CN115299463B - Application of yellow microsporidian fungus in promoting plant growth and improving plant stress resistance - Google Patents

Application of yellow microsporidian fungus in promoting plant growth and improving plant stress resistance Download PDF

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CN115299463B
CN115299463B CN202210962001.9A CN202210962001A CN115299463B CN 115299463 B CN115299463 B CN 115299463B CN 202210962001 A CN202210962001 A CN 202210962001A CN 115299463 B CN115299463 B CN 115299463B
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徐利剑
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Heilongjiang University
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Abstract

An application of yellow microsporidian fungus in promoting plant growth and improving plant stress resistance, belonging to the technical field of biological control and biological bacterial fertilizer. In order to further excavate the new application of Microsporum album (Parametarhiziumchangbiense) and Microsporum dahliae (Parametarhizium hingganense), the invention carries out a phosphorus dissolving test on two Microsporum fungi, tests the influence of spore suspension of the two Microsporum fungi on plant growth, plant salt-tolerant alkalinity and plant pathogen resistance, discovers that the Microsporum dahliae and the Microsporum dahliae have better phosphorus dissolving activity, has promoting effects on the growth of mung beans and rice, can improve the capability of the mung beans to resist salt-tolerant stress, and can obviously inhibit the growth of Rhizoctonia solani, sunflower septoria, beet cercospora and Fusarium moniliformis, and improves the capability of the mung beans to resist damping-off and rice sheath blight. Both the Bai Huangwei spore bacteria and the chrysosporium dahuricum have application prospects as microbial fertilizers and bactericides.

Description

Application of yellow microsporidian fungus in promoting plant growth and improving plant stress resistance
Technical Field
The invention belongs to the technical fields of biological control, biological bacterial fertilizer, mycorrhizal fungi and plant biostimulant, and in particular relates to application of yellow microsporidian fungi in promoting plant growth and improving plant stress resistance.
Background
The crop diseases and insect pests are important threats for agricultural production, effective control means are needed, in various control methods, biological control is expected because of the advantage of no environmental pollution, the interrelation among biological species is utilized, one type of organisms is used for inhibiting the other type of organisms, and the environment-friendly control characteristics are incomparable with non-biological control disease and insect pest methods such as chemical pesticides and the like. Fungi are important natural resources, and various biocontrol fungi are currently applied to the field of agricultural pest control. For example: entomopathogenic Beauveria bassiana (Beauveria spp.), metarhizium anisopliae (Metarhizium spp.); paecilomyces lilacinus (Purpureocillium lilacinum), a nematode pathogen, propionibacterium (Pochonia spp.); trichoderma reesei (Thricoderma spp.) and the like. Biological control mechanisms of fungi on plant diseases include: re-hosting of pathogens, production of antibacterial metabolites, preempting ecological niches and nutrients, promotion of plant growth, induction of plant disease resistance, and the like.
In a previous study, two new species of Clavicepitaceae (Clavicitaceae) were isolated and cultured from northeast forest litter, identified as new genus, and found that the two new species could be used as entomopathogenic bacteria for controlling Aphis bifidus (Monolepta hieroglyphica), bulbophyllum viridis (Callosobruchus chinensis) and Aphis zea (Rhopalosiphum maidis). The conidium of the fungus is obviously smaller than other similar ergot fungi, the colony of the fungus is white to yellow, yellow pigment can be produced, the morphological characteristics of the fungus are different from those of other ergot fungi, and the fungus can form a high-support single-line group by combining with the genetic analysis of a polygene system. This genus is designated as Microsporum chrysosporium (Parametarhizium) (Latin Wen Xueming has been registered on the Index Fungorum, mycoBank, NCBI, et al website). The phialides of the spore-producing structures of the two new species are respectively cylindrical and inverted pear-shaped, and are respectively named as Microsporum albophora (P.chanbase) and Microsporum (P hingganense) according to the source of the samples.
