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A method for auxiliary identification of wheat canopy temperature traits and its special primer set

A wheat, specific primer technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve problems such as deviation, canopy temperature vegetation coverage index, and high technical requirements for the determination process.

Active Publication Date: 2022-05-10
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the physiological traits of drought resistance are easily affected by the environment, and the determination process requires high technology
For example, the temperature of the canopy is greatly affected by the planting density and vegetation coverage index, and the measurement may cause a deviation of more than 1°C due to different infrared projection angles.

Method used

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  • A method for auxiliary identification of wheat canopy temperature traits and its special primer set
  • A method for auxiliary identification of wheat canopy temperature traits and its special primer set
  • A method for auxiliary identification of wheat canopy temperature traits and its special primer set

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1. Acquisition of SNP marker AX94392210 of wheat canopy temperature-related genes

[0075] First, the acquisition of DH groups

[0076] Zhongmai 895 is a semi-winter multi-spike type mid-late-maturing cultivar that was bred by the Institute of Crop Science, Chinese Academy of Agricultural Sciences and Cotton Research Institute, Chinese Academy of Agricultural Sciences, with Zhoumai 16 as the female parent and Liken 4 as the male parent. It has the characteristics of long functional period, strong tillering ability and fast grouting speed. In September 2009, it passed the approval of the southern part of the national Huanghuai wheat area. In the three-year variety comparison test and field demonstration from 2013 to 2015, Zhongmai 895 showed the characteristics of high yield and wide adaptability, disease resistance and lodging resistance, and high temperature resistance in the later stage of grain filling.

[0077] Yangmai 16 is the variety with the largest pl...

Embodiment 2

[0103] Example 2, the establishment of the method

[0104] 1. Extract the genomic DNA of the leaves of the wheat to be tested, and dilute to obtain a template solution. The DNA concentration in the template solution was approximately 20 ng / μL.

[0105] 2. Carry out KASP.

[0106] Primer working solution: Take primer A, primer B and primer C, add sterile ultrapure water to 100 μL, and mix thoroughly. In the primer working solution, the concentrations of primer A, primer B and primer C were all 100 μM.

[0107] KASP 2×Master Mix is ​​a product of LGC Corporation (Item No. KBS-1016-002). KASP 2×Master Mix contains fluorescent probe A, fluorescent probe B, quenching probe A, quenching probe B, high-fidelity Taq enzyme, dNTP, Mg 2+ Wait. The sequence of fluorescent probe A is 5'-GAAGGTGACCAAGTTCATGCT-3', and the 5' end is connected with a FAM fluorescent group. The sequence of fluorescent probe B is 5'-GAAGGTCGGAGTCAACGGATT-3', and the 5' end is connected with a HEX fluoresce...

Embodiment 3

[0120] Embodiment 3, application KASP primer set detects the family in the DH population

[0121] Wheat to be tested: 14 families in the DH population obtained in Example 1.

[0122] 1. Detect the genotype of the wheat to be tested based on the specific SNP, the method is the same as in Example 2.

[0123] 2. Detect the canopy temperature of the wheat to be measured.

[0124] The results are shown in Table 1.

[0125] Table 1 Segregation of AX94392210 marker allelic variation in Yangmai 16 / Zhongmai 895DH population

[0126]

[0127] The canopy temperature of CC genotype wheat was lower than that of AC / AA genotype wheat.

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Abstract

The invention discloses a method for assisting identification of wheat canopy temperature traits and a special primer set thereof. The present invention provides a specific primer set, which consists of primer A shown in sequence 1, primer B shown in sequence 2 and primer C shown in sequence 3. The invention also protects a method for identifying the canopy temperature trait of the wheat to be tested, comprising the following steps: detecting the genotype of the wheat to be tested based on a specific SNP; the canopy temperature of the CC genotype wheat is lower than that of the AA genotype wheat. The invention also protects a method for identifying the canopy temperature traits of the wheat to be tested, which includes the following steps: using the genomic DNA of the wheat to be tested as a template, using the primer set to perform KASP, performing fluorescence scanning, and determining that the wheat to be tested is based on the specificity The genotype of SNP, the canopy temperature of CC genotype wheat was lower than that of AA genotype wheat. The invention can be used for screening wheat with excellent canopy temperature properties, and plays an important role in cultivating drought-resistant wheat varieties.

Description

technical field [0001] The invention relates to a method for assisting identification of wheat canopy temperature traits and a special primer set thereof. Background technique [0002] Canopy temperature, vegetation cover index and chlorophyll content are important physiological traits related to drought resistance in wheat. The canopy temperature refers to the average surface temperature of stems, leaves and other organs at different heights of the crop canopy. Under drought stress, the crop canopy absorbs solar radiation, some of which are used for photochemical reactions in photosynthesis, some are used for radiation fluorescence, and the remaining part is converted into heat energy, which increases the temperature of the plant canopy. Green leaves can use more solar radiation for photochemical reactions in photosynthesis, release less heat, and the canopy temperature is significantly lower than that of high temperature-sensitive wheat, resulting in higher biomass and yi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 肖永贵杨梦娇武玉莹何中虎夏先春
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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