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Soybean aphid-resistant gene as well as molecular marker and application thereof

A molecular marker and anti-aphid technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of low efficiency, long cycle, difficult breeding, etc., and achieve the effects of improving screening efficiency, shortening the breeding period, and high reliability

Pending Publication Date: 2022-05-06
INST OF CROP SCI CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Local varieties such as Qingpi Pingding, Duludou and wild soybean 85-32 have been screened and identified in China. These resources are difficult to be directly used in breeding due to poor agronomic traits
Traditional breeding methods are limited by long periods and low efficiency. However, molecular marker-assisted breeding technology uses the characteristics of close linkage between molecular markers and genes that determine target traits. By detecting molecular markers, the existence of target genes can be detected, which can significantly Improve breeding efficiency
However, there are relatively few domestic studies on soybean aphid resistance, lack of excellent resistance resources and molecular markers that can be used for breeding, and the discovered resistance resources cannot be well utilized.

Method used

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  • Soybean aphid-resistant gene as well as molecular marker and application thereof
  • Soybean aphid-resistant gene as well as molecular marker and application thereof
  • Soybean aphid-resistant gene as well as molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Population construction and phenotyping

[0061] 1. Raw materials: Founder's moss soybean (sourced from the National Crop Germplasm Bank, unified number ZDD00236), judged as aphid resistance level 1 according to the resistance grading standard in Table 2, and the resistance does not depend on the number of soybean aphids.

[0062] Table 2 Grading standards for identification and grading of resistance to soybean aphids

[0063]

[0064]

[0065] 2. Use Fangzheng moss-eating bean to construct a population, and locate the aphid-resistant gene by map-based cloning: use Fangzheng moss-eating bean and the highly susceptible aphid variety Beifeng No. 9 to cross ( figure 1 ), and then self-crossed to construct a genetic segregation population resistant to susceptible aphids. The results of resistance identification showed that F 1 Plants showed resistance to aphids, F 2 The segregation ratio of aphid-resistant plants and aphid-susceptible plants in the generation popula...

Embodiment 2

[0067] Localization of resistance genes and determination of linked molecular markers

[0068] Using the BSA (cluster segregation analysis) method, the DNA of aphid-resistant and susceptible plants was mixed to construct a mixed pool, and the 580 pairs of SSR primers covering the whole soybean genome in the SoyBase database were screened to have polymorphisms between parents and meet extreme mixed pools According to regular primers, three markers, Satt114, Sct_033 and Satt_335, were found to be resistance-linked markers. Use these 3 linked markers and encrypt other markers to identify F 2 Generation plants and by F 2 F 5 The genotype of the generation plants was combined with the phenotype for initial mapping, and the initial mapping interval was between markers M1133 and M1184, with a total of 1.01Mb. In order to further narrow the interval, the encryption mark identifies F 5:6 The genotype of the generation plant, combined with the phenotype data, screened the exchanged ...

Embodiment 3

[0075] Validation of molecular markers in progeny populations

[0076] Genomic DNA from the leaves of Fangzheng Moshidou, Beifeng 9 and the offspring of the group were extracted by CTAB method and PCR amplified. The system is 20μl, including 1μl of genomic DNA, the concentration is 2mmol·L -1 2 μl each of the upstream and downstream primers, ddH 2 O 4 μl, 2хM5 HiPerplus Taq HiFi PCR Mix (with blue dye) 10 μl (Beijing Jumei Biotechnology Co., Ltd., Cat. No. MF002-plus-01). The PCR program was denaturation at 95°C for 3 min, denaturation at 95°C for 25 s, annealing at 58°C for 25 s, extension at 72°C for 10 s, and 34 cycles; extension at 72°C for 5 min, and finally storage at 4°C. The above PCR reaction was amplified on a PCR thermal cycler from ABI (Applied Biosystems, USA).

[0077] The amplified sequences labeled SNPM20 and SNPM80 were detected by 1% agarose gel electrophoresis. After obtaining a band consistent with the size of the target band, the PCR product was sequenc...

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Abstract

The invention provides a soybean aphid-resistant gene as well as a molecular marker and application thereof, and relates to the technical field of molecular marker-assisted breeding. The soybean aphid-resistant gene is located at the physical positions of 30,700,000-30,900,000,000 on the No.13 chromosome of a soybean genome, and a molecular marker and a corresponding molecular marker detection method are designed based on the soybean aphid-resistant gene. The molecular markers are used for auxiliary selection, so that the screening efficiency is greatly improved, and the breeding period of breeding aphid-resistant varieties is shortened.

Description

technical field [0001] The invention belongs to the technical field of molecular marker-assisted breeding, and in particular relates to a soybean aphid-resistant gene, its molecular marker and its application. Background technique [0002] Soybean aphids (Aphisglycines Matsμmura) are one of the main pests of soybeans, widely distributed in the main soybean producing areas of my country, and affect the yield and quality of soybeans by sucking photosynthetic products of soybean phloem and spreading plant viruses. Soybean aphids often gather on the tender stems and young leaves of plants, causing the chlorophyll to disappear in the damaged parts of the plants, and the leaves are curled. In severely damaged plants, the stems and leaves are curled, yellowed, the plants are short, and the branches and pods are reduced, thus affecting soybeans. Yield. When the aphids occur seriously at the seedling stage, the whole plant may die. If the control measures are not taken in time when ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12Q1/6895C12N15/11C12Q1/686
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q1/6858C12Q2531/113C12Q2565/125
Inventor 邱丽娟郭勇杨静刘广阳王秀君
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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