Disclosure of Invention
The invention relates to a new species of Clavipitaceae found and established in 2021 for both Microsporum longum (Parametarhizium changbaiense) and Microsporum chrysogenum (Parametarhizium hingganense), which are found and established in the nomenclature article (Gao S, meng W, zhang L, yue Q, zheng X and Xu L (2021) Parametarhizium (Clavicipitaceae) gen. Nov. With Two New Species as a Potential Biocontrol Agent Isolated From Forest Litters in Northeast China. Front. Microbiol.12: prior to the publication of 627044. Doi:10.3389/fmib.2021.627244, the great Bai Huangwei spore fungus was named Metarhizium anisopliae (Metarhizium changbaiensis) (deposited at China general microbiological culture Collection center (CGMCC), the deposited address was CGMCC No.19143, the deposited date was 2019 12 months 05), the great 35 spore fungus was named Metarhizium anisopliae (Metarhizium hingganensis) (deposited at China General Microbiological Collection Center (CGMCC), the deposited address was CGMCC No.19144, and the deposited date was 2019 12 months 05). The MycoBank accession number of the strain Bai Huangwei is MB 837522, the DNA sequence thereof is GenBank accession number is MN589741 (ITS), MN589994 (LSU), MN590231 (SSU), MN908589 (TEF), MT921830 (TUB), MN917168 (RPB1α), MT921829 (RPB2α), the MycoBank accession number of the strain Microsporum is MB 837523, the DNA sequence thereof is GenBank accession number is MN055703 (ITS), MN061635 (LSU), MN055706 (SSU), MN065770 (TEF), MN061672 (TUB), MN917170 (RPB1α), MT939494 (RPB2α).
In order to further excavate new applications of Microsporum albopictus (P.chanbase) and Microsporum dahuricum (P.hingganense), the invention provides an application of Microsporum fungus in improving the utilization rate of indissoluble phosphorus in soil, wherein the Microsporum fungus is Microsporum albopictus (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense).
The invention also provides application of the yellow microsporidian fungus in promoting plant growth, wherein the yellow microsporidian fungus is Microsporum albolbosum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense).
The invention also provides application of the yellow microsporidian fungus in improving the salt tolerance of plants, wherein the yellow microsporidian fungus is Microsporum albolbosum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense).
The invention also provides application of the yellow microsporidian fungus in improving the alkali resistance of plants, wherein the yellow microsporidian fungus is Microsporum albolbosum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense).
The invention also provides an application of the yellow microsporidian fungus in inhibiting the activity of pathogenic fungi causing plant diseases, wherein the yellow microsporidian fungus is Microsporum albolbosum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense).
Further defined, the pathogenic fungi causing plant disease include rhizoctonia solani (Rhizoctonia solani), septoria sunflower (Septoria helianthi), cercospora betana (Cercospora beticola) and fusarium moniliforme (Fusarium moniliforme).
The invention also provides a bacterial fertilizer which contains Microsporum albopictum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense) or both Microsporum albopictum (Parametarhizium changbaiense) and Microsporum dahuricum (Parametarhizium hingganense); the bacterial fertilizer can improve the utilization rate of indissolvable phosphorus in soil, promote plant growth, help plants to withstand salt stress, help plants to withstand alkali stress and help plants to withstand plant diseases.
Further defined, the plant disease is a plant disease caused by rhizoctonia solani (Rhizoctonia solani) or aschersonia sunflower (Septoria helianthi) or cercospora beet (Cercospora beticola) or fusarium moniliforme (Fusarium moniliforme).
The invention also provides a mycorrhizal fungi which contains Microsporum albopictum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense) or contains Microsporum albopictum (Parametarhizium changbaiense) and Microsporum dahuricum (Parametarhizium hingganense); the mycorrhizal fungi can improve the utilization rate of indissolvable phosphorus in soil, promote plant growth, help plants to withstand salt stress, help plants to withstand alkali stress and help plants to withstand plant diseases.
Further defined, the plant disease is a plant disease caused by rhizoctonia solani (Rhizoctonia solani) or aschersonia sunflower (Septoria helianthi) or cercospora beet (Cercospora beticola) or fusarium moniliforme (Fusarium moniliforme).
The invention also provides a plant biological hormone which contains Microsporum albopictum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense) or both Microsporum albopictum (Parametarhizium changbaiense) and Microsporum dahuricum (Parametarhizium hingganense); the plant biological hormone can improve the utilization rate of insoluble phosphorus in soil, promote plant growth, help plants to withstand salt stress, help plants to withstand alkali stress and help plants to withstand plant diseases.
