CN112746053A - Gliocladium roseum NF-06 and application of conidia in inhibition of fusarium graminearum subcapsule shell - Google Patents
Gliocladium roseum NF-06 and application of conidia in inhibition of fusarium graminearum subcapsule shell Download PDFInfo
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- CN112746053A CN112746053A CN202011431302.6A CN202011431302A CN112746053A CN 112746053 A CN112746053 A CN 112746053A CN 202011431302 A CN202011431302 A CN 202011431302A CN 112746053 A CN112746053 A CN 112746053A
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Abstract
The invention relates to the technical field of microorganisms, in particular to gliocladium roseum NF-06 and application of conidia in inhibiting fusarium graminearum subcapsule shells. The gliocladium roseum NF-06 conidia prepared by the method is sprayed under the large background of returning the total amount of crop straws to the field, so that the generation of fusarium graminearum ascocarp is inhibited, the initial infection source of wheat scab is reduced, and the green prevention and control purpose of 'prevention and forward movement comprehensive prevention and control' of the wheat scab is achieved. The inhibition rate of the invention for inhibiting the fusarium graminearum ascocarp is high, 80-90%, and the invention is safe and pollution-free.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to gliocladium roseum NF-06 and application of conidia in inhibiting fusarium graminearum subcapsule shells.
Background
The pathogen of wheat scab is mainly Fusarium graminearum (Fusarium graminearum), which is a filamentous ascomycete fungus that can be divided into two stages according to its growth and development: asexual and sexual periods. Conidia are produced in asexual period; the spores produced during sexual periods are ascospores. Researches show that fusarium graminearum overwinter on disease plant residues such as corn, wheat and rice in the form of saprophytic hypha, and the generation and maturation of fusarium graminearum ascocarp are facilitated in warm and humid days in spring. In the flowering period of wheat, a large amount of viscous ascospores are strongly ejected from ascospores in ascospores mature on the surfaces of plant disease residues and spread to host plants along with media such as wind, rainwater, insects and the like, the ascospores are used as a primary infection source to be infected into ear tissues such as wheat or anthers, lemma or glume of other host plants to be attached to form hypha, and the hypha invades the host tissues and grows and expands in cells. Thus, the number of mature ascospores and their release of ascospores play a crucial role in the cycle of fusarium graminearum infestation.
The method controls the initial infection source of the wheat scab, reduces the base number of bacteria sources, and has important significance for preventing and controlling the occurrence of the wheat scab. However, the traditional control strategy of wheat scab is mainly chemical control, which is mainly carried out in the heading and flowering period of wheat, especially in the severe epidemic years of wheat scab, the chemical control effect and the toxin reducing effect are not ideal, and more importantly, the chemical pesticide is used in successive years, so that the generation of fusarium graminearum resistance is caused, the quality of wheat and the safety of agricultural ecological environment are seriously influenced, and the wheat production faces huge pressure.
Therefore, a method which is safe, does not affect the ecological environment and has high inhibition rate of inhibiting the ascochyta of fusarium graminearum is urgently needed in the prior art to control the wheat scab.
Disclosure of Invention
The invention aims to provide application of gliocladium roseum NF-06 and conidia in inhibition of fusarium graminearum subcapsule shells, and the inhibition rate of the gliocladium roseum NF-06 and conidia on the fusarium graminearum subcapsule shells is 80-90%.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of gliocladium roseum NF-06 conidia, which comprises the following steps:
(1) inoculating gliocladium roseum NF-06 into a spore production culture medium, and culturing to obtain a culture solution containing conidia;
(2) and filtering the culture solution containing the conidia, centrifuging the filtrate, and taking the precipitate to obtain the NF-06 conidia.
Preferably, the spore-forming medium is a CMC liquid medium.
Preferably, the culture conditions in step (1) are: the culture temperature is 23-27 ℃; the rotating speed of the shaking table is 170-180 rpm; the culture time is 45-51 h.
The invention also provides application of the NF-06 conidium or the gliocladium roseum NF-06 prepared by the method in inhibiting the fusarium graminearum ascocarp.
The invention also provides application of the NF-06 conidium or gliocladium roseum NF-06 prepared by the method in inhibiting wheat scab caused by fusarium graminearum.
Preferably, the NF-06 conidia is prepared at a concentration of 1 × 105～1×107The cfu/mL suspension is sprayed on crop straws for use.
