NL2035080B1 - Alternaria gaisen and its application in biological weeding - Google Patents
Alternaria gaisen and its application in biological weeding Download PDFInfo
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- 241000412368 Alternaria gaisen Species 0.000 title claims abstract description 24
- 238000009333 weeding Methods 0.000 title abstract description 11
- 241000196324 Embryophyta Species 0.000 claims abstract description 52
- 239000004009 herbicide Substances 0.000 claims abstract description 19
- 240000003065 Malva verticillata Species 0.000 claims abstract description 14
- 235000002384 Malva verticillata Nutrition 0.000 claims abstract description 14
- 240000006122 Chenopodium album Species 0.000 claims abstract description 12
- 235000009344 Chenopodium album Nutrition 0.000 claims abstract description 12
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 16
- 241000131458 Elsholtzia Species 0.000 claims description 13
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- 238000000855 fermentation Methods 0.000 claims description 12
- 230000004151 fermentation Effects 0.000 claims description 12
- 235000007320 Avena fatua Nutrition 0.000 claims description 11
- 244000075850 Avena orientalis Species 0.000 claims description 10
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- 230000002363 herbicidal effect Effects 0.000 claims description 9
- 240000006928 Persicaria lapathifolia Species 0.000 claims description 6
- 239000002207 metabolite Substances 0.000 claims description 6
- 241000219318 Amaranthus Species 0.000 claims description 3
- 240000005979 Hordeum vulgare Species 0.000 abstract description 6
- 244000098338 Triticum aestivum Species 0.000 abstract description 6
- 240000006677 Vicia faba Species 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 6
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- 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
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P13/00—Herbicides; Algicides
- A01P13/02—Herbicides; Algicides selective
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- 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
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- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Mycology (AREA)
- Pest Control & Pesticides (AREA)
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Abstract
Disclosed are Alternaria gaisen and its application in biological weeding, and relates to the 5 technical field of biological weeding. The Alternaria gaisen GD-011 is preserved in the China General Microbiological Culture Collection Center on April 6‘“, 2023, with a preservation number of CGMCC No.40558. The invention also discloses an application of the Alternaria gaisen GD- 011 in biological weeding or preparation of biological herbicides. The present invention aims to screen a strain of Alternaria gaisen for biological weed control in broad-leaved weeds such as 10 Gal/um, Chenopodium album, Malva verticillata, etc. in H. vulgare, T. aestivum, B. napus, V. faba, and P. sativum fields. The invention may effectively and safely control and prevent most broad- leaved weeds, and has the advantages of low cost, no pollution, low residue and environmental friendliness.
Description
ALTERNARIA GAISEN AND ITS APPLICATION IN BIOLOGICAL WEEDING
The invention relates to the technical field of biological weeding, and in particular to
Alternaria gaisen and its application in biological weeding.
Weeds have caused great losses to agricultural production. At present, chemical herbicides are mainly used to control weeds in agriculture. However, the application of chemical herbicides has potential serious problems such as environmental pollution and threat to food safety. Biological herbicide provides an effective way to solve the problem. Biological herbicide is to control weeds by using natural pathogenic bacteria of weeds, which has the advantages of high control efficiency, lasting control effect and no environmental problems. It is an important direction of new herbicide research to develop microbial herbicides by using weed pathogenic microorganisms and active substances in their fermentation broth or metabolites as lead compounds. It is urgent to develop new herbicides by using new organisms such as microorganisms.
In recent 20 years, some microbial herbicides have been successfully developed abroad, and certain economic benefits have been achieved. Bialaphos is the first metabolite of actinomycetes developed as a commercial herbicide. Bialaphos is a non-selective herbicide that may kill monocotyledonous and dicotyledonous plants, and it has been sold in many places.
However, at present, the ability to create biological pesticides is weak, and there are few new varieties. It is urgent to change the pattern of long-term single production of chemical herbicides in the existing herbicide industry and form a new biological herbicide industry. Therefore, it is necessary to screen more kinds of potential strains.
