilute the extracted genomic DNA to a concentration of 50ng / μL for later use.

[0043] Step 2: Detection of genetically modified components in pollen - fluorescent quantitative PCR. The primers and probes for the quantitative PCR reaction are shown in Table 2. The PCR reaction system is 20 μL, including 10 μL of TaqMan Universal PCR Master Mix, 0.4 μmol / L of upstream and downstream primers, 0.4 μmol / L of probe, and 2 μL of template DNA. Make up to 20 μL with water. In the blank control, ultrapure water was used instead of template DNA. PCR amplification reaction adopts two-step method, pre-denaturation at 95°C for 3min, 1 cycle; denaturation at 95°C for 30s, annealing at 60°C for 30s, 40 cycles.

[0044]Result...

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Abstract

The invention provides a method for collecting to-be-screened transgenic oilseed rape pollen in a large scale by utilizing bees, and belongs to the technical field of transgenic plant detection. The method comprises the steps as follows: a beehive is prepared, a plurality of honeycombs with queen bees are arranged in the beehive, and a pollen remover is installed at an entrance of the beehive; the beehive is placed in an oilseed rape field at the full-bloom stage, a pollen receiving box is placed below the pollen remover of the beehive, and worker bees start to take pollen; and the pollen in the pollen receiving box is collected after the pollen taking lasts for 4-6 hours. The method for large-scale screening of the transgenic components of the oilseed rape planted in the field by utilizing the bees is simple and convenient to operate, the labor cost can be reduced, and meanwhile, compared with a common multi-point sampling detection method, the method disclosed by the invention is accurate and reliable in detection result, wider in coverage and better in randomness.

Description

technical field [0001] The invention belongs to the technical field of transgenic plant detection, and in particular relates to a method for large-scale collection of transgenic rape pollen to be screened by bees. Background technique [0002] In the supervision of GMOs, large-scale screening of GMO ingredients in key units, key regions, and key crops is a necessary means. Sampling of crops in the field generally uses multi-point sampling and mixed extraction. According to the method stipulated in "National Standards of the People's Republic of China Ministry of Agriculture Announcement No. 2031-19-2013, Sampling for Detection and Sampling of Genetically Modified Plants and Their Product Components", according to the sampling plan and quantity, a single plant sample was drawn using the diagonal method, and each plant was sampled At the same time, use a puncher to punch holes in the leaves of the plants or cut the leaves of the plants with scissors to obtain a duplicate samp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K59/00
CPCA01K59/00
Inventor 曾新华吴刚袁荣王淼刘芳王盼睇朱莉李晓飞李均
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI