CN105349483A - Alternaria alternata sporulation culture medium and preparing method thereof - Google Patents
Alternaria alternata sporulation culture medium and preparing method thereof Download PDFInfo
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- CN105349483A CN105349483A CN201510901620.7A CN201510901620A CN105349483A CN 105349483 A CN105349483 A CN 105349483A CN 201510901620 A CN201510901620 A CN 201510901620A CN 105349483 A CN105349483 A CN 105349483A
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- substratum
- spore
- concentration
- brown spot
- spot pathogen
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Abstract
The invention discloses an alternaria alternata sporulation culture medium and a preparing method thereof. The alternaria alternata sporulation culture medium is prepared from yeast powder with the concentration of 4-10 g/L, NaNO3 with the concentration of 4-12 g/L, KH2PO4 with the concentration of 0.5-2.5 g/L, KCl with the concentration of 0.2-1.0 g/L, MgSO4 with the concentration of 0.1-0.3 g/L, glycerinum with the concentration of 10-25 ml/L, agar with the concentration of 14.5-20 g/L and the balance deionized water. The amount of all the components are weighed according to the matching, the deionized water is added to of the scale mark indicating 1 L in a volumetric flask, and the alternaria alternata sporulation culture medium is obtained by carrying out sterilization for 15-30 min at the temperature of 121 DEG C-130 DEG C.
Description
Technical field
The present invention relates to biological technical field, relate in particular to a kind of tobacco brown spot pathogen and produce spore substratum, also relate to the preparation method that this tobacco brown spot pathogen produces spore substratum simultaneously.
Background technology
Alternaria alternate [
alternariaalternata(Frises) Keissler] be the tobacco growing middle and later periods leaf diseases the most serious occurs, multiple positions such as tobacco leaf, stem, petiole, bennet, capsule can be endangered, but to endanger blade, cause quality of tobacco to decline, lose baking and be worth.Tobacco brown spot pathogen conidium at pathogen transmission, infect, cause a disease in play an important role, conidium can propagate on the tobacco leaf in field by air-flow or rainwater, when there being free water droplet, conidium can sprout at 4-8h and produce germ tube and the epidermic cell invading tobacco leaf.Form scab in 4-5 days, conidium is propagated by air-flow in scab generation afterwards, carries out repeatability infect the tobacco of field growing.At present the seed selection of flue-cured tobacco brown spot resistant variety, tobacco bred red star Disease Resistance Identification and evaluate chemical control red-star like disease effect test in often need the conidium of a large amount of artificial culture as the research material of inoculation material or other experiment, and conidial source and sprout effect and will directly affect the accuracy of test-results.
The product spore that in prior art, the normal PDA of employing substratum carries out tobacco brown spot pathogen is cultivated, research finds too low by the spore quantity of the acquisition of PDA culture medium culturing, the research of being correlated with is carried out in morbidity only under the natural condition of dependence field, is difficult to advance the seed selection of Alternaria alternate resistant variety fast, is difficult to advance the screening of red-star like disease pesticide control.Therefore in order to meet the seed selection of Alternaria alternate resistant variety in agriculture production, the red star Disease Resistance Identification of tobacco bred and evaluate the requirement of chemical control red-star like disease drug effect, and the plantation of tobacco bred and popularization, it is a very crucial index that the output improving tobacco brown spot pathogen spore carries out artificial inoculation.
Summary of the invention
The object of the invention is to overcome above-mentioned shortcoming and the one that provides to produce a large amount of form fast consistent, sprout effective, that germination rate is high conidial tobacco brown spot pathogen and produce spore substratum.
Another object of the present invention is to provide this tobacco brown spot pathogen to produce the preparation method of spore substratum.
A kind of tobacco brown spot pathogen of the present invention produces spore substratum, comprises following raw material:
4 ~ 10g/L yeast powder, 4 ~ 12g/LNaNO
3, 0.5 ~ 2.5g/LKH
2pO
4, 0.2 ~ 1.0g/LKCl, 0.1 ~ 0.3g/LMgSO
4, 10 ~ 25mL/L glycerine, 14.5 ~ 20g/L agar, all the other are deionized water.
A kind of tobacco brown spot pathogen of the present invention produces the preparation method of spore substratum, comprising:
Take the amount of each composition by said ratio, add deionized water constant volume to 1L, 121 ~ 130 DEG C of sterilizing 15 ~ 30min, obtain tobacco brown spot pathogen and produce spore substratum.