The objective of the invention is to provide A/fernaria gaisen and its application in biological weeding, so as to solve the problems existing in the prior art. A/ternaria gaisen GD-011 may effectively and safely control and prevent most broad-leaved weeds, and has the advantages of low cost, no pollution, low residue and environmental friendliness, and may be used for biological weeding.
In order to achieve the above objective, the present invention provides the following schemes.
The invention provides Alternaria gaisen GD-011, wherein it is preserved in the China
General Microbiological Culture Collection Center on April 6", 2023, with a preservation address of No.3, No.1 Yard of Beichen West Road, Chaoyang District, Beijing, and a preservation number of CGMCC No.40558.
The invention also provides an application of the Alternaria gaisen GD-011 in biological weeding or preparation of biological herbicides.
The invention also provides a biological herbicide, including at least one of a strain, fermentation broth or metabolite of Alternaria gaisen GD-011.
The invention also provides a biological weeding method, including a step of applying the strain, fermentation broth or metabolite of Alfernaria gaisen GD-011 to weed plants.
Further, the weed plants are broad-leaved weeds.
Further, the broad-leaved weeds include Malva verticillata, Elsholtzia densiflora, Herba
Polygoni Avicularis, Galium, Polygonum lapathifolium, Chenopodium album, Amaranthus retrofiexus, wild oat and penny cress.
The invention discloses the following technical effects.
Firstly, the invention uses microbial resources to control weeds, which has the advantages of less impact on crops, less negative environmental effects, high safety and the like, conforms to the development of sustainable agriculture, and has been highly valued by countries all over the world. The evaluation of microbial herbicides is theoretically based on two criteria, namely, effectiveness (efficacy) and specificity (safety). Weed biocontrol bacteria have two characteristics: strong pathogenicity to target weeds and relative safety to crops.
Secondly, the present invention aims to screen a strain of Alternaria gaisen for biological weed control in broad-leaved weeds such as Galium, Chenopodium album, Malva verticillata, etc. in H. vulgare, T. aestivum, B. napus, V. faba, and P. sativum fields. The invention may effectively and safely control and prevent most broad-leaved weeds, and has the advantages of low cost, no pollution, low residue and environmental friendliness.
In order to explain the embodiment of the invention or the technical scheme in the prior art more clearly, the drawings used in the embodiment will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the invention. For ordinary technicians in the field, other drawings may be obtained according to these drawings without making creative efforts.
FIG. 1 shows pathogenicity of strain GD-011 to leaves in vitro; A. Chenopodium album; B.
Elsholtzia densiflora; C. Polygonum lapathifolium, D. Malva verticillata; E. Amaranthus retroflexus; F. Galium; G. wild oats; H. penny cress; |. Herba Polygoni Avicularis.
FIG. 2 shows pathogenicity of the strain GD-011 to potted weeds; A. Chenopodium album; B.
Elsholtzia densiflora; C. Polygonum lapathifolium, D. Malva verticillata; E. Amaranthus retroflexus; F. Galium; G. wild oats; H. penny cress; |. Herba Polygoni Avicularis.
FIG. 3 shows safety of the strain GD-011 to crops; A: V. faba; B: P. sativum; C: B. napus; D: H. vulgare, E: T. aestivum.
FIG. 4 shows morphological characteristics of the strain GD-011; A, D: morphology of the strain
GD-011 on PDA plate: B: conidiophore; C and E - F: conidia.
FIG. 5 is a phylogenetic tree of the GD-011 based on rDNA-ITS, EF-1a and Alt a1 gene sequences.
FIG. 6 shows characteristics of invasion of the strain GD-011 into a leaf tissue of Elsholtzia densiflora by scanning electron microscope; A: not vaccinated; B - C: inoculation for 1-2 days; D - E: inoculation for 3 - 4 days; F-G: inoculation for 5 - 6 days; H: inoculation for 7 days; where s: stomata; H: hyphae; ih: invasive hyphae; and td: tissue destruction.
Various exemplary embodiments of the present invention will now be described in detail, which should not be regarded as a limitation of the present invention, but rather as a more detailed description of certain aspects, characteristics and embodiments of the present invention.
