CN104480060B - A kind of Rapid Sporulation method of colletotrichum gloeosporioides Penz - Google Patents
A kind of Rapid Sporulation method of colletotrichum gloeosporioides Penz Download PDFInfo
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- CN104480060B CN104480060B CN201410712920.6A CN201410712920A CN104480060B CN 104480060 B CN104480060 B CN 104480060B CN 201410712920 A CN201410712920 A CN 201410712920A CN 104480060 B CN104480060 B CN 104480060B
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- mycelium
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Abstract
The invention discloses a kind of Rapid Sporulation method of anthrax bacteria, it including the activation of colletotrichum gloeosporioides Penz bacterial strain, Mycelium culture, blow mycelia, aseptic water washing, dry, step etc. constant temperature illumination cultivation, the rapid induction big volume production spore of anthrax-bacilus mycelium in 24~48 hours, the sporulation quantity of unit area is up to 7.8 × 107~6.5×109Individual/cm2.Spore can rapidly be produced with rapid induction colletotrichum gloeosporioides Penz mycelium by the method, be that good basis is established in the forms such as follow-up spore shape, appresorium and Pathogenicity experiment, substantially reduce experimental period, save the researcher substantial amounts of quality time.
Description
Technical field
The invention belongs to technical field of life science, and in particular to a kind of Rapid Sporulation method of colletotrichum gloeosporioides Penz, it is applicable
In kept dry, activation etc. colletotrichum gloeosporioides Penz under state mycelial Rapid Sporulation, generally in 24~48 h
Rapid to produce spore, spore amount every square centimeter is up to 7.8 × 107 ~ 6.5×109It is individual.
Background technology
Colletotrichum gloeosporioides Penz host range is quite varied, can infect various hosts such as peach, nectarine, apple, grape, mango and pears
Cause anthracnose.Between colletotrichum gloeosporioides Penz bacterial strain in terms of morphological feature, pathogenic, molecular genetic level, trophosome compatibility group etc.
All have larger variation, bacterium colony can for canescence to Dark grey, aerial hyphae is flat, felted or cotton-shaped, conidiophore
Generally colourless, conidium is unit cell, khaki, cylindrical shape or club shape, two ends blunt round, is typically sized to 12~26
3.5~5 μm of μ m.Research on colletotrichum gloeosporioides Penz spore is extremely widespread, and its classification to anthrax-bacilus, pathogenic etc. are ground
Study carefully particularly important, and induce and produce the key link that spore then turns into research, how quickly to obtain a large amount of spores in very great Cheng
Progress of research and success or failure are decide on degree.
The content of the invention
The purpose of the present invention is to produce spore using simple and feasible method rapid induction colletotrichum gloeosporioides Penz mycelium, so as to add
Good basis is established in fast follow-up spore, appresorium and pathogenic correlative study, and for vast research work has been saved largely
Quality time.
In order to realize the above object technical scheme is as follows：
Mycelium with the colletotrichum gloeosporioides Penz of PDA solid medium cultures is the basis for producing spore, by scraping mycelia, sterilized water
Rinsing, drying and illumination cultivation measure induction mycelium sprout rapidly a large amount of spores under the h of illumination 24~48.
Specifically include following steps：
（1）The colletotrichum gloeosporioides Penz for separating or activated is transferred on PDA plate again, is placed in 28 DEG C of constant incubators
The h of illumination cultivation 24~48；
（2）When bacterium colony 3~4.5cm long to a diameter of, in superclean bench with aseptic cotton carrier by center a diameter of 2
~ 3.5cm aerial hyphaes remove, and with sterilized water that surface washing is clean, and back-off dries naturally；
（3）Flat board after drying continues illumination cultivation 24 ~ 48 during 28 DEG C of constant incubator is laid in after closeing the lid
h；
（4）Orange-red pycnidia is seen in planar surface after the h of illumination 24 ~ 48, with sterilized water by conidium
Device is washed down and racked, and conidium is then automatically discharged into water；20 mL are settled to, 20 μ L are taken and is seen to blood counting chamber
Examine the sporulation quantity of simultaneously unit of account area.
PDA solid culture based formulas：Potato 200g, the g of glucose 20, the g of agar 20, are settled to 1 L.
A small amount of orange-red pycnidia can be seen in planar surface after general illumination 24h, will be mitogenetic with sterilized water
Spore device is washed down and racked, and conidium is then automatically discharged into water；Pycnidia showed increased after illumination 48h, uses
Appropriate amounts of sterilized water by all pycnidias wash it is lower after be settled to 20mL, take 20 μ L and observed to blood counting chamber and unit of account
The sporulation quantity of area.
Described glue spore anthrax sporulation quantity is up to 7.8 × 107 ~ 6.5×109Individual/cm2。
The beneficial effects of the invention are as follows：It is quick by making colletotrichum gloeosporioides Penz mycelium in 24~48h by light stimulation
A large amount of mitogenetic big devices of spore and conidium are produced, whole experimental period is substantially reduced, manpower and materials and scientific research is effectively saved
A large amount of quality time of worker.
Specific embodiment
Embodiment 1
（1）The colletotrichum gloeosporioides Penz of separation is transferred on PDA plate again, illumination training in 28 DEG C of constant incubators is placed in
Support 24 h；
（2）Whne bacterium colony is long be about 3 cm to diameter when, in superclean bench with aseptic cotton carrier by center a diameter of 2
Cm aerial hyphaes remove, and with sterilized water by surface washing 3 times, back-off dries naturally；
（3）Flat board after drying continues the h of illumination cultivation 24 during 28 DEG C of constant incubator is laid in after closeing the lid
Left and right；
（4）A small amount of orange-red pycnidia is seen in planar surface after the h of illumination 24, with sterilized water by mitogenetic spore
Sub- device is washed down and smashed up, and conidium is then automatically discharged into water, is settled to 20 mL, takes 20 μ L to blood counting chamber
Observe the sporulation quantity of simultaneously unit of account area.
