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Water hyacinth fungal biocontrol agent-eichhornia crassipes alternaria alternata

A technology for biocontrol of fungi and water hyacinth, applied in the biological control of weeds and microorganisms, can solve the problems of no research and application of germs, economic losses, non-target biosafety of chemical preparations and drug residues, etc.

Inactive Publication Date: 2009-08-19
SOUTHWEST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Water hyacinth (Eichhornia crassipes), also known as water lily, is native to South America and is one of the top ten harmful weeds in the world. The most serious damage is Zhejiang, Fujian, Guangdong, Yunnan and Taiwan, mainly causing pollution and blockage of rivers, lakes, reservoirs and other water areas, hindering waterway transportation and interfering with the normal operation of hydropower stations, causing huge economic losses every year
[0003] At present, water hyacinth is mainly controlled by manual salvage and chemical herbicide control at home and abroad. Although some results have been achieved, the control effects of the two measures are very limited. Salvage can only remove water hyacinth plants, and the seeds left in the water body can still Germinate and quickly form new weed strains; and after using chemical herbicides to kill weeds, plant residues can organicize water bodies and cause environmental pollution. Chemical agents themselves also have drug residues and non-target biosafety, etc. Environmental Pollution
Therefore, in recent years, people have carried out explorations on the biological control of weeds. In the 1960s, while studying and utilizing herbivorous insects such as water hyacinth weevil, many countries have also done a lot of investigation and research work on water hyacinth pathogens. , and has made some progress: more than 60 kinds of pathogenic fungi infecting water hyacinth have been reported in the world, and Cercospora rodmanii (Cercospora rodmanii) and Alternaria eichhorniae (Alternaria eichhorniae) that have been screened in South Africa and other countries ) has a strong inhibitory effect on water hyacinth, and the technology of controlling water hyacinth by Cercospora naumanii has been protected by international patents, but the control effect and application range of this pathogen on weeds are quite limited, and it has not been seen in the field of water hyacinth. Reports applied in other countries or regions
[0004] In my country, some people have investigated and studied water hyacinth diseases and pathogenic fungi, and found that the pathogenic fungi of water hyacinth mainly include Alternaria tenuissima, Cladosporium oxysporum, and Pseudomonas arseni. (Pestalotiopsis adusta), Monosporium eichhorniae and Thanatephorus cucumeris, etc., but these fungi have very limited pathogenic effects on water hyacinth and have no practical application value. A detailed report on further research and application of

Method used

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  • Water hyacinth fungal biocontrol agent-eichhornia crassipes alternaria alternata
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Examples

Experimental program
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Effect test

Embodiment 1

[0023] Isolation, purification and screening of pathogenic bacteria

[0024] (1) Potato sucrose agar medium (PSA) for fungal isolation and culture, its formula is 200 g of peeled fresh potatoes, 20 g of sucrose and agar each, and tap water is added to 1000 ml. First add water to boil the potatoes, filter through double-layer gauze, add sucrose and agar, add appropriate amount of water to 1000ml, stir well, put it into a triangular flask, put it in a sterilizer for sterilization, and place it. Heat and melt before use, pour into sterilized petri dishes or test tubes to make flat or slant medium for fungal isolation and culture or strain preservation.

[0025](2) Separation, cultivation and purification: collect water hyacinth plant samples with typical disease symptoms under natural conditions, rinse and cut off the small pieces (2mm) of the tissue at the junction of leaf disease spots and healthy parts, and put them into 5% sodium hypochlorite solution After surface disinfect...

Embodiment 2

[0029] Pathogen culture and inoculation condition test

[0030] (2) Temperature test: On the PDA plate cultivated for 5 days, use a puncher to take the mycelia block of the AFT001 strain with a diameter of 0.5 cm, inoculate it on the PDA medium, and set it at 5°C, 10°C, 15°C, and 20°C respectively. , 25°C, 30°C, 35°C and 40±0.5°C 8 temperature treatments, each treatment was repeated 3 times. They were cultured in 8 light incubators with the required temperature set in advance, and the colony size was measured daily by the vertical cross method, once every 24 hours, 5 times in a row, and the data was recorded. The pH of the PDA medium is about pH7.0. Therefore, the optimal growth temperature of Alternaria was determined.

[0031] (2) pH experiment: PDA medium was used in the experiment, and 11 pH treatments were set up. The pH value of the culture medium was adjusted to 4.0, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, 9.0, 10.0, 11.0 and 12.0 (determined with an electronic pH meter) with ...

Embodiment 3

[0036] Pathogen crude toxin isolation

[0037] (1) Preparation of pathogen culture filtrate: Pour 250ml of potato sucrose (PS) culture solution into 500ml Erlenmeyer flasks respectively, and sterilize for later use. Select the colonies that grow well after 6-7 days of cultivation, use a puncher to take a Φ6mm-sized bacterial cake, inoculate it into a triangular flask containing 250ml of PS culture solution, inoculate 8 pieces per bottle, and place it in a 25°C incubator with 12h light / day Under the conditions of static culture for 20 days. On the ultra-clean workbench, filter with qualitative filter paper (medium speed) to obtain the culture filtrate. Thus prepare enough pathogenic culture filtrate for toxin separation and extraction.

[0038] (2) Extraction and activity test of toxin: choose organic solvent petroleum ether, chloroform, phenol and ethyl acetate etc. with different polarities to extract the culture filtrate respectively. Extracted 3 times, then combined the ...

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Abstract

A water hyacinth fungal biocontrol agent, Alternaria crassipies, is separated from the natural environment and obtained. Spores are easily produced by the fungus under ultraviolet irradiation, the optimum culture medium for bacterial colony growth is Potato Sucrose Agar (PSA), the optimum temperature is 25 to 30 DEG C, and the optimum pH is 7.0 to 10; and the optimum conditions of fungal infection pathopoiesia are that moisturizing is carried out for 24 hours in dark light after inoculation and the temperature keeps at 28 to 30 DEG C during infection period. The thalli preparation and toxin liquid have very strong effects of pathopoiesia and grass killing on malignant ruderal water hyacinth, thereby being capable of producing environment-friendly mycoherbicide by the technology of biomimetic synthesis for zymogen or toxin and having potential business development using value. Pathopoiesia crude toxin is obtained from fungus culture fluid through extraction with ethyl acetate and reduced pressure distillation. The thalli suspension and the toxin liquid have no adverse influence on 20 kinds of important crops such as rice and the like.

Description

technical field [0001] The invention relates to the biological control of weeds and the field of microorganisms, in particular to a water hyacinth biocontrol fungus—Alternaria crassipes. Background technique [0002] Water hyacinth (Eichhornia crassipes), also known as water lily, is native to South America and is one of the top ten harmful weeds in the world. The most serious damage is Zhejiang, Fujian, Guangdong, Yunnan and Taiwan, mainly causing pollution and blockage of rivers, lakes, reservoirs and other water areas, hindering waterway transportation and interfering with the normal operation of hydropower stations, causing huge economic losses every year . [0003] At present, water hyacinth is mainly controlled by manual salvage and chemical herbicide control at home and abroad. Although some results have been achieved, the control effects of the two measures are very limited. Salvage can only remove water hyacinth plants, and the seeds left in the water body can stil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/04A01P13/00C12R1/645
Inventor 罗化东谭万忠殷利利孙现超王利军周君唐秀丽
Owner SOUTHWEST UNIV