CN114933974A - Trichoderma pseudokoningii C5-9 and application thereof in antagonism of botrytis cinerea - Google Patents
Trichoderma pseudokoningii C5-9 and application thereof in antagonism of botrytis cinerea Download PDFInfo
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Abstract
The invention discloses Trichoderma pseudokoningii (Trichoderma koningiensis) C5-9 with the preservation number of CGMCC No.40015, which is preserved by the common microorganism center of China Committee for culture Collection of microorganisms with the preservation date of 2021, 12 months and 22 days. The trichoderma pseudokoningii C5-9 has a direct antagonistic effect on Botrytis cinerea, and the trichoderma pseudokoningii C5-9 can be applied to prevention and treatment of Botrytis cinerea diseases caused by infection of Botrytis cinerea.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to trichoderma pseudokoningii C5-9 and application thereof in antagonism of botrytis cinerea.
Background
Botrytis caused by botrytis cinerea is the most serious fungal disease worldwide, and can infect more than 200 plants including vegetables and fruits such as tomatoes, cucumbers, strawberries and the like. Botrytis cinerea is a dead body nutritional pathogenic fungus and can produce various phytotoxins and cell wall hydrolases to kill plant host cells. Botrytis cinerea has various infection modes, and sclerotium survives on plant residues for a long time and generates drug resistance, so that the prevention and control of gray mold worldwide are difficult. Statistically, the cost for controlling gray mold is more than one billion yuan each year in the world, and even then, the disease still causes billions of economic losses to agriculture. Among the technologies for controlling plant diseases, biological control is a good measure for sustainable development to gradually replace chemical control.
Trichoderma has antagonistic action on Botrytis cinerea in different degrees, and the biological control mechanism of Trichoderma is divided into a direct biocontrol mechanism and an indirect biocontrol mechanism, wherein the former mainly refers to heavy parasitism, antibiotic and nutrition competition involved in the direct action process of Trichoderma and Botrytis cinerea, and the latter is that Trichoderma controls Botrytis cinerea by inducing plant to generate systemic resistance. Therefore, the screening of trichoderma with development and utilization values has important theoretical and practical significance for industrial and agricultural production. The application range of the trichoderma preparation is wider, but the current trichoderma preparation has certain self-defects. The optimal growth temperature of the trichoderma is 25-35 ℃, the growth and spore production are inhibited at low temperature, and the screening of the strains is carried out at normal temperature. The gray mold disease is a medium-low temperature disease, the temperature of 20-23 ℃ is the optimum temperature, and the gray mold disease can grow under the condition of lower temperature. The growth moderate temperature of the trichoderma is different from the growth moderate temperature of the gray mold, so that the control effect of the gray mold generated under the conditions of low temperature and haze in winter is poor, and the further improvement of the biocontrol effect of the trichoderma preparation and the further widening of the application field are limited. At present, the study on trichoderma in low-temperature areas at home and abroad is few. Therefore, the development of the low-temperature trichoderma preparation is the key for solving the development bottleneck of the current trichoderma preparation, particularly, the northern part of China has obvious seasonal variation and low temperature, and the common trichoderma has little effect on resisting gray mold. Therefore, the trichoderma which can be rapidly proliferated under the low-temperature condition and has strong ability to resist botrytis cinerea is found, and the trichoderma has very important significance for efficiently preventing and treating the botrytis cinerea.
Disclosure of Invention
The invention aims to provide trichoderma koningii C5-9 aiming at the technical defects in the prior art.
The invention also aims to provide application of the trichoderma pseudokoningii C5-9 in antagonism of botrytis cinerea.
The technical scheme adopted for realizing the purpose of the invention is as follows:
trichoderma pseudokoningii C5-9 with preservation number of CGMCC No. 40015.
In another aspect of the invention, the application of the trichoderma pseudokoningii C5-9 in antagonism of botrytis cinerea is also included.
In another aspect of the invention, the application of the trichoderma pseudokoningii C5-9 in preventing and treating Botrytis cinerea diseases caused by infection of Botrytis cinerea (Botrytis cinerea) is also included.
