CN113308382A - Biocontrol bacterium preparation for enhancing stress resistance of crops - Google Patents
Biocontrol bacterium preparation for enhancing stress resistance of crops Download PDFInfo
- Publication number
- CN113308382A CN113308382A CN202110530490.6A CN202110530490A CN113308382A CN 113308382 A CN113308382 A CN 113308382A CN 202110530490 A CN202110530490 A CN 202110530490A CN 113308382 A CN113308382 A CN 113308382A
- Authority
- CN
- China
- Prior art keywords
- bacillus licheniformis
- fermentation
- powder
- trichoderma longibrachiatum
- fermentation liquor
- 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.)
- Pending
Links
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
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
-
- 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
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention provides a biocontrol bacterium preparation for enhancing stress resistance of crops, which consists of trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris fermentation liquor, wherein the total viable count of the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris is more than or equal to 5107cfu/ml; the ratio of viable count of Trichoderma longibrachiatum, Bacillus licheniformis and Gliocladium incarnatum is (0.1-10): 0.1-10), and fermentation medium for preparing Bacillus licheniformis fermentation liquid is added with 0.1-0.5 wt% radix astragali powder and 0.1-0.5 wt% Notoginseng radix powder. The biocontrol bacterium preparation disclosed by the invention comprises active ingredients with different action mechanisms, can effectively adjust the stress resistance of plants under the synergistic action, improves the stress resistance of the plants, and belongs to green, environment-friendly and non-toxic active ingredients with double biological regulation functions of pesticide effect and fertilizer effect.
Description
Technical Field
The invention relates to a biocontrol bacterium preparation, in particular to a biocontrol bacterium preparation for enhancing stress resistance of crops.
Background
Corn is an important food economic crop all over the world, the total yield of the corn is second to that of wheat, the corn is second to the wheat, the corn is an important food crop in China, and the corn has an important strategic position in food safety. The growth of the corn is often interfered by the external environment, and drought is one of the main factors influencing the growth of the corn at present. It is estimated that corn is reduced by 15% -20% annually due to drought, and this trend increases as drought climates become more frequent and severe. Therefore, how to improve the drought resistance of corn becomes a focus of attention.
Disclosure of Invention
The invention aims to overcome the defects in the prior art, provides a biocontrol bacterium preparation for enhancing the stress resistance of crops, and can effectively improve the drought resistance of corn.
The invention provides a biocontrol bacterium preparation for enhancing stress resistance of crops, which consists of trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris fermentation liquor, and in the biocontrol bacterium preparation, the total viable count of the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris is more than or equal to 5 multiplied by 107cfu/ml; the viable count ratio of the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris is (0.1-10): 0.1-10), wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor is added with 0.1-0.5 wt% of astragalus powder and 0.1-0.5 wt% of pseudo-ginseng powder.
Preferably, 0.3 wt% of astragalus powder and 0.2 wt% of panax notoginseng powder are added into the fermentation medium for preparing the bacillus licheniformis fermentation liquor.
Preferably, the ratio of viable count of the trichoderma longibrachiatum, the bacillus licheniformis and the saccharum arenosum is (4-9): 4-9.
The present invention is not particularly limited to the fermentation medium for preparing the fermentation broth of trichoderma longibrachiatum, and any medium can be used as long as it can perform fermentation culture of trichoderma longibrachiatum, and those skilled in the art can select the medium according to the nature of the strain.
The present invention is not particularly limited to the fermentation medium for preparing sacculus terreus fermentation broth, and any medium may be used as long as it can perform fermentation culture on sacculus terreus, and those skilled in the art can select the medium according to the nature of the strain.
The fermentation medium for preparing the bacillus licheniformis fermentation liquor is prepared by adding astragalus powder and pseudo-ginseng powder into a basic fermentation medium, and the basic fermentation medium is not particularly limited in the invention, so long as the fermentation medium can perform fermentation culture on the bacillus licheniformis, and the fermentation medium can be reasonably selected by a person skilled in the art according to the strain property.
