Brassica napus drought-tolerant gene, and molecular marker and application thereof

A drought-tolerant technology of Brassica napus, applied in the direction of DNA/RNA fragments, microbial measurement/inspection, recombinant DNA technology, etc., can solve the problems of inability to use molecular markers, labor and time-consuming, time-consuming, etc.

Inactive Publication Date: 2019-07-19
SOUTHWEST UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The field direct identification method is limited to identification in arid, semi-arid areas or dry seasons, which is limited by the external environment; while the identification of dry sheds and pots is limited by the facility space, it is not conducive to large-scale materials in the same period of time, that is, in the same environment. Drought tolerance sex identification
Furthermore, identification of a single indicator lacks accuracy, while identification of multiple indicators is complex, time-consuming, labor-intensive and destructive to materials
[0004] Rapeseed is an important oil crop in my country. The current main planted Brassica napus requires a lot of water throughout the growing season and has poor adaptability to drought.
The drought tolerance of Brassica napus is a complex comprehensive trait controlled by multiple genes and easily affected by environmental factors. However, in recent years, the research on drought stress in Brassica napus has mainly focused on the mining of QTL, significant association markers and candidate genes. At present, no alleles with significant drought tolerance have been obtained, which cannot be used for the application of molecular marker-assisted selection in the identification of germplasm resources and the selection of drought-tolerant varieties
[0005] In the process of identification of drought-tolerant germplasm resources of Brassica napus and selection of new varieties, conventional identification methods are mainly used, but conventional identification methods are labor-intensive, time-consuming, inefficient, destructive to materials, and susceptible to external environment and subjective Awareness influence and accuracy are poor, which seriously restricts the breeding of drought-tolerant varieties

Method used

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  • Brassica napus drought-tolerant gene, and molecular marker and application thereof
  • Brassica napus drought-tolerant gene, and molecular marker and application thereof
  • Brassica napus drought-tolerant gene, and molecular marker and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Genome-wide association analysis of seed germination rate (GP) and germination index (GI) of Brassica napus under drought stress

[0039] The 520 germplasm resources used in this example were jointly collected by Chinese rapeseed breeding and scientific research units. For each germplasm resource, 50 plump seeds were selected and sterilized with 4% NaClO solution, and evenly placed in a 0.9 cm diameter petri dish covered with filter paper. For the first time, 30ml 20% PEG6000 solution was added to the drought stress treatment, and 30ml ddH was added to the control 2 o 2 , then add 15ml 20% PEG6000 solution to the drought stress treatment, add 15ml ddH to the control 2 o2 , continued until the 7th day. Carry out 3 repeated experiments, investigate and count germination rate (GP) and germination index (GI). Combined with the genotype data obtained by 60K microarray analysis, the association analysis was carried out based on the MLM model, and the SNPs signi...

Embodiment 2

[0044] Embodiment 2: Haplotype analysis and drought tolerance phenotype identification:

[0045] According to the 18 SNPs detected by resequencing, haplotype analysis was performed, and a total of 5 main haplotypes (Hap, Haplotype) were obtained ( figure 2 a), comparing the GP and GI of these haplotypes under drought stress, it was found that the GP and GI of Hap4 were significantly higher than those of Hap3 and Hap5 (p figure 2 b,c). 20% PEG6000 was used to simulate stress, and different lines of five haplotypes were selected for verification. The results showed that under the treatment of PEG6000, the GP and GI of different haplotype materials were consistent with the previous research results, and the seeds of Hap4 had the highest GP and GI , followed by Hap1, while the GP and GI of Hap3 and Hap5 haplotype materials were lower under PEG6000 treatment ( figure 2 d, e). In addition, the results of comparing the drought tolerance of different haplotype materials at the see...

Embodiment 3

[0046] Example 3: Using BnA.NF-YA7 to target dCAPS molecular markers for drought tolerance identification

[0047] A screening method for drought-tolerant varieties of Brassica napus, comprising the steps of:

[0048] 1) Based on the base site located at 19159584bp on the Brassica napus A09 chromosome, use the dCAPSFinder 2.0 online website to design primers and develop dCAPS markers in combination with the restriction endonuclease NdeI;

[0049] 2) DNA of multiple strains is extracted, and after the following identification, if Brassica napus has the gene sequence described in SEQ ID NO.1, it is a drought-tolerant Brassica napus.

[0050] Specifically, in the step 2), PCR amplification, enzyme digestion and electrophoresis are performed after the DNA is extracted. The PCR amplification procedure is as follows: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30s, annealing at 55°C for 30s, and renaturation at 72°C 1min, 35 cycles; 72°C extension for 10min, 12...

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Abstract

The present invention discloses a brassica napus gene and an application thereof. A drought-tolerant related gene BnA.NF-YA7 having functions in different growth stages of brassica napus is screened,and a dCAPS label designed according to SNP loci of the BnA.NF-YA7 gene can accurately identify drought-tolerant materials and has certain versatility, wide adaptability and timeliness. The brassica napus drought-tolerant gene can be used for brassica napus drought-tolerant germplasm resource identification and molecular marker-assisted selection, and lays a foundation for breeding materials or varieties with drought-tolerant and good agronomic and quality traits.

Description

technical field [0001] The invention relates to a drought-tolerant gene of Brassica napus and its molecular marker and application. Background technique [0002] The drought tolerance phenotype of crops is determined by their own physiological resistance and structural characteristics, as well as the degree of cooperation between the rhythm of growth and development and changes in agroclimatic factors. It is related to the period of occurrence, intensity and duration, and is a complex comprehensive trait. [0003] The most commonly used crop drought tolerance identification technology is field direct identification. In addition, there are also dry shed water control identification and pot tests to control soil moisture content. Identification indicators mainly include yield, morphology, physiological and biochemical indicators, etc. In addition, the total level of drought tolerance of materials is obtained based on the measured data of multiple indicators, or the drought to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 刘列钊李阳阳王嘉唐章林周清元卢坤林呐李加纳
Owner SOUTHWEST UNIVERSITY
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