Further defined, the plant disease is a plant disease caused by rhizoctonia solani (Rhizoctonia solani) or aschersonia sunflower (Septoria helianthi) or cercospora beet (Cercospora beticola) or fusarium moniliforme (Fusarium moniliforme).
The invention also provides a bactericide, which is characterized by comprising Microsporum albopictum (Parametarhizium changbaiense) or Microsporum dahuricum (Parametarhizium hingganense) or both Microsporum albopictum (Parametarhizium changbaiense) and Microsporum dahuricum (Parametarhizium hingganense).
Further defined, the germicides are capable of inhibiting rhizoctonia solani (Rhizoctonia solani), aschersonia sunflower (Septoria helianthi), cercospora betaensis (Cercospora beticola) and fusarium moniliforme (Fusarium moniliforme).
The invention has the beneficial effects that:
the invention further digs the functions of the two strains of yellow microsporidian fungi, and discovers that the two strains of yellow microsporidian fungi have the following functions:
1. both Bai Huangwei and Microsporum dahuricum can be used in the preparation of insoluble phosphorus Ca 3 (PO 4 ) 2 Can produce phosphate-dissolving ring and has better phosphate-dissolving activity.
2. As a result of treating seeds of mung beans and rice respectively by using spore fungus suspensions of Microsporum albus and Microsporum dahuricum, the spore fungus suspensions of Microsporum dahuricum and Microsporum dahuricum are found to have promotion effects on growth of mung beans and rice.
3. The spore bacterial suspension of Microsporum albus and Microsporum dahuricum is used for respectively treating the seeds of mung beans and rice, and the result shows that the Microsporum album and Microsporum dahuricum can form mycorrhizal shapes with the roots of mung beans and rice.
4. The spore bacterial suspensions of the Microsporum albus and Microsporum dahuricum are used for respectively soaking mung bean seeds and carrying out salt stress on mung beans, and the Bai Huangwei spore bacteria and the Microsporum dahuricum are found to be capable of reducing the damage of the salt stress to the growth of the mung beans and improving the salt stress resistance of the mung beans.
5. The spore bacterial suspensions of the Microsporum albus and Microsporum dahuricum are used for respectively soaking mung bean seeds and carrying out alkali stress on mung beans, and the Bai Huangwei spore bacteria and the Microsporum dahuricum are found to be capable of reducing the damage of alkali stress to the growth of the mung beans and improving the alkali stress resistance of the mung beans.
6. In-vitro antibacterial tests prove that the growth of rhizoctonia solani, the sunflower septoria, the fusarium moniliforme and the tail-producing spore of beet can be obviously inhibited by both the long Bai Huangwei spore fungus and the yellow microsporium dahliae, and in-vivo disease resistance tests prove that the long Bai Huangwei spore fungus and the yellow microsporium dahliae have better prevention and control effects on mung bean damping-off (Rhizoctonia solani) and rice sheath blight (Rhizoctonia solani).
