Rubber tree colletotrichum drug-resistant strain HcgHNQZ1736 and application thereof in drug resistance research

A technology of anthrax bacteria and drug resistance, applied in the direction of application, fungicides, fungi, etc., can solve the problems of increasing difficulty in anthrax control, limiting rubber anthrax bacteria compound species, and polluting the environment

Active Publication Date: 2020-12-04
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Claims
  • Application Information

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Problems solved by technology

However, traditional morphology, cultural characteristics and ITS have limited the understanding of rubber anthracnose complex species. For a long time, many scholars have agreed that rubber tree anthracnose is mainly caused by glyospora anthracnose, and a few have oxysporum anthracnose. It is divided into two populations, C.gloeosporioides and C.acutatum, but with the development of taxonomy and the improvement of analysis methods, multi-locus-based phylogenetic analysis has helped scientists discover and clarify more compound species of rubber tree anthracnose fungus, and Realize good cultivation of natural rubber trees
[0005] In addition, in the environment where chemical control is still the main method to deal with anthracnose of rubber trees, long-term application of pesticides not only pollutes the environment, but also easily makes pathogenic bacteria resistant to pesticides. This situation increases the difficulty of anthracnose control. To effectively prevent and control anthracnose, it is particularly important to deepen the understanding of the characteristics of different pathogenic bacteria, and to grasp the dynamics of pathogenic bacteria in rubber planting areas in real time

Method used

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  • Rubber tree colletotrichum drug-resistant strain HcgHNQZ1736 and application thereof in drug resistance research
  • Rubber tree colletotrichum drug-resistant strain HcgHNQZ1736 and application thereof in drug resistance research
  • Rubber tree colletotrichum drug-resistant strain HcgHNQZ1736 and application thereof in drug resistance research

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Rubber tree anthracnose sample collection, separation, purification and preservation

[0054] From 2017 to 2018, disease samples were collected from the main rubber planting areas in Hainan Province. The collected disease samples were separated and purified indoors for pathogenic bacteria. The method is as follows: wash the collected diseased leaves with clean water and dry the surface water with filter paper. Use 75% ethanol to disinfect the leaf surface again; use sterilized scissors to cut about 3 × 3mm leaf tissue at the junction of disease and health, use 70% ethanol for 30 seconds in the workbench, and 0.1% mercuric chloride for 2 minutes. Bacterial water was treated for 3 times; dry the water with sterilized filter paper and inoculate it on a PDA plate, cultivate it in a 28°C incubator for 5 days, and then use a sterilized inoculation needle to pick a single mycelia and inoculate it on a new PDA plate above to purify the strain. Koch's law was used to ...

Embodiment 2

[0056] Embodiment 2 morphology observation

[0057] After the test strain was cultured on the PDA plate for 7 days, observe the colony morphology, promote the sporulation of the strain through AEA medium and SNA medium, observe the microscopic morphology of conidia, conidiophores and appresses under a microscope, and record the size; Refer to "Fungal Taxonomy" by Shao Liping, "General Phytopathology" by Xu Zhigang, etc., "Handbook of Fungal Identification" by Wei Jingchao, etc., and "Plant Pathogenic Mycology" by Zhang Zhongyi, etc. to carry out preliminary morphological classification and identification of strains .

[0058] The results of cultivation and observation of bacterial strains HcgHNQZ1736 and MeCkYN1705 showed that ( figure 2 ): The aerial hyphae on the PDA medium is felt-like, the mycelium is flat, the edges are neat, and the shape is round. The growth rate of the colony in the 28°C incubator is about 10.5±1.5mm / d. It is milky white to milky yellow, and the bac...

Embodiment 4

[0085] Example 4 Determination of basic biological characteristics

[0086] (1) Determination of suitable temperature for growth

[0087] The tested strains were inoculated on PDA plates, and cultured in the dark at 10°C, 15°C, 20°C, 25°C, 28°C, 30°C, 32°C and 35°C for 5 days, and the experiment was repeated three times. Measure and record the diameter of colony growth by cross method, and determine the suitable temperature range for the growth of pathogenic bacteria.

[0088] Table 4 below, Figure 5 and Figure 6 As shown, the growth temperature range of the tested strains HcgHNQZ1736 and MeCkYN1705 is 15-32°C; 28°C is the optimum growth temperature range of the colonies, and the colony growth rate is the fastest, about 11.7mm / d; when the temperature reaches below 10°C and 35°C Above, the colony stops growing completely.

[0089] Table 4 Mycelium growth in different temperature environments

[0090]

[0091] Note: The letters after the values ​​in the table are the r...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to karst colletotrichum and application thereof in the field of rubber anthracnose research. The karst colletotrichum is found on rubber trees in Hainan Province for the first time; and a strain obtained through separation is C.karstii. Biological characteristic determination finds that the optimumgrowth temperature of the strain is 28 DEG C; the lethal temperature is 35 DEG C; the optimum growth pH is 6; light and dark alternation is beneficial to colony growth; and pectin and peptone are respectively an optimal carbon source and an optimal nitrogen source. Pathogenicity evaluation finds that the strain has relatively strong pathogenicity on four main recommended varieties including PR107,RRIM600, hot research 7-33-97 and Dafeng 95; a drug resistance evaluation result shows that the strain HcgHNQZ1736 has no drug resistance to a sterol demethylation inhibitor (DMIs) bactericide prochloraz-manganese chloride complex, and has high drug resistance to a benzimidazole bactericide carbendazim.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a karst anthracnose fungus and its application in the field of rubber anthracnose research. Background technique [0002] Natural rubber, steel, coal, and petroleum are known as the world's four major industrial raw materials, and natural rubber is the only renewable resource among them. Rubber tree is an important tropical economic crop and the main production carrier of natural rubber (cis-polyisoprene). It plays an important role in national economy and national defense construction, and has important economic value. [0003] Plant anthracnose is caused by fungal infection of the genus Colletotrichum Corda. Plants that can become hosts of this type of fungus are widely distributed. Plants suitable for fungal reproduction in tropical, subtropical, and temperate regions are found. Among them More than 600 kinds of plants, such as food, fruits and v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12Q1/04C12Q1/18A01G7/06A01G13/00A01N47/18A01N47/38A01P3/00C12R1/645
CPCC12N1/14C12Q1/04C12Q1/18A01G13/00A01G7/06A01N47/38A01N47/18C12R2001/645C12N1/145
Inventor 郑肖兰刘先宝李博勋时涛蔡吉苗冯艳丽郑行凯黄贵修
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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