Functional marker associated with drought resistance of rape in seedling stage and application of functional marker

A technology for drought resistance and rapeseed seedlings, which is applied in the field of rapeseed breeding and molecular biology, can solve the problems of drought resistance identification, such as heavy workload, long cycle, and easy to be affected by the environment, so as to improve breeding efficiency, facilitate operation, and facilitate popularization and application Effect

Inactive Publication Date: 2016-08-24
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Another object of the present invention is to provide the application of a functional marker highly correlated with drought resistance of rapeseed in rapeseed drought resistance breeding. This method effectively solves the problems of large drought resistance identification workload, long period, and problems in conventional breeding methods. Susceptible to problems such as environmental influences, the accuracy rate is as high as 92.5%

Method used

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  • Functional marker associated with drought resistance of rape in seedling stage and application of functional marker
  • Functional marker associated with drought resistance of rape in seedling stage and application of functional marker
  • Functional marker associated with drought resistance of rape in seedling stage and application of functional marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A preparation method of functional marker EE476451-465 associated with drought resistance in rapeseed seedling stage (Lu Guangyuan et al., Associative transcriptomics study dissects the genetic architecture of seed glucosinolate content in Brassicanapus, DNA Research, 2014, 21:613-625), the steps are:

[0034] In this example, taking 101 rapeseed varieties with different genetic sources and large differences in drought resistance as an example, a natural population was constructed, and the method for obtaining molecular markers associated with drought resistance was described in detail, as follows:

[0035] (1) Group construction:

[0036] 101 rapeseed varieties with different genetic origins were used as research materials (published, see literature for details: Lu Guangyuan et al., Associative transcriptomics study dissects the genetic architecture of seed glucosinolate content in Brassica napus, DNA Research, 2014, 21(6):613- 625), grown to 1-2 true leaves in a green...

Embodiment 2

[0066] The application of a functional molecular marker highly correlated with drought resistance at seedling stage of rapeseed in drought-resistant breeding comprises the following steps:

[0067] Select 20 parts of drought-resistant variety materials (the average drought survival rate is 93%) from 101 varieties resources in Example 1, and 20 parts of drought-sensitive materials (the average drought survival rate is 30%), using specific PCR Mark EE476451-465 for identification, the steps are as follows:

[0068] (1) extracting the DNA of the rapeseed strain;

[0069] (2) Perform PCR amplification with functional marker EE476451-465, and the primer sequence is:

[0070] Forward primer EE476451-89F: 5′-TGAGGCTGATAGGATACTGG-3′

[0071] Reverse primer EE476451-554R: 5'-CCACGGGCACCTACATTA-3';

[0072] (3) PCR reaction system: the total volume is 10 μL, and the specific components are as follows:

[0073] DNA template (25ng / μl)

1.0 μL

Forward primer (50ng / μl...

Embodiment 3

[0085] The application of a functional marker highly correlated with drought resistance at seedling stage of rape in drought-resistant breeding of rapeseed comprises the following steps:

[0086] (1) Using the drought-resistant rape variety 'Apex' (derived from Examples 1 and 2) and the poor drought-resistant rape variety 'Zhongshuang 5' (bred by the Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, the seeds can be purchased in the market) to construct the F 2 The isolated populations are used as research materials. Samples are taken at the seedling stage, and the detection range is further expanded by using the functional marker EE476451-465. The detection method is the same as that in Example 2.

[0087] (2) It is predicted that the drought resistance of individual plants with 465-bp specific band type (positive) will be significantly improved, while the drought resistance of individual plants without 465-bp specific fragments (negative) will be ...

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Abstract

The invention discloses a functional marker associated with drought resistance of rape in the seedling stage and application of the functional marker. The sequence of the functional marker is as shown in SEQ ID NO:1, and the molecular weight is 465 bp. Application of the functional marker in drought-resistant seedling breeding of rape includes the steps that firstly, the DNA of a rape plant is extracted; secondly, PCR amplification is conducted, wherein the primer sequence F is 5'-TGAGGCTGATAGGATACTGG-3', and the primer sequence R is 5'-CCACGGGCACCTACATTA-3'; thirdly, after the amplification product is subjected to electrophoretic separation with agarose gel with the concentration being 1.5%, it is predicted that draught resistance of the rape plant is good when a specific band of the functional marker with the molecular weight being 465 bp occurs, and it is predicted that draught resistance of the rape plant is poor when the specific band of the functional marker with the molecular weight being 465 bp lacks. The new genetic marker is provided for genetic improvement of drought resistance of the rape in the seedling stage, the problems that a conventional seedling breeding method is large in drought resistance identification workload, the period is long and the method is influenced by the environment easily are solved, selection efficiency of drought resistance seedling breeding of rape is remarkably improved, and the seedling breeding progress of new varieties is accelerated.

Description

technical field [0001] The invention belongs to the fields of rapeseed breeding and molecular biology, and more specifically relates to a functional marker associated with rape seedling drought resistance, and also relates to the application of the functional marker in rapeseed drought-resistant breeding. Background technique [0002] Drought and lack of water resources are the biggest constraints on agricultural production in the world, and the loss of crop yield due to drought exceeds the sum of other natural disasters. Rapeseed is the most important winter oil crop in my country. It is mainly planted in the Yangtze River Basin. However, in recent years, seasonal droughts have occurred frequently in this area, seriously affecting the yield of rapeseed. Studies have shown that rapeseed seedling stage is one of the most sensitive periods of drought, which can cause 41% yield loss after drought (Hashem et al. Drought stress effects on seed yield, yield attributes, growth, cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 陆光远黄倩赵永国张学昆乔醒
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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