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Ustilago maydis composition and application thereof

A kind of smut and a pair of technology, applied in the biological field, can solve the problems of poor test accuracy, consistency and repeatability, uncertain results of artificial inoculation, corn plant rot, etc., to achieve strong pathogenicity, Ease of harvesting and transportation, high inoculation success rate

Active Publication Date: 2018-10-26
INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent "A Production Method and Product of Corn Mushrooms" (CN10540409593A) discloses a method of injecting an aqueous solution mixed with corn smut chlamydospores or basidiospores into the top of corn ears for inoculation to produce corn mushrooms. Although this method is simple to operate, However, because chlamydospores are easily mixed with other pathogenic bacteria and are not easy to find, direct inoculation with smut chlamydospore aqueous solution often causes the whole corn plant to rot and cause the test to fail; In order to reproduce in combination, two kinds of sexually compatible basidiospores must be mixed and inoculated together into the plant to be successful. However, the existing method is to directly make the basidiospore mixture after the chlamydospore germination into the inoculum for inoculation. The randomness of the mixture does not necessarily guarantee that the inoculum contains two sexually compatible haploid strains, which leads to great uncertainty in the results of artificial inoculation and the accuracy, consistency and repeatability of the test. very bad

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  • Ustilago maydis composition and application thereof
  • Ustilago maydis composition and application thereof
  • Ustilago maydis composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 Isolation, screening and identification of Ustilago maize haploid strain of the present invention

[0039] Proceed as follows:

[0040](1) Collect teliospores and prepare teliospore suspension: Naturally air-dried corn smut galls were collected in Hainan in December 2016, and a small amount of teliospores were directly picked from the corn smut galls and placed in a sterilized 2ml centrifuge tube, then add 1ml of sodium hypochlorite solution with a weight percentage of 1%, mix it upside down for 2min, centrifuge at 12000rpm for 1min, remove the supernatant, add sterilized water to wash the precipitate 3 times, and finally dilute the precipitate with sterile water. Promptly get teliospore suspension, its final concentration is 10 3 spores / ml.

[0041] (2) Teliospore culture: Take 200 μl of the teliospore suspension obtained in step (1) and spread it on PDA medium, and cultivate it at 25° C. for 2 days, and then small colonies visible to the naked eye can b...

Embodiment 2

[0050] Embodiment 2: the screening test of different maize smut fungus combinations

[0051] (1) Propagation of haploid strains: No. 3, No. 6, No. 7, No. 8 and No. 11 five kinds of bacterial strains of the haploid strains obtained in Example 1 are uniformly coated on different PDA plate culture medium respectively , cultured at 25°C in the dark for 3 days to obtain haploid cells.

[0052] (2) Preparation of haploid strain inoculation mother solution: use sterilized glass slides to scrape off all the thallines obtained in step (1) from the PDA plate medium and place them on the seed plate according to 1 petri dish (the diameter of the petri dish is 9cm). The thalline of the bacterium was diluted with the ratio of 1L sterilized water to obtain the inoculation mother solution of No. 3, No. 6, No. 7, No. 8 or No. 11 haploid strains.

[0053] (3) According to the mating types of each haploid strain in Example 1, the haploid strains of different mating types are combined in pairs, ...

Embodiment 3

[0060] Example 3 Taxonomic identification of haploid strains UMO1 and UMO2 of the present invention

[0061] Extract the genomic DNA of UM01 or UM02, use the genomic DNA of UM01 or UM02 bacterial strains as a template, and use fungal universal primers ITS1 and ITS4 to perform PCR amplification on the ITS segment of the fungal genome. The primer sequence is:

[0062] ITS1: 5'-TCCGTAGGTGAACCTGCGG-3';

[0063] ITS4: 5'-TCCTCCGCTTATTGATATGC-3'.

[0064] The PCR reaction system (25 μL), wherein: ITS1 (10 μmol / L) 0.5 μL, ITS4 (10 μmol / L) 0.5 μL, Es Taq MasterMix (Kang Wei Century) 12.5 μL, template 2 μL, add ddH 2 0 to 25 μL. The PCR reaction program: pre-denaturation at 95°C for 5 min, denaturation at 95°C for 30 s, annealing at 56°C for 45 s, extension at 72°C for 1 min, 35 cycles; final extension at 72°C for 10 min, storage at 4°C. Take 5 μ L of the amplified product and carry out 1% agarose gel electrophoresis, detect the size of the PCR product under an ultraviolet transillu...

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Abstract

The invention discloses a Ustilago maydis composition. The Ustilago maydis composition comprises a haploid strain UM01 and a haploid strain UM02, wherein the preservation number of the UM01 is CGMCC No. 15263, and the preservation number of the UM02 is CGMCC No. 15387. The ustilago maydis composition is highly pathogenic, high in vaccination success rate, accurate and reliable in identification results of resistance to ustilago maydis, and good in repeatability and consistency; high yield of corn mushroom is achieved when the ustilago maydis composition is applied to the production of the cornmushrooms, the corn mushrooms are wrapped in bracts, and convenience in harvesting and transporting is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a combination of smut and the application of the smut combination. Background technique [0002] Corn smut (or corn smut) is a corn disease caused by corn smut (Ustilago maydis, or called corn smut, corn smut, etc.), mainly occurs in warm and dry areas, And the spring sowing area is more serious than the summer sowing area, the general incidence rate is 5-10%, and serious ones can cause 30-80% yield loss, which is a problem to be solved urgently. [0003] In addition to causing harm to corn, the galls formed on the ears of corn after being infected by smut fungus are also a delicious food. The Aztecs in Mexico have eaten them for more than 600 years. They are called "Mexican truffles." "In our country, it is called corn mushroom, corn black mold, corn smut, corn black mold, stick bag, rentou, black rice or black gallbladder, etc. Corn mushrooms can be eaten raw or stir-fried, and t...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/18A01G18/00C12R1/645
CPCA01G18/00C12N1/14C12Q1/18C12N1/145C12R2001/645G01N2333/37
Inventor 石洁张家齐郭宁刘树森金戈张海剑杨硕
Owner INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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