Drawings
FIG. 1 is a graph showing the results of detecting the phosphorus-solubilizing activity of a fungus belonging to the genus Microsporum; wherein A in FIG. 1 is a phosphorus-dissolving activity detection result diagram of Microsporum albolabrium, and B in FIG. 1 is a phosphorus-dissolving activity detection result diagram of Microsporum dahuricum;
FIG. 2 is a graph showing the results of microscopic observation of symbiosis of yellow microsporidian fungi and mung bean roots; wherein, A and B in FIG. 2 are respectively controls under fluorescence and non-fluorescence, C and D in FIG. 2 are respectively graphs of the root symbiosis of Green Fluorescent Protein (GFP) labeled long Bai Huangwei spore bacterial strain and mung bean under fluorescence and non-fluorescence, E and F in FIG. 2 are respectively graphs of the root symbiosis of GFP labeled Khingei Microsporum strain and mung bean under fluorescence and non-fluorescence;
FIG. 3 is a graph showing the results of microscopic observation of symbiosis of Microsporum fungi and rice roots; wherein, A and B in FIG. 3 are respectively controls under fluorescence and non-fluorescence, C and D in FIG. 3 are respectively graphs of the symbiotic condition of the yellow microsporium GFP strain and the rice root under fluorescence and non-fluorescence, E and F in FIG. 3 are respectively graphs of the symbiotic condition of the yellow microsporium GFP strain and the rice root under fluorescence and non-fluorescence;
FIG. 4 is a graph showing the effect of Microsporum fungus on the salt stress resistance of plants; wherein A in FIG. 4 is a graph of the effect of yellow microsporidian fungus on the growth of the leaves and roots of mung bean plants under the stress of 60mM salt solution, and B in FIG. 4 is a graph of the statistical results of the above-ground fresh weight and the underground fresh weight of mung bean plants under the stress of different concentrations of salt solution;
FIG. 5 is a graph showing the effect of Microsporum fungus on the alkali stress resistance of plants; wherein A in FIG. 5 is a graph of the effect of yellow microsporidian fungus on the growth of the leaves and roots of mung bean plants under the stress of 60mM alkali solution, and B in FIG. 5 is a graph of the statistical results of the overground fresh weight and underground fresh weight of mung bean plants under the stress of different concentrations of alkali solution;
FIG. 6 is a graph showing the results of an in vitro antimicrobial test of a fungus of the genus Microsporum; wherein A1, A2, A3 and A4 in FIG. 6 are respectively graphs of the results of in vitro antibacterial tests of Microsporum candidum, septoria sunflower, cercospora betana and Fusarium moniliforme, and B1, B2, B3 and B4 in FIG. 6 are respectively graphs of the results of in vitro antibacterial tests of Microsporum candidum, septoria sunflower, cercospora betana and Fusarium moniliforme;
FIG. 7 is a graph showing the effect of yellow microspora fungi on controlling green bean damping-off; wherein, A in figure 7 is the prevention and treatment effect diagram of Microsporum albolabrim on mung bean damping-off, B in figure 7 is the prevention and treatment effect diagram of Microsporum dahuricum on mung bean damping-off, and C in figure 7 is a control group;
FIG. 8 is a graph showing the effect of yellow microspora fungi on controlling rice damping-off; wherein A in FIG. 8 is a control group, B in FIG. 8 is a graph of the control effect of Microsporum albolabrim on rice damping-off, and C in FIG. 8 is a graph of the control effect of Microsporum dahuricum on rice damping-off.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described in detail with reference to the following detailed description and the accompanying drawings. The experimental methods used in the examples below were conventional, and the materials, reagents, methods and apparatus used, unless otherwise indicated, were all conventional in the art and commercially available to those skilled in the art.
The test plant pathogenic bacteria related to the invention are as follows:
rhizoctonia solani (Rhizoctonia solani) with hypha fusion AG-1 and strain collection number of CCTCC AF 2021031. Acremonium Helianum FXL003 (Septoria helianthi), cercospora betana FWZ004 (Cercospora beticola) and Fusarium moniliforme FXL011 (Fusarium moniliforme) were both stored in the university of Heilongjiang plant pathology laboratory.
Example 1: phosphorus-solubilizing Activity of yellow microsporidian fungi
The phosphorus-dissolving medium was prepared according to the following formula: 20g glucose, 8g Ca 3 (PO 4 ) 2 ，5g MgCl 2 ，0.24g MgSO 4 ，0.2g KCl，0.1g NH 4 (SO 4 ) 2 Constant volume to 1L with water, packaging into triangular flask containing agar, shaking, sterilizing at 121deg.C for 20min under moist heat, shaking, and pouring into flat plate. The microbial cakes of Microsporum albopictum and Microsporum dahuricum are inoculated to the center of the flat plate respectively, and the growth condition of fungi and the phosphorus dissolution condition around the colony are observed and measured every day.
As shown in FIG. 1, both the Bai Huangwei and the Microsporum chrysogenum can be used in the presence of insoluble phosphorus Ca 3 (PO 4 ) 2 Obvious phosphate solubilizing circles can be seen at the edge of colony growth, indicating that Bai Huangwei spore bacteria and Microsporum dahuricum have phosphate solubilizing activity.
