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KASP molecular marker of wheat powdery mildew resistance gene Pm21 derived from haynaldia villosa and application of KASP molecular marker

A powdery mildew resistance gene and molecular marker technology, which is applied in the field of biotechnology and genetic breeding, can solve the problems affecting the efficient utilization of Pm21 gene, and achieve the effects of accelerating the utilization of high-efficiency breeding, high-throughput detection and tracking, and improving the utilization of transgenics

Active Publication Date: 2020-06-05
YANTAI UNIV
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Problems solved by technology

However, Pm21 is a powdery mildew resistance gene with important breeding value. Although it has been cloned, there are no reports of KASP markers available for breeding, especially the reports of functional KASP markers, which seriously affects the efficient use of Pm21 gene in wheat genetic improvement.

Method used

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  • KASP molecular marker of wheat powdery mildew resistance gene Pm21 derived from haynaldia villosa and application of KASP molecular marker

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Embodiment 1

[0029] Example 1 Wheat Powdery Mildew Resistance Gene Pm21 The acquisition and application of KASP molecular marker KASP-Pm21

[0030] 1. Preparation of DNA amplification template

[0031] wheat powdery mildew resistance Pm21 The vector of the gene is the vector constructed in He et al. (2018), which is ready for use. The rest of the DNA amplification templates were extracted by CTAB method from 1 T. villosa and 2 Pm21 Transgenic lines, 8 carrying Pm21 6VS translocation lines and 20 do not carry Pm21 The genomic DNA of the one-leaf stage leaves of common wheat, the specific operation steps are as follows:

[0032] (1) Cut fresh leaves and freeze and grind them with liquid nitrogen. Take 0.5g of ground powder for each sample in a 2mL centrifuge tube;

[0033] (2) Add 600 μL of 65°C preheated CTAB extract to each sample, and bathe in 65°C water bath for 45 minutes;

[0034] (3) Cool the centrifuge tube to room temperature, add the same amount of chloroform-isoamyl alcohol...

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Abstract

The present invention discloses a KASP molecular marker of a wheat powdery mildew resistance gene Pm21 derived from haynaldia villosa and an application of the KASP molecular marker. The KASP molecular marker is KASP-Pm21 and primers of the KASP molecular marker KASP-Pm21 comprise a haynaldia villosa allelotype upstream primer KASP-Pm21-F, a wheat allelotype upstream primer KASP-Pm21-H and a shared downstream primer KASP-Pm21-C; a nucleotide sequence of the haynaldia villosa allelotype upstream primer KASP-Pm21-F is shown in SEQ ID NO:1; a nucleotide sequence of the wheat allelotype upstream primer KASP-Pm21-H is shown in SEQ ID NO:2; and a nucleoside sequence of the shared downstream primer KASP-Pm21-C is shown in SEQ ID NO:3. The provided KASP molecular marker KASP-Pm21 of the wheat powdery mildew resistance gene Pm21 derived from the haynaldia villosa can realize efficient tracking and detection of the Pm21 gene and is applied to molecular marker assisted selection breeding of the wheat powdery mildew resistance Pm21 gene.

Description

technical field [0001] The invention relates to the fields of biotechnology and genetic breeding, in particular to a wheat powdery mildew resistance gene derived from A. Pm21 KASP molecular markers and their applications. Background technique [0002] Wheat (Triticum aestivum L., 2n = 42, AABBDD) is one of the most important food crops in my country, and maintaining high and stable yields of wheat is very important to ensure national food security. Among the many factors affecting the yield and quality of wheat, the occurrence of diseases is very important. Wheat powdery mildew is a leaf disease caused by powdery mildew brucei, which can generally cause 10-15% yield loss of wheat and affect wheat quality to a certain extent (Singh et al. Disease impacton wheat yield potential and prospects of genetic control. Annu Rev Phytopathol 2016, 54, 303–322.). In order to prevent and control wheat powdery mildew, several measures such as field agricultural management, spraying pest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6895C12Q1/6851C12Q2600/13C12Q2531/113
Inventor 马朋涛何华纲宋建成杜文晓王文瑞贾梦淑张旭梁萧武莉茹
Owner YANTAI UNIV
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