CN105132294B - A kind of alternaric bacteria and its application in dendrobium candidum disease-resistanting leaf spot - Google Patents
A kind of alternaric bacteria and its application in dendrobium candidum disease-resistanting leaf spot Download PDFInfo
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- CN105132294B CN105132294B CN201510624581.0A CN201510624581A CN105132294B CN 105132294 B CN105132294 B CN 105132294B CN 201510624581 A CN201510624581 A CN 201510624581A CN 105132294 B CN105132294 B CN 105132294B
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- dendrobium candidum
- alternaric bacteria
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Abstract
The invention discloses a kind of alternaric bacteria, its Classification And Nomenclature is Deuteromycotina Alternaria alternaric bacteria (Alternaria alternata), bacterial strain number is JSNL005 2, it has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.11312, and preservation date is September in 2015 11.Alternaric bacteria JSNL005 2 of the present invention is notable in raising Growth of Dendrobium candidum disease-resistanting leaf spot ability etc. action effect；And can manually cultivate, reproduction speed is fast, is convenient for large-scale production, has good development prospect.
Description
Technical field
The invention belongs to microbial technology field, and in particular to a kind of alternaric bacteria and its in dendrobium candidum disease-resistanting leaf spot
Application.
Background technology
Dendrobium candidum is the perennial herbaceous plant that grows nonparasitically upon another plant of orchid family Dendrobium, and research shows, is sprouted from its seed to the strong of seedling
Kang Shengchang, without the participation for not needing symbiotic effects.Because dendrobium officinale resource is on the verge of exhaustion, at present, the city of dendrobium candidum
Field demand is mainly by artificial cultivation, but artificial growth Growth of Dendrobium candidum speed is slow, and average product is low, and disease occurs more tight
Weight.At present, the preventing and treating of disease still relies primarily on agricultural chemicals.From the angle preserved the ecological environment with food security, iron sheet stone
Dry measure used in former times should reduce the amount of application of agricultural chemicals, chemical fertilizer as far as possible as rare Chinese medicine.Mycorhiza is mycorrhizal fungi and plant roots in soil
It is the homobium with specific modality 26S Proteasome Structure and Function formed, is a kind of symbiosis of generally existing in nature.Orchid family
Plant is stronger to the dependence of mycorhiza, and mycorrhizal fungi and the conditional formation homobium of root system of plant, are being physiologically a phases
The entirety mutually coordinated, and during common evolutionary, form specific morphological feature.Mycorrhizal fungi and host plant it is mutual
Relation be in most cases it is mutually beneficial, mycorrhizal fungi obtained out of plant growth necessary to nutriment, and plant by with
Symbiotic effects are combined, and required nutrient and moisture etc. also can be preferably obtained from soil.Mycorhiza resource is rationally utilized, improves iron
The resistance against diseases of the skin stem of noble dendrobium, the amount of application of chemical pesticide is reduced to a certain extent, this also complies with the sustainable development war of agricultural
Slightly.Therefore, bush mycorrhiza agent correlative study has turned into an important directions of dendrobium candidum industry.
The content of the invention
It is true that the technical problems to be solved by the invention are to provide a kind of to dendrobium candidum resistant effect significant effect mycorhiza
Bacterium --- Deuteromycotina Alternaria alternaric bacteria (Alternaria alternata).
The present invention also technical problems to be solved are to provide application of the above-mentioned fungi in dendrobium candidum resistance against diseases is improved.
In order to solve the above technical problems, the technical solution adopted by the present invention is as follows：
A kind of alternaric bacteria, its Classification And Nomenclature are Deuteromycotina Alternaria alternaric bacteria (Alternaria
Alternata), bacterial strain number is JSNL005-2, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center (CGMCC), deposit number are CGMCC No.11312, and preservation date is September in 2015 11.Preservation address is Beijing
No. 3 Institute of Microorganism, Academia Sinica of institute of Chaoyang District North Star West Road 1, postcode 100101.
The above-mentioned bacterial strains JSNL005-2 upgrowth situations that are in October, 2013 in Jurong City of Jiangsu Province dendrobium candidum plant area compared with
Separation obtains on good plant, and the bacterial strain is not notable to dendrobium candidum growth-promoting functions.But the anti-of dendrobium candidum can be obviously improved
Sick ability.