It should be understood that the terms described in the present invention are only for describing specific embodiments, and are not intended to limit the present invention. In addition, as for the numerical range in the present invention, it should be understood that every intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Intermediate values within any stated value or stated range and every smaller range between any other stated value or intermediate values within the stated range are also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.
Unless otherwise stated, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which the present invention relates. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe methods and/or materials related to the documents. In case of conflict with any incorporated documents, the contents of this specification shall prevail.
Without departing from the scope or spirit of the invention, it is obvious to those skilled in the art that many modifications and changes may be made to the specific embodiments of the specification of the invention. Other embodiments derived from the description of the present invention will be apparent to the skilled person. The specification and examples of this application are only exemplary.
As used herein, the terms “including”, “comprising”, “having”, “containing”, etc. are all open terms, and they mean including but not limited to.
Embodiment 1 1. Materials and methods 1.1 Test strain
Alternaria gaisen GD-011 is isolated from alfalfa roots which are naturally susceptible to diseases, and is named Alternaria gaisen. It is preserved in the China General Microbiological
Culture Collection Center, with a preservation address of No.3, No.1 Yard of Beichen West
Road, Chaoyang District, Beijing, and a preservation number of CGMCC No.40558. 1.2 Test weeds and crops
Common broad-leaved weeds in fields in Qinghai province: Chenopodium album,
Elsholitzia densiflora, Malva verticillata, Polygonum lapathifolium, Amaranthus retroflexus, wild oats, penny cress, Herba Polygoni Avicularis and Galium; main crops in Qinghai province: H. vulgare, T. aestivum, B. napus, V. faba and P. sativum. 1.3 Test design and method (1) Pathogenicity of biocontrol bacteria to weed leaves
Leaves of weeds with normal leaf colour (Chenopodium album, Elsholtzia densiflora, Malva verticillata, Polygonum lapathifolium, Amaranthus retroflexus, wild oats, penny cress, Herba
Polygoni Avicularis and Galium) are collected from the experimental field, and brought indoors for surface washing, disinfected with 75% alcohol for 30 s, washed with sterile water for 3 times, naturally dried, and placed on a petri dish (d = 9 cm) padded with sterile filter paper, with 3 - 4 pieces of filter paper per dish; and sterile water wets the filter paper to provide a moist environment. Hyphae pieces {(d = 8 mm) are taken from edges of colonies cultured for 7 days and inoculated to the front of leaves. Taking inoculated sterile PDA medium pieces as control, each treatment is repeated three times. They are cultured in an incubator with light for 12 h and dark for 12 h (12L : 12D) at (25 + 1)°C. After 7 days, the lesion area is measured, and the lesion area is 1/4 x length x width x 3.14. (2) Pathogenicity of biocontrol bacteria to potted weeds and crop safety (D Pathogenicity of potted weeds
Hyphae pieces are inoculated into PDB culture medium (250 mL/bottle), 5 pieces per bottle, and shaken and cultured for 120 h at 25°C and 180 r/min. After the fermentation broth is filtered with four layers of sterilized gauze to obtain the fermentation filtrate, it is inoculated on the normally growing potted weed plants with 4-7 leaves for 3 days by spray inoculation, and the inoculation amount is 25 mL/pot. After inoculated weed plants are cultured in plastic bags for 24 h, they are placed in an artificial climate box at 25-30°C and L/D=12 h//12 h, and each treatment is repeated for 3 times. The plants inoculated with sterile PDB medium are used as control. After 7 days, the incidence of inoculated weeds is observed and the incidence rate and fresh weight control effect are calculated.