For example, the composition of the basal fermentation medium may be: 2-4 wt% of corn flour, 2-4 wt% of bean cake powder, 2-4 wt% of bran powder and K2HPO4 2-2.5wt％、KH2P04 0.5-1wt％、MgSO40.05-0.1 wt%, peanut oil 0.1-0.5 wt%, and the balance water, starting at ph7.5, then the fermentation medium for preparing a bacillus licheniformis broth in the present invention may be composed of: 2-4 wt% of corn flour, 2-4 wt% of bean cake powder, 2-4 wt% of bran powder and K2HPO4 2-2.5wt％、KH2P040.5-1wt％、MgSO40.05-0.1 wt%, peanut oil 0.1-0.5 wt%, astragalus root powder 0.1-0.5 wt%, notoginseng powder 0.1-0.5 wt%, and water in balance, with an initial pH of 7.5.
In a particular embodiment of the invention, the fermentation medium from which the Bacillus licheniformis broth is prepared comprises: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P040.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.3 wt%, panax notoginseng powder 0.2 wt%, and the balance water, starting at ph 7.5.
In a second aspect of the invention there is provided a biocontrol agent as described in the first aspect of the inventionThe preparation method of the agent comprises the following steps: respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the trichoderma longibrachiatum fermentation liquor, the bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to obtain the trichoderma longibrachiatum fermentation liquor; wherein the total viable count of Trichoderma longibrachiatum, Bacillus licheniformis and saccule mycoderma is not less than 5 × 107cfu/ml; the viable count ratio of the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris is (0.1-10): 1-10): 0.1-10), and 0.1-0.5 wt% of astragalus powder and 0.1-0.5 wt% of panax notoginseng powder are added into a fermentation culture medium for preparing the bacillus licheniformis fermentation liquor.
Preferably, 0.3 wt% of astragalus powder and 0.2 wt% of panax notoginseng powder are added into the fermentation medium for preparing the bacillus licheniformis fermentation liquor.
Preferably, the ratio of viable count of the trichoderma longibrachiatum, the bacillus licheniformis and the saccharum arenosum is (4-9): 4-9.
The medium for seed culture is not particularly limited herein, and any medium may be used for the Trichoderma longibrachiatum as long as it can perform seed culture on the Trichoderma longibrachiatum, any medium may be used for Bacillus licheniformis as long as it can perform seed culture on the Bacillus licheniformis, and any medium may be used for sacculus terrestris as long as it can perform seed culture on the sacculus terrestris, and those skilled in the art can select the medium appropriately according to the nature of the species.
The present invention is not particularly limited to the fermentation medium for preparing the fermentation broth of trichoderma longibrachiatum, and any medium can be used as long as it can perform fermentation culture of trichoderma longibrachiatum, and those skilled in the art can select the medium according to the nature of the strain.
The present invention is not particularly limited to the fermentation medium for preparing sacculus terreus fermentation broth, and any medium may be used as long as it can perform fermentation culture on sacculus terreus, and those skilled in the art can select the medium according to the nature of the strain.
The fermentation medium for preparing the bacillus licheniformis fermentation liquor is prepared by adding astragalus powder and pseudo-ginseng powder into a basic fermentation medium, and the basic fermentation medium is not particularly limited in the invention, so long as the fermentation medium can perform fermentation culture on the bacillus licheniformis, and the fermentation medium can be reasonably selected by a person skilled in the art according to the strain property.
For example, the composition of the basal fermentation medium may be: 2-4 wt% of corn flour, 2-4 wt% of bean cake powder, 2-4 wt% of bran powder and K2HPO4 2-2.5wt％、KH2P04 0.5-1wt％、MgSO40.05-0.1 wt%, peanut oil 0.1-0.5 wt%, and the balance water, starting at ph7.5, then the fermentation medium for preparing a bacillus licheniformis broth in the present invention may be composed of: 2-4 wt% of corn flour, 2-4 wt% of bean cake powder, 2-4 wt% of bran powder and K2HPO4 2-2.5wt％、KH2P040.5-1wt％、MgSO40.05-0.1 wt%, peanut oil 0.1-0.5 wt%, astragalus root powder 0.1-0.5 wt%, notoginseng powder 0.1-0.5 wt%, and water in balance, with an initial pH of 7.5.
In a particular embodiment of the invention, the fermentation medium from which the Bacillus licheniformis broth is prepared comprises: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P040.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.3 wt%, panax notoginseng powder 0.2 wt%, and the balance water, starting at ph 7.5.
