CN104195057A - Umbelopsis nana and applications thereof in preventing leaf spot disease of dendrobium officinale - Google Patents
Umbelopsis nana and applications thereof in preventing leaf spot disease of dendrobium officinale Download PDFInfo
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- CN104195057A CN104195057A CN201410459653.6A CN201410459653A CN104195057A CN 104195057 A CN104195057 A CN 104195057A CN 201410459653 A CN201410459653 A CN 201410459653A CN 104195057 A CN104195057 A CN 104195057A
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Abstract
The invention discloses umbelopsis nana, the classification and name of the umbelopsis nana are zygomycota mucorales umbelopsis nana, the strain number is JSNL001-3, and the umbelopsis nana is currently collected in China General Microbiological Culture Collection Center (CGMCC) at 15th August, 2014, with the collection number of CGMCC No.9565. The umbelopsis nana JSNL001-3 is remarkable in effect on improving the leaf spot disease during the growth of dendrobium officinale; the umbelopsis nana can be artificially cultured, is fast in breeding speed and convenient for large-scale production, and has excellent development and application prospects.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to the mould and application in Herba Dendrobii disease-resistanting leaf spot of a kind of short and small umbrella.
Background technology
Herba Dendrobii is the perennial herbaceous plant that grows nonparasitically upon another plant of the orchid family Dendrobium, and research shows that the healthy growth from its seed germination to seedling needs the participation of symbiosis fungi invariably.Because wild Herba Dendrobii resource is on the verge of exhaustion, at present, the market requirement of Herba Dendrobii is mainly by artificial culture, but artificial growth Growth of Dendrobium candidum speed is slow, and mean yield is low, and disease occurs comparatively serious.At present, the control of disease still mainly relies on agricultural chemicals.From preserving the ecological environment and the angle of food safety, Herba Dendrobii, as rare Chinese medicine, should reduce the amount of application of agricultural chemicals, chemical fertilizer as far as possible.Mycorhiza is the symbiote with specific modality structure and function that in soil, mycorrhizal fungi and root system of plant form, and is the ubiquitous a kind of commensalism of occurring in nature.Orchid is stronger to the dependency of mycorhiza, and the conditional formation symbiote of mycorrhizal fungi and root system of plant is an entirety of mutually coordinating, and in common process of evolving, forms specific morphological specificity on physiology.The mutual relationship of mycorrhizal fungi and host plant is in most cases mutually beneficial, and mycorrhizal fungi obtains the essential nutritive substance of growth in plant materials, and plant is by being combined with symbiosis fungi, also can better from soil, obtain required nutrient and moisture etc.Rationally utilize mycorhiza resource, improve the resistance against diseases of Herba Dendrobii, reduce to a certain extent the amount of application of chemical pesticide, this also meets the agriculture strategy of sustainable development.Therefore, bush mycorrhiza agent correlative study has become an important directions of Herba Dendrobii industry.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of to Herba Dendrobii resistant effect effect mycorrhizal fungi significantly---the short and small umbrella of the mould genus of Zygomycotina mucorales umbrella shape mould (Umbelopsis nana).
The technical problem that the present invention also will solve is to provide above-mentioned fungi in the application improving in Herba Dendrobii resistance against diseases.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
A kind of short and small umbrella is mould, its Classification And Nomenclature is the short and small umbrella of the mould genus of Zygomycotina mucorales umbrella shape mould (Umbelopsis nana), bacterial strain number is JSNL001-3, now be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), deposit number is CGMCC No.9565, and preservation date is on August 15th, 2014.Preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, postcode 100101.
Above-mentioned bacterial strains JSNL001-3 separates and obtains on the good plant of upgrowth situation in October, 2010 in Yueqing City of Zhejiang Province Herba Dendrobii is planted district, and this bacterial strain is not remarkable to Herba Dendrobii growth-promoting functions.But can obviously improve the resistance against diseases of Herba Dendrobii.
The colony characteristics of short and small umbrella mould (Umbelopsis nana) JSNL001-3 is as follows:
Morphological observation shows: on PDA puts down a rebellion, cultivate, mycelia is sparse, is wheel line shape, invests base thing surface; Tow sides are canescence (Fig. 1).Amplification ITS district complete sequence also checks order, and the 16sRNA complete sequence that pcr amplification obtains is as shown in SEQ ID No:1.
The mould application in Herba Dendrobii disease-resistanting leaf spot of above-mentioned short and small umbrella is also within protection scope of the present invention.
Concrete application method is: mould short and small umbrella (Umbelopsis nana) JSNL001-3 is prepared into solid fungicide, be placed in Herba Dendrobii seedling cultivation matrix surface, the solid fungicide inoculum size of every clump of Herba Dendrobii seedling is 5g, use after microbial inoculum, Relative water content maintains more than 60% 10 days, to guarantee that mycelia survives in the substrate.
Wherein, described solid fungicide prepares as follows: taking cotton seed hulls bacterium slag as culture medium, ammonium nitrate as external source nitrogenous source, sucrose be external source carbon source, be prepared into solid medium, sterilizing, inoculate short and small umbrella mould (Umbelopsis nana) JSNL001-3, under 28 DEG C, 12h illumination+12h dark, pH6.5 condition, cultivate 25～35 days, cultured microbial inoculum is cut into 1cm
3the block of size, for the inoculation of Herba Dendrobii plant bush mycorrhiza agent.
