Comprehensive prevention and treatment method for iron deficiency of under-forest polygonatum sibiricum

Abstract

The invention relates to the technical field of ecological cultivation of traditional Chinese medicinal materials, in particular to a comprehensive prevention and treatment method for iron deficiency of under-forest polygonatum sibiricum. The bacillus subtilis, the bacillus pumilus, the bacillus tequilensis, the bacillus mycoides and the bacillus amyloliquefaciens are combined with a compound microbial agent and a pseudomonas fluorescens agent to release iron carriers; the symptoms that young leaves of the under-forest polygonatum sibiricum are yellowish white, oil stain-shaped yellowish-brown spots appear on the leaves, tender tips are withered, internodes are lengthened, lodging is prone to occurring and the like are fundamentally solved; polygonatum sibiricum is used as a material, a three-step comprehensive strategy is provided for physiological diseases caused by iron element deficiency during ecological cultivation in a forest, and the symptom of element deficiency is eliminated, so that synthesis of chloroplast cystosomal membranes and chlorophyll of plants of Polygonatum sibiricum under the forest is recovered, and photosynthesis is recovered to a normal state; the chlorophyll fluorescence kinetic parameters show a healthy state.

Description

Technical Field

[0001] The invention relates to the technical field of ecological cultivation of Chinese medicinal materials, and in particular to a comprehensive prevention and control method for the rhizosphere soil microenvironment of iron deficiency disease of Polygonatum truncatum ecologically planted under forests. Background Art

[0002] Iron is an essential trace element in the growth and development of plants. It participates in many metabolic reactions including photosynthesis. As the main influencing factor determining the activity of various enzymes in the cytochrome structure-porphyrin ring synthesis pathway, it plays an important role in respiration and photosynthesis, the two main sources of cellular energy, and affects many physiological processes such as nitrogen metabolism, organic acid metabolism, carbohydrate metabolism and protoplasm characteristics.

[0003] Iron deficiency can lead to the obstruction of photosynthetic electron transfer, photosynthesis and chloroplast synthesis, causing young leaves to show typical iron deficiency chlorosis. Iron plays an important role in the synthesis and functional improvement of chlorophyll precursors. Most of the iron in plant cells is evenly distributed on the thylakoid membrane and has a good correlation with the chlorophyll content. Therefore, when plants are iron deficient, the thylakoids are prone to disintegration and the chlorophyll content is reduced.

[0004] The effective iron content in soil is mainly affected by various environmental factors such as soil parent material, redox potential, pH, organic matter, and microorganisms. The pH value has the greatest impact on the iron content. Due to the high oxygen content on the earth's surface, iron is gradually converted into hydroxyl oxide polymers with extremely low solubility. The concentration of soluble iron in soil with low pH is relatively large, and the minimum solubility of iron oxide in the pH range of 7.5-8.5 is only 10 -10 mol / L, which is far from meeting the needs of normal growth and development of plants and microorganisms.

[0005] In the distribution area of ​​Polygonatum cyrtonema, there are a large number of plots with neutral or slightly alkaline soil. Since Polygonatum cyrtonema is a medicinal and edible plant with multiple physiological and health benefits and rich nutrients, its shade-tolerant and perennial characteristics give it the natural endowment of forest granaries and grain storage in the ground. Therefore, the understory ecological cultivation of Polygonatum cyrtonema has been widely carried out in Northeast China, North China, and Shandong, Henan, Shaanxi, Hubei and other provinces. In some plain ecological forests, iron deficiency often occurs during the vigorous growth season, and the damage to young leaves and apical meristems is particularly serious. Simple fertilization can only partially alleviate the symptoms, and cannot fundamentally solve the problem of negative impact on the growth and development of Polygonatum cyrtonema cultivated under the forest ecology. Therefore, it is urgent to provide a comprehensive prevention and control method for iron deficiency of Polygonatum cyrtonema under the forest. Summary of the invention

[0006] The present invention provides a comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forests, comprising the following steps S1, regulating the pH value of rhizosphere soil: biochar based on forest cultivation residues is acidified with wood vinegar and then applied to the rhizosphere soil.

[0007] S11. The tree waste obtained from forest cultivation is crushed into particles with a particle size of 2 mm, and the particles are placed in a pyrolysis carbonization furnace (4) and anaerobically pyrolyzed at 520° C. for 2 h under nitrogen conditions to produce garden biochar C0.

