CN117551560A - Tobacco endophytic fungus 001I-14 with remarkable drought resistance effect and application thereof - Google Patents
Tobacco endophytic fungus 001I-14 with remarkable drought resistance effect and application thereof Download PDFInfo
- Publication number
- CN117551560A CN117551560A CN202311511095.9A CN202311511095A CN117551560A CN 117551560 A CN117551560 A CN 117551560A CN 202311511095 A CN202311511095 A CN 202311511095A CN 117551560 A CN117551560 A CN 117551560A
- Authority
- CN
- China
- Prior art keywords
- tobacco
- fungus
- strain
- endophytic
- remarkable
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 131
- 241000233866 Fungi Species 0.000 title claims abstract description 49
- 230000000694 effects Effects 0.000 title claims description 7
- 244000061176 Nicotiana tabacum Species 0.000 title abstract description 3
- 241000208125 Nicotiana Species 0.000 claims abstract description 128
- 241000196324 Embryophyta Species 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 15
- 230000001580 bacterial effect Effects 0.000 claims abstract description 13
- 230000009418 agronomic effect Effects 0.000 claims abstract description 9
- 239000002028 Biomass Substances 0.000 claims abstract description 7
- 239000002689 soil Substances 0.000 claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 241001515917 Chaetomium globosum Species 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 16
- 238000011282 treatment Methods 0.000 claims description 14
- 241000221955 Chaetomium Species 0.000 claims description 12
- 239000008223 sterile water Substances 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 9
- 230000012010 growth Effects 0.000 claims description 9
- 238000000227 grinding Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 239000001965 potato dextrose agar Substances 0.000 claims description 8
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 5
- 241000894006 Bacteria Species 0.000 claims description 4
- 238000012408 PCR amplification Methods 0.000 claims description 4
- 238000005520 cutting process Methods 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 4
- 229960002523 mercuric chloride Drugs 0.000 claims description 4
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 229960005322 streptomycin Drugs 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- 108091023242 Internal transcribed spacer Proteins 0.000 claims description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 2
- 244000061456 Solanum tuberosum Species 0.000 claims description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 2
- 241001572088 Thrixopelma pruriens Species 0.000 claims description 2
- 238000009395 breeding Methods 0.000 claims description 2
- 230000001488 breeding effect Effects 0.000 claims description 2
- 235000019504 cigarettes Nutrition 0.000 claims description 2
- 239000008121 dextrose Substances 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 239000011159 matrix material Substances 0.000 claims description 2
- 230000000877 morphologic effect Effects 0.000 claims description 2
- 150000007523 nucleic acids Chemical group 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 238000012163 sequencing technique Methods 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 239000000758 substrate Substances 0.000 claims description 2
- 230000008641 drought stress Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 230000035882 stress Effects 0.000 abstract description 2
- 241000238631 Hexapoda Species 0.000 abstract 1
- 241000607479 Yersinia pestis Species 0.000 abstract 1
- 201000010099 disease Diseases 0.000 abstract 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract 1
- 229910001385 heavy metal Inorganic materials 0.000 abstract 1
- 244000020518 Carthamus tinctorius Species 0.000 description 8
- 235000003255 Carthamus tinctorius Nutrition 0.000 description 8
- 241000589158 Agrobacterium Species 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 241000588914 Enterobacter Species 0.000 description 2
- 241000187809 Frankia Species 0.000 description 2
- 241000520272 Pantoea Species 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 230000036579 abiotic stress Effects 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 235000013479 Amaranthus retroflexus Nutrition 0.000 description 1
- 240000005674 Ceanothus americanus Species 0.000 description 1
- 235000014224 Ceanothus americanus Nutrition 0.000 description 1
- 235000001904 Ceanothus herbaceus Nutrition 0.000 description 1
- 241000589323 Methylobacterium Species 0.000 description 1
- 244000118351 Pluchea indica Species 0.000 description 1
- 241000209504 Poaceae Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 241000223260 Trichoderma harzianum Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004790 biotic stress Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 210000002377 thylakoid Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/45—Tobacco
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses tobacco endophytic fungi 001I-14 with remarkable drought resistance and application thereof. Tobacco is one of important cash crops in China, plays a vital role in the economic development of China, but in recent years, tobacco planting production is often limited by multiple factors such as plant diseases and insect pests, heavy metal stress, drought stress and the like, so that tobacco plants grow slowly in a seedling stage or a field stage, and the production quality of tobacco leaves is seriously threatened. The 1 endophytic fungus bacterial liquid separated from the cloud tobacco 87 is poured on the upper part and root soil of tobacco seedlings of different varieties, so that the biomass and agronomic characters of the tobacco seedlings can be remarkably improved.