Alternaric bacteria (Alternaria alternata) JSNL005-2 colony characteristicses are as follows：
Morphological observation shows：Cultivated on PDA plate, mycelia is sparse, and secretly to black, bacterium colony is in villiform, more
For grey black to black, substratess surface (Fig. 1) is invested.Amplification ITS area's complete sequences are simultaneously sequenced, and PCR expands what is obtained
16sRNA complete sequences such as SEQ ID No：Shown in 1.
Application of the above-mentioned alternaric bacteria in dendrobium candidum disease-resistanting leaf spot is also within protection scope of the present invention.
Specifically application process is：Alternaric bacteria (Alternaria alternata) JSNL005-2 is prepared into solid
Microbial inoculum, dendrobium candidum seedling cultivation matrix surface is placed in, the solid fungicide inoculum concentration of every clump of dendrobium candidum seedling is 5g, using bacterium
After agent, Relative water content maintains more than 60% 10 day, to ensure that mycelia survives in the substrate.
Wherein, described solid fungicide is prepared as follows obtaining：It is by culture matrix, ammonium nitrate of cotton seed hulls bacteria residue
External source nitrogen source, sucrose are external source carbon source, are prepared into solid medium, sterilizing, inoculation alternaric bacteria (Alternaria
Alternata) JSNL005-2, cultivated 30~35 days under the conditions of 28 DEG C, 12h illumination+12h dark, pH6.5, will be cultured
Microbial inoculum cuts into 1cm3The block of size, the inoculation for dendrobium candidum plant bush mycorrhiza agent.
Beneficial effect：Compared with prior art, it is an advantage of the invention that：
(1) alternaric bacteria (Alternaria alternata) JSNL005-2 to dendrobium candidum resistant effect effect more
Significantly, there is certain effect to the use for reducing chemical pesticide, and ensure the important means of edible safety.
(2) alternaric bacteria (Alternaria alternata) JSNL005-2 condition of culture is simple, easily preserves, and is easy to
Industrialized production, there is good development prospect.
Brief description of the drawings
Fig. 1 is JSNL005-2 bacterium colony culture photos.
Fig. 2 is the disease-resistanting leaf spot capability analysis of JSNL005-2 Mycorrhizal dendrobium candidums.
Embodiment
According to following embodiments, the present invention may be better understood.It is however, as it will be easily appreciated by one skilled in the art that real
Apply the content described by example and be merely to illustrate the present invention, without should be also without limitation on sheet described in detail in claims
Invention.
Embodiment 1：
Resistance against diseases action effects point of alternaric bacteria (Alternaria alternata) JSNL005-2 to dendrobium candidum
Analysis.
(1) be external source nitrogen source using cotton seed hulls bacteria residue as culture matrix, ammonium nitrate, sucrose be external source carbon source, be prepared into solid
Culture matrix loads in autoclave bags, is sterilized 20 minutes under 121 DEG C and 101.3 kPas of condition of high voltage.Treat culture matrix
Aseptically Arbuscular Mycorrhizal Fungi fungi JSNL005-2 after cooling, is then manually cultivated.Specific condition of culture is 28 DEG C,
12h illumination+12h is dark, pH6.5, cultivates 30~35 days.Cultured microbial inoculum is processed into 1cm3The block of size, is placed in
Dendrobium candidum seedling cultivation matrix surface, the solid fungicide inoculum concentration of every clump of dendrobium candidum seedling is 5g, after microbial inoculum, matrix
Relative water content maintains more than 60% 10 day, to ensure that mycelia survives in the substrate.
(2) Leaf blotch pathogeny by ultraviolet light and dark processing culture in PDA culture medium, germ spore is made and hangs
Supernatant liquid, it is diluted to 2 × 105Individual/mL.
(3) it will under equal conditions be managed, be chosen for the 5th, top of examination plant leaf for examination dendrobium candidum plant,
Wound is manually manufactured with needle point method, in every plant of the blade surface sprinkling pathogen spore suspension 2ml/ of manufacture wound, greenhouse
Temperature is 30-35 DEG C, and humidity 80%, every kind of processing is 100 plants, and sets 3 groups of repetitions.
(4) occurring degree of leaf spot, the statistics incidence of disease, disease index are monitored after being inoculated with daily.
