Pair of ustilago esculenta for successfully achieving artificial breeding of normal zizania caduciflora and application of pair of ustilago esculenta

A technology of black powder fungus and water jasmine, which can be used in applications, fungi, gardening tools/equipment, etc., and can solve problems such as wild water jasmine pregnancy

Active Publication Date: 2019-02-22
CHINA JILIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing artificial breeding technology can only make the wild Zizania conceive the gray Zizania

Method used

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  • Pair of ustilago esculenta for successfully achieving artificial breeding of normal zizania caduciflora and application of pair of ustilago esculenta
  • Pair of ustilago esculenta for successfully achieving artificial breeding of normal zizania caduciflora and application of pair of ustilago esculenta
  • Pair of ustilago esculenta for successfully achieving artificial breeding of normal zizania caduciflora and application of pair of ustilago esculenta

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Experimental program
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Effect test

Embodiment 1

[0013] Embodiment 1: wild rice smut ( Ustilago esculenta ) Itd1 gene cloning

[0014] 1. Cloning of the Itd1 gene: Through the analysis of the transcriptome data of normal Zizania and Glycyrrhizae, combined with the third-generation sequencing data of the Ustilago smut genome, it was predicted that the Ustilago smut gene locus related to teliospore formation was g1449. Using the ORF Finder function of NCBI, the open reading frame (ORF) sequence of g1449 gene was predicted, and the length of the open reading frame was found to be 2670bp. Ustilago smut ( Ustilago esculenta ) haploid strain UET1 and Ustilago smut ( Ustilago esculenta ) The genomic DNA of the haploid strain UET2 was used as a template, and the primer Itd1-F / R was designed to amplify the Itd1 gene sequence, and the primer Itd1-UF / UR was amplified Itd1 The upstream promoter sequence of the gene, the primer Itd1-DF / DR amplifies the downstream promoter sequence of the Itd1 gene. The total RNA of the two stra...

Embodiment 2

[0025] Embodiment 2: wild rice smut ( Ustilago esculenta ) haploid strain UET1△Itd1 and Ustilago smut ( Ustilago esculenta ) Obtaining of haploid strain UET2△Itd1

[0026] 1. Cloning of upstream and downstream knockout vectors: Using the principle of homologous recombination, using the strain UET1 as a template, use primer Itd1-U2 / U3 to amplify the upstream knockout fragment, and use primer Itd1-D1 / D2 to amplify the downstream knockout fragment , using the plasmid pUM1507 as a template, use primer hyg-F / 3 to amplify the upstream segment of hygromycin, use primer hyg-4 / R to amplify the downstream segment of hygromycin, and fuse the upstream segment of the gene with the upstream segment of hygromycin PCR method, and use the primer Itd1-U2 / hyg3 to obtain the upstream knockout long fragment, connect it to the T vector, and transform Escherichia coli; use the method of fusion PCR for the downstream fragment of the gene and the downstream fragment of hygromycin, and use the primer ...

Embodiment 3

[0048] Embodiment 3: the artificial inoculation method that makes Zizania zizania plant pregnant Zizania

[0049] 1) The smut fungus ( Ustilago esculenta ) haploid strain UET1△Itd1 and Ustilago smut ( Ustilago esculenta ) The haploid strain UET2△Itd1 was shaken in YEPS liquid medium at 25-30℃ to OD 600 0.8-1.0, the cells were collected by centrifugation, and diluted with 0.5×YEPS liquid medium to a final concentration of OD 600 2.0-3.0 bacteria solution, mix the bacteria solution two by two for inoculation;

[0050] 2) Cultivate seedlings of the wild water pipe roots with more than 3 complete internodes, and cultivate them in the greenhouse for 15-20 days. Inoculate the tubular roots with more than 3 small seedling plants that have germinated, and pierce the base of the seedlings ;

[0051] 3) Inject the mixed bacterial liquid obtained in step 1) into the tubular root with a syringe until the bacterial liquid overflows;

[0052] 4) Trim the treated inoculated seedlings...

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Abstract

The invention discloses a pair of ustilago esculenta for successfully achieving artificial breeding of normal zizania caduciflora and application of the pair of ustilago esculenta, and belongs to thefield of biotechnology. The pair of ustilago esculenta comprises an ustilago esculenta haploid strain UET1 delta Itd1 and an ustilago esculenta haploid strain UET2 delta Itd1, wherein the ustilago esculenta haploid strain UET1 delta Itd1 has the preservation number of CGMCC No. 16723; and the ustilago esculenta haploid strain UET2 delta Itd1 has a preservation number of CGMCC No. 16724. Accordingto the invention, the sexual compatible ustilago esculenta haploid strain UET1 delta Itd1 and ustilago esculenta haploid strain UET2 delta Itd1 are obtained by an artificial breeding method; the pairof ustilago esculenta can be inoculated in zizania aquatica to successfully produce normal zizania caduciflora, thereby providing a new idea for the artificial breeding of zizania caduciflora.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pair of wild smut fungus which successfully realizes the artificial breeding of Zizania and normal Zizania 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 fertiliser is the result of the interaction between Zizania plants and the smut fungus that specifically parasitizes in the body. So far, the planting and breeding of Zizania zizania still adopts the method of separating and screening Zizania pier, which requires a lot of manpower and material resources. Therefore, artificial inoculation is the development direction of Zizania zizania breeding. However, the existing artificial breeding technology can only make the wild Zizania conceive the gray Zizania. Contents of the invention...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01G7/06C12R1/645
CPCA01G7/06C12N1/14C12N1/145C12R2001/645
Inventor 张雅芬叶子弘殷淯梅夏文强俞晓平崔海峰
Owner CHINA JILIANG UNIV
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