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Method for breeding zizania latifolia plant seedlings

A technology of plants and wild rice stems, applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve problems such as long feedback time, unfavorable decision-making of wild wild rice seedlings, and influence on wild wild rice seedlings, to achieve Effects of reducing economic losses and optimizing the process of Zizania seedlings

Pending Publication Date: 2022-05-06
CHINA JILIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The feedback time of the whole process is too long, which is not conducive to the decision-making of Zizania seedlings, and the quality of the previous batch of seedlings will affect the next Zizania plant seedlings

Method used

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  • Method for breeding zizania latifolia plant seedlings
  • Method for breeding zizania latifolia plant seedlings
  • Method for breeding zizania latifolia plant seedlings

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation, identification and plant infection of smut fungus.

[0028] (1) Experimental materials

[0029] The normal Zizania and Gray Zizania of different varieties were collected from Jinhua, Yuyao, Jiaxing, Lishui, Taizhou and Suzhou, Jiangsu. YEPS medium (containing 10g / L yeast extract, 20g / L peptone and 20g / L sucrose) was purchased from Beijing Suolaibao Biotechnology Co., Ltd. Ampicillin and kanamycin were purchased from Shanghai Sangon Biotechnology Co., Ltd.

[0030] (2) Haploid isolation and identification of Ustilago smut

[0031] a. Pick out the gray spore piles in the fleshy stems of normal Zizania zizania and gray zizania respectively, and dilute to 1×10 with sterile water. 4 cells / mL, spread evenly on YEPS solid medium, and culture at 28°C;

[0032] b. Rinse the culture plate with sterile water after 24 hours, and dilute the mixture to a cell concentration of 1×10 3 cells / mL, transferred to the micro clean bench;

[0033] c. Take 1 μL of the diluted ...

Embodiment 2

[0045] Resequencing analysis of Ustilago smut.

[0046] (1) Experimental materials

[0047] The plant genome extraction kit was purchased from Tiangen Biotechnology Co., Ltd. The normal Zizania and Gray Zizania of different varieties were collected from Jinhua, Yuyao, Jiaxing, Lishui, Taizhou and Suzhou, Jiangsu. The samples of Ustilago smut strains were isolated, identified and preserved by Zhejiang Key Laboratory of Biometrics, Inspection and Quarantine Technology, China Jiliang University.

[0048] (2) DNA extraction, sequencing and sequence analysis

[0049] Using the Plant Genomic DNA Extraction Kit (Tiangen, DP305), the operation was performed according to the instruction manual, and finally the DNA was dissolved in 50 μL sterile water.

[0050] A total of 20 species of Zizania zizania were collected in Zhejiang, Jiangsu and other places, and 36 strains of Ustilago smut were isolated. Genome resequencing data were obtained through high-throughput next-generation seque...

Embodiment 3

[0055] Detection of smut fungus content in different parts of Zizania zizania plants.

[0056] (1) Experimental materials

[0057] The upstream and downstream primers and probes were synthesized by Nanjing GenScript Biotechnology Co., Ltd.; sterile nuclease-free water, PerfectStart II Probe qPCR SuperMix were purchased from Beijing Quanshijin Biotechnology Co., Ltd.; the plant genome extraction kit was purchased from Tiangen Biology Technology Co., Ltd.

[0058] (2) Sample collection and DNA extraction

[0059] The seedlings of normal Zizania and Glycyrrhizae were selected for seedling cultivation, and the roots, stem nodes and shoot tips of the plants were collected at regular intervals after seedling cultivation to extract total DNA.

[0060] Using the Plant Genomic DNA Extraction Kit (Tiangen, DP305), the operation was performed according to the instruction manual, and finally the DNA was dissolved in 50 μL sterile water.

[0061] (3) Fluorescent quantitative PCR detecti...

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Abstract

The invention discloses a method for breeding zizania latifolia plant seedlings. The method comprises the following steps: extracting DNA (Deoxyribose Nucleic Acid) of a to-be-detected sample containing Ustilago esculenta as an amplification template; a fluorescent quantitative PCR (Polymerase Chain Reaction) method is used for detection, and a primer pair used for detection comprises a forward primer with a nucleotide sequence as shown in SEQ ID No.1 and a reverse primer with a nucleotide sequence as shown in SEQ ID No.2; the nucleotide sequences of the probes used for detecting the MT type Ustilago esculenta and the T type Ustilago esculenta are as shown in SEQ ID No.3 and SEQ ID No.4; predicting the phenotype of the zizania latifolia plant in the mature period according to the ratio of the content of the T-type zizania latifolia to the content of the MT-type zizania latifolia, and breeding zizania latifolia plant seedlings. By analyzing the content of T-type and MT-type zizania esculenta in stem node or stem tip tissues of zizania aquatica plants, whether grey zizania aquatica or male zizania aquatica can be formed in the future can be effectively predicted, so that the zizania aquatica seedling raising process is optimized, and economic losses are reduced.

Description

technical field [0001] The invention relates to the field of molecular biology technology, in particular to a method for seedling selection of Zizania zizania plants. Background technique [0002] Zizania as a kind of aquatic vegetable has a history of more than 2,000 years in China. It is widely planted, especially in Jiangsu and Zhejiang regions. It is the second largest aquatic vegetable in my country after lotus root. Ustilago fungus infection is a key factor in inducing plant stem swelling to form edible fleshy stems. There are three phenotypes of normal Zizania, gray Zizania and male Zizania in field production. What people eat daily is the white fleshy stem of the plant, that is, normal Zizania zizania; the smut smut fungus forms a large number of black teliospores in the succulent stem of gray zizania, which can cause allergic pneumonia and is not suitable for consumption; male zizania does not contain wild wild rice powder fungus, the stem cannot expand to form an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6895C12Q1/6851C12Q2531/113C12Q2561/101C12Q2563/107
Inventor 叶子弘夏文强李士玉汤近天崔海峰张雅芬
Owner CHINA JILIANG UNIV
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