Description
Technical Field
The invention relates to the field of tobacco endophytic fungi with drought resistance, in particular to tobacco endophytic fungi 001I-14 with obvious drought resistance and application thereof.
Background
Plant endophytes are a group of microorganisms that promote plant growth, assist their host plants in combating severe environmental stresses, produce secondary metabolites similar to the host plants, and induce the accumulation of plant secondary metabolites, and generally include endophytes, and endophytes.
Currently, over 130 endophytes have been found in a large number of crops, medicinal plants and commercial crops such as fruit trees, covering 60 genera. Common endophytes include Agrobacterium (Agrobacterium), enterobacter (Enterobacter), bacillus (Bacillus), pseudomonas (Pseudomonas), bacillus (Methylobacterium) and Pantoea (Pantoea) and Agrobacterium (Agrobacterium), most of which are Frankia (Frankia) and most of which are grasses, medicinal plants, marine plants, etc., but tobacco is still less.
The endophytes live in tobacco bodies for a long time and co-evolve, and research of Hui Feiqiong and the like shows that under drought stress, the inoculation of Pityrosporum indicum can reduce the damage degree of tobacco cell membranes, accumulate more cell permeation regulating substances, and improve the expression of related drought resistance genes in tobacco, so that the drought resistance of the tobacco is enhanced. Hamilton et al believe that endophytic fungi can relieve abiotic stress by changing the oxidation resistance of a host, under drought stress, the endophytic fungi colonize the host, symbiotic with the host plant, and the fungi interact with the plant to produce alkaloids, improve the oxidation resistance of the plant, and relieve biotic and abiotic stress from the environment. Sun et al believe that under drought stress, plant thylakoid membrane-associated Ca2+ receptors, CASmRNA levels and CAS protein numbers colonized by endophytic fungi P.indica are increased, increasing plant drought stress tolerance. Studies by Shukla et al also found that endophytic fungi T.harzianum colonized plants not only reduced proline, MDA and H2O2 levels in the plants, but also increased phenolic and MSI levels in the plants. Therefore, the tobacco endophytic fungi are a new way for improving the stress resistance of tobacco leaves and improving the quality of tobacco leaves.
In our study on endophytic fungus flora of Yunnan tobacco dominant variety Yunyan 87, 103 endophytic fungi are separated from field picked Yunyan 87 tobacco strains, 52 endophytic fungi belong to 29 genera. Wherein, the upper leaf has 10 genera and 11 endophytes, the middle leaf has 7 genera and 8 endophytes, the lower leaf has 6 genera and 8 endophytes, the stem has 9 genera and 11 endophytes, the crude root has 12 genera and 21 endophytes, and the fibrous root has 5 genera and 7 endophytes. Through screening, 1 drought-resistant endophytic fungi in total obviously promote tobacco seedling biomass and character indexes of different tobacco varieties.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide tobacco endophytic fungi 001I-14 with remarkable drought resistance and application thereof; provides a new method for improving the yield and quality of tobacco leaves.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
tobacco endophytic fungi 001I-14 with remarkable drought resistance;
the method is characterized in that: the tobacco endophytic fungi are as follows: chaetomium fungus (Chaetomium globosum) 001I-14;
the strain is obtained through the following steps:
A. materials: dividing the cloud tobacco 87 plants collected from the field into six parts, namely an upper leaf, a middle leaf, a lower leaf, a tobacco stem, a thick root and a fibrous root;
B. and (3) disinfection: upper leaf, middle leaf, lower leaf: soaking in 75% alcohol for 30 s-0.1% mercuric chloride for 15 s-washing with sterile water for 3 times; tobacco stems: wiping epidermis-remaining cortex and center column with 75% alcohol; coarse root, fibrous root: soaking in 75% alcohol for 60 s-0.1% mercuric chloride for 20 s-washing with sterile water for 3 times, and scribing the sterile water after the last washing on a flat plate as a control to check whether the sample is thoroughly disinfected or not;
C. inoculating and separating: adding sterile water into the tobacco leaf part for grinding treatment, and uniformly laying grinding liquid and residues on the surface of a culture medium; cutting the stem part of the cigarette into small pieces with the diameter of 5-8 mm by an inoculating knife, and laying the small pieces on the surface of a culture medium; cutting the thick root part into small sections with the diameter of 3-5 mm by an inoculating knife, and laying the section on the surface of a culture medium downwards; adding sterile water into the fibrous root part for grinding treatment, uniformly laying grinding fluid and residues on the surface of a culture medium, placing all materials into a constant temperature incubator at 28 ℃ for culturing for 5-7 days, and observing the growth condition of bacterial colonies; because of different propagation speeds of different fungi, the strain with faster growth is preferentially separated and purified, and the strain with slower growth is separated and purified after the strain is stabilized.