In a third aspect, the invention provides the use of the biocontrol agent of the first aspect of the invention for enhancing stress resistance of crops.
Preferably, the biocontrol bacterial preparation is used for a base fertilizer when crops are planted.
When the biocontrol bacterium preparation is used, the biocontrol bacterium preparation can be used independently or together with other fertilizers, the base fertilizer is ploughed after the surface of the land is spread, the biocontrol bacterium preparation can also be used as the base fertilizer for furrow application when sowing, and the using amount of the biocontrol bacterium preparation per mu is 1-2 kilograms.
The biocontrol bacteria preparation disclosed by the invention comprises the effective components of trichoderma longibrachiatum and fermentation metabolites thereof, bacillus licheniformis and fermentation metabolites thereof, glomus sacculus and fermentation metabolites thereof, astragalus membranaceus fermentation metabolites after bacillus licheniformis fermentation, and panax notoginseng fermentation metabolites after bacillus licheniformis fermentation, and belongs to the active components with different action mechanisms.
Detailed Description
The invention will be better understood by reference to the following examples. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
In a specific embodiment of the present invention, the present invention is not particularly limited to a culture medium for seed culture, but any medium may be used for trichoderma longibrachiatum as long as it can perform seed culture on trichoderma longibrachiatum, any medium may be used for bacillus licheniformis as long as it can perform seed culture on bacillus licheniformis, and any medium may be used for sacculus terrestris as long as it can perform seed culture on sacculus terrestris, and those skilled in the art can select the medium appropriately according to the nature of the species.
In the embodiment of the present invention, the present invention is not particularly limited to the fermentation medium, and any medium may be used for the Trichoderma longibrachiatum as long as it can ferment and culture the Trichoderma longibrachiatum, and any medium may be used for the sacculus terrestris as long as it can ferment and culture the sacculus terrestris, and those skilled in the art can appropriately select the medium according to the nature of the species.
The fermentation medium for preparing the bacillus licheniformis fermentation liquor is prepared by adding astragalus powder and pseudo-ginseng powder into a basic fermentation medium, and the basic fermentation medium is not particularly limited in the invention, so long as the fermentation medium can perform fermentation culture on the bacillus licheniformis, and the fermentation medium can be reasonably selected by a person skilled in the art according to the strain property.
Example 1
Respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the Trichoderma longibrachiatum fermentation liquor, the Bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to ensure that the viable count of the Trichoderma longibrachiatum is 4 multiplied by 107cfu/ml, viable count of 9 × 107cfu/ml, viable count of sacculus terrestris 4X 107cfu/ml。
Wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor contains: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.3 wt%, panax notoginseng powder 0.2 wt%, and the balance water, starting at ph 7.5.
Example 2
Respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the Trichoderma longibrachiatum fermentation liquor, the Bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to ensure that the viable count of the Trichoderma longibrachiatum is 9 multiplied by 107cfu/ml, viable count of Bacillus licheniformis 4 × 107cfu/ml, viable count of sacculus terrestris 9 × 107cfu/ml。
Wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor contains: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus root powder 0.3 wt%, notoginseng powder 0.2 wt%, andand the balance water, starting at pH 7.5.
Example 3
Respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the Trichoderma longibrachiatum fermentation liquor, the Bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to ensure that the viable count of the Trichoderma longibrachiatum is 4 multiplied by 107cfu/ml, viable count of 9 × 107cfu/ml, viable count of sacculus terrestris 4X 107cfu/ml。
Wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor contains: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.1 wt%, panax notoginseng powder 0.5 wt%, and the balance water, starting at ph 7.5.
Example 4
Respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the Trichoderma longibrachiatum fermentation liquor, the Bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to ensure that the viable count of the Trichoderma longibrachiatum is 9 multiplied by 107cfu/ml, viable count of Bacillus licheniformis 4 × 107cfu/ml, viable count of sacculus terrestris 9 × 107cfu/ml。
Wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor contains: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.5 wt%, panax notoginseng powder 0.1 wt%, and the balance water, starting at ph 7.5.
