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Combination of a pair of maize smut fungus and its application

A kind of smut fungus, a pair of technology, applied in the direction of application, fungi, instruments, etc., can solve the problems of poor test accuracy, consistency and repeatability, uncertain results of artificial inoculation, corn plant rot, etc. Achieve the effects of strong pathogenicity, easy harvesting and transportation, and high success rate of inoculation

Active Publication Date: 2021-06-29
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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  • Combination of a pair of maize smut fungus and its application
  • Combination of a pair of maize smut fungus and its application
  • Combination of a pair of maize smut fungus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Separation and Screening and Identification of White Polygezide Straisage of Obusic Black Fungus

[0039] Perform as follows:

[0040](1) Collecting winter spores and preparing a Winter Spore suspension: In December 2016, the natural air-dried corn tumor flavor is collected in Hainan, directly picking a small amount of winter spores directly from the cornoma black powder. Place in sterilization. In the 2ml centrifuge tube, then 1 ml of sodium hypochlorite solution was added in weight percent, and centrifuged at 2 min, 12000 rpm, 1 min, to the supernatant, add sterilization water to clean the precipitation 3, and finally use sterile water to dilute the precipitate. That is, the petrospore suspension, its final concentration is 10 3 Spore / ml.

[0041] (2) Winter aprotic culture: Take 200 μL of the Winter Spore suspension obtained from step (1) to PDA medium, cultured for 2 days at 25 ° C, which can be observed to observe the fine colony visible to the naked eye.

...

Embodiment 2

[0050] Example 2: Screening assay for combination of different maize black powder

[0051] (1) The propagation of the monocrete strain: 5 of the monocretetic strains obtained in Example 1 were evenly applied to different PDA plate media, respectively. 3 days were cultured at 25 ° C, dark conditions to obtain a haploid.

[0052] (2) Preparation of the inoculus of the monocrete strain: Steps (1) from the PDA plate culture medium were scraped from the PDA plate culture medium with a sterilized slide, and in accordance with 1 petri dish (9 cm in diameter of the culture dish). The bacteria were diluted with 1 L of sterilization of water, respectively, with a peripheral strain of 3, 6, 7, 8 or 11, respectively.

[0053] (3) According to the mating type of each haploid strain in Example 1, two two-two combinations of different interstitial monoclonal strains, that is, the second type of loftter strain prepared by step (2). The mother liquor was mixed according to the mass ratio of 1: 1, ...

Embodiment 3

[0060] Example 3 Classification and identification of haplot strains UM01 and UM02 of the present invention

[0061] The genomic DNA of UM01 or UM02 was extracted, the genomic DNA of UM01 or UM02 strain was a template, and the fungi germic primer ITS1 and ITS4 were used for PCR amplification, the primer sequence as:

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

[0063] ITS4: 5'-tcctccgcttttgatatgc-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 12.5μL, Template 2μL, DDH 2 O-25 μL. The PCR reaction procedure: 95 ° C pre-degeneration is 5 min, 95 ° C denaturation 30s, 56 ° C for 1 min, 35 cycles at 72 ° C, 35 cycles; final 72 ° C for 10 min, 4 ° C. 5 μl of the amplified product was taken for 1% agarose gel electrophoresis, and the PCR product size was detected under the UV transmitter, and the PCR amplification product was directly sent to Biological Engineering (Shanghai) Co., Ltd. to obtain the ITS sequen...

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Abstract

The invention discloses a pair of Ustilago maydis combination, including Ustilago maydis haploid strain UMO1 and Ustilago maydis haploid strain UMO2; wherein the preservation number of UMO1 is CGMCC No.15263; the deposit number of the UM02 is CGMCC No.15387. The maize smut combination of the present invention has strong pathogenicity, high inoculation success rate, accurate and reliable identification results of resistance to maize smut resistance of maize varieties, and good repeatability and consistency; when used in the production of maize mushrooms, not only The corn mushroom yield is high, and the corn mushroom is wrapped in bracts, which is convenient for harvesting and transportation.

Description

Technical field [0001] The present invention relates to the field of biotechnology, and in particular, a combination of jade black powder, further involves the use of the combination of jade black powder. Background technique [0002] Corn tumor black powder (or corn black powder disease) is corn disease caused by jade black powder (ustilago maydis, or corn black powder, cornoma black powder, etc.) mainly occurs in warm dry areas. And the spring is seriously involved in the summer birthplace, and the general incidence rate is 5 ~ 10%, which is a serious resulting loss of 30 ~ 80%, is an urgent problem to be solved. [0003] In addition to the harm to corn, the bacteria formed on the cobfur infected with corn fruit is still a delicious food, and the Aztec people in Mexico eat it has been more than 600 years, known as "Mexico Truffle." ", In my country, called corn mushrooms, corn, corn tumor, corn, corn mash, base bag, head, umi or black gallbladder." Corn mushrooms can be eaten, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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