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Method for assessing weedization gene flow risk of transgenic rice and application thereof

A technology of transgenic rice and gene drift, applied in the direction of biochemical equipment and methods, applications, rice cultivation, etc., to achieve the effects of ensuring accuracy, saving detection time and economic costs, and evaluating scientifically and accurately

Active Publication Date: 2020-05-12
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0012] Aiming at the problem that the existing detection and evaluation technology for the risk of transgenic gene drift only considers positive drift, which almost underestimates half of the risk, the present invention provides a method and application for assessing the risk of gene drift in transgenic rice weeds, and designs transgenic rice pollen Forward drift to weedy rice and reverse drift test of weedy rice pollen to transgenic rice, resistance detection and red skin gene Rc detection, to assess the risk of drift

Method used

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  • Method for assessing weedization gene flow risk of transgenic rice and application thereof
  • Method for assessing weedization gene flow risk of transgenic rice and application thereof
  • Method for assessing weedization gene flow risk of transgenic rice and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] A kind of detection method of weedy rice to transgenic rice gene drift rate (reverse gene drift rate), comprises the following steps:

[0082] (1) Random sampling method was adopted to extract 20000 seeds from the harvested transgenic rice seeds.

[0083] (2) Randomly sample 60 grains among the 20,000 grain samples taken, and perform dehulling and grinding, and pass through a 60-mesh sieve;

[0084] Take 0.5mL sample (0.342~0.377g) for DNA extraction;

[0085] The extracted DNA with a concentration of 39.8-77.7 μg / mL is detected for the Rc gene, and whether there are 104bp and 118bp amplified fragments in the sample is used to determine whether there are hybrids formed by the drift from weedy rice to transgenic rice; if there is 104bp and 118bp amplified fragments, then there is a rice seed gene drift in this group of rice;

[0086] The Rc gene amplification primer sequence is:

[0087] Upstream primer: 5′-TTACAGGGGAGCAGAAACA-3′;

[0088] Downstream primer: 5'-TCTTT...

Embodiment 2

[0097] Example 2 Simulating the spatial distribution pattern of the hybrids formed after the transgenic rice resistance gene drift and the hybrids formed after the pollen of weedy rice drifted into the transgenic rice in the transgenic rice progeny seeds

[0098] Conventional rice was used as the experimental material, and rice of the same size was selected to be dyed with red ink to replace transgenic rice. According to the mixing ratio (weight ratio) of dyed rice and non-dyed rice 2.5%, 1.25%, 0.8%, 0.625%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.05%, 0.025%, the total amount is 10kg, Simulate the factors that may lead to the mixing of the two kinds of rice during harvesting, transportation and storage. After the mixed rice was turned over 30 times, samples were taken. The sample size accounted for 20% of the total, that is, 2kg. Spread the sampled rice flat, every 10cm×10cm area, a total of 50 grids, count the number of dyed rice in this area. Four replicates were set for each gen...

Embodiment 3

[0103] Example 3 Determination of the Optimum Sampling Quantity for Transgenic Rice Gene Drift Rate Detection

[0104] After mixing dyed rice and non-dyed rice according to the proportion (weight ratio) of 0.025%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, the simulated process of harvesting, transportation and storage may result in two kinds of rice Mixing factors, turn the mixed rice 30 times, according to the measured thousand-grain weight of the rice seeds (26.1123+0.0820g), randomly weigh 5000 grains, 10000 grains, 20000 grains, 30000 grains, 40000 grains from a total of 10kg of rice grains, 50000 seeds with a weight of 50000 grains, and the number of dyed rice was counted, repeated 16 times, and then the deviation coefficient between the simulated drift rate of dyed rice and non-dyed rice and the drift rate obtained by sampling was less than 0.05 as the standard, and determined under different mixing ratios. the optimal sampling size. Under different mixing ratios of dyed ric...

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Abstract

The invention discloses a method for assessing a weedization gene flow risk of transgenic rice and application thereof. The method comprises the following steps: performing mixed cropping by intercropping or surrounding cropping, respectively harvesting offspring seeds of transgenic rice and weedy rice, performing forward flow detection and seedling resistance screening, and detecting a resistancegene of surviving plants; and performing reverse flow detection on a red husk Rc gene in seeds. According to detection rates of the resistance gene and the Rc gene, a gene flow rate of the transgenicrice is calculated, and a gene flow risk is assessed. The method overcomes the defect that the actual flow risk is underestimated in the past since only the forward flow risk is considered and the reverse flow risk is ignored, and thus, the pollen-mediated gene flow risk of the transgenic rice can be assessed more accurately, thereby providing technical supports for environmental safety evaluation and management in the transgenic rice commercialization process.

Description

technical field [0001] The invention belongs to the field of biosafety assessment, and in particular relates to an assessment method and application of the risk of gene drift of weedy transgenic rice. Background technique [0002] Genetically modified crops have been widely planted and have brought huge benefits to society. At the same time, there are also potential ecological risks. A large number of studies have shown the risk of genetic drift from transgenic crops to conventional varieties and wild relatives mediated by pollen. Although the commercial planting of transgenic rice in the world is still in its infancy, but there is no commercial planting in my country, the resistance gene of transgenic rice can drift into conventional rice through pollen, contaminating conventional rice seeds. Moreover, the resistance gene of transgenic rice can also escape to wild relative weeds through pollen drift-mediated gene flow (Gene flow), so that the resistance gene can continue a...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895A01H1/02A01G22/22
CPCC12Q1/6895A01H1/02A01G22/22C12Q2600/13
Inventor 强胜宋小玲戴伟民张晶旭王敏李雷
Owner NANJING AGRICULTURAL UNIVERSITY
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