Further, in the medium, solid culture: potato Dextrose Agar (PDA) (46 g/L) is used, streptomycin (10 mg/L) is added to prevent bacteria from breeding, the culture temperature is 28 ℃, and the culture time is 5-7 days;
and (3) inoculating the separated endophytic fungi of different types into a PDA culture medium for purification, performing spot connection purification for 2-3 times until colonies are single, and screening to obtain 1 strain of functional endophytic fungi with drought resistance.
Further, PCR amplification of rDNAITS was performed on the strain with fungal universal primers ITS1/ITS4, sequencing was performed on the ITS PCR amplification product, BLAST searching was performed on the obtained sequence in the GenBank nucleic acid sequence database, the chaetomium fungus (Chaetomium globosum) 001I-14 found that the ITS region of the test strain was the highest with the reported homology, and the species of 1 endophyte was determined in combination with morphological identification.
The tobacco endophytic fungi are as follows:
the chaetomium fungus (Chaetomium globosum) 001I-14, the colony growing on the substrate is in a dark green velvet shape. Hyphae are white, grow rapidly and spread rapidly.
Further, the preparation method of the tobacco endophytic fungi 001I-14 bacterial liquid comprises the following steps: inoculating the chaetomium fungus 001I-14 into a liquid culture medium, shake culturing by a shaking table at a rotating speed of 220rpm and a culturing temperature of 30 ℃ for 48-72 hours for later use; liquid medium: potato dextrose broth, wherein potato extract powder 5g/L, dextrose 20g/L, streptomycin 10mg/L.
Application of tobacco endophytic fungi 001I-14 with remarkable drought resistance; the bacterial liquid is poured into the ground parts and root matrix soil of tobacco seedlings of different varieties, so that the drought resistance is improved.
Further, the bacterial solutions are applied when 5-6 true leaves grow out of the tobacco seedlings, and the biomass and agronomic characters of the tobacco seedlings are detected after two weeks.
Further, the biomass and agronomic traits to be detected include: relative water content and agronomic traits.
Compared with the prior art, the invention has the beneficial effects that:
tobacco endophytic fungi 001I-14 with remarkable drought resistance and application thereof, wherein the fungi are inoculated into four tobacco seedlings of different varieties of safflower Dajinyuan, yunyan 87, K326 and Yunyan 97, and the drought resistance of the tobacco seedlings is further verified according to the judgment of 7 indexes such as biomass of the tobacco seedlings, agronomic characters and the like, so that basic support is provided for further application in production.
Drawings
FIG. 1 tobacco seedling tray and seeding mode;
FIG. 2 morphology of endophytic fungi from Yunyan 87;
FIG. 3 is a diagram of fungal fermentation in Yunyan 87;
FIG. 4 drought resistance of the tobacco endophytic fungi of the invention to the growth of different varieties of tobacco seedlings;
FIG. 5 shows the growth of tobacco seedlings inoculated with endophytes compared to tobacco seedlings not inoculated with endophytes.
Detailed Description
The technical scheme of the invention is further described in detail below with reference to the attached drawings and the detailed description:
as shown in fig. 1-5:
example 1: culturing plant materials;
according to the invention, four different varieties of tobacco coated seeds of safflower Dajinyuan, yunyan 87, K326 and Yunyan 97 are selected, sowed on a 3 x 4 small seedling tray (as shown in figure 2), 3 seeds are sowed in each hole, nutrition soil for seedling culture is subjected to high-pressure sterilization treatment, light and dark alternate culture is carried out in an incubator, light and culture is carried out for 16 hours, and the light intensity is 18000LX and 28 ℃; culturing in darkness for 8h at 16 ℃; nutrient solution is added into the bottom of the seedling raising tray, and the nutrient solution needs to be supplemented every two days.
