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Ustilago maydis haploid strain UM01 and use thereof

A technology of smut and haploid of maize, which is applied in the biological field, can solve the problems of poor test accuracy, consistency and repeatability, uncertain results of artificial inoculation, rot of corn plants, etc., so as to achieve convenient harvesting and High success rate of transportation and inoculation, good reproducibility

Active Publication Date: 2018-10-23
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 haploid strain UM01 and use thereof
  • Ustilago maydis haploid strain UM01 and use thereof
  • Ustilago maydis haploid strain UM01 and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Isolation, screening and identification of haploid strains of Ustilago zeae of the present invention

[0039] Proceed as follows:

[0040] (1) Collect the teliospores and prepare the teliospore suspension: Naturally air-dried smut spore galls were collected in Hainan in December 2016, and a small amount of teliospores were directly picked from the smut spore galls and placed in a sterile In a 2ml centrifuge tube, add 1ml of 1% by weight sodium hypochlorite solution, mix by inversion for 2min, centrifuge at 12000rpm for 1min, remove the supernatant, add sterile water to wash the precipitate 3 times, and finally dilute the precipitate with sterile water. Get a teliospore suspension with a final concentration of 10 3 Spores / ml.

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

[0042] (3) M...

Embodiment 2

[0051] Example 2 Classification and identification of the haploid strain UM01 of the present invention

[0052] (1) The morphological characteristics of the strain: the spores of this strain are short rod-shaped, without septa, and 15-25 μm in length.

[0053] (2) The culture characteristics of the strain: On the PDA plate medium, the colony is milky white, with wrinkles on the surface, and moist, similar to yeast colonies, but when picked, it is more viscous than bacterial and yeast colonies.

[0054] (3) Molecular identification of the haploid strain UM01 of the present invention:

[0055] Using the genomic DNA of the UM01 strain as a template, the fungal genome ITS segment was amplified by PCR using the fungal universal primers ITS1 and ITS4. The primer sequence is:

[0056] ITS1: 5’-TCCGTAGGTGAACCTGCGG-3’;

[0057] ITS4: 5’-TCCTCCGCTTATTGATATGC-3’.

[0058] The PCR reaction system (25μL), in which: ITS1 (10μmol / L) 0.5μL, ITS4 (10μmol / L) 0.5μL, Es Taq MasterMix (Kangwei Century) 12.5μL...

Embodiment 3

[0060] Example 3 Preparation of mother liquor for inoculation of haploid strain UM01 of the present invention

[0061] Proceed as follows:

[0062] (1) Strain activation: inoculate the low-temperature stored UM01 strain on a PDA plate medium (the preparation method is: 200g potatoes are peeled and cut into small pieces, boiled in 800ml distilled water for 20 minutes, and filtered with 4 layers of gauze, Add 20g glucose and 20g agar, boil and dilute to 1000ml, sterilize at 121°C for 20min), culture at 25°C in the dark for 1 day to obtain activated strains.

[0063] (2) Bacterial cell preparation: Use an inoculating needle to pick an appropriate amount of the UM01 activated strain obtained in step (1) and evenly spread it on a PDA plate medium, culture at 25° C. and dark conditions for 3 days to obtain UM01 cell.

[0064] (3) Preparation of inoculation mother solution: Use sterile glass slides to scrape all the bacteria obtained in step (2) from the PDA plate medium, and use the bacteri...

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Abstract

The invention discloses an Ustilago maydis haploid strain UM01. The Ustilago maydis haploid strain UM01 has a preservation number of CGMCC No. 15263. Through combination of the Ustilago maydis haploidstrain UM01 and a haploid strain matching with the mating type, the pathogenicity is strong and a success rate of inoculation is high. The Ustilago maydis haploid strain UM01 produces accurate and reliable identification results of resistance to Ustilago maydis-caused diseases and has good consistency. In corn mushroom production, a corn mushroom yield is high and the corn mushrooms are easy to collect and transport.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a haploid strain UM01 of smut fungus, and also relates to the use of the strain. Background technique [0002] Corn smut (or corn smut) is a corn disease caused by Ustilago maydis (or called smut fungus, corn smut, etc.), which mainly occurs in warm and dry areas. In addition, the disease in spring sowing areas is more serious than in summer sowing areas. The general incidence rate is 5-10%, and serious cases can cause 30-80% yield loss, which is an urgent problem to be solved. [0003] In addition to causing harm to corn, the fungus galls formed on the ears of corn ears after infection by smut fungus is also a delicious food. The Aztecs in Mexico have eaten it for more than 600 years and are called "Mexican truffles". "In our country, it is called corn mushroom, corn black mold, corn smut fungus, corn black mold, cornbun, nuntou, black gall, etc. Corn mushrooms can be eaten ra...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/18A01G18/00A01G7/06C12R1/645
CPCA01G7/06A01G18/00C12N1/14C12Q1/18C12N1/145C12R2001/645
Inventor 张家齐石洁金戈杨硕张海剑郭宁刘树森陈丹
Owner INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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