Example 2: effect of yellow microspora fungi on plant growth
(1) Cultivation of yellow microsporidian fungi and preparation of spore suspension
Two yellow microsporidian fungi were inoculated onto 9cm PDA plates, incubated at 25℃for 14 days, sterilized 0.05% Tween 20 solution was added, spores were scraped off, and the spore suspension was filtered through sterile medical gauze (hyphae were filtered off). Then the spore concentration was measured by microscope and hemocytometer, and the spore suspension was diluted to a spore concentration of 1X 10 8 And (3) one/mL for later use.
(2) Treatment of plant seeds with spore suspensions
Mung bean: cleaning mung bean seeds with sterile water, airing, respectively soaking the mung bean seeds in the spore suspension obtained in the step (1) and in a 0.05% Tween 20 solution (contrast), accelerating germination at 25 ℃, planting the mung bean seeds in soil sterilized at the high temperature of 121 ℃ for 40min after the mung bean seeds germinate (humus soil: vermiculite=1:1, v/v), and embedding the mung bean seeds in the depth of 2-3 cm. Culturing under 16h illumination and 8h darkness at room temperature. .
Rice: washing rice seeds with sterile water, soaking until the rice seeds are exposed to white, transferring the rice seeds into the spore suspension obtained in the step (1) and sterile water (contrast), and continuously soaking for 3 days under the conditions of 16h illumination, 8h darkness and 28 ℃. Then the rice seeds are transferred into a 96-well rice cultivation box (the box body is black and light-proof 270X 245X 180 mm) filled with Yoshida nutrient solution, and liquid cultivation is carried out under the conditions of 16h illumination, 8h darkness and 28 ℃.
The spore suspension treatment plant seed test, comprising control, microsporum albophora treatment, microsporum dahuricum treatment, was repeated for more than 3 rounds of treatment with 20 to 40 plants per round (same round, same number of individual treatment plants).
(3) Plant growth assay
Plants of each treatment group were assayed for growth when the groups to be treated produced a significant difference in phenotype from the control group, i.e., 21 days after spore suspension treatment. For mung bean plants, measuring plant height, root length, fresh weight, root area and leaf area, wherein the leaf area and root area are obtained by imaging the 1 st, 2 nd and 3 rd true leaves and complete roots by using Image J1.8.0 software, and measuring total leaf area and root area; measuring plant height and root length by using a ruler; fresh weight is measured by washing mung bean plants clean and wiping off water, and weighing with an analytical balance. For rice plants, the plant height, root length, overground fresh weight and underground fresh weight are measured, wherein the plant height and root length are measured by using a ruler, the overground fresh weight and the underground fresh weight are obtained by washing rice plants cleanly and wiping off water, then dividing the plants into overground parts and underground parts, and respectively weighing the fresh weights of the overground parts and the underground parts by using an analytical balance.
The statistical mung bean growth index is shown in table 1 when the mung bean potted plant grows for 21 days. Compared with a control group which is not treated by spore suspensions of the yellow microsporidian fungi, plants treated by spore suspensions of the two yellow microsporidian fungi show obvious increase in plant height, root length, fresh weight, root area and leaf area, and the two yellow microsporidian fungi have obvious promotion effect on the growth of mung beans.
TABLE 1 influence of Microsporum albopictum on mung bean plant growth index by Microsporum dahuricum
When the rice is liquid cultured for 21 days, various growth indexes of the rice are counted as shown in table 2. Compared with a control group which is not treated by the spore suspension of the yellow microsporidian fungi, the rice plants treated by the spore suspension of the yellow microsporidian fungi show obvious increase in plant height, root length, overground fresh weight and underground fresh weight, and the rice plants treated by the spore suspension of the yellow microsporidian fungi show obvious increase in overground fresh weight and underground fresh weight, so that the two yellow microsporidian fungi have obvious promotion effect on the growth of rice.