Example 2: endophytic fungus liquid pouring application
Separating and purifying endophytic fungi to obtain 1.5cm 2 Inoculating bacterial blocks with the left and right sizes into 100ml of liquid culture medium, filling the culture medium into a 250ml conical flask, shake culturing in a shaking table at 30 ℃ and rotating at 220rpm for 48-72 hours; when the tobacco seedlings grow to 5-6 true leaves, the overground part and root soil of the tobacco seedlings are subjected to watering treatment by taking bacterial liquid, one tobacco seedling is reserved in each hole of a seedling raising tray before the watering treatment, the consistency of the growth vigor of each tray of tobacco seedlings is ensured, 3ml of bacterial liquid is taken for each hole to be watered on the tobacco seedlings, and the bacterial liquid is uniformly shaken before the taking; after the bacteria are poured, the transparent isolation cover is covered on the upper part of the seedling raising tray, and the transparent isolation cover is taken down after three days to continuously culture in the incubator, namely, culture solution and water are not added in the whole process; and a control tobacco seedling is additionally arranged, and only a liquid culture medium with the same volume is poured on the control tobacco seedling, and no bacteria liquid is poured. When the tobacco seedlings grow to 7 true leaves, the relative water content and agronomy of the tobacco seedlings are improvedAnd measuring indexes such as properties.
Example 3:
3.1 Effect of Chaetomium fungus (Chaetomium globosum) 001I-14 and other different strains on the relative moisture content of different tobacco seedling varieties.
The results show that the relative water content of the cloud tobacco 87 control tobacco seedlings and the cloud tobacco 87 tobacco seedlings of the C.globosum fungi are respectively 33.49 percent to 50.46 percent, and the relative water content of the cloud tobacco 87 tobacco seedlings after endophytic fungi are poured is obviously higher than that of the control tobacco seedlings and is respectively 16.97 percent higher than that of the control tobacco seedlings.
The relative water content of the cloud tobacco 97 control tobacco seedling and the cloud tobacco 97 tobacco seedling with the C.globosum three fungi applied is 32.73 percent and 52.08 percent. The relative water content of the cloud tobacco 87 tobacco seedlings after the endophytic fungi are poured is obviously higher than that of the control tobacco seedlings, and is respectively 19.35 percent higher than that of the control tobacco seedlings.
The relative moisture content of tobacco K326 control seedlings and tobacco K326 seedlings on which three fungi of C.globosum were applied was 40.12% and 62.16%. The relative water content of the tobacco K326 tobacco seedlings after being applied with endophytic fungi is obviously higher than that of the control tobacco seedlings, and is 18.17 percent, 18.02 percent and 22.04 percent respectively higher than that of the control tobacco seedlings.
The relative water content of the safflower Dajinyuan control tobacco seedling and the safflower Dajinyuan tobacco seedling which is coated with C.globosum fungus is 34.37 percent and 53.17 percent respectively. The relative water content of the safflower Dajinyuan tobacco seedlings after being sprayed with endophytic fungi is obviously higher than that of the control tobacco seedlings by 18.80 percent.
3.2 Effect of Chaetomium fungus (Chaetomium lobosum) 001I-14 Strain on agronomic traits of different tobacco seedling varieties.
The results show that the plant heights of the cloud tobacco 87 control tobacco seedlings and the cloud tobacco 87 tobacco seedlings on which the C.globosum fungi are poured are respectively 1.25cm and 5.41cm, and are 2.66cm and 4.16cm higher than the control tobacco seedlings; the maximum leaf length is 6.92cm and 6.96cm respectively; the maximum leaf width is 3.48cm and 4.33cm respectively; the root length is 8.28cm and 7.95cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference.
The plant heights of the cloud tobacco 97 control tobacco seedlings and the cloud tobacco 97 tobacco seedlings on which the C.globosum fungi are poured are respectively 1.25cm and 4.63cm; the maximum leaf length is 7.34cm and 7.47cm respectively; the maximum leaf width is 3.20cm and 4.15cm respectively, the root length is 8.06cm and 7.86cm respectively, and the tobacco seedlings treated in two different modes have no obvious difference.