The present invention compared with prior art, has obvious beneficial effect, as can be known from the above technical solutions: tobacco brown spot pathogen of the present invention produces spore substratum by yeast powder, NaNO
3, KH
2pO
4, KCl, MgSO
4, glycerine, deionized water composition, this formula is that contriver goes through the several years, and experiment obtains repeatedly, and confirm that alternaria altemata strain bacterial strain aerial hyphae on this substratum is few through experiment, sporulation quantity is high.Produce spore media surface and can produce a large amount of form very fast unanimously, sprout effective, that germination rate is high conidium.
Embodiment
embodiment 1
Tobacco brown spot pathogen produces a preparation method for spore substratum, comprising:
Take 4g yeast, 4.5gNaNO
3, 1.8gKH
2pO
4, 0.7gKCl, 0.10gMgSO
4, 10mL glycerine, 14.5g agar, adds deionized water constant volume to 1L, 121 DEG C of sterilizing 15min, obtains tobacco brown spot pathogen and produces spore substratum.
Example: obtained tobacco brown spot pathogen is produced spore substratum and melts, getting 15-20mL, to pour diameter into be in the culture dish of 9mm, after substratum cooled and solidified, by activated tobacco brown spot pathogen (
a.alternata) strain bacterium C2(provides by Guizhou Province Tabacco Science and Technology Institute) make bacterium dish with 5mm punch tool, bacterium dish is moved to and produces spore media surface.After connecing bacterium, cultivate under 28 DEG C of dark conditions, can be observed the form of Alternaria alternate bacteria strain on this substratum after 3-7d, aerial hyphae is few, and in light brown, sporulation quantity is large.On PDA substratum, bacterial strain aerial hyphae clearly, and in white, sporulation quantity is few.Produce spore media surface and can produce a large amount of form very fast unanimously, sprout effective, that germination rate is high conidium.The sporulation quantity cultivated after 4d is 1.53 × 10
7individual/cm
2, produce spore successful and be better than PDA substratum (0/cm
2).After cultivation 10d, producing spore substratum sporulation quantity is (2.73 × 10
8individual/cm
2), PDA substratum sporulation quantity is (4.09 × 10
5individual/cm
2), this sporulation quantity can reach 667.5 times of equivalent PDA substratum.
embodiment 2
Tobacco brown spot pathogen produces a preparation method for spore substratum, comprising:
Take 7.5g yeast, 8gNaNO
3, 2.0gKH
2pO
4, 0.8gKCl, 0.20gMgSO
4, 20mL glycerine, 16g agar, adds deionized water constant volume to 1L, 125 DEG C of sterilizing 20min, obtains tobacco brown spot pathogen and produces spore substratum.
Example: obtained tobacco brown spot pathogen is produced spore substratum and melts, getting 15-20mL, to pour diameter into be in the culture dish of 9mm, after substratum cooled and solidified, by activated tobacco brown spot pathogen (
a.alternata) strain bacterium C6(provides by Guizhou Province Tabacco Science and Technology Institute) make bacterium dish with 5mm punch tool, bacterium dish is moved to and produces spore media surface.After connecing bacterium, cultivate under 25 DEG C of dark conditions, can be observed the form of Alternaria alternate bacteria strain on this substratum after 3-7d, aerial hyphae is few, and in light brown, sporulation quantity is large.On PDA substratum, bacterial strain aerial hyphae clearly, and in white, sporulation quantity is few.Produce spore media surface and can produce a large amount of form very fast unanimously, sprout effective, that germination rate is high conidium.The sporulation quantity cultivated after 4d is 2.42 × 10
7individual/cm
2, produce spore successful and be better than PDA substratum (0/cm
2).After cultivation 10d, producing spore substratum sporulation quantity is (2.86 × 10
8individual/cm
2), PDA substratum sporulation quantity is (5.24 × 10
5individual/cm
2), this sporulation quantity can reach 545.8 times of equivalent PDA substratum.
embodiment 3
Tobacco brown spot pathogen produces a preparation method for spore substratum, comprising:
Take 10g yeast, 12gNaNO
3, 2.5gKH
2pO
4, 1.0gKCl, 0.30gMgSO
4, 25mL glycerine, 20g agar, adds deionized water constant volume to 1L, 130 DEG C of sterilizing 30min, obtains tobacco brown spot pathogen and produces spore substratum.
