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A method of artificial inoculation of smut fungus that successfully makes Zizania zizania plants pregnant

A technology of artificial inoculation and black powder fungus, which is applied in the directions of botanical equipment and methods, horticultural methods, plant cultivation, etc., and can solve the problem of no artificial inoculation method of wild black powder fungus.

Active Publication Date: 2019-01-11
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no effective artificial inoculation method for Ustilago smut

Method used

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  • A method of artificial inoculation of smut fungus that successfully makes Zizania zizania plants pregnant
  • A method of artificial inoculation of smut fungus that successfully makes Zizania zizania plants pregnant
  • A method of artificial inoculation of smut fungus that successfully makes Zizania zizania plants pregnant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: wild rice smut ( Ustilago esculenta ) haploid strain UET1 and Ustilago smut ( Ustilago esculenta ) for screening of haploid strain UET2, such as figure 1 shown.

[0023] 1. Collect teliospores: from Longjiao No. 2 gray bamboo ( figure 1 A) Pick teliospores directly.

[0024] 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.

[0025] 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...

Embodiment 2

[0041] Embodiment 2: the selection of inoculation method, inoculation object and inoculation time

[0042] Ustilago smut ( Ustilago esculenta ) haploid strain UET1 and Ustilago smut ( Ustilago esculenta ) The haploid strain UET2 was shaken in YEPS liquid medium at 25-30°C 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 The bacterial liquid is 2.0-3.0, and the two bacterial liquids are mixed to obtain a mixed bacterial liquid.

[0043] Taking the tubular root of wild asparagus collected from Wujiang as the inoculation object, according to the growth of its buds and leaves, it is divided into three types: bud stage, small seedling stage and seedling stage. 3-5 complete internodes were reserved; 7 inoculation methods (Table 1) were used for artificial inoculation at the end of March and the end of April. In October, a unified observation found that: only the wild Zizania at the young ...

Embodiment 3

[0047] Embodiment 3: Verification test

[0048] In order to further confirm that the artificial inoculation of Ustilago sativa is the cause of the pregnancy of Zizania zizizaziza, the mixed infection method of strains UET1-EGFP and UET2-EGFP overexpressing EGFP was used to infect wild wild plants at the young seedling stage at the end of March using the fifth inoculation procedure. Zizania, Zizania japonica was successfully impregnated. At the same time, after tissue sectioning of Zizania zizania, the teliospores and hyphae of teliospores and hyphae had green fluorescent signals produced by overexpression of EGFP. EGFP of Ustilago smut ( figure 2 ).

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Abstract

The invention provides an Ustilago esculenta artificial inoculation method capable of realizing successful stem-swelling of a water bamboo plant, belonging to the technical field of artificial inoculation of water bamboo. According to the invention, by inoculating Ustilago esculenta haploid strain UET1 and Ustilago esculenta haploid strain UET2 selected from Ustilago esculenta in wild water bamboo, stem-swelling of the wild water bamboo is successfully realized. The method provided by the invention has simple operation, is effective, and provides a new direction for seed breeding of the water bamboo.

Description

technical field [0001] The invention belongs to the field of artificial inoculation of wild rice stems, and in particular relates to an artificial inoculation method of smut fungus for successfully fertilizing wild rice stems. 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. 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 dire...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01G7/06A01G22/00
CPCA01G7/06A01G22/00
Inventor 张雅芬叶子弘俞晓平崔海峰
Owner CHINA JILIANG UNIV
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