CN101372670A - In Vitro inoculation method for powdery mildew - Google Patents
In Vitro inoculation method for powdery mildew Download PDFInfo
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- CN101372670A CN101372670A CNA2008100619317A CN200810061931A CN101372670A CN 101372670 A CN101372670 A CN 101372670A CN A2008100619317 A CNA2008100619317 A CN A2008100619317A CN 200810061931 A CN200810061931 A CN 200810061931A CN 101372670 A CN101372670 A CN 101372670A
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- powdery mildew
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Abstract
The invention discloses an method for inoculating grape powdery mildew in vitro, which comprises the following steps: leaves in vitro are selected: the third to the sixth leaves below the tips of grapes are picked; a strain is selected: fresh grape powdery mildew spores are selected; medicament is prepared: 10-90mg/L of benzimidazole solution is prepared; the leaves are treated: after cleaning, the petioles of the leaves are twisted with aseptic cotton and dropped with the prepared benzimidazole solution; and inoculation is carried out: the grape powdery mildew is directly inoculated on the leaves in vitro treated by the benzimidazole solution. The method takes the benzimidazole as a green-keeping agent, and directly inoculates the grape powdery mildew on the leaves in vitro; the erysiphe cichoracearum of the leaves in vitro is not affected by the benzimidazole, thus solving the problem of culture and preservation of the grape powdery mildew.
Description
(1) technical field
The present invention relates to a kind of inoculation method of grape powdery mildew.
(2) background technology
Grape powdery mildew belongs to obligate parasite, can only on fresh and alive organization material, cultivate and preservation at present, so the cultivation of grape powdery mildew and preserve very difficult.Now under the technical qualification, grape powdery mildew only exists on the plant of grape growth and survival, when infecting the blade of grapevine seedling and fruit, it just falls ill, and it is full sub to produce fresh white powder germ, just can carry out experimental studies such as diseases prevention, breeding for disease resistance, be subjected to the influence of natural condition big, lifetime is short, be not easy control and preservation, experimental study is made troubles.
(3) summary of the invention
In order to overcome the deficiency that existing grape powdery mildew can be cultivated and therefore preservation makes troubles to experimental study on fresh and alive organization material, the invention provides a kind of in vitro inoculation method of grape powdery mildew.
The technical scheme that the present invention solves its technical problem is: a kind of in vitro inoculation method of grape powdery mildew comprises the steps: the selection of excised leaf: the 3rd～6 the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 10～90mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Preferably, the concentration of described benzimidazole solution is 50～70mg/L, and when the concentration of benzimidazole solution was 50～70mg/L, it is best that it protects green effect, can protect green more than 22 days.
Preferably, the excised leaf of harvesting is the 3rd a following leaf of grape tip point, and the Powdery Mildew morbidity of the 3rd excised leaf that grape tip point is following is best.
Beneficial effect of the present invention is: adopt benzimidazole to use as green-preserving agent, grape powdery mildew is seeded on the excised leaf, benzimidazole does not have influence to excised leaf generation Powdery Mildew, has solved the problem of the cultivation and the preservation of grape powdery mildew.
(4) embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Embodiment one
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 3rd the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 60mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment two
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 4th the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 10mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment three
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 5th the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 90mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment four
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 6th the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 70mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment five
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 3rd the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 50mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment six
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 4th the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 20mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment seven
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 5th the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 40mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Embodiment eight
A kind of in vitro inoculation method of grape powdery mildew comprises the steps:
The selection of excised leaf: the 3rd the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification: select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament: compound concentration is the benzimidazole solution of 80mg/L;
The processing of blade: the fresh grape leave that will gather, behind aseptic water washing three times, with soaking 5 minutes in the 0.5%NaCIO solution, use aseptic water washing again three times, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculate: dry after writing brush is sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
Claims (3)
1. the in vitro inoculation method of a grape powdery mildew is characterized in that comprising the steps:
The selection of excised leaf:
The 3rd～6 the fresh blade that grape-picking tip point is following;
Selecting for use of bacterial classification:
Select the full son of uncinula necator bacterium fresh in the wine-growing garden for use;
The preparation of medicament:
Compound concentration is the benzimidazole solution of 10～90mg/L;
The processing of blade:
With the fresh grape leave of gathering, behind aseptic water washing three times,, use aseptic water washing again three times with soaking 5 minutes in the 0.5%NaCIO solution, drain and place sterile petri dish, twine aseptic cotton on the petiole of blade, add and drip the benzimidazole solution for preparing;
Inoculation:
Dry after writing brush sterilized in 70% alcohol, with the white powder germ on the grape leave of infection Powdery Mildew, gently brush with writing brush, direct inoculation to above-mentioned on the excised leaf of benzimidazole solution-treated, placing 25 ℃, RH is 85% growth cabinet, carry out 16 hours light every day and cultivate, 8 hours dark cultivations.
2. the in vitro inoculation method of grape powdery mildew as claimed in claim 1, it is characterized in that: the concentration of described benzimidazole solution is 50～70mg/L.
3. the in vitro inoculation method of grape powdery mildew as claimed in claim 1 or 2 is characterized in that: the excised leaf of harvesting is the 3rd a following leaf of grape tip point.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100619317A CN101372670A (en) | 2008-05-27 | 2008-05-27 | In Vitro inoculation method for powdery mildew |
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| CNA2008100619317A CN101372670A (en) | 2008-05-27 | 2008-05-27 | In Vitro inoculation method for powdery mildew |
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| CN101372670A true CN101372670A (en) | 2009-02-25 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102972302A (en) * | 2012-12-27 | 2013-03-20 | 新疆农业科学院哈密瓜研究中心 | Method for preserving melon powdery mildew fungus |
| CN109439551A (en) * | 2018-12-17 | 2019-03-08 | 靖西市秀美边城农业科技有限公司 | A method of using Isolated leaf inoculation grey leaf spot disease of maize and morbidity |
-
2008
- 2008-05-27 CN CNA2008100619317A patent/CN101372670A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102972302A (en) * | 2012-12-27 | 2013-03-20 | 新疆农业科学院哈密瓜研究中心 | Method for preserving melon powdery mildew fungus |
| CN109439551A (en) * | 2018-12-17 | 2019-03-08 | 靖西市秀美边城农业科技有限公司 | A method of using Isolated leaf inoculation grey leaf spot disease of maize and morbidity |
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Open date: 20090225 |