Example 5
Long branches of treeRespectively inoculating trichoderma, bacillus licheniformis and sacculus exigua into the culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus exigua fermentation liquor; then mixing the Trichoderma longibrachiatum fermentation liquor, the Bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to ensure that the viable count of the Trichoderma longibrachiatum is 1 multiplied by 106cfu/ml, viable count of Bacillus licheniformis 1 × 108cfu/ml, viable count of sacculus terrestris of 1 × 106cfu/ml。
Wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor contains: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.3 wt%, panax notoginseng powder 0.2 wt%, and the balance water, starting at ph 7.5.
Example 6
Respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the Trichoderma longibrachiatum fermentation liquor, the Bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to ensure that the viable count of the Trichoderma longibrachiatum is 4 multiplied by 107cfu/ml, viable count of 9 × 107cfu/ml, viable count of sacculus terrestris 4X 107cfu/ml。
Wherein the fermentation medium for preparing the bacillus licheniformis fermentation liquor contains: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO4 2.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, and the balance water, starting at pH 7.5.
Example 7: stress resistance test
Corn is planted in a greenhouse (the temperature is 27 +/-2 ℃), the experimental group adopts the biocontrol bacterium preparation prepared in the embodiment 1 to spread the soil surface according to the dosage of 2 kg/mu and then ploughs the base fertilizer, takes the mixed fermentation liquor of the embodiment 6 in addition, centrifuges the mixed fermentation liquor at 4000r/min for 30min, filters the supernatant fluid by a 0.22 mu L membrane, filters the thallus to obtain the sterile fermentation supernatant fluid which is used as a control group, spreads the soil surface according to the dosage of 2 kg/mu and then ploughs the base fertilizer, and adopts clear water for the blank group. After growing for 21 days under natural illumination, drought stress treatment is carried out, and watering is stopped for 9 days. And (3) freezing the corn leaves on the 9 th day after drought stress by using liquid nitrogen, and storing the frozen corn leaves in a refrigerator at the temperature of-80 ℃ for subsequent determination of related physiological indexes. The amount of matrix and moisture used for each individual experimental material remained consistent.
(1) Phenotype
And (4) selecting the corn plants in the seedling stage of 21 days, stopping watering, and carrying out drought stress for 9 days. As can be seen from the actual growth results of the maize plants, after drought stress, the plants in the blank group and the plants in the control group show obvious water deficiency phenotypes such as wilting, inhibited growth and the like.
(2) Proline and soluble sugar content
After the plants are stressed by drought and the like, proline and soluble sugar are synthesized in vivo to maintain the osmotic form of the cells of the plants, so that the aims of protecting the cells and maintaining osmotic balance are fulfilled, and therefore, the content of the proline and the soluble sugar indirectly reflects the drought stress resistance of the plants.
And (3) proline content determination:
the proline content was analyzed by ninhydrin colorimetry. 0.5g of each of the plant leaves treated differently is cut into pieces and placed in a large test tube, 5mL of 3% sulfosalicylic acid solution is added, and the mixture is leached in a boiling water bath for 10 min. Taking out the test tube, cooling to room temperature, sucking 2mL of supernate, adding 2mL of glacial acetic acid and 3mL of 2.5% acid ninhydrin color development solution, heating in a boiling water bath for 40min, finding out the concentration of proline in the sample from the standard curve, and calculating the proline content according to the following formula: proline [ mu.g.g ]-1(fresh weight or Dry weight)]＝[(C·V0/V)/W]N, wherein C is the proline content in the sample; v0Is the total volume of the sample extracting solution; v is the volume aspirated during the assay; w is the sample weight; n is the dilution factor.
And (3) soluble sugar content determination:
the content of soluble sugar is measured by anthrone colorimetry, and the specific method comprises the following steps: weighing 0.1-0.2 g of sample, adding 1mL of distilled water, grinding into homogenate, pouring into a centrifuge tube with a cover, carrying out water bath at 95 ℃ for 10min (tightly covering to prevent water loss), cooling, centrifuging at 8000g and 25 ℃ for 10min, taking supernatant into a 10mL test tube, adding distilled water to constant volume of 10mL, and shaking uniformly for later use. 0.5mL of the sample extract was aspirated into a 20mL graduated tube, and 1.5mL of distilled water was added. Adding 0.5mL of anthrone ethyl acetate reagent and 5mL of concentrated sulfuric acid into a test tube in sequence, fully oscillating, immediately putting the test tube into a boiling water bath, accurately keeping the temperature for 1min tube by tube, taking out, naturally cooling to room temperature, taking a blank as a reference, and measuring the absorbance at the wavelength of 620 nm.