In another aspect of the invention, the fermentation product of the trichoderma pseudokoningii C5-9 is also included.
In the above technical scheme, the fermentation product is prepared by the following method: inoculating Trichoderma pseudokoningii C5-9 spore in PDB liquid culture medium to make its final concentration be 1 x 10 4 -1*10 6 cfu/mL, carrying out shake culture at 16 ℃ for 72-96h, and then centrifuging to collect fermentation liquor.
In the technical scheme, the fermentation liquor is prepared into dry powder by using a vacuum freeze dryer, the freeze-drying temperature is-40 ℃ to-55 ℃, and the freeze-drying time is 36h to 42 h. Dissolving the dry powder in water, and filtering with 0.22 μm microporous membrane to obtain sterile fermentation product.
In another aspect of the invention, the application of the fermentation product in antagonism of botrytis cinerea is also included.
In another aspect of the invention, the application of the fermentation product in preventing and treating gray mold disease caused by Botrytis cinerea infection is also included.
Compared with the prior art, the invention has the beneficial effects that:
1. the Trichoderma pseudokoningii C5-9 provided by the invention can be rapidly propagated in a low-temperature environment of 16 ℃, and the inhibition rate of the antagonistic culture of a PDA culture medium on botrytis cinerea is 59.7%, which shows that the Trichoderma pseudokoningii C5-9 has a direct antagonistic effect on the botrytis cinerea.
2. The invention also provides a fermentation product obtained by Trichoderma koningii C5-9 in a PDB fermentation medium. The inhibition rate of the fermentation product obtained by 96-hour fermentation on botrytis cinerea is 64%, which shows that the trichoderma pseudokoningii C5-9 produces high-efficiency antibiotic substances for antagonizing botrytis cinerea in the fermentation culture medium.
Drawings
FIG. 1 is a phylogenetic tree constructed by strain C5-9 based on ITS sequence homology
FIG. 2 shows the results of the plate confrontation experiment of example 1 and control 1 in example 1. In the example group 1, the upper pellet was Trichoderma pseudokoningii C5-9, the lower pellet was Botrytis cinerea, and the lower pellet was Botrytis cinerea. In fig. 2, A, C and B, D show the front and back of the plate, respectively. The arrows indicate the growth diameter of Botrytis cinerea measured in both groups.
FIG. 3 is the growth of Botrytis cinerea hyphae of example group 2 and control group 2 in example 2. As can be seen from FIG. 3, the fermentation product of Trichoderma pseudokoningii C5-9 inhibited the growth of Botrytis cinerea hyphae.
Detailed Description
The present invention will be described in further detail with reference to specific examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The following examples refer to media and test strains of botrytis cinerea:
PDA medium (1L): 200g of potato, 20g of glucose, 15g of agar and 1000mL of water, and sterilizing at 115 ℃ for 20min at a natural pH value for later use.
PDB liquid medium (1L): 200g of potato, 20g of glucose and 1000mL of water, natural pH value, and sterilizing at 115 ℃ for 20min for later use.
The Botrytis cinerea strain is a published genome data strain and is numbered as Botrytis cinerea, B05.10.
Example 1
The screening method of Trichoderma pseudokoningii C5-9 comprises the following steps:
collecting surface soil samples from the south hillside of the semiwall mountain in Chengdong county of Hebei, diluting and plating, and separating strains by PDA culture medium. Weighing 10g of soil sample, dissolving in 90mL of sterile water, fully oscillating for 30min at 180r/min on an oscillator, standing for 10min, and gradually diluting to 10 degrees with sterile water -2 -10 -4 Draw 100. mu.L of each dilution gradient of 10 -3 And 10 -4 The diluted solution is evenly coated on a PDA culture medium, cultured in a constant temperature incubator at 16 ℃, and screened for trichoderma strains with better low-temperature adaptability. As a result, 64 Trichoderma strains with different colony morphologies were co-isolated, and purified by streaking a plurality of times to obtain pure cultured strains, which were then stored at 4 ℃.