The plant heights of a tobacco K326 control tobacco seedling and a tobacco K326 tobacco seedling poured with C.globosum fungi are respectively 1.60cm and 6.43cm, wherein the plant height of the tobacco K326 tobacco seedling poured with C.globosum is obviously higher than that of the control tobacco seedling and is 4.83cm higher than that of the control tobacco seedling, and the plant heights of the rest tobacco seedlings treated by the methods have no obvious difference; the maximum leaf length is 7.58cm and 7.93cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference; the maximum leaf width is 3.61cm and 4.38cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference; the root length is 8.93cm and 8.92cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference.
The plant heights of the safflower Dajinyuan control tobacco seedlings and the safflower Dajinyuan tobacco seedlings which are sprayed with the C.globosum fungi are respectively 1.60cm and 5.12cm, and the plant heights of the safflower Danyuan tobacco seedlings which are sprayed with the C.globosum are obviously higher than that of the control tobacco seedlings and are respectively 3.52cm higher than that of the control tobacco seedlings; the maximum leaf length is 7.34cm and 7.62cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference; the maximum leaf width is 3.52cm and 3.90cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference; the red root length is 7.61cm and 8.10cm respectively, and the tobacco seedlings treated by the two different treatments have no obvious difference.
The foregoing is merely illustrative of specific embodiments of the present invention, and the scope of the invention is not limited thereto, but any changes or substitutions that do not undergo the inventive effort should be construed as falling within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope defined by the claims.
Claims (7)
1. Tobacco endophytic fungi 001I-14 with remarkable drought resistance, which is characterized in that:
the method is characterized in that: the tobacco endophytic fungi are as follows: chaetomium fungus (Chaetomium globosum) 001I-14;
the strain is obtained through the following steps:
A. materials: dividing the cloud tobacco 87 plants collected from the field into six parts, namely an upper leaf, a middle leaf, a lower leaf, a tobacco stem, a thick root and a fibrous root;
B. and (3) disinfection: upper leaf, middle leaf, lower leaf: soaking in 75% alcohol for 30 s-0.1% mercuric chloride for 15 s-washing with sterile water for 3 times; tobacco stems: wiping epidermis-remaining cortex and center column with 75% alcohol; coarse root, fibrous root: soaking in 75% alcohol for 60 s-0.1% mercuric chloride for 20 s-washing with sterile water for 3 times, and scribing the sterile water after the last washing on a flat plate as a control to check whether the sample is thoroughly disinfected or not;
C. inoculating and separating: adding sterile water into the tobacco leaf part for grinding treatment, and uniformly laying grinding liquid and residues on the surface of a culture medium; cutting the stem part of the cigarette into small pieces with the diameter of 5-8 mm by an inoculating knife, and laying the small pieces on the surface of a culture medium; cutting the thick root part into small sections with the diameter of 3-5 mm by an inoculating knife, and laying the section on the surface of a culture medium downwards; adding sterile water into the fibrous root part for grinding treatment, uniformly laying grinding fluid and residues on the surface of a culture medium, placing all materials into a constant temperature incubator at 28 ℃ for culturing for 5-7 days, and observing the growth condition of bacterial colonies; because of different propagation speeds of different fungi, the strain with faster growth is preferentially separated and purified, and the strain with slower growth is separated and purified after the strain is stabilized.
2. The tobacco endophytic fungus 001I-14 having a remarkable drought-resistant effect according to claim 1, wherein:
in the culture medium, solid culture: potato Dextrose Agar (PDA) (46 g/L) is used, streptomycin (10 mg/L) is added to prevent bacteria from breeding, the culture temperature is 28 ℃, and the culture time is 5-7 days;
and (3) inoculating the separated endophytic fungi of different types into a PDA culture medium for purification, performing spot connection purification for 2-3 times until colonies are single, and screening to obtain 1 strain of functional endophytic fungi with drought resistance.
3. The tobacco endophytic fungus 001I-14 having a remarkable drought-resistant effect according to claim 1, wherein: the endophytic fungi identification specifically comprises the following steps: performing rDNA ITS PCR amplification on the strain by using fungus universal primers ITS1/ITS4, sequencing the ITS PCR amplification products, performing BLAST search on the obtained sequence in a GenBank nucleic acid sequence database, finding out that ITS regions of the test strain are the highest in homology with reported strains respectively by chaetomium fungus (Chaetomium globosum) 001I-14, and determining the strain of 1 endophyte by combining morphological identification;
the 1 tobacco endophytic fungus:
the chaetomium fungus (chaetomium lobosum) 001I-14 grows on the substrate in a dark green velvet shape, and the mycelium is white, grows rapidly and spreads rapidly.