Fresh weight effect (%0) = Control fresh weight — Processed fresh weight | 100%
Control fresh weight
Incidence rate = _ Number of diseased leaves of diseased leaves x 100%
Number of leaves investigat ed
Disease index = umber of diseased leaves x Number of corresponding levels
Total number of leaves investigated x Number of highest levels © Crop safety
Five main crops in Qinghai, V. faba, P. sativum, H. vulgare, T. aestivum and B. napus, are 5 planted in pots with d = 12 cm and cultured indoors. After dilution, the fermentation broth of the strain is inoculated on crop plants with 3-6 leaf stages according to the above weed pathogenicity method. Each treatment is repeated 3 times, and the crops inoculated with sterile
PDB medium are used as control. After 7 days, the incidence is investigated. The safety evaluation criteria of crops are as follows: NS means that the plants have no susceptibility (no disease spots, normal growth); LS indicates light susceptibility (scattered patches on the leaves, and slightly controlled growth and development); MS indicates moderate susceptibility (1/5 - 1/4 leaf area appears diseased spots, and the growth is inhibited); SS indicates severe susceptibility (a large number of plants die and their growth and development are seriously controlled). (3) Identification of biocontrol bacteria © Morphological identification
The strain is placed on PDA plate and cultured in an incubator at 25°C with L/D=12 h//12 h alternately. The growth rate, colony shape and colour change are observed, and the morphology of hyphae and spores is observed under the optical microscope. Preliminary identification is carried out in combination with Fungus Identification Manual. @ Molecular identification and phylogenetic tree construction
The genomic DNA of the strain is extracted by CATB method. PCR amplification is performed with universal primers ITS1 and ITS4, EF1-728F and EF1-986R, Alt-for and Alt-rev (Table 1), all of which are synthesized by Shanghai Sangon Biotech Co., Ltd. The PCR reaction systems are all (25 pL): 0.5 pL of forward and reverse primers (10 mal/l) respectively, 0.5 HL of
DNA template, 2.5 pL of 10xPCR buffer, 2.5 pL of Taq enzyme and 0.2 pL of dd HzO. The PCR cycle settings are as follows: pre-denaturing at 94°C for 5 min, denaturing at 94°C for 45 s, annealing at 55°C for 45 s, extending at 72°C for 1 min, 30 cycles, extending at 72°C for 10 min, and performing heat preservation at 4°C. After the amplification is completed, electrophoresis detection is carried out, and the purified product is recovered by using SanPrep column DNA J gel recovery kit (SK8131, Shanghai Sangon), and sent to Shanghai Sangon for two-way sequencing. The nucleotide sequence obtained by sequencing is compared and analysed with the sequence in GenBank by BLAST, the sequences are compared and analysed by Clustal X 1.8 and MEGAX 7.0, and the multiple sequences are clustered and analysed by neighbour- joining (NJ). It is tested by bootstrap method, which is repeated for 1000 times to analyse the genetic relationship between the strain and different strains of the same genus.
Table 1 Primer information used
Primers Sequence (5'—3% Type
ITS1 TCCGTAGSTGAACCTGCGG Universal
ITS4 TCCTCCGCTTATTGATATGC primers
EF1-728F CATCGAGAAGTTCGAGAAGG Universal
EF1-986R TACTTGAAGGAACCCTTACC primers
Alt-for ATGCAGTTCACCACCATCGC Universal
Alt-rev ACGAGGGTGAYGTAGGCGTC primers 1.4 Scanning electron microscope is used to observe the infection process of the strain on the leaves of Elsholtzia densiflora
Filter paper is laid in a sterilized Petri dish (® = 90 mm) and E/sholtzia densiflora leaves are placed. The filter paper is soaked with sterile water to provide a moist environment, and a bacterial cake (® = 8 mm) is placed in the centre of the leaves with a punch, and a sterile PDA bacterial cake is used as a control. Each treatment and control are repeated for 3 times. The experiment is carried out at 25°C-28°C. After inoculation, samples are taken every day to observe the invasion process of hyphae for 7 days.