Among the strains obtained above, C5-9 is a strain classified and named as Trichoderma pseudokoningii (Trichoderma koningiensis), which has been preserved in China general microbiological culture Collection center (CGMCC) in 22 months and 12 months in 2021, the preservation number is CGMCC No.40015, and the preservation address is as follows: the institute of microbiology, national academy of sciences No. 3, Xilu No. 1, Beijing, Chaoyang, Beijing.
The trichoderma C5-9 is subjected to molecular biological identification. Genomic DNA of the strain was extracted, ITS regions (internal transcribed spacer regions) were amplified using primers ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS6 (5'-GAAGGTGAAGTCGTAACAAGG-3'), and the amplified fragments were subjected to Sanger sequencing. The sequencing results were as follows:
AGGGGTTCGAGGGTTGAAATGACGCTCGGACAGGCATGCCCGCCAGAATACTGGCGGGCGCAATGTGCGTTCAAAGATTCGATGATTCACTGAATTCTGCAATTCACATTACTTATCGCATTTCGCTGCGTTCTTCATCGATGCCAGAACCAAGAGATCCGTTGTTGAAAGTTTTGATTCATTTTGAATTTTTGCTCAGAGCTGTAAGAAATAACGTCCGCGAGGGGACTACAGAAAGAGTTTGGTTGGTCCCTCCGGCGGGCGCCTGGTTCCGGGGCTGCGACGCACCCGGGGCGTGACCCCGCCGAGGCAACAGTTTGGTATGGTTCACATTGGGTTTGGGAGTTGTAAACTCGGTAATGATCCCTCCGCTGGTTCACCAACGGAGACCTTGTTACG。
the obtained ITS sequence of the C5-9 strain is compared with the ITS sequence of a Trichoderma model strain published by an NCBI database, a phylogenetic tree is constructed by using MEGA software, the result is shown in figure 1, the C5-9 and Trichoderma pseudokoningii have the latest genetic relationship and gather in the same branch, and the C5-9 strain screened by the research is determined to be Trichoderma pseudokoningiensis (Trichoderma koningiopsis).
Example 2
The antagonistic ability of the trichoderma pseudokoningii obtained in example 1 against botrytis cinerea was examined by a plate-confronted culture method.
Example set 1: respectively inoculating botrytis cinerea and the trichoderma pseudokoningii C5-9 obtained in the embodiment 1 to positions, which are 1cm away from two ends, on a PDA culture medium with the diameter of 90mm, so that the trichoderma pseudokoningii forms trichoderma blocks with the diameter of 0.5cm, the botrytis cinerea forms botrytis cinerea blocks with the diameter of 0.5cm, and the trichoderma blocks and the botrytis cinerea blocks are arranged along the radial direction of the PDA culture medium;
control group 1: inoculating Botrytis cinerea on PDA culture medium with diameter of 90mm at a distance of 1cm from the edge to form Botrytis cinerea blocks with diameter of 0.5 cm. Each of example group 1 and control group 1 above was subjected to 3 parallel experiments. The example group 1 and the control group 1 were cultured at 16 ℃ for 14d to observe the growth inhibition of the trichoderma pseudokoningii C5-9 on botrytis cinerea, and the colony diameter (R) of the botrytis cinerea was measured to calculate the inhibition rate. As shown in FIG. 2, in the absence of Trichoderma pseudokoningii C5-9, the entire plate was covered with Botrytis cinerea in control group 1 when cultured at 16 ℃ for 14 days. While the growth of Botrytis cinerea in example group 1 was significantly inhibited by Trichoderma pseudokoningii C5-9, FIG. 1A shows that Trichoderma pseudokoningii C5-9 completely covered the Botrytis cinerea colonies.
The diameters of the botrytis cinerea colonies in fig. 1 were measured, and the average of the diameters of the botrytis cinerea colonies obtained in 3 example groups 1 and the average of the diameters (R) of the botrytis cinerea colonies obtained in 3 control groups 1 are shown in table 1. The colony diameter of Botrytis cinerea (3.06. + -. 0.03cm) in example group 1 was significantly shorter than that of the Botrytis cinerea (7.60. + -. 0.04cm) in control group 1.