The invention provides application of gliocladium roseum NF-06 and conidia in inhibiting fusarium graminearum subcapsule shells, and compared with the prior art, the gliocladium roseum NF-06 and conidia have the following beneficial effects:
(1) the gliocladium roseum NF-06 has the effect of efficiently inhibiting the ascocarp of fusarium graminearum, and the bacteriostasis rate is 80-90 percent;
(2) the application of the invention changes the chemical prevention and control of wheat scab only in the period of wheat full spike and flowering at present, the straw is returned to the field by spraying the conidia of the gliocladium roseum, and the fusarium graminearum cyst shell is inhibited by the gliocladium roseum, thereby achieving the purpose of green prevention and control of wheat scab.
Drawings
FIG. 1 is a diagram of Fusarium graminearum culture.
FIG. 2 is a graph showing the inhibitory effect of NF-06 on Fusarium graminearum in the medium.
FIG. 3 is a graph showing the effect of NF-06 on Fusarium graminearum in field.
Deposit description
The NF-06 is classified and named as gliocladium roseum, the Latin is named as Clonostachys rosea, the preservation unit is China general microbiological culture Collection center (CGMCC), the address is No. 3 of Xilu No.1 of Beijing Kogyo Beichen, the preservation date is 2018, 08 and 07 days, and the preservation number is CGMCC No. 16262.
Detailed Description
The invention provides a preparation method of gliocladium roseum NF-06 conidia, which comprises the following steps:
(1) inoculating gliocladium roseum NF-06 into a spore production culture medium, and culturing to obtain a culture solution containing conidia;
(2) and filtering the culture solution containing the conidia, centrifuging the filtrate, and taking the precipitate to obtain the NF-06 conidia.
In the invention, the classification of NF-06 is named as gliocladium roseum, the Latin is named as Clinostachy rosea, the preservation unit is China general microbiological culture Collection center (CGMCC), the address is No. 3 of No.1 Siro of Beijing Kogyang area, the preservation date is 2018, 08 and 07 days, and the preservation number is CGMCC No. 16262.
In the invention, the spore-forming culture medium is a CMC liquid culture medium.
In the invention, the CMC liquid culture medium comprises the following components in parts by mass: 13-17 g/L of CMC, 0.8-1.2 g/L of Yeast Extract and KH2PO4 0.8～1.2g/L，KCl 0.8～1.2g/L， MgSO4·7H2O 0.4～0.6g/L，NH4NO30.8-1.2 g/L; preferably 15g/L of CMC, 1.0g/L of Yeast Extract, KH2PO4 1.0g/L，KCl 1.0g/L，MgSO4·7H2O 0.5g/L，NH4NO3 1.0g/L。
In the invention, the culture temperature of the conidia obtained by the culture is 23-27 ℃, and preferably 25 ℃; the rotating speed of the shaking table is 170-180 rpm, preferably 175 rpm; the culture time is 45-51 h, preferably 48 h.
In the invention, the conidium culture solution is filtered by using a sterilized filter cloth.
The invention also provides application of the NF-06 conidia or the NF-06 gliocladium roseum prepared by the preparation method of the NF-06 conidia in inhibiting the ascocarp of fusarium graminearum.
The invention also provides application of the NF-06 conidia or the NF-06 gliocladium roseum prepared by the preparation method of the NF-06 conidia in inhibiting wheat scab caused by fusarium graminearum.
In the present invention, the NF-06 conidia are prepared to have a concentration of 1X 105～1×107The cfu/mL suspension is sprayed on crop straws for use, and the optimal concentration is 1 multiplied by 106cfu/mL of suspension.
In the embodiment of the invention, the preparation method of the carrot culture medium comprises the following steps: cleaning and peeling carrots, cutting the carrots into small blocks, putting the small blocks into water with the mass-volume ratio of 240-260 g/500ml, sterilizing for 8-12 min, juicing, adding water, adding agar with the mass-volume ratio of 18-22 g/1000ml, and sterilizing for 25-35 min at the high temperature of 121 ℃ by moist heat; preferably, carrot is firstly added into water with the mass volume ratio of 250g/500ml, sterilized for 10min, juiced, added with water, added with agar with the mass volume ratio of 20g/1000ml, and sterilized by moist heat at the high temperature of 121 ℃ for 30 min.
