Phytophthora nicotianae fermentation liquid pathogenic active ingredient and preparation method thereof

A technology of Tobacco black shank and black shank bacteria, which is applied in the field of pathology, can solve problems such as the unsatisfactory development of tobacco black shank disease, and achieve the effects of strong pathogenic ability, reliable results and good stability

Active Publication Date: 2017-07-04
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] With the continuous development of the tobacco industry in the direction of green and pollution-free, the prevention and control of tobacco black shank only stays at a reasonable rotation as the main, ridge cultivation and chemical control as supplementary comprehensive prevention and control measure

Method used

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  • Phytophthora nicotianae fermentation liquid pathogenic active ingredient and preparation method thereof
  • Phytophthora nicotianae fermentation liquid pathogenic active ingredient and preparation method thereof
  • Phytophthora nicotianae fermentation liquid pathogenic active ingredient and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0038] Example 1: Pathogenic effect of sterile fermentation broth of Black Shank

[0039] (1) Pathogenic effect of sterile fermentation broth of Black Shank cultured on V8 medium

[0040] by figure 1 It can be seen that compared with the control (CK) (CK is cultured for 0 days), the fermentation broth obtained after 5 days of shaking culture of Black Shank has caused the leaf injection area to appear slightly sunken with white circles. After a period of shaking culture, After the 7th day, the fermented liquid was taken so that the injection area of ​​the leaf was sunken and dotted. But generally speaking, no obvious HR necrotic spots appeared in the tobacco leaves.

[0041] (2) The pathogenic effect of the aseptic fermentation broth of Black Shank cultured on carrot medium

[0042] by figure 2 It can be seen that the sterile fermentation broth of Phytophthora nigricans obtained after 5 days of culture is the same as the control (CK), and there is no HR reaction. After 6 days of cult...

Example Embodiment

[0054] Example 2: Determination of the pathogenicity of the active ingredients in the fermentation broth of Black Shank

[0055] From Table 2 and Image 6 It can be seen that the ultrafiltrate treated with 5KDa microfiltration hollow cellulose membrane and 3KDa ultrafiltration hollow cellulose membrane of sterile fermentation broth of Black Shank can make tobacco leaves produce obvious HR necrotic spots, with an average area of ​​lesions. Value up to 0.87cm 2 , Compared with the lesion area after the original solution injected into the leaf, the mean value increased by 0.32cm 2 , The difference between the two reached a significant level of 5%; after the ultrafiltrate was processed by the open chromatography column, the 70%, 80% and 95% ethanol eluates obtained can make the tobacco leaf produce HR reaction. The HR reaction of the filtrate, the necrotic spots caused by the ethanol eluate are more obvious, the color is darker, slightly yellow, and the average area of ​​the lesion is...

Example Embodiment

[0058] Example 3 Obtainment of anther mutants against black shank

[0059] (1) Screening comparison of the optimal selection pressure of the anti-black shank stock solution of anthers and the active ingredients of the fermentation broth of the present invention

[0060] ① Tobacco high-potassium strain HKDN-2

[0061] First, from Figure 7 As shown in Table 3, the germination rate of HKDN-2 anthers gradually decreases with the increase of the concentration of active ingredients in the fermentation broth of Black Shank of the present invention; compared with the control group (CK, 0%), it contains 1% of the original The germination rate of the anthers in the culture medium with the active ingredient concentration of the black shank fermentation broth of the invention is 12.0%, and there is no significant difference between the two; when the active ingredient concentration of the black shank fermentation broth of the invention is 2%, the anther germination rate is significantly lower C...

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Abstract

The invention relates to a phytophthora nicotianae fermentation liquid pathogenic active ingredient and a preparation method thereof. The method comprises the following steps that (1) phytophthora nicotianae hyphae are subjected to shaking bed oscillation culture in a culture medium; sterile fermentation liquid is collected; centrifugation is performed; supernate is filtered and degermed for use; (2) the degermed phytophthora nicotianae sterile fermentation liquid obtained in the step (1) is subjected to membrane filtration separation through a microfiltration hollow cellulose membrane and an ultrafiltration hollow cellulose membrane to obtain clear and transparent ultrafiltration liquid; ultrafiltration filter liquid is fed into a column to be subjected to macroporous resin enrichment; then, ingredients adsorbed on the resin can be flushed down by ethyl alcohol; ethyl alcohol elution liquid, i.e., the phytophthora nicotianae fermentation liquid pathogenic active ingredient, is obtained. The phytophthora nicotianae fermentation liquid pathogenic active ingredient and the preparation method have the advantages that the operation is simple; the result is reliable; the pathogenic capability of the phytophthora nicotianae fermentation liquid pathogenic active ingredient is high; the stability is high; the phytophthora nicotianae fermentation liquid pathogenic active ingredient can be efficiently used for screening phytophthora nicotianae resistance varieties.

Description

technical field [0001] The invention belongs to the field of pathology, and in particular relates to a pathogenic active component of fermented liquor of tobacco black shank bacteria and a preparation method thereof. Background technique [0002] Tobacco black shank was first discovered in Java, Indonesia. It was first reported in 1950 that the disease occurred in the Huanghuai smoking area of ​​my country, and it has been harmful for a long time and the condition has become more and more serious. At present, except for the sporadic incidence of tobacco black shank in the northeast smoking areas, tobacco black shank disease is common in other smoking areas in the country, causing serious economic losses every year, second only to viral diseases. [0003] At present, the commonly used chemical agents for the prevention and treatment of tobacco black shank mainly include Ruifu mold (metalaxyl) wettable powder, Bordeaux mixture, thiophanate-methyl, dikexone wettable powder, an...

Claims

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Application Information

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IPC IPC(8): C12P1/02C12R1/645
CPCC12P1/02C12N1/145C12R2001/645
Inventor 戴林建陈武钟军周田
Owner HUNAN AGRICULTURAL UNIV
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