The regression equation determined under standard conditions is: y is 4.275 x-0.07; wherein x is standard substance concentration (mg/mL), and y is light absorption value.
Calculating according to the fresh weight of the sample: soluble sugar (mg/g fresh weight) [ (Δ a +0.07)/4.275 × V1]/(W×V1/V2) 2.34 × (Δ a + 0.07)/W; wherein, V1: adding sample volume, mL; v2: adding the volume of the extracting solution, mL; w: fresh weight of sample, g.
The results of the measurement of the proline and soluble sugar contents of the blank plant, the control plant and the experimental plant are shown in table 1, and it can be found from the results that the proline content and the soluble sugar content of the experimental plant are higher than those of the blank plant and the control plant, and the results also show that the experimental plant has higher capacity of accumulating osmoregulation substances such as proline and soluble sugar than those of the blank plant and the control plant and has stronger drought stress resistance.
(3) Malondialdehyde content
Plant tissues are damaged under the adverse circumstances and often generate membrane lipid peroxidation, and malondialdehyde is a final decomposition product of the membrane lipid peroxidation, and the content of malondialdehyde can reflect the degree of the plant damaged by the adverse circumstances.
Determination of malonaldehyde content:
taking 0.2g of each of the plant leaves treated differently, shearing, putting into a mortar, adding 2mL of 10% TCA and a small amount of quartz sand, grinding into homogenate, and adding 8mFurther grinding the L TCA, transferring the homogenate into a centrifuge tube, and centrifuging at 4000rpm for 10min to obtain a supernatant which is an extracting solution. Absorbing 2mL of supernatant, adding 2mL of 0.6% TBA solution, mixing uniformly, boiling for 15min in a boiling water bath, cooling, and centrifuging for 1 time. And taking the supernatant, and respectively measuring the absorbance values at the wavelengths of 450nm, 532nm and 600 nm. The concentration of MDA in the extract was determined according to C ═ 6.45(D532-D600) -0.56D450, and the content of MDA (μmol g) in the sample was calculated from the fresh weight of the plant tissue-1) (ii) MDA concentration (. mu. mol. L)-1) X volume of extract (L)/fresh weight of plant tissue (g).
The results of the malondialdehyde content measurements of the blank, control and experimental plants are shown in table 1, and it can be seen from the results that the malondialdehyde content of the experimental plants is lower than that of the blank and control plants, which also indicates that the experimental plants are more tolerant than the blank and control plants under drought stress.
TABLE 1
The same stress resistance test was carried out using the biocontrol agent of examples 2-5, and the results were similar to those of example 1.
The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.
Claims (8)
1. The biocontrol bacterium preparation for enhancing stress resistance of crops is characterized by comprising trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris fermentation liquor, wherein the total viable count of the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris is more than or equal to 5 multiplied by 107cfu/ml; the trichoderma longibrachiatum,The ratio of viable count of Bacillus licheniformis and Sphaerotheca fuliginea is (0.1-10): 0.1-10), wherein the fermentation medium for preparing the Bacillus licheniformis fermentation liquid is added with 0.1-0.5 wt% of radix astragali powder and 0.1-0.5 wt% of Panax notoginseng powder.
2. The biocontrol microbial agent of claim 1, wherein 0.3 wt% of astragalus powder and 0.2 wt% of panax notoginseng powder are added to the fermentation medium for preparing the bacillus licheniformis fermentation broth.
3. The biocontrol bacterial preparation of claim 1, wherein the fermentation medium for preparing the Bacillus licheniformis fermentation broth comprises: 2-4 wt% of corn flour, 2-4 wt% of bean cake powder, 2-4 wt% of bran powder and K2HPO4 2-2.5wt％、KH2P040.5-1wt％、MgSO40.05-0.1 wt%, peanut oil 0.1-0.5 wt%, astragalus root powder 0.1-0.5 wt%, notoginseng powder 0.1-0.5 wt%, and water in balance, with an initial pH of 7.5.