Described glue spore anthrax sporulation quantity is up to 7.8 × 107Individual/cm2。
Embodiment 2
（1）The colletotrichum gloeosporioides Penz that will have been activated is transferred on PDA plate again, is placed in illumination in 28 DEG C of constant incubators
Cultivate 48 h；
（2）When bacterium colony 4.5 cm long to a diameter of, in superclean bench with aseptic cotton carrier by center a diameter of 3.5
Cm aerial hyphaes remove, and with sterilized water that surface washing is clean, and back-off dries naturally；
（3）Flat board after drying continues the h of illumination cultivation 48 during 28 DEG C of constant incubator is laid in after closeing the lid；
（4）Orange-red pycnidia is seen in planar surface after the h of illumination 48, with sterilized water by pycnidia
Wash down and smashed up, conidium is then automatically discharged into water, be settled to 20 mL, take 20 μ L and observed to blood counting chamber
And the sporulation quantity of unit of account area.
Described glue spore anthrax sporulation quantity is up to 6.5 × 109Individual/cm2。
Embodiment 3
（1）The colletotrichum gloeosporioides Penz that will have been activated is transferred on PDA plate again, is placed in illumination in 28 DEG C of constant incubators
Cultivate 36 h；
（2）When bacterium colony 4 cm long to a diameter of, in superclean bench with aseptic cotton carrier by center a diameter of 2.5
Cm aerial hyphaes remove, and with sterilized water by surface washing 2 ~ 3 times, back-off dries naturally；
（3）Flat board after drying continues the h of illumination cultivation 36 during 28 DEG C of constant incubator is laid in after closeing the lid；
（4）Orange-red pycnidia is seen in planar surface after the h of illumination 36, with sterilized water by pycnidia
Wash down and racked, conidium is then automatically discharged into water, be settled to 20 mL, take 20 μ L and observed to blood counting chamber
And the sporulation quantity of unit of account area.
Described glue spore anthrax sporulation quantity is up to 3.6 × 108Individual/cm2。
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent with
Modification, should all belong to covering scope of the invention.
Claims (2)
1. a kind of Rapid Sporulation method of glue spore anthrax, it is characterised in that：With the colletotrichum gloeosporioides Penz of PDA solid medium cultures
Mycelium is the basis for producing spore, and mycelium is induced in illumination by striking off mycelia, aseptic water washing, drying and illumination cultivation measure
A large amount of spores are sprouted under 24~48 hs rapidly；
Mainly include the following steps that：
（1）To separate or the colletotrichum gloeosporioides Penz that has activated be transferred on PDA culture medium flat board again, be placed in 28 DEG C it is incubated
Illumination cultivation 24-48h in case；
（2）When bacterium colony 3~4.5cm long to a diameter of, in superclean bench with aseptic cotton carrier by mid-diameter be 2 ~ 3.5cm
Aerial hyphae remove, with sterilized water by surface washing 2 ~ 3 times, flat board back-off is dried naturally；
（3）Flat board after drying continues illumination cultivation 24 ~ 48 during 28 DEG C of constant incubator is laid in after closeing the lid again
h；
（4）Orange-red pycnidia is seen in planar surface after the h of illumination 24 ~ 48, pycnidia is washed with sterilized water
Descend and smashed up, conidium is then automatically discharged into water, is settled to 20 mL, is taken 20 μ L and is observed simultaneously to blood counting chamber
The sporulation quantity of unit of account area.
2. the Rapid Sporulation method of glue spore anthrax according to claim 1, it is characterised in that：Described glue spore anthrax produces spore
Measure is 7.8 × 107~6.5×109Individual/cm2。
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| CN106754623A (en) * | 2017-01-10 | 2017-05-31 | 广西大学 | A kind of anthrax bacteria for efficiently avoiding polluting is conidial to prepare cultural method |
| CN106754423A (en) * | 2017-01-10 | 2017-05-31 | 广西大学 | The standing form and its preparation and application method of a kind of anthrax bacteria lab material |
| CN108048388A (en) * | 2018-01-24 | 2018-05-18 | 四川农业大学 | A kind of colletotrichum gloeosporioides Penz produces spore method |
| CN109122727B (en) * | 2018-07-16 | 2021-02-19 | 华南农业大学 | Application of colletotrichum gloeosporioides BWH-1 in preparation of herbicide |
| CN108570442B (en) * | 2018-07-24 | 2021-05-28 | 湖北省农业科学院经济作物研究所 | Method for rapidly inducing spore production of anthrax |
| CN108998404B (en) * | 2018-09-13 | 2022-02-25 | 安徽农业大学 | Method for inducing anthrax bacteria to produce conidia |
| CN109355207B (en) * | 2018-12-07 | 2021-10-08 | 山东省农业科学院植物保护研究所 | Liquid fermentation culture medium for producing spores from anthrax |
| CN110878280B (en) * | 2019-12-12 | 2021-11-09 | 北京市农林科学院 | Method for inducing cacao trichoderma to quickly generate conidia |
| CN112175838B (en) * | 2020-09-19 | 2022-03-18 | 福建省农业科学院植物保护研究所 | Culture medium for inducing and destroying anthrax bacteria to produce spores, preparation method and method for inducing and producing spores |
| CN112375728B (en) * | 2020-10-23 | 2022-05-17 | 浙江省农业科学院 | New application of nano titanium dioxide |
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| CN1284558A (en) * | 2000-09-04 | 2001-02-21 | 南京农业大学 | Psorospermial anthracin strain and its application method in biological herbicide |
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