[0008] S12, soak the biochar C0 in 0.5 mol·L -1 FeCl3 and 0.5 mol·L -1 In the FeSO4 solution, the Fe:C mass ratio is 0.56, and the mass volume ratio of biochar C0:solution is 1:8. After stirring for 30 minutes until uniform, the solution is placed in a pyrolysis carbonization furnace (4) and calcined at 430°C for 2 hours to obtain modified biochar C1.

[0009] S13, dilute the wood vinegar stock solution 10 times, mix the modified biochar C1 and the diluted wood vinegar stock solution in a ratio of 1g:10ml, stir thoroughly, let it stand for 2 days for acidification, and obtain the acidified modified biochar. In early spring, dig a 10cmmm shallow trench and apply it to the rhizosphere of Polygonatum cyrtonema under the forest. Irrigate with 5L of the mixed solution per square meter, and use 1500L per mu of Polygonatum cyrtonema under the forest.

[0010] S2. Effective supply of iron: Apply chelated iron organic fertilizer to the rhizosphere soil of the Polygonatum cyrtonema under the forest that has been improved in step S1, and spray the leaves with thin chelated iron organic fertilizer several times to quickly relieve the iron deficiency symptoms of the Polygonatum cyrtonema plants.

[0011] S21, crush the horseshoe slices, pass them through a 40-mesh sieve, take 12 kg and put them into a corrosion-resistant container, add 4.6 kg of water (pH 7.2), and mix in 48 g of Actinomadura keratinilytica (EU637009, strain WCC-2265T), wherein the live bacteria content is 2×10 10 CFU / g, stir evenly, spray water once every 12 hours at 42-45°C, keep it moist, and treat the stubborn keratin by biodegradation for 5 days; then add 30L of 1.0mol / L sulfuric acid, stir evenly with a glass rod, and react for 12h; evaporate and deacidify in a water bath (≤90°C) and concentrate to 10L, slowly inject it into another anti-corrosion container containing 20L water, evaporate and deacidify again, and do this operation 3 times to obtain the hydrolyzed amino acid solution of horseshoe.

[0012] S22, crush 260kg sesame cake into pieces with a diameter of less than 1cm, put 60kg millet bran into a fermentation tank with a length of 300cm, a width of 200cm and a depth of 90cm, add 240L of water and stir evenly; slowly inject the water solution of hydrolyzed amino acids from horseshoe palm, stir evenly, add the composite bacterial agent, mix and stir evenly, so that the bacterial count of the total material after mixing is not less than 700,000 / gram; regularly record the central temperature of the fertilizer pile and the ambient temperature, turn the pile every 2 days, and add water to keep the moisture content of the whole composting process at 62%. The fertilizer composting time is 30 days. After the composting is completed, accurately weigh and uniformly sample more than 5kg to determine the moisture content, protein N, total N, mineral nitrogen (NH4 + +NO3 - ), total amino acids, and organic C content, and calculated the maturity and protein N degradation rate of cake fertilizer under different bacterial agent treatments, where the composite bacterial agent included one or more of yeast, Bacillus subtilis, black mold, and Pseudomonas.

[0013] S23, add 22kg of ferrous sulfate, add a large amount of water to completely dissolve, finally the liquid level reaches 50cm, stir evenly, and chelate for one week to obtain chelated iron organic fertilizer.

[0014] S24. From late April to early May, take 100L of fermented fertilizer and dilute it 10 times, and spray it on the leaves 4 times, with an interval of 7-15 days between two times; take 900L of fermented fertilizer and dilute it 5 times, and use it with drip irrigation to the roots of Polygonatum cyrtonema under the forest. Each batch of fermented fertilizer can meet the usage of 10,000 square meters, or 1 hectare.

[0015] S3, high-yield siderophore bacterial agent secretes siderophores;

[0016] S31. Preparation of Bacillus composite bacterial agent

[0017] S311. Preparation of cow dung biochar: After collecting fresh cow dung, clean out the stones and other debris therein, spread the cow dung on a clean large core board, the lower layer is a flat cement floor, and air-dry it under full light and natural ventilation conditions to a moisture content of less than 18%, spread the dried cow dung at 105°C and dry it for 3 hours for preheating, put the preheated cow dung into a pyrolysis carbonization furnace 4, control the heating rate to 100°C·h-′, and when the pyrolysis temperature reaches 380°C, nitrogen is blown into the pyrolysis carbonization furnace 4 through the air inlet 3 by a nitrogen source 1, and the pyrolysis time is 3 hours under anoxic conditions; after closing the pyrolysis carbonization furnace 4, cool it to below 80°C, cool the pyrolyzed cow dung for standby use, put the cow dung biochar prepared by pyrolysis into a pulverizer for crushing, and pass it through a 40-mesh sieve to obtain cow dung biochar.

