Indel molecular marker for identifying rape female sterile germplasm resource mutants and application of Indel molecular marker

A molecular marker and mutant technology, applied in the field of rape biology, can solve the problems of complexity, backward research on female sterility mechanism, and difficulty in female sterility phenotype, achieving good repeatability, improving breeding efficiency, and broadening the selection range. Effect

Active Publication Date: 2022-03-25
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004]The occurrence of abnormal male organs can improve the seed setting rate by supplementing foreign pollen, but the developmental defects of female organs cannot be rescued
[0005]Because the pheno

Method used

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  • Indel molecular marker for identifying rape female sterile germplasm resource mutants and application of Indel molecular marker
  • Indel molecular marker for identifying rape female sterile germplasm resource mutants and application of Indel molecular marker
  • Indel molecular marker for identifying rape female sterile germplasm resource mutants and application of Indel molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Acquisition of Indel molecular markers for identification of rapeseed germplasm mutants:

[0042] 1. Acquisition and phenotype of female sterile germplasm mutants in rapeseed

[0043] The applicant purchased rapeseed 9538R from Zhongken Jinxiu Huanong Wuhan Science and Technology Co., Ltd., and the applicant found that the rapeseed was a female sterile mutant. When blooming normally, compared with normal single plant ( figure 1 Middle A, E), the siliques of mutants are shorter ( figure 1 Middle A), seed setting rate reduced by 93.4% ( image 3 , Figure 4 ). When the mutants were used as pollen donors, the seed-setting rate was in the normal range; when the mutants were used as pollen receivers, the pollen of the conventional cultivars 'westar' and 'Zhongshuang11' were spread on the stigma, and the seed-setting rates were also very high. Low. The male organ of the mutant was no different from that of the wild type; the pollen viability of the mutant was 97.45%, wh...

Embodiment 2

[0066] The use and determination of Indel molecular marker primers for the identification of rapeseed germplasm mutants:

[0067] Step 1.CTAB method extracts the DNA of the plant to be tested;

[0068] Step 2. Use the DNA obtained in step 1 as a template, add two positive control DNAs (9538R) and two negative control DNAs (ZS11) to each plate, and use specific molecular markers to carry out PCR amplification. The primers are as shown in Example 1 , specifically: ID4-62 and SS3-C6-13.

[0069] The reaction system for PCR amplification is 10ul, including 10X Taq buffer 1ul, 0.5U / ul Taq enzyme 1ul, 10mMdNTP 0.5ul, 50ng / ul DNA template 1ul, 10uM forward and reverse primers 0.5ul each, ultrapure water 5.5ul.

[0070] The PCR amplification program was 94°C for 5min, 94°C for 30s, 60°C for 30s-0.5°C / c, 72°C for 30s, 9X, 94°C for 30s, 56°C for 30s, 72°C for 30s, 29X, 72°C for 10min, and 25°C for 5min.

[0071] Step 3. The PCR amplification product is detected, and the band type of Z...

Embodiment 3

[0073] The application of Indel molecular marker primers for the identification of rapeseed germplasm resource mutants in the screening of rapeseed seed setting rate is as follows:

[0074] (1) Planting F from Wuhan 2 4 female sterile mutants and 4 normal plants were selected from the segregation population (parent ZS11 x 9538R), and the number of grains per silique of 10 consecutive siliques in the middle of the main stem of each plant was counted.

[0075] (2) The genotype distribution of four molecular markers significantly associated with female sterility in 4 female sterile materials and 4 normal materials was examined.

[0076] The results show that the combination of molecular markers provided by the present invention can completely distinguish these 8 parts of materials, and the number of grains per silique of 10 consecutive siliques in the middle part of the main stem of the female sterile material is about 0.95, while that of normal plants is in the middle part of th...

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Abstract

The invention belongs to the field of oilseed rape biology, and particularly relates to an Indel molecular marker for identifying oilseed rape female sterile germplasm resource mutants and application of the Indel molecular marker. The applicant discovers a female sterile brassica napus, and genetic analysis shows that the female sterile character is controlled by two recessive nuclear genes, one of the recessive nuclear genes is located in the ChrA07 30-kb interval and is closely linked with the recessive nuclear genes is ID4-62 and ID4-66, and the other of the recessive nuclear genes is located in the ChrA07 30-kb interval and is closely linked with the recessive nuclear genes. The other one is located in a ChrC06 67-kb interval, and markers closely linked with the other one are SS3-C6-9 and SS3-C6-13. The primer group ID4-62 and the primer group SS3-C6-13 are used as an identification combination, the female sterile material of the brassica napus can be identified, and a material basis is provided for studying a plant female gametogenesis mechanism and applying female sterile characters by exploring the material.

Description

technical field [0001] The invention belongs to the field of rapeseed biology, and in particular relates to an Indel molecular marker for identification of rapeseed female sterile germplasm resource mutants and its application. Background technique [0002] Brassica napus is an important oilseed crop in the world, which originated about 7500 years ago from the spontaneous hybridization of Chinese cabbage and Brassica oleracea (Chalhoub et al., 2014). Heterosis is a biological phenomenon ubiquitous in nature, and it has gradually become an effective measure to increase crop yield in recent years (Kempe & Gils, 2011; Wang & Deng, 2018). Crop yield is generally composed of the number of spikes per unit area, the number of grains per spike, and the weight of grains (Chen et al., 2007). At the same time, the seed setting rate of plants is also one of the important factors that constitute yield (Zuo and Li, 2014; Xiang et al ., 2019). There are many factors affecting the seed se...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11A01H1/04
CPCC12Q1/6895A01H1/04C12Q2600/156C12Q2600/13
Inventor 涂金星秦培高金祥
Owner HUAZHONG AGRI UNIV
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