After hyphae pieces of the strain are inoculated for 1-7 days, 3-6 leaf segments (0.5-1.0 cm) are prepared with fresh shaving blades of each inoculated leaf, and are fixed in 2.5% {volume/volume) glutaraldehyde in 0.1 M phosphate buffer (pH 7.2) under vacuum for 2 hours at room temperature, and in the same fixed buffer at 4°C. The leaf samples are washed three times with 0.1 M phosphate buffer (pH 7.2), and dehydrated for 30 minutes by fractional ethanol series (70%, 80%, 90% and 100%) in each gradient. The samples are dried with liquid CO: at a critical point. The fixing material is coated with a layer of 10 nm gold/palladium and observed by scanning electron microscope. 1.5 Statistical analysis
Excel and SPSS 25.0 are used for statistical analysis of the experimental data, and one- way analysis of variance (ANOVA) and least significant difference (LSD) are used to compare the difference significance test. 2. Result analysis 2.1 Pathogenicity of Alternana gaisen GD-011 to weeds, crop safety and strain identification 2.1.1 Pathogenicity of Alternaria gaisen GD-011 on weed leaves
As shown in FIG. 1, seven days after inoculation with GD-011 bacterial cake, the leaves in vitro are damaged, and the leaves of Chenopodium album, Polygonum lapathifolium and Malva verticillata are yellow and green, and hyphae penetrated and grew on the opposite side of the leaves at the inoculation site. E/sholtzia densiflora, Galium and Amaranthus retroffexus have gray hyphae, and there are withering symptoms in the later stage. Wild oats, penny cress and
Herba Polygoni Avicularis have faded, turned yellow and turned black after 7 days of inoculation. The pathogenicity of GD-011 hyphae to different weeds in vitro is as follows: Malva verticillata > Elsholtzia densiflora > Herba Polygoni Avicularis > Galium > Polygonum
J/apathifolium > Chenopodium album > Amaranthus retroflexus > wild oats > penny cress. 2.1.2 Pathogenicity of Alternaria gaisen GD-011 on potted weeds
After GD-011 fermentation products are sprayed for 7 days, the incidence rate and fresh weight control effect of Malva verticillata reach 94.83% and 90.81%, and the incidence rates of
Chenopodium album, Elsholtzia densiflora, Polygonum lapathifolium, Galium and penny cress are 75.5%, 80.87 and 77 respectively, showing that the leaves are curly and the lower leaves are wilting and yellow. After 7 days, the leaves of Malva verticillata lose their green, necrosis and shedding; the leaves of wild oats turn yellow, and 2/3 of them have died, with the incidence rate of 82.13% respectively. The leaves of Amaranthus retroflexus and Herba Polygoni
Avicularis are spotted sporadically, showing that the stems and leaves are black and withered, with the incidence rates of 56.2% and 42.07%. The disease indexes show that the fermentation filtrate of the strain is most sensitive to Malva verticillata, Galium and wild oats. After 7 days, it is found that the inoculated weeds do not revive, and the disease worsens until the whole pot of weeds die (FIG. 2 and Table 2).
Table 2 Pathogenicity of fermentation filtrate of strain GD-011 to different weeds
Weed species Incidence rate/% Disease index Fresh weight control effect/% "Chenopodium album 755%237b 614%3.14c 60.2:265bc
Elsholtzia densiflora 80.87+2.38ab 81.87+2.96b 83.9+1.63a
Polygonum fapathifolium 77.13+4.70ab 59.23+1.81cd 52.47+2.36cd
Malva verticillata 94.83+1.66a 94.47+2.38a 90.81+1.414a
Amaranthus retroflexus 56.2+6.18cd 43.412.50e 45.2712.43d
Galium 68.5113.05bc 76.1+5.78b 66.513.67b
Wild oats 82.13+1.38ab 84.03+3.29ab 71.8£7.52b
Penny cress 41.2310.54d 48.7313.19de 44.4313.38d
Herba Polygoni Avicularis 42.0713.24d 42.1712.22e 46.911.15d 2.1.3 Safety of Alternaria gaisen GD-011 on crops
The fermentation broth of strain GD-011 is not pathogenic to pea, V. faba and T. aestivum, and the growth and plant height of crops are not affected compared with the blank control plants, and they grow healthily, showing no response (NS). Rape has slight pathogenicity, 5% leaves have black spots on the leaf edge, and plants have wilting phenomenon, showing mild reaction (LS). It is slightly pathogenic to H. vulgare, and a small number of leaves have sporadic spots on the veins, and the leaves are yellow, showing a slight reaction (LS) as a whole (FIG. 3). 2.1.4 Morphological identification of Alternana gaisen GD-011