Beneficial effect: compared with prior art, advantage of the present invention is:
(1) short and small umbrella mould (Umbelopsis nana) JSNL001-3 is comparatively remarkable to Herba Dendrobii resistant effect effect, and the use that reduces chemical pesticide is had to certain effect, is also the important means that ensures edible safety.
(2) short and small umbrella mould (Umbelopsis nana) JSNL001-3 culture condition simply, easily preserve, be easy to suitability for industrialized production, there is good development prospect.
Brief description of the drawings
Fig. 1 is that JSNL001-3 bacterium colony is cultivated photo.
Fig. 2 is the disease-resistanting leaf spot capability analysis of JSNL001-3 bacterium Mycorrhizal Herba Dendrobii.
Embodiment
According to following embodiment, the present invention may be better understood.But, those skilled in the art will readily understand, the described content of embodiment is only for the present invention is described, and should also can not limit the present invention described in detail in claims.
Embodiment 1:
Short and small umbrella mould (Umbelopsis nana) JSNL001-3 analyzes the resistance against diseases action effect of Herba Dendrobii.
(1) taking cotton seed hulls bacterium slag as culture medium, ammonium nitrate is as external source nitrogenous source, sucrose are external source carbon source, be prepared into solid culture matrix and pack in autoclaving bag, sterilizing 20 minutes under the condition of high voltage of 121 DEG C and 101.3 kPas.After culture medium is cooling under aseptic condition Arbuscular Mycorrhizal Fungi fungi JSNL001-3, then carry out artificial culture.Concrete culture condition is 28 DEG C, 12h illumination+12h dark, pH6.5, cultivates 25～35 days.Cultured microbial inoculum is processed into 1cm
3the block of size, is placed in Herba Dendrobii seedling cultivation matrix surface, and the solid fungicide inoculum size of every clump of Herba Dendrobii seedling is 5g, uses after microbial inoculum, and Relative water content maintains more than 60% 10 days, to guarantee that mycelia survives in the substrate.
(2) cultivate the leaf spot pathogenic bacteria on PDA substratum by UV-light and dark processing, make germ spore suspension, be diluted to 2 × 105/mL.
(3) will under equal conditions manage for examination Herba Dendrobii plant, all choose for the 5th, plant top of examination leaf, manually manufacture wound with needle punching, spray the every strain of pathogenic bacteria spore suspension 2ml/ at the blade surface of manufacturing wound, temperature room temp is 30-35 DEG C, humidity 95%, every kind of processing is 100 strains, and establishes 3 groups of repetitions.
(4) the rear occurring degree of monitoring leaf spot every day of inoculation, statistics sickness rate, disease index.
Diseased plant grade scale: severity Scaling mainly occurs Lesion number and diameter according to inoculation blade tow sides, just determine grade scale and be:
0: normal;
I: tow sides have a small amount of scab;
II: tow sides have scab, onset area accounts for the blade total area below 1/4;
III: tow sides have scab, onset area accounts for the 1/4-1/2 of the blade total area;
IV: tow sides have scab, onset area accounts for blade total area 1/2-3/4;
IV: tow sides have scab, onset area accounts for the blade total area more than 3/4.
Susceptible index=∑ (diseased plant numbers at different levels × this level represents numerical value) × 100/ (total strain number × highest level represents numerical value);
Susceptible rate=(strain tree is set/inoculates in susceptible strain) × 100%.
The results are shown in Figure 2.
Claims (4)
1. a short and small umbrella is mould, its Classification And Nomenclature is the short and small umbrella of the mould genus of Zygomycotina mucorales umbrella shape mould (Umbelopsis nana), bacterial strain number is JSNL001-3, now be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.9565, and preservation date is on August 15th, 2014.
2. the mould application in Herba Dendrobii disease-resistanting leaf spot of short and small umbrella claimed in claim 1.
3. application according to claim 2, it is characterized in that: mould short and small umbrella (Umbelopsis nana) JSNL001-3 is prepared into solid fungicide, be placed in Herba Dendrobii seedling cultivation matrix surface, the solid fungicide inoculum size of every clump of Herba Dendrobii seedling is 5g, use after microbial inoculum, Relative water content maintains more than 60% 10 days, to guarantee that mycelia survives in the substrate.
4. application according to claim 3, it is characterized in that, described solid fungicide prepares as follows: taking cotton seed hulls bacterium slag as culture medium, ammonium nitrate as external source nitrogenous source, sucrose be external source carbon source, be prepared into solid medium, sterilizing, inoculate short and small umbrella mould (Umbelopsis nana) JSNL001-3, under 28 DEG C, 12h illumination+12h dark, pH6.5 condition, cultivate 25～35 days, cultured microbial inoculum is cut into 1cm
3the block of size, for the inoculation of Herba Dendrobii plant bush mycorrhiza agent.
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Cited By (1)
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| CN105132294B (en) * | 2015-09-25 | 2018-03-30 | 江苏农林职业技术学院 | A kind of alternaric bacteria and its application in dendrobium candidum disease-resistanting leaf spot |
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