4. The biocontrol bacterial preparation of claim 3, wherein the fermentation medium for preparing the Bacillus licheniformis fermentation broth comprises: 3 wt% of corn flour, 3 wt% of bean cake powder, 3 wt% of bran powder and K2HPO42.15wt％、KH2P04 0.8wt％、MgSO40.075 wt%, peanut oil 0.3 wt%, astragalus powder 0.3 wt%, panax notoginseng powder 0.2 wt%, and the balance water, starting at ph 7.5.
5. The biocontrol bacterial preparation of claim 1, wherein the viable count ratio of the trichoderma longibrachiatum, the bacillus licheniformis and the glomus marmoreus is (4-9): 4-9.
6. A method for preparing the biocontrol bacterial preparation of any one of claims 1-5, comprising the steps of: respectively inoculating the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris on the ground surface into a culture solution for liquid seed culture and liquid fermentation culture to obtain trichoderma longibrachiatum fermentation liquor, bacillus licheniformis fermentation liquor and sacculus terrestris on the ground surface; then mixing the trichoderma longibrachiatum fermentation liquor, the bacillus licheniformis fermentation liquor and the sacculus terrestris fermentation liquor uniformly to obtain the trichoderma longibrachiatum fermentation liquor;
wherein the total viable count of Trichoderma longibrachiatum, Bacillus licheniformis and saccule mycoderma is not less than 5 × 107cfu/ml; the viable count ratio of the trichoderma longibrachiatum, the bacillus licheniformis and the sacculus terrestris is (0.1-10): 1-10): 0.1-10), and 0.1-0.5 wt% of astragalus powder and 0.1-0.5 wt% of panax notoginseng powder are added into a fermentation culture medium for preparing the bacillus licheniformis fermentation liquor.
7. Use of the biocontrol bacterial preparation of any one of claims 1-5 for enhancing stress resistance of crops.
8. The use according to claim 7, wherein the biocontrol bacterial formulation is used in a base fertilizer at the time of crop planting.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110530490.6A CN113308382A (en) | 2021-05-14 | 2021-05-14 | Biocontrol bacterium preparation for enhancing stress resistance of crops |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110530490.6A CN113308382A (en) | 2021-05-14 | 2021-05-14 | Biocontrol bacterium preparation for enhancing stress resistance of crops |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113308382A true CN113308382A (en) | 2021-08-27 |
Family
ID=77373236
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110530490.6A Pending CN113308382A (en) | 2021-05-14 | 2021-05-14 | Biocontrol bacterium preparation for enhancing stress resistance of crops |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113308382A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114287448A (en) * | 2022-01-06 | 2022-04-08 | 烟台大学 | Compound medicament for improving heat resistance of Lanzhou lily and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102432399A (en) * | 2011-09-16 | 2012-05-02 | 北京平安福生物技术研究所有限公司 | Biological organic fertilizer for improving stress resistance and output of paddy rice and preparation method thereof |
| CN102533579A (en) * | 2010-12-30 | 2012-07-04 | 中国科学院沈阳应用生态研究所 | Culture medium for producing spores, and optimizing process and application thereof |
| CN108641966A (en) * | 2018-05-16 | 2018-10-12 | 海南大学 | A kind of microorganism Biological agents |
| CN109234173A (en) * | 2018-11-02 | 2019-01-18 | 河北省科学院生物研究所 | A kind of long shoot trichoderma, its solid microbial and application |
| CN110627554A (en) * | 2018-06-22 | 2019-12-31 | 潍坊市云涛有机肥料有限公司 | Meat and bone meal organic fertilizer and preparation process thereof |
| CN111733080A (en) * | 2019-03-22 | 2020-10-02 | 安琪酵母股份有限公司 | Trichoderma solid preparation, preparation method and application thereof |
-
2021
- 2021-05-14 CN CN202110530490.6A patent/CN113308382A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533579A (en) * | 2010-12-30 | 2012-07-04 | 中国科学院沈阳应用生态研究所 | Culture medium for producing spores, and optimizing process and application thereof |
| CN102432399A (en) * | 2011-09-16 | 2012-05-02 | 北京平安福生物技术研究所有限公司 | Biological organic fertilizer for improving stress resistance and output of paddy rice and preparation method thereof |