TABLE 1
| Group of | Diameter of colony of Botrytis cinerea R (unit: cm) | Inhibition rate |
| Example set 1 | 3.06±0.03* | 59.7％ |
| Control group 1 | 7.60±0.04 | - |
Represents that the diameters of the botrytis cinerea bacterial plaques of the example groups are remarkably different from those of the control group by Student's t-test analysis, and P is less than 0.05.
The inhibition ratio (%) - (R1-R2)/R1 × 100%, R1 being the colony diameter of botrytis cinerea obtained in control 1; r2 is the colony diameter of Botrytis cinerea obtained in example group 1.
Namely, the inhibition rate of the low-temperature Trichoderma pseudokoningii (Trichoderma koningiensis) with the preservation number of CGMCC No.40015 to botrytis cinerea is 59.7%, and the low-temperature Trichoderma pseudokoningiensis can antagonize botrytis cinerea through direct action mechanisms such as heavy parasitism, antibiotics and nutritional competition.
Example 3
Inoculating Trichoderma pseudokoningii C5-9 spore in 100mL PDB liquid medium to make its final concentration 1 x 10 6 cfu/mL, shaking culture at 16 ℃ for 96h, and centrifuging to collect fermentation liquor. Freezing the fermentation liquid at-40 deg.C to-55 deg.C for 36-42 h to obtain 1.5g dry powder, dissolving in water, filtering with 0.22 μm microporous membrane, and preparing into 0.3g/mL sterile fermentation product.
PDA medium plates (example 2) containing a final concentration of 7.5mg/mL of sterile fermentation product were prepared, and PDA medium plates without fermentation product were used as a control (control 2). 5 μ L of inoculum with a concentration of 1 x 10 6 cfu/mL of Botrytis cinerea spores at the central positions of example group 2 and control group 2, after 2d, 3d, and 4d of culture at 16 ℃, the diameters (R') of the Botrytis cinerea plaques were measured, respectively. As a result, as shown in FIG. 3, the growth of the plaque of Botrytis cinerea in example 2 was significantly inhibited after 2d, 3d and 4d cultivation at 16 ℃ as compared with control 2.
The inhibition rate was calculated by measuring and averaging the diameters (R') of the bacterial plaque of Botrytis cinerea in 3 parallel experiments of example group 2 and control group 2, and the results are shown in Table 2.
TABLE 2
It is shown that the diameter of the botrytis cinerea plaque of example group 2 was significantly different from that of control group 2 by Student's t-test analysis, P <0.05.
The inhibition ratio (%) - (R1'-R2')/R1 'x 100%, R1' being the diameter of the botrytis cinerea bacterial plaque obtained in control 2; r2' is the diameter of the Botrytis cinerea plaque obtained in example group 2.
Example 2 the diameter of the botrytis cinerea plaque is significantly different from that of the control group 2, and the inhibition rates of the fermentation product generated after the Trichoderma pseudokoningii (Trichoderma koningiensis) C5-9 is fermented for 96h in the PDB culture medium on the botrytis cinerea are 64.3%, 54.6% and 51.7% at 2d, 3d and 4d respectively, which shows that the Trichoderma pseudokoningiensis (Trichoderma koningiensis) with the preservation number of CGMCC No.40015 can generate the antibiotic substance for antagonizing the botrytis cinerea.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, many modifications and adaptations can be made without departing from the principle of the present invention, and such modifications and adaptations should also be considered as the scope of the present invention.
SEQUENCE LISTING
<110> North China university of Engineers
<120> Trichoderma pseudokoningii C5-9 and application thereof in antagonism of Botrytis cinerea
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 399
<212> DNA
<213> Trichoderma koningiopsis
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tgatccctcc gctggttcac caacggagac cttgttacg 399
Claims (8)
1. Trichoderma pseudokoningii C5-9 is characterized in that the preservation number is CGMCC No. 40015.
2. The trichoderma pseudokoningii C5-9 antagonistic botrytis cinerea strain as claimed in claim 1.
3. The use of trichoderma pseudokoningii C5-9 according to claim 1 for controlling Botrytis cinerea diseases caused by infection with Botrytis cinerea (Botrytis cinerea).
