A kind of wild rice smut haploid strain uemt2 and its application

A haploid and mushroom black technology, applied in the fields of application, fungi, plant cultivation, etc., can solve the problem of no haploid strains, etc., and achieve the effect of improving environmental adaptability and controlling the mating time

Active 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

In nature, Ustilago smut exists as diploid teliospores or dikaryotic hyphae, and there is no haploid strain

Method used

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  • A kind of wild rice smut haploid strain uemt2 and its application
  • A kind of wild rice smut haploid strain uemt2 and its application
  • A kind of wild rice smut haploid strain uemt2 and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: bacterial strain screening identification method

[0021] 1. Collect teliospores: from the normal Zizania longzia No. 2 ( figure 1 A) Pick teliospores directly.

[0022] 2. Teliospore suspension: pile the picked teliospores into 5 mL sterile water, filter through 4 layers of sterile gauze to obtain a relatively pure teliospore suspension, and finally dilute the teliospore suspension with sterile water Dilute to a final concentration of 10 3 spores / mL. Teliospores form as figure 1 Shown in B.

[0023] 3. Teliospore culture: Take 100 µL of teliospore suspension and spread it on the basidiospore isolation medium, and culture it at 28°C for about 60 hours, finely dispersed single colonies can be seen with the naked eye ( figure 1 C) as standard. Basidiospore isolation medium (1 L) formula is: K 2 HPO 4 1g, MgSO 4 ·7H 2 O 0.5 g, FeSO 4 ·7H 2 O 0.01g, KCl 0.5 g, glucose 18 g, (NH 4 ) 2 SO 4 5.28 g, 10 g of agar.

[0024] 4. Single spore isol...

Embodiment 2

[0045] Embodiment 2: artificial fertilized bamboo shoot test

[0046] The vector of overexpressing EGFP green fluorescent protein was transformed into Ustilago sativa ( Ustilago esculenta ) haploid strain UEMT2 and its sex-compatible strains (select No. 2 and No. 3 in Example 1) to overexpress EGFP green fluorescent protein, and the obtained overexpression strains can be detected by fluorescence microscopy, and successfully overexpress EGFP The strains with green fluorescent protein can see the green fluorescent signal under the fluorescent microscope, such as figure 2 shown. Then carry out artificial inoculation experiment, concrete artificial inoculation process is as follows:

[0047] 1) Shake two haploid strains of smut to OD in YEPS liquid medium at 28°C 600 was 1.0, centrifuged to collect the bacteria, diluted with 0.5×YEPS liquid medium to a final concentration of OD 600 The bacterial solution is 3.0, and the two bacterial solutions are mixed for inoculation;

[0...

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Abstract

The invention relates to a haploid strain UEMT2 of smut smut and its application, belonging to the field of biotechnology. In the present invention, smut fungus ( Ustilago esculenta ) haploid strain UEMT2, the preservation number is: CGMCC No.11841. Zizania artificial breeding requires the co-infection of two sexually compatible haploid strains. This strain can be used as the mother strain of Zizania artificial breeding and after screening to obtain its sexually compatible haploid strains, it can be achieved by artificial inoculation. Artificially pregnant watermelon. At the same time, the haploid strain can be genetically engineered to provide basic materials for the subsequent improvement of Zizania varieties, such as improving the environmental adaptability of Zizania, controlling the time of mating and so on.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a haploid strain UEMT2 of smut smut and its application. Background technique [0002] Ustilago rugosa belongs to the Basidiomycotina genus Ustilago, is a typical dimorphic fungus, and is closely related to Ustilago zea. So far, Zizania is the only known host, and Zizania gestation is the result of the interaction between the specific parasitic Ustilago smut and Zizania plants. Up to now, the planting and breeding of Zizania still adopts the method of separation and screening of Zizania pier, which requires a lot of manpower and material resources, and there are multiple physiological races of Zizania smut, which may be mixed in Zizania plants, resulting in different traits of Zizania varieties. Stable with obvious degradation. Therefore, artificial inoculation is the development direction of Zizania zizania breeding. [0003] Studies have shown that the mixed inoculat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A01G7/06A01G22/00A01H1/00C12R1/645
CPCA01G7/06A01H1/00A01G22/00C12N1/145C12R2001/645
Inventor 叶子弘俞晓平张雅芬崔海峰
Owner CHINA JILIANG UNIV
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