KASP mark for detecting powdery mildew resistant gene of aegilops tauschii and application

A technology of powdery mildew resistance gene and goat grass, which is applied in the field of wheat genetics and molecular breeding, can solve the problems that are difficult to apply in breeding, and achieve the effect of simple operation and improved efficiency

Active Publication Date: 2018-02-02
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And some powdery mildew resistance genes, such as: Pm12, Pm13, Pm16, Pm18, Pm20 , although it shows high resistance to wheat powd

Method used

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  • KASP mark for detecting powdery mildew resistant gene of aegilops tauschii and application
  • KASP mark for detecting powdery mildew resistant gene of aegilops tauschii and application
  • KASP mark for detecting powdery mildew resistant gene of aegilops tauschii and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Construction and fine positioning of mapping populations

[0028] 1.1 Materials

[0029] The disease-resistant A. tachyphylla 2147 was used as the parent to cross the susceptible A. tachyphyllum AL8 / 78 to obtain F 1 , 4299 strains of F 2 , further self-intersection, get the corresponding F 2:3 family line;

[0030] 1.2 Method

[0031] 1.2.1F 2 Identification of Powdery Mildew Resistance in Population Seedling Stage

[0032] Will F 2 The populations were planted in small plastic pots of 6cm×6cm and placed in light incubators. The conditions were set as follows: temperature 19°C, humidity 80%, light 2000LUX, 12h; temperature 17°C, humidity 80%, darkness, 12h. Inoculate with E09 strain at the one-heart-one-leaf stage. The inoculation method is the sweeping method. Investigate the condition after 7-10 days. Disease resistance classification adopts 6-level classification method:

[0033] Type 0, immunity, no lesions on plants;

[0034] 0; type, near imm...

Embodiment 2

[0060] Example 2 Ml2147 Map-based cloning and development of functional markers

[0061] 2.1 Materials

[0062] Using the 9 key recombinants screened in the process of fine mapping, we further designed markers and encrypted the linkage map.

[0063] 2.2 Method

Embodiment 1）

[0065] 2.2.2 Developing markers, encrypting fine-positioning linkage map (see Example 1 for specific methods of developing and screening markers)

[0066] 2.2.3 Develop feature flags

[0067] (1) After finding a candidate gene, design primers to amplify the full length of the gene based on the gene sequence information 2147-qc-1 , the upstream sequence of the primer 2147-qc-1 F is: CGGATGGACGCGCCTTCG, downstream sequence 2147-qc-1 R is: GTACTCTAGTGGCACCTTAAACCGG. The gene fragments in 180 natural populations of A. tachyphylla were amplified, sequenced and compared.

[0068] The specific steps for amplifying the gene are as follows:

[0069] a. take wheat or goat grass DNA as PCR amplification template, with 2147-qc-1 As primers, carry out PCR amplification, and the reaction system is 50 μl. The above reaction system specifically includes: 50-100ng / μl DNA 4μl, 25μl 2×GC buffer (containing Mg 2+ ), 4 μl ldNTP, 1.0 μl each of upper and lower primers, 1 μl LA-Taq DNA po...

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Abstract

The invention relates to the field of wheat genetics and molecular breeding thereof, in particular to development and application of a function mark of a powdery mildew resistant gene of aegilops tauschii. On the basis that the powdery mildew resistant gene M12147 of aegilops tauschii is subjected to map position clone, special structure sequences of candidate genes are used, a function mark SDAU-kasp2147 is developed, and verification is conducted by using 180 or more parts of aegilops tauschii natural germplasm and genetic offspring materials. The result proves that the function mark has theadvantage that the function mark is quick, direct and high in flux, is high in specificity, can complete screening and phenotype prediction work of the disease-resistant gene in mass materials in a short time, and can effectively serve for breeding work of wheat powdery mildew resistance.

Description

technical field [0001] The invention relates to the field of wheat genetics and its molecular breeding, in particular to a functional marker for detecting the powdery mildew resistance gene of A. tachyphylla and its application. Background technique [0002] Wheat powdery mildew is caused by wheat powdery mildew ( Blumeriagraminisf.sp.tritici ) is one of the most important wheat diseases in the world, seriously affecting the quality and yield of wheat. In recent years, with the adjustment of the farming system and the continuous change of the climate, the disease has become more and more serious in several major wheat regions such as Southwest my country, North China, Northwest China and Northeast China. Therefore, the control of wheat powdery mildew is of great significance to improve wheat quality and increase wheat yield. [0003] At present, there are more than 70 wheat powdery mildew resistance genes that have been officially named internationally. With the variation...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 孔令让王宏伟唐恒胡立芹宋时洋
Owner SHANDONG AGRICULTURAL UNIVERSITY
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