TABLE 2 influence of Microsporum albopictum on growth index of rice plants by Microsporum dahuricum
Example 3: symbiotic conditions of yellow microsporidian fungi and plant roots
The symbiotic observation method comprises the following steps: the pCAMBIA1307-eGFP plasmid is transformed into the Microsporum fungus by using an agrobacterium-mediated fungus genetic transformation method to obtain the GFP-marked Microsporum fungus. According to the method for treating mung beans and rice by using yellow microsporidian fungi, which is described in the embodiment 2, mung beans and rice are treated by using GFP-marked yellow microsporidian fungi spore liquid, germinated mung beans and rice seeds are transferred to a Yoshida liquid culture medium for culture after the treatment is finished, root observation is carried out every other day, and the observation method is as follows:
1) Drawing materials: root sampling was performed every other day starting on day 3 after inoculation until day 21.
2) Fixing: the roots were cut into root sections of about 2cm, placed in FAA fixative and fixed overnight at 4 ℃.
3) And (3) transparent treatment: washing the fixed root section with distilled water until the root section is odorless, soaking the root section in 100g/L KOH solution, boiling the root section in a water bath at 90 ℃ until the solution is transparent, timely replacing the solution if the solution changes color in the middle, washing the root section with distilled water until the solution is colorless after the root section is boiled to be transparent, and storing the washed root section in the distilled water.
4) And (3) tabletting: 60% of glycerin was added dropwise to the center of the slide, the roots after the transparent treatment were grasped with forceps and spread in glycerin, and the cover glass was covered.
5) And (5) microscopic examination: root section slide was observed with a fluorescence microscope.
Microscopic observation shows that both the Bai Huangwei and the Microsporum chrysogenum can be symbiotic with the roots of mung beans and rice (see FIG. 2 and FIG. 3).
Example 3: effect of Microsporum fungi on salt stress resistance of plants
The preparation method of the suspension of spore of fungus belonging to genus Xanthomonas and the method of treating mung bean seed with the suspension of spore of fungus belonging to genus Xanthomonas are as shown in (1) and (2) in example 2. When the mung bean grows to 4-5cm, subjecting the mung bean to salt stress treatment, wherein the salt solution is prepared from NaCl, na 2 SO 4 In the (molar ratio) configuration of (9:1), concentration gradients of 0mmol/L, 15mmol/L, 30mmol/L, 45mmol/L and 60mmol/L are set, 50mL of saline solution is fixedly poured into each mung bean pot every day, the saline solution is ensured to exist in the pot all the time, the plant is always in a salt stress state, and phenotypic analysis is carried out after 10 days of stress.
The results are shown in fig. 4, and it can be known from fig. 4 that both the chrysosporium dahuricum and the chrysosporium longum can alleviate the damage caused by salt stress to plants to different degrees, under salt stress, when obvious wilting occurs in a control group, mung beans treated by the chrysosporium fungi still keep healthy growth state, and the chrysosporium dahuricum is significantly superior to the control and the chrysosporium longum in terms of improving the fresh weight on the ground and the fresh weight under the ground.
Example 4: effect of Fusarium fungi on alkali stress resistance of plants
The preparation method of the suspension of spore of fungus belonging to genus Xanthomonas and the method of treating mung bean seed with the suspension of spore of fungus belonging to genus Xanthomonas are as shown in (1) and (2) in example 2. When the mung bean grows to 4-5cm, performing alkali stress treatment on the mung bean, and pressing the alkali solution into NaHCO (sodium bicarbonate) 3 :Na 2 CO 3 The preparation method comprises the following steps of (1) configuring the solution with the concentration gradient of 0mmol/L, 15mmol/L, 30mmol/L, 45mmol/L and 60mmol/L, and pouring 50mL of alkaline solution into each pot of mung bean every day to ensure that the alkaline solution exists in the pot all the time, so that the plant is always in an alkaline stress state, and performing phenotype analysis after 10 days of stress.
The results are shown in FIG. 5, and it is known from FIG. 5 that both Microsporum dahuricum and Microsporum albus can alleviate the damage of alkali stress to plants to different degrees. Under the alkali stress, the control group grows slowly, and mung beans treated by the yellow microsporidian fungi still keep a continuous growth state.