Diseased plant grade scale：Mainly there is Lesion number and diameter according to inoculation blade tow sides in severity Scaling, just fixed
Grade scale is：
0：Normally；
Ⅰ：Tow sides have a small amount of scab；
Ⅱ：Tow sides have scab, and onset area accounts for the blade gross area below 1/4；
Ⅲ：Tow sides have scab, and onset area accounts for the 1/4-1/2 of the blade gross area；
Ⅳ：Tow sides have scab, and onset area accounts for blade gross area 1/2-3/4；
Ⅳ：Tow sides have scab, and onset area accounts for the blade gross area more than 3/4.
(total strain number × highest level represents number to disease-sensitive index=∑ (the diseased plant numbers at different levels × level represents numerical value) × 100/
Value)；
Susceptible gene=(susceptible strain tree/inoculation strain tree) × 100%.
As a result Fig. 2 is seen.
Claims (4)
1. a kind of alternaric bacteria, its Classification And Nomenclature is Deuteromycotina Alternaria alternaric bacteria (Alternaria
Alternata), bacterial strain number is JSNL005-2, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center, deposit number are CGMCC No.11312, and preservation date is September in 2015 11.
2. application of the alternaric bacteria in dendrobium candidum disease-resistanting leaf spot described in claim 1.
3. application according to claim 2, it is characterised in that：By alternaric bacteria (Alternaria alternata)
JSNL005-2 is prepared into solid fungicide, is placed in dendrobium candidum seedling cultivation matrix surface, the solid bacterium of every clump of dendrobium candidum seedling
Agent inoculum concentration is 5g, and after microbial inoculum, Relative water content maintains more than 60% 10 day, with ensure mycelia in the substrate into
It is living.
4. application according to claim 3, it is characterised in that described solid fungicide is prepared as follows obtaining：With
Cotton seed hulls bacteria residue is culture matrix, ammonium nitrate is external source nitrogen source, sucrose is external source carbon source, is prepared into solid medium, sterilizing, is connect
Kind alternaric bacteria (Alternaria alternata) JSNL005-2, under the conditions of 28 DEG C, 12h illumination+12h dark, pH6.5
Culture 30~35 days, 1cm is cut into by cultured microbial inoculum3The block of size, for dendrobium candidum plant bush mycorrhiza agent
Inoculation.
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| CN105349440B (en) * | 2015-12-18 | 2019-05-24 | 浙江省农业科学院 | For cultivating the culture medium containing Specific amino acid of dendrobium candidum tar spot bacterium |
| CN105420121B (en) * | 2015-12-18 | 2019-05-24 | 浙江省农业科学院 | For cultivating the culture medium containing special sugar source of dendrobium candidum tar spot bacterium |
| CN107548986B (en) * | 2017-08-30 | 2021-06-08 | 浙江工业大学 | Dendrobium officinale planting method capable of realizing high yield of dendrobine under salt stress |
| CN112280821A (en) * | 2020-10-28 | 2021-01-29 | 钱康(北京)科技有限公司 | Method for artificially synthesizing variety |
Citations (3)
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|---|---|---|---|---|
| CN102732471A (en) * | 2012-07-23 | 2012-10-17 | 江苏农林职业技术学院 | Bacillus subtilis and application thereof in preventing and treating plant leaf spot diseases |
| CN104195057A (en) * | 2014-09-10 | 2014-12-10 | 江苏农林职业技术学院 | Umbelopsis nana and applications thereof in preventing leaf spot disease of dendrobium officinale |
| CN104195055A (en) * | 2014-09-10 | 2014-12-10 | 江苏农林职业技术学院 | Fusarium oxysporum and application thereof in resisting leaf spot disease of dendrobium officinale |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732471A (en) * | 2012-07-23 | 2012-10-17 | 江苏农林职业技术学院 | Bacillus subtilis and application thereof in preventing and treating plant leaf spot diseases |
| CN104195057A (en) * | 2014-09-10 | 2014-12-10 | 江苏农林职业技术学院 | Umbelopsis nana and applications thereof in preventing leaf spot disease of dendrobium officinale |
| CN104195055A (en) * | 2014-09-10 | 2014-12-10 | 江苏农林职业技术学院 | Fusarium oxysporum and application thereof in resisting leaf spot disease of dendrobium officinale |
Non-Patent Citations (1)
| Title |
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| 石斛兰镰刀菌叶斑病的生物防治研究;程萍等;《中国农学通报》;20080930(第09期);357-361页 * |
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