The preparation method of 0.1 percent of Tween-20 comprises the following steps: 100 mu L of Tween-20 stock solution is added with distilled water to 100 ml and evenly mixed, and then the mixture is subjected to moist heat sterilization at the high temperature of 121 ℃ for 20 min.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
In an incubator at 25 ℃, fusarium graminearum is overgrown on a carrot culture medium with a diameter of 6cm plastic plate, sterile 0.1% Tween-20 (Tween-20) is added, and an aerial hypha is pressed down by a sterilization spoon to be attached to the surface of the culture medium, and the result is shown in figure 1.
Inoculating the separated gliocladium roseum NF-06 into a CMC spore production culture medium, culturing for 2 days, filtering with sterilized filter cloth to obtain filtrate, and centrifuging the filtrate at 3000rpm for 4min to obtain precipitate, i.e. gliocladium roseum NF-06 conidia. Preparing gliocladium roseum NF-06 conidia with sterile water to obtain a concentration of 1 × 105cfu/mL of suspension.
10 mu L of the obtained NF-06 conidia suspension is sprayed into a fusarium graminearum plate culture medium to be used as a treatment group, a group without spraying the NF-06 conidia suspension is used as a control group, and 3 repeats are arranged in each group. In 25 ℃ incubator, black light: dark-12 h: 12h conditioned medium, and after 4 days, the number of shells of Fusarium graminearum produced in the treated and control groups was observed, and the results are shown in FIG. 2 and Table 1. In which the formation of the ascochyta graminis shell was observed in the control and treated groups at 24h, 48h, 72h and 96h, respectively, and the results are shown in FIG. 3.
The preparation of the CMC liquid medium comprises the following steps: according to 15g/L CMC, 1.0g/L Yeast Extract, KH2PO4 1.0g/L，KCl 0.8g/L，MgSO4·7H2O0.5g/L，NH4NO31.2g/L, adding distilled water to a constant volume of 1L, and sterilizing at 121 deg.C for 20 min.
Carrot culture medium: cleaning carrot, peeling, cutting into small pieces, adding 250g into 500mL of distilled water, sterilizing for 10min, squeezing, adding distilled water to 1000mL, adding 20g of agar, and sterilizing at 121 deg.C for 30 min.
The preparation method of 0.1 percent of Tween-20 comprises the following steps: 100 mu L of Tween-20 stock solution is added with distilled water to 100 ml and evenly mixed, and then the mixture is subjected to moist heat sterilization at the high temperature of 121 ℃ for 20 min.
TABLE 1 number of Fusarium graminearum cysts in culture medium for treatment and control groups
Example 2
The isolated Gliocladium roseum NF-06 was formulated into conidia concentration of 1X 10 in the manner of example 16cfu/mL of suspension.
Referring to CN110741889A, a method for predicting the initial infection source abundance of wheat scab in example 1, corn straws are prepared, fusarium graminearum pathogens are selected as inoculation strains, spore suspension of the inoculation strains is prepared, and the corn straws are inoculated. Growing straw inoculated with fusarium graminearum in an incubator at 25 ℃ for 24 hours, and then, enabling NF-06 conidium suspension to grow according to the proportion of 1 × 106cfu/mL was sprayed on maize straws inoculated with Fusarium graminearum as a treatment group, straws not sprayed with NF-06 conidia suspension were used as a control group, and 3 replicates were set for each group. And respectively putting the straws of the treatment group and the control group again into the incubator at 25 ℃ for continuous culture for 6-7 days. After the cultivation is finished, the straws are taken out, the straw control is carried out by referring to the straw-inoculated wheat field placing step in the embodiment 1, and the straws are fixed on the ground surface of the wheat field. And (3) investigating the quantity of the fusarium graminearum ascochyta shells of the treatment group and the control group 10-15 days before wheat blossom. The results are shown in Table 2.
The method for counting the fusarium graminearum ascochyta comprises the steps of using a dissecting mirror, selecting 6 points before counting each straw, wherein the area of each point is 0.1923cm2And 3 straws are calculated in each group, 30 or 40 times of dissecting mirror is used, the number of the ascocarp shells in the selected area is counted, and then the number is converted into the number of the ascocarp shells on the surfaces of the straws.
