Molecular marker for authenticating zizania latifolia turcz. phenotype characteristics, and application and acquisition method thereof

A technology for molecular marker and water bamboo separation, applied in the field of molecular biology, can solve the problems of decreased taste, quality of water bamboo, inedible, etc., and achieves the effect of eliminating interference and shortening the research period.

Inactive Publication Date: 2019-04-26
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result of the accumulation of a large number of gray spores in the stems of gray water bamboo, the taste and quality of water bamboo shoots are reduced, and they are generally inedible.

Method used

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  • Molecular marker for authenticating zizania latifolia turcz. phenotype characteristics, and application and acquisition method thereof
  • Molecular marker for authenticating zizania latifolia turcz. phenotype characteristics, and application and acquisition method thereof
  • Molecular marker for authenticating zizania latifolia turcz. phenotype characteristics, and application and acquisition method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: Classification of test materials and phenotypic characteristics

[0047] The following test materials come from Jinhua Academy of Agricultural Sciences, Zhejiang Province, Jinyun County Agricultural Bureau, Lishui City, Zhejiang Province, Yuyao City Agricultural Science Research Institute, Ningbo City, Zhejiang Province, Suzhou Academy of Agricultural Sciences, Jiangsu Province, and Tongxiang City, Jiaxing City, Zhejiang Province. Dongjia Zizania Cooperative, Shengzhou Puhui Vegetable Cooperative in Shaoxing City, Zhejiang Province, Vegetable Research Institute of Huangyan District, Taizhou City, Zhejiang Province, and School of Life Sciences, China Jiliang University, Hangzhou City, Zhejiang Province. Samples were collected in autumn 2016, autumn 2017, summer 2018, and autumn 2018.

[0048] Table 1 Variety, phenotype and source of tested samples

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[0050]

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[0052]

Embodiment 2

[0053] Example 2: Population Analysis

[0054] 1) Genomic DNA of Ustilago smut was extracted by CTAB method, the DNA concentration was measured with a nucleic acid micrometer, and the DNA quality was detected by agarose gel electrophoresis. with ddH 2 O was diluted to 100ng / μL and stored at -20°C.

[0055] 2) Send the DNA of 47 Zizania zizania varieties with different phenotypes to next-generation resequencing. The reference genome is the MT-type genome of Ustilago sativa, WGS INSDC: JTLW00000000.1.

[0056] 3) Sequencing data were compared to the reference genome using GATK, SAMTOOLS, and BWA software to obtain SNP data files. A total of 321659 SNP sites were obtained.

[0057] 4) All samples of Ustilago smut samples participating in the SNP scanning were numbered into two types, A and B, which represent two types of normal Zizania and gray Zizania respectively. By extracting the Genotype data from the SNP scanning results, output the SNP data of A and B into SNP c a / b f...

Embodiment 3

[0058] Embodiment 3: SNP site identification

[0059] 1) Among the 1330 screened SNP sites, one SNP site was selected at every interval of 20 as the verification site. At the same time, due to the quality of the reference genome sequencing, it was necessary to exclude the SNP sites in the relatively late contig of the reference genome. A total of 39 SNP sites that could be used for verification were screened out, and relevant primers were designed.

[0060] 2) AS-PCR (allele-specific PCR), AS-PCR uses the principle that the base mismatch at the 3' end of the primer can interrupt the PCR extension program and cause no amplification product. important role. In this example, 40 pairs of AS-PCR primers are designed, and all primers are attached in Table 2

[0061] Table 2 Allele-specific PCR primer sequence list

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[0065]

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[0067] 3) In this example, 8 different phenotypes of the same species of Zizania smut DNA were used as tem...

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Abstract

The invention discloses a molecular marker for authenticating zizania latifolia turcz. phenotype characteristics, and application and an acquisition method thereof, and belongs to the technical fieldof the molecular biology. The molecular marker for authenticating zizania latifolia turcz. phenotype characteristics comprises UeG1, UeG2, UeG3, UeG4, UeG5 and UeG6. Through a SNP (single nucleotide polymorphism) marker, grey zizania latifolia turcz. is screened, normal zizania latifolia turcz. can be selected in advance, a workload is reduced, and screening efficiency is improved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker for identifying phenotypic characteristics of Zizania zizania and its application and acquisition method. Background technique [0002] Water bamboo (Zizania latifolia Turcz.) is a perennial herbaceous plant of Gramineae, native to my country and Southeast Asia, and is the second largest aquatic vegetable in my country. Zizania has been cultivated in my country for a long time. Before the Western Zhou Dynasty, the seeds of Zizania were eaten as food. Later, it was found that the galls formed by the infection of the stems by Ustilago esculenta were not only edible, but also the meat was tender and delicious. It is good, but it is eaten as a vegetable, and it has been widely planted as a vegetable since the end of the Tang Dynasty. In addition to being a vegetable, Zizania can also be used medicinally, which has the effects of diuresis, quen...

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 叶子弘张雅芬葛鑫涛夏文强俞晓平崔海峰
Owner CHINA JILIANG UNIV
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