| CN108641966A (en) * | 2018-05-16 | 2018-10-12 | 海南大学 | A kind of microorganism Biological agents |
| CN110627554A (en) * | 2018-06-22 | 2019-12-31 | 潍坊市云涛有机肥料有限公司 | Meat and bone meal organic fertilizer and preparation process thereof |
| CN109234173A (en) * | 2018-11-02 | 2019-01-18 | 河北省科学院生物研究所 | A kind of long shoot trichoderma, its solid microbial and application |
| CN111733080A (en) * | 2019-03-22 | 2020-10-02 | 安琪酵母股份有限公司 | Trichoderma solid preparation, preparation method and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| BAKHSHANDEH 等: "Plant growth promoting microorganisms can improve germination, seedling growth and potassium uptake of soybean under drought and salt stress", 《PLANT GROWTH REGULATION》 * |
| BEGUM 等: "Improved Drought Tolerance by AMF Inoculation in Maize (Zea mays) Involves Physiological and Biochemical Implications", 《PLANTS（BASEL）》 * |
| VARDHARAJULA 等: "Drought-tolerant plant growth promoting Bacillus spp.: effect on growth, osmolytes, and antioxidant status of maize under drought stress", 《JOURNAL OF PLANT INTERACTIONS》 * |
| 刘润进 等: "菌根真菌与植物抗逆性研究进展", 《菌物研究》 * |
| 张骕 等: "豇豆叶片生防细菌的鉴定及其拮抗作用测定", 《分子植物育种》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114287448A (en) * | 2022-01-06 | 2022-04-08 | 烟台大学 | Compound medicament for improving heat resistance of Lanzhou lily and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105145603B (en) | Compound wheat seed soaking agent and preparation method thereof, application method and application | |
| JP7393824B2 (en) | Organic cultivation method of red cedar | |
| CN112806370A (en) | Agricultural composition containing 5-aminolevulinic acid and dihydrojasmonic acid propyl ester | |
| CN107018903A (en) | A kind of implantation methods of dendrobium candidum | |
| CN107041271A (en) | A kind of implantation methods of walnut | |
| CN109275485A (en) | A kind of high viability oil tea implantation methods | |
| CN113308382A (en) | Biocontrol bacterium preparation for enhancing stress resistance of crops | |
| KR100555077B1 (en) | Ferment solution of plants and method for fertilizer application or killing insects using the solution | |
| CN106172523B (en) | A kind of Green Water spike of rice sprouts inhibitor and its preparation method and application | |
| CN105255744B (en) | A kind of lucidum strain purification and rejuvenation cultural method and its culture medium | |
| CN107114175A (en) | A kind of method for culturing seedlings for improving Exocarpium Citri Rubrum survival rate | |
| Salamon et al. | Agro-ecological cultivation, secondary metabolite characteristics and microbiological tests of lemon balm (Melissa officinalis)–the variety Citronella | |
| CN114190131B (en) | Selenium-rich silicon foliar spraying agent special for tomatoes and preparation method and application thereof | |
| CN109418266A (en) | One plant growth regulators, its application method and purposes | |
| CN108250001A (en) | A kind of disease-resistant growth-promoting formula fertilizer of photinia glabra and preparation method thereof | |
| CN110117205B (en) | Germanium-rich nutrient solution for increasing germanium content of rapeseeds and preparation method and application thereof | |
| CN102960350B (en) | Application of pyrroloquinoline quinone (PQQ) active materials in aspect of preventing and treating bacterial wilt of plants | |
| CN107912264A (en) | A kind of Orchis planting special soil and preparation method thereof | |
| CN113749130A (en) | Sugarcane biological seed stem coating agent and preparation method thereof | |
| CN107624464A (en) | The method for the crops enriched using far infrared cultivation nutrient composition | |
| Kabir et al. | Vegetative Propagation of Punica granatum by Stem Cuttings Using Non-Mist Propagator | |
| Latunra et al. | Effect of green bean sprout extract on in vitro shoot multiplication of taro Colocasia esculenta L. var. antiquorum | |
| EP4296251A1 (en) | Fertilizer preparation process, fertilizer therefor, and fertilizer composition | |
| CN107114386B (en) | The application of cupreol and preparation | |
| CN107434508A (en) | Application of the Luohanguo's residue shoot vegetable root in elm mushroom compost is made |
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 |