TABLE 2 Fusarium graminearum cyst Shell counts in treatment and control groups
The embodiment shows that the invention provides the application of the gliocladium roseum NF-06 and the conidium in inhibiting the fusarium graminearum ascocarp. As can be seen from Table 1, the inhibition rate of the treatment group on the Fusarium graminearum ascocarp in the culture medium can reach more than 95%. As can be seen from Table 2, the inhibition rate of the treatment group on the Fusarium graminearum ascocarp in the field can reach 80-90%, and the treatment group is safe and does not influence the ecological environment.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (6)
1. A preparation method of gliocladium roseum NF-06 conidia is characterized by comprising the following steps:
(1) inoculating gliocladium roseum NF-06 into a spore production culture medium, and culturing to obtain a culture solution containing conidia;
(2) and filtering the culture solution containing the conidia, centrifuging the filtrate, and taking the precipitate to obtain the NF-06 conidia.
2. The method for preparing the gliocladium roseum NF-06 conidia according to claim 1, wherein the sporulation medium is a CMC liquid medium.
3. The method for preparing the gliocladium roseum NF-06 conidia according to the claim 2, characterized in that the culture conditions of the step (1) are as follows: the culture temperature is 23-27 ℃; the rotating speed of the shaking table is 170-180 rpm; the culture time is 45-51 h.
4. An application of NF-06 conidium or gliocladium roseum NF-06 prepared by the method of any one of claims 1 to 3 in inhibiting the fusarium graminearum ascocarp.
5. An application of NF-06 conidia or gliocladium roseum NF-06 prepared by the method of any one of claims 1 to 3 in inhibiting wheat scab caused by fusarium graminearum.
6. Use according to claim 4 or 5, wherein the NF-06 conidia are produced at a concentration of 1 x 105～1×107The cfu/mL suspension is sprayed on crop straws for use.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6495133B1 (en) * | 1998-09-30 | 2002-12-17 | Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture & Agri-Food Canada | Gliocladium roseum strains useful for the control of fungal pathogens in plants |
| US20160007613A1 (en) * | 2014-07-14 | 2016-01-14 | Adjuvants Plus Usa, Inc. | Clonostachys rosea Inoculated Plant Materials with Fungicides and Adjuvants |
| CN106085928A (en) * | 2016-08-29 | 2016-11-09 | 河北省农林科学院植物保护研究所 | A kind of bacillus subtilis and microbial bacterial agent thereof and application |
| CN109762743A (en) * | 2019-01-30 | 2019-05-17 | 河南省农业科学院植物保护研究所 | Gliocladium roseum, its solid fermentation microbial inoculum and its application |
| CN109843068A (en) * | 2016-06-08 | 2019-06-04 | 免疫生物防治法国公司 | The cell extract of one or more microalgaes of cross anastomosis is used for plant and cultivates the antifungal and/or bactericidal active purposes of the fungi of seed, oomycetes and/or pathogenic bacteria |
-
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- 2020-12-09 CN CN202011431302.6A patent/CN112746053B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6495133B1 (en) * | 1998-09-30 | 2002-12-17 | Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture & Agri-Food Canada | Gliocladium roseum strains useful for the control of fungal pathogens in plants |
| US20160007613A1 (en) * | 2014-07-14 | 2016-01-14 | Adjuvants Plus Usa, Inc. | Clonostachys rosea Inoculated Plant Materials with Fungicides and Adjuvants |
| CN109843068A (en) * | 2016-06-08 | 2019-06-04 | 免疫生物防治法国公司 | The cell extract of one or more microalgaes of cross anastomosis is used for plant and cultivates the antifungal and/or bactericidal active purposes of the fungi of seed, oomycetes and/or pathogenic bacteria |
| CN106085928A (en) * | 2016-08-29 | 2016-11-09 | 河北省农林科学院植物保护研究所 | A kind of bacillus subtilis and microbial bacterial agent thereof and application |
| CN109762743A (en) * | 2019-01-30 | 2019-05-17 | 河南省农业科学院植物保护研究所 | Gliocladium roseum, its solid fermentation microbial inoculum and its application |
Non-Patent Citations (3)
| Title |
|---|
| MING-MING ZHAI ET AL.: "Isolation of Secondary Metabolites from the Soil-Derived Fungus Clonostachys rosea YRS-06, a Biological Control Agent, and Evaluation of Antibacterial Activity", 《J. AGRIC. FOOD CHEM.》 * |
| 张洁等: "粉红螺旋聚孢霉NF−06固体发酵条件优化及对南方根结线虫的防治效果", 《中国生物防治学报》 * |
| 杨蕊等: "粉红粘帚霉对玉米茎基腐病的抑菌防病作用", 《河南科技学院学报（自然科学版）》 * |
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