Method for reproducing verticillum lecanii and recovering vigor of verticillum lecanii by using larvas of argyrogramma agnata
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A technology of Verticillium leucoidans and Spodoptera frugiperda is applied in the biological field to achieve the effect of restoring vitality and infectivity
Inactive Publication Date: 2015-08-19
YUNNAN AGRICULTURAL UNIVERSITY
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Literature search did not find any reports on technical solutions for restoring the vitality and infectivity of Verticillium lecanii
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Embodiment 1
[0034] Embodiment 1. method is as follows:
[0035] 1. Rearing the intermediate host Silverworm: Collect healthy larvae of Silverleaf from the wild, and use Eupatorium adenophora under the environment of room temperature 24°C, relative humidity 77%, and light conditions L:D=16:8. The dry powder is the main raw material of the silver leafworm semi-artificial diet (according to the patent document ZL201410125557.8), and the larvae are reared for 4 days before use.
[0036] 2. Preparation of Verticillium lecanii bacterial classification:
[0037] (1), cultivating Verticillium lecanii: Inoculate Verticillium lecanii KMZW-1 bacterial strain on the PDA medium, put into a constant temperature incubator at 25±0.5°C and cultivate for 10 days;
[0038] (2), prepare the conidia liquid of Verticillium lecanii: get the KMZW-1 bacterium colony that grows well on the PDA medium, wash down and filter the spores with sterile water, quantify the amount of Verticillium lecanii in the filtrate ...
Embodiment 2
[0043] Embodiment 2. method is as follows:
[0044] 1. Rearing the intermediate host Silverworm: collect healthy larvae of Silverleaf from the wild, and use Eupatorium adenophora under the environment of room temperature 26°C, relative humidity 75%, and light conditions L:D=16:8. The dry powder is the main raw material of the silver leafworm semi-artificial diet (according to the patent document ZL201410125557.8), and the larvae are reared for 7 days before use.
[0045] 2. Preparation of Verticillium lecanii bacterial classification:
[0046] (1), cultivating Verticillium lecanii: Inoculate Verticillium lecanii KMZW-1 bacterial strain on the PDA medium, put into a constant temperature incubator at 25 ± 0.5°C and cultivate for 8 days;
[0047] (2), prepare the conidia liquid of Verticillium lecanii: get the KMZW-1 bacterium colony that grows well on the PDA medium, wash down and filter the spores with sterile water, quantify the amount of Verticillium lecanii in the filtrate ...
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Abstract
The invention provides a method for reproducing verticillum lecanii and recovering vigor of the verticillum lecanii by using larvas of argyrogramma agnata, and belongs to the biotechnology. The method comprises the following steps of feeding the argyrogramma agnata; culturing the verticillum lecanii; preparing conidium liquid of the verticillum lecanii; preparing the prepared conidium liquid into suspension liquid; uniformly spraying the suspension liquid to the larvas of the argyrogramma agnata until surfaces of the larvas are wet; placing the larvas in a sterile artificial climate room and culturing the larvas until hyphae grow on the larvas fully; collecting the larvas; dewatering and drying the larvas; and storing the larvas in a refrigerator of which the temperature is minus 95 DEG C to minus 78 DEG C. In a reproduction process of the verticillum lecanii reproduced by the method, the vigor and the infection ability of strain spores can be kept for a long time. Moreover, by the method, the vigor and the infection ability of strains of which the infection ability and the vigor are reduced severely can be recovered.
Description
technical field [0001] The present invention belongs to biotechnology. Background technique [0002] Verticillium lecanii (Zimm) is a common entomogenous fungus, belonging to the fungal phylum Deuteromycetes subphylum Hyphospora Hyphospora. The fungus has a wide geographical distribution and a wide range of hosts. It has good parasitic effects on agricultural pests such as aphids, whiteflies, scales, thrips and harmful mites. It is a relatively mature biological pesticide. In the previous research process, Verticillium lecanii often reproduced on a large scale in the form of solid or liquid medium. Verticillium lecanii KMZW-1 is a strain with high toxicity against fly insects screened by the patent applicant and obtained the patent (ZL200810233784.7). After 4 to 5 generations, the viability of the spores obviously began to decline. The study found that other Verticillium lecanii also had the problem of the ability to infect the host and the viability of the spores decrease...
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