4. The tobacco endophytic fungus 001I-14 having a remarkable drought-resistant effect according to claim 1, wherein:
the preparation method of the tobacco endophytic fungi 001I-14 bacterial liquid comprises the following steps: inoculating the chaetomium fungus 001I-14 into a liquid culture medium, shake culturing by a shaking table at a rotating speed of 220rpm and a culturing temperature of 30 ℃ for 48-72 hours for later use; liquid medium: potato dextrose broth, wherein potato extract powder 5g/L, dextrose 20g/L, streptomycin 10mg/L.
5. Application of tobacco endophytic fungi 001I-14 with remarkable drought resistance; the method is characterized in that the bacterial liquid is poured into the ground parts and root matrix soil of different varieties of tobacco seedlings, so that the drought resistance is improved.
6. The use according to claim 5; the method is characterized in that the bacterial liquid is applied when 5-6 true leaves grow out of tobacco seedlings, and the biomass and agronomic characters of the tobacco seedlings are detected after two weeks.
7. The use according to claim 6; the method for detecting biomass and agronomic traits according to claim 5, comprising: relative water content and agronomic traits.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311511095.9A CN117551560A (en) | 2023-11-14 | 2023-11-14 | Tobacco endophytic fungus 001I-14 with remarkable drought resistance effect and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311511095.9A CN117551560A (en) | 2023-11-14 | 2023-11-14 | Tobacco endophytic fungus 001I-14 with remarkable drought resistance effect and application thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117551560A true CN117551560A (en) | 2024-02-13 |
Family
ID=89815965
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311511095.9A Withdrawn CN117551560A (en) | 2023-11-14 | 2023-11-14 | Tobacco endophytic fungus 001I-14 with remarkable drought resistance effect and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117551560A (en) |
-
2023
- 2023-11-14 CN CN202311511095.9A patent/CN117551560A/en not_active Withdrawn
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107446847B (en) | Bacillus belgii GT11 and application thereof | |
CN109749943A (en) | A kind of Trichoderma asperellum and its application | |
CN110669691B (en) | Bacillus megaterium for preventing and treating plant nematode diseases and application thereof | |
CN117305130A (en) | Tobacco endophytic fungus 001I-21 with remarkable drought resistance effect and application thereof | |
CN116875471A (en) | Tobacco endophytic fungus X-5 with remarkable drought resistance effect and application thereof | |
CN109749953B (en) | Bacillus cereus, microbial inoculum and preparation method and application thereof | |
CN116121082A (en) | Smoke tube bacterium MR-8 with drought resistance function and application thereof | |
CN109536425A (en) | A kind of Te Jila bacillus and its application | |
CN111363691B (en) | Paenibacillus polymyxa and application thereof | |
CN116024106A (en) | Tobacco endophytic fungus capable of remarkably promoting tobacco seedling growth and application thereof | |
CN111849856A (en) | Indoca chlamydospore, P.indoca spore bacterial agent and preparation method thereof | |
CN103937684B (en) | A kind of from Eucalypt soil screening press down grass fungus | |
CN116676234A (en) | Salt-tolerant bacillus JK-25, microbial inoculum, preparation method and application thereof | |
CN114480143B (en) | Trichoderma harzianum M6 for preventing and treating sclerotinia sclerotiorum of sunflower and application thereof | |
CN1234846C (en) | Strain of pseudomonas for promoting growth of paddy as well as preventing and curing sheath blight of rice | |
CN102007869B (en) | Ectomycorrhizal fungi efficiently symbiotic with pinus koraiensis | |
CN117551560A (en) | Tobacco endophytic fungus 001I-14 with remarkable drought resistance effect and application thereof | |
CN111187724B (en) | Endophytic fungus with dark blueberry root and application thereof | |
CN116445294A (en) | Tobacco seedling drought-resistant endophytic fungus W10 | |
CN112625954A (en) | Pseudomonas CM11 and application thereof | |
LU501276B1 (en) | Culture Method and Application of Botrytis Cinerea with Herbicidal Effect | |
CN116179370A (en) | Tobacco endophytic fungus for promoting tobacco seedling growth and application thereof | |
CN117025409A (en) | Tobacco endophytic fungus H-10 for promoting tobacco seedling growth | |
CN113854318B (en) | Application of Pityrosporum indicum in improving drought resistance of Mao She seeds | |
CN115927051B (en) | Biocontrol bacterium, biocontrol compound microbial inoculum and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20240213 |