GD-011 colony is white at the beginning on PDA plate, and gradually develops into olive or dark green velvet in the later stage, with neat edges. The aerial hyphae are dense. The hyphae are colorless and septate. Conidia are brown or dark brown, oval, oval, rod-shaped or inverted pear-shaped, with 2-5 transverse septa and 0-3 mediastinum. The septa slightly overflow and contract, and the beak is cylindrical or conical. According to the culture and morphological characteristics of the strain, the pathogen is initially identified as Alternaria sp (FIG. 4). 2.1.5 Molecular biological identification of Alternaria gaisen GD-011
The rDNA-ITS, EF-1a and Alt a 1 gene sequences of strain GD-011 are amplified by PCR, and three gene fragments with lengths of 533 bp, 264 bp and 477 bp are obtained. According to the sequence of the strain, the phylogenetic tree of the strain is constructed with Ulocladium consortiale as the outer group (FIG. 5). It is found that GD-011 and Alternaria gaisen are clustered together on the phylogenetic tree, and the support rate is 99%. Alternaria gaisen may be accurately separated from other species through these three gene sequences, showing good conservation on the whole. According to the phylogenetic analysis and morphological characteristics of three gene sequences of GD-011, strain GD-011 is identified as Alternaria gaisen. 2.1.6 Observation on the pathogenic process of Elsholtzia densiflora by strain GD-011
As may be seen from FIG. 8, the ultrastructure of Elsholtzia densiflora leaves is affected by
GD-011 strain. The cell structure of uninoculated healthy E/sholízia densiflora leaves is normal and the tissues are arranged neatly (A in FIG. 6). After 1-2 days of inoculation, hyphae penetrate into stomata and little hyphae are formed around them (B and C in FIG. 6). After 3-4 days of inoculation, a large number of hyphae attaches to the tissue surface, and the leaf tissue is damaged (D and E in FIG. 8). After 5-6 days, as hyphae form a fungus net and parasitize the tissue surface, the parasitic tissue of hyphae absorbs nutrients, and the plant tissue becomes diseased (F and G in FIG. 6). With the infection of a large number of hyphae, the tissue surface appears obvious destruction. After 7 days, the hyphae grow vigorously all over the tissue surface, and the tissue metabolism of Elsholtzia densiflora leaves is disordered, and the infected cells gradually die (H in FIG. 6).
The above-mentioned embodiments only describe the preferred mode of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, all kinds of modifications and improvements made by ordinary technicians in the field to the technical scheme of the present invention should fall within the protection scope determined by the claims of the present invention.
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| US20080280761A1 (en) * | 2005-01-28 | 2008-11-13 | Sheng Qiang | Biological Control Of Weeds Using The Metabolites Of Alternaria Alternata |
| CN105861318A (en) * | 2016-04-13 | 2016-08-17 | 青海省农林科学院 | Alterneria tenuissima and application thereof in biological weeding |
| CN107969423A (en) * | 2017-11-22 | 2018-05-01 | 青海省农林科学院 | A kind of toVerticilliumdahliaActivitie Activitie S of Relative bacterial strain wettable powder and preparation method thereof |
| CN114806896A (en) * | 2022-06-06 | 2022-07-29 | 青岛农业大学 | Alternaria alternata, herbicide and application thereof |
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| US20080280761A1 (en) * | 2005-01-28 | 2008-11-13 | Sheng Qiang | Biological Control Of Weeds Using The Metabolites Of Alternaria Alternata |
| CN105861318A (en) * | 2016-04-13 | 2016-08-17 | 青海省农林科学院 | Alterneria tenuissima and application thereof in biological weeding |
| CN107969423A (en) * | 2017-11-22 | 2018-05-01 | 青海省农林科学院 | A kind of toVerticilliumdahliaActivitie Activitie S of Relative bacterial strain wettable powder and preparation method thereof |
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