4. A fermentation product of trichoderma pseudokoningii C5-9 of claim 1.
5. The fermentation product of claim 4, produced by a process comprising: inoculating Trichoderma pseudokoningii C5-9 spore in PDB liquid culture medium to make its final concentration be 1 x 10 4 -1*10 6 cfu/mL, carrying out shake culture at 16 ℃ for 72-96h, and then centrifuging to collect fermentation liquor.
6. The fermentation product of claim 5, wherein the fermentation broth is prepared into a dry powder using a vacuum freeze dryer at a freeze drying temperature of-40 ℃ to-55 ℃ for a freeze drying time of 36h to 42 h. Dissolving the dry powder in water, and filtering with 0.22 μm microporous membrane to obtain sterile fermentation product.
7. Use of a fermentation product according to any one of claims 4 to 6 for antagonising botrytis cinerea.
8. Use of a fermentation product according to any one of claims 4 to 6 for controlling gray mold disease caused by an infestation with Botrytis cinerea (Botrytis cinerea).
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117305135A (en) * | 2023-10-30 | 2023-12-29 | 六盘水师范学院 | Trichoderma pseudokoningii T0027 and application thereof in preventing and treating soft rot of kiwi fruits |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104059860A (en) * | 2014-07-04 | 2014-09-24 | 江苏丘陵地区镇江农业科学研究所 | Trichoderma koningiopsis TZ-11 bacterial strain and application thereof |
| CN105036916A (en) * | 2015-07-15 | 2015-11-11 | 黄冈师范学院 | Compound microbial fertilizer for control of persimmon gray mold and preparation method thereof |
| CN105586274A (en) * | 2016-02-02 | 2016-05-18 | 华中农业大学 | Trichoderma koningiopsis T-51 strain and application thereof in growth promotion of tomatoes and biological prevention and control of Botrytis cinerea Pers |
| CN105695342A (en) * | 2016-03-16 | 2016-06-22 | 中国农业科学院油料作物研究所 | Trichoderoma koningii TC-72 and application of trichoderoma koningii TC-72 to biological control of aspergillus flavus |
| CN105802859A (en) * | 2016-05-03 | 2016-07-27 | 唐山市畜牧水产品质量监测中心 | Trichoderoma koningii TG-102 and application of trichoderoma koningii TG-102 to prevention and control of corn aflatoxin pollution |
-
2022
- 2022-05-11 CN CN202210508946.3A patent/CN114933974A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104059860A (en) * | 2014-07-04 | 2014-09-24 | 江苏丘陵地区镇江农业科学研究所 | Trichoderma koningiopsis TZ-11 bacterial strain and application thereof |
| CN105036916A (en) * | 2015-07-15 | 2015-11-11 | 黄冈师范学院 | Compound microbial fertilizer for control of persimmon gray mold and preparation method thereof |
| CN105586274A (en) * | 2016-02-02 | 2016-05-18 | 华中农业大学 | Trichoderma koningiopsis T-51 strain and application thereof in growth promotion of tomatoes and biological prevention and control of Botrytis cinerea Pers |
| CN105695342A (en) * | 2016-03-16 | 2016-06-22 | 中国农业科学院油料作物研究所 | Trichoderoma koningii TC-72 and application of trichoderoma koningii TC-72 to biological control of aspergillus flavus |
| CN105802859A (en) * | 2016-05-03 | 2016-07-27 | 唐山市畜牧水产品质量监测中心 | Trichoderoma koningii TG-102 and application of trichoderoma koningii TG-102 to prevention and control of corn aflatoxin pollution |
Non-Patent Citations (1)
| Title |
|---|
| 杨合同, 北京：中国大地出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117305135A (en) * | 2023-10-30 | 2023-12-29 | 六盘水师范学院 | Trichoderma pseudokoningii T0027 and application thereof in preventing and treating soft rot of kiwi fruits |
| CN117305135B (en) * | 2023-10-30 | 2024-03-19 | 六盘水师范学院 | Trichoderma pseudokoningii T0027 and application thereof in preventing and treating soft rot of kiwi fruits |
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