Example 5: anti-phytopathogenic fungal activity of yellow microsporidian fungi
(1) In vitro antibacterial test
The anti-phytopathogenic fungi activity of the yellow microspora fungi was tested by the counter culture assay: the cakes of Microsporum albopictus and Microsporum dahuricum were inoculated onto the PDA culture plates, respectively, and the cakes were about 3cm from the left edge and about 4.5cm from the upper and lower edges. Then, the plant pathogenic bacteria (Rhizoctonia solani, septoria sunflower, cercospora beet and Fusarium moniliforme) cakes were inoculated to the right side of the Microsporum flavum cake by about 3cm, respectively. The growth of pathogenic fungi was observed daily. Both the Bai Huangwei and the Microsporum dahliae were found to have activity in inhibiting the growth of Rhizoctonia solani, septoria sunflower, cercospora betas and Fusarium moniliforme (see FIG. 6).
(2) In vivo antibacterial test of Rhizoctonia solani by yellow microsporidiana
The preparation method of the suspension of spore of fungus belonging to genus Xanthomonas and the method of treating mung bean and rice seed with the suspension of spore of fungus belonging to genus Xanthomonas are as shown in (1) and (2) in example 2.
The culture and inoculation modes of rhizoctonia solani are as follows:
inoculating rhizoctonia solani on mung beans: the rhizoctonia solani bacterial cake is inoculated on a PDA culture plate, and is cultured at 25 ℃ until hyphae are fully paved on the PDA culture plate, and bacterial colonies are beaten into bacterial cakes with the diameter of 6mm by using a puncher. When the plant height of the potted mung bean (comprising the steps of controlling, inoculating the yellow microspore fungus for treatment and inoculating the yellow microspore fungus for treatment) is about 4-5cm, inoculating a rhizoctonia solani cake at a position 2cm away from the seedling in the flowerpot, and burying the cake to a depth of 1cm in soil.
Rice inoculation rhizoctonia solani: preparing fungus-grafting sticks with beech straight grain bark, cutting bark into fungus-grafting sticks with length of 1cm and width of 0.2mm by scissors, and sterilizing at 121deg.C for 20min under moist heat. Inoculating rhizoctonia solani bacterial cake onto PDA culture medium plate, placing sterilized bacterial inoculating sticks around bacterial cake with tweezers to make bacterial inoculating stick direction perpendicular to mycelium growth direction, facilitating mycelium winding, and culturing at room temperature until mycelium is completely covered with bacterial inoculating sticks. Inoculating bacterial strain with hypha (with sterile bacterial strain control) in the three-leaf stage of rice. When in inoculation, the leaves of the rice basal part are gently poked by hands, the bacteria-inoculating sticks are gently inserted by forceps, and the normal plant morphology of the rice is not affected after the bacteria-inoculating sticks are inserted.
And (3) disease resistance related index statistics:
after most plants of the control (plants not treated by the yellow microsporidian fungus spores) are obviously ill, the plant height, fresh weight and root length of mung bean and rice plants are measured, and the wilting index, the disease incidence, the disease index and the control effect are counted. The specific formula is as follows:
when mung bean plants of the control group are ill and most plants fall, the resistance of the mung bean plants to diseases caused by rhizoctonia solani can be remarkably improved by treating the chrysosporium fungi (see figure 7), and the Bai Huangwei spore bacteria and the chrysosporium dahuricum can be found to reduce wilting index, disease incidence and disease index, and have higher control effect on diseases caused by rhizoctonia solani (see table 3). By detecting the content of Jasmonic Acid (JA) and Salicylic Acid (SA) in mung bean plants of each group, it is found that the Microsporum flavum can induce plants to accumulate a large amount of Jasmonic Acid (JA) and Salicylic Acid (SA), and induce the plants to generate broad-spectrum resistance. Compared with the JA content (3.83+/-0.2 ng/g fresh weight) of the control root, the JA content in the mung bean treated by the long Bai Huangwei spore fungus (18.6+/-0.52 ng/g fresh weight) and the mung bean treated by the micro spore fungus from Khingan (14.58+/-0.29 ng/g fresh weight) is obviously increased. The SA content in the mung bean treated with Bai Huangwei spore bacteria (4.44+ -0.19 ng/g fresh weight) was slightly and significantly increased compared with the SA content in the control root (3.35+ -0.12 ng/g fresh weight). The SA content (26.92+/-1.21 ng/g fresh weight) of the mung bean leaves treated by the Bai Huangwei spore fungus is obviously higher than that of the control leaves (17.13+/-0.46 ng/g fresh weight).