Example: obtained tobacco brown spot pathogen is produced spore substratum and melts, getting 15-20mL, to pour diameter into be in the culture dish of 9mm, after substratum cooled and solidified, by activated tobacco brown spot pathogen (
a.alternata) strain bacterium C8(provides by Guizhou Province Tabacco Science and Technology Institute) make bacterium dish with 5mm punch tool, bacterium dish is moved to and produces spore media surface.After connecing bacterium, cultivate under 25 DEG C of dark conditions, can be observed the form of Alternaria alternate bacteria strain on this substratum after 3-7d, aerial hyphae is few, and in light brown, sporulation quantity is large.On PDA substratum, bacterial strain aerial hyphae clearly, and in white, sporulation quantity is few.Produce spore media surface and can produce a large amount of form very fast unanimously, sprout effective, that germination rate is high conidium.The sporulation quantity cultivated after 4d is 2.64 × 10
7individual/cm
2, produce spore successful and be better than PDA substratum (0/cm
2).After cultivation 10d, producing spore substratum sporulation quantity is (3.86 × 10
8individual/cm
2), PDA substratum sporulation quantity is (5.15 × 10
5individual/cm
2), this sporulation quantity can reach 749.5 times of equivalent PDA substratum.
The above, it is only preferred embodiment of the present invention, not any pro forma restriction is done to the present invention, anyly do not depart from technical solution of the present invention content, the any simple modification done above embodiment according to technical spirit of the present invention, equivalent variations and modification, all still belong in the scope of technical solution of the present invention.
Claims (2)
1. tobacco brown spot pathogen produces a spore substratum, comprises following raw material:
4 ~ 10g/L yeast powder, 4 ~ 12g/LNaNO
3, 0.5 ~ 2.5g/LKH
2pO
4, 0.2 ~ 1.0g/LKCl, 0.1 ~ 0.3g/LMgSO
4, 10 ~ 25mL/L glycerine, 14.5 ~ 20g/L agar, all the other are deionized water.
2. a kind of tobacco brown spot pathogen as claimed in claim 1 produces the preparation method of spore substratum, comprising:
Take the amount of each composition by said ratio, add deionized water constant volume to 1L, 121 ~ 130 DEG C of sterilizing 15 ~ 30min, obtain tobacco brown spot pathogen and produce spore substratum.
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| CN201510901620.7A CN105349483A (en) | 2015-12-09 | 2015-12-09 | Alternaria alternata sporulation culture medium and preparing method thereof |
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| CN105349483A true CN105349483A (en) | 2016-02-24 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110622720A (en) * | 2019-10-14 | 2019-12-31 | 贵州省烟草科学研究院 | Inoculation method of alternaria alternate |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104017735A (en) * | 2014-05-28 | 2014-09-03 | 云南省烟草农业科学研究院 | Alternaria alternata medium and preparation method thereof |
-
2015
- 2015-12-09 CN CN201510901620.7A patent/CN105349483A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104017735A (en) * | 2014-05-28 | 2014-09-03 | 云南省烟草农业科学研究院 | Alternaria alternata medium and preparation method thereof |
Non-Patent Citations (7)
| Title |
|---|
| 于莉等: "烟草赤星病菌生物学特性的研究", 《吉林农业大学学报》 * |
| 姚玉霞等: "烟草赤星病发病程度与烟叶内总氮含量关系的初报", 《吉林农业大学学报》 * |
| 张乐等: "烟草赤星病菌(Alternaria alternata)营养生理研究", 《中国农学通报》 * |
| 方中达: "《植病研究方法》", 31 December 1998, 中国农业出版社 * |
| 杨玉范等: "烟草赤星病菌（Alternaria alternata(Fr.)Keisskr)的生长和孢子产生条件及寄主范围的研究", 《吉林农业科学》 * |
| 马贵龙等: "不同生育期、氮肥水平和品种对烟草赤星病菌抗性组分的影响", 《吉林农业大学学报》 * |
| 马贵龙等: "烟草赤星病菌分生孢子产生条件的研究", 《吉林农业大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110622720A (en) * | 2019-10-14 | 2019-12-31 | 贵州省烟草科学研究院 | Inoculation method of alternaria alternate |
| CN110622720B (en) * | 2019-10-14 | 2021-08-24 | 贵州省烟草科学研究院 | Inoculation method of alternaria alternate |
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Application publication date: 20160224 |
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