TABLE 3 activity of yellow microspora fungi against mung bean diseases
When large-area moire spots appear on the disease of the rice plants in the control group, it can be observed that the yellow microsporidian fungi can help the rice to resist diseases, reduce the spot area and reduce the disease degree (see figure 8). The Bai Huangwei spore bacteria and the Microsporum dahuricum can reduce the influence of rhizoctonia solani on the growth of rice, and the fresh weight on the ground are obviously improved compared with the control (see Table 4). The rhizoctonia solani can obviously reduce the fresh weight of the overground part and the underground part of rice while causing the disease spots, and can obviously relieve the influence of the rhizoctonia solani on the fresh weight after being treated by the yellow microsporidian fungi.
TABLE 4 Effect of yellow microspora fungi on Rhizoctonia solani (Rhizoctonia solani)
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (3)

1. The application of the yellow microsporidian fungus in inhibiting the activity of pathogenic fungi causing plant diseases is characterized in that the yellow microsporidian fungus is yellow microspora longum @ or the likeParametarhizium changbaiense) Or Microsporum chrysogenumParametarhizium hingganense) The method comprises the steps of carrying out a first treatment on the surface of the The pathogenic fungi causing plant diseases are rhizoctonia solani @Rhizoctonia solani) Or Fusarium moniliformeFusarium moniliforme）。
2. The application of the bacterial fertilizer in preventing and controlling plant diseases is characterized in that the bacterial fertilizer contains Microsporum longumParametarhizium changbaiense) Or Microsporum chrysogenumParametarhizium hingganense) Or contains Microsporum albolbostemma simultaneouslyParametarhizium changbaiense) And Microsporum chrysosporium of XinganParametarhizium hingganense) The method comprises the steps of carrying out a first treatment on the surface of the The bacterial fertilizer can prevent and treat rhizoctonia solaniRhizoctonia solani) Or Fusarium moniliformeFusarium moniliforme) Caused plant diseases.
3. The application of the bactericide in inhibiting the activity of pathogenic fungi causing plant diseases is characterized in that the bactericide contains Microsporum longumParametarhizium changbaiense) Or Microsporum chrysogenumParametarhizium hingganense) Or contains Microsporum albolbostemma simultaneouslyParametarhizium changbaiense) And Microsporum chrysosporium of XinganParametarhizium hingganense) The method comprises the steps of carrying out a first treatment on the surface of the The pathogenic fungi causing plant diseases are rhizoctonia solani @Rhizoctonia solani) Or Fusarium moniliformeFusarium moniliforme）。
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA3149337A1 (en) * 2018-09-14 2020-03-19 Bionema Ltd Insect-pathogenic fungus, spores, composition and use of same
CN112940944A (en) * 2020-08-28 2021-06-11 黑龙江大学 Entomopathogenic fungi and screening method and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA3149337A1 (en) * 2018-09-14 2020-03-19 Bionema Ltd Insect-pathogenic fungus, spores, composition and use of same
CN112940944A (en) * 2020-08-28 2021-06-11 黑龙江大学 Entomopathogenic fungi and screening method and application thereof

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
Parametarhizium hingganense, a Novel Ectomycorrhizal Fungal Species, Promotes the Growth of Mung Beans and Enhances Resistance to Disease Induced by Rhizoctonia solani;Ying Gao等;《Fungi》;第1-16页 *
兴 安 落 叶 松 林 下凋落物真菌的抗植物病原菌活性;吴彤等;《植 物 病 理 科 技 创 新 与 绿 色 防 控》;第72页 *
大兴安岭森林凋落物的活性真菌及其代谢产物研究;邱天艺等;《中国农学通报》;第38卷(第18期);122-127 *
戴氏绿僵菌抗微生物潜力的评估;肖代敏;肖建辉;张志敏;孙中华;;中药材(06);第952-957页 *
来自大兴安岭凋落物的绿僵菌及其相近 菌属真菌的生物活性;高思禹等;《植物保护》;第47卷(第5期);第19-27页 *

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