[0018] S312. Preparation of fermentation broth: Pick single colonies of Bacillus tekila, Bacillus mycoides, Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus brevis and inoculate them in a conical flask filled with LB medium, and culture them at 30°C and 180 r / min for 12 h as seed liquid. Inoculate them in a fermenter filled with LB medium with an inoculum size of 2%, and continue to culture at 30°C and 180 r / min for 24 h to obtain fermentation broth.

[0019] S313, preparation of bacterial powder: the fermentation liquid cultured in the fermentation tank was taken out, centrifuged at 5000rpm for 10min, the supernatant was removed, the protective agent maltodextrin was added at a ratio of 20:3, oscillated for 30s, and placed on a shaker at a speed of 150rpm for incubation at 30°C for 30min, spray dried to obtain bacterial powder; the bacterial powder and cow dung biochar were uniformly mixed at a volume ratio of 23:77 to obtain a Bacillus composite bacterial agent; the bacterial agent prepared by the formula had a spore content of 3.9×10 8 CFU·g -1 , moisture content (w) ≤ 0.9%, fineness ≥ 99%, wetting time is 33.0s, suspension rate reaches 81.0%, and all indicators meet national standards.

[0020] S32, Preparation of Pseudomonas fluorescens Agent

[0021] S321. Activate Pseudomonas fluorescens on a KB plate at 28°C for 24 hours. After activation, pick a single colony with an inoculation loop, inoculate it into 5 mL of liquid seed KB culture medium, and culture it in a shaker at 28°C and 150 r / min for 24 hours. Then inoculate it into solid seed KB culture medium and culture it at 28°C for 24 hours. Pick a ring of the cultured first-level seeds with an inoculation loop, inoculate it into 50 mL of liquid seed KB culture medium, and culture it in a shaker at 28°C and 150 r / min for 24 hours to obtain second-level seeds.

[0022] S322, the fermentation medium was first treated with 1g 8-hydroxyquinoline to remove trace iron, the fermentation medium composition was: 1.5g glycerol, 1.0g hydrolyzed casein, 2.5g wok sulfate, 2.5g dipotassium hydrogen phosphate and 7.0g glucose, pH was 6.5, the culture conditions were bottling volume 200 / 500 (mL), i.e. 500ml culture bottle was filled with 200ml culture medium, the secondary seed inoculation amount was 2%, the optimal culture time was 48h, Fe 3+ The concentration is 0.8 mg / L, and the final viable count is 5.0×10 9 CFU·g -1 .

[0023] S323. Under sterile conditions, collect the fluorescent Pseudomonas cells in the logarithmic growth phase by centrifugation and suspend them in fresh liquid seed KB medium to make the number of viable cells reach 5.0×109 CFU·g -1 The above, then add sodium carboxymethyl cellulose according to the volume ratio of bacterial liquid: stabilizer = 10:1, mix thoroughly and mix with a wetting agent, a dispersant, a protective agent and a carrier, wherein the wetting agent is sodium dodecyl sulfate (9%), the dispersant is sodium tripolyphosphate (9%), the protective agent is humic acid (1.2%), and the carrier is diatomaceous earth with a particle size of 4 mm (52.8%). After standing at room temperature for 48 hours, package and store at room temperature. When used, the plate count method is used to detect the number of live bacteria in the fluorescent Pseudomonas agent.

[0024] S33. Detection of the ability of strains to produce iron carriers. The strains (Bacillus composite bacteria and Pseudomonas fluorescens) were activated and cultured on NB plates for 24 hours. Single colonies were picked up with toothpicks and spotted on CAS plates. After culturing at 28°C for 2 days, single colonies were picked up and inoculated into MSA culture medium. The culture was shaken at 28°C and 150 r / min for 2 days. The bacterial suspension was centrifuged at 10000 r / min for 10 min. An equal volume of bacterial culture supernatant was mixed with CAS detection solution. The mixture was protected from light for 1 hour. The absorbance As was detected at a wavelength of 680 nmm. At the same time, the absorbance Ar of the reaction between the blank culture medium and the equal volume of CAS detection solution was determined. The iron carrier production ability of the strain was analyzed by As / Ar. As / Ar was between 0-1. The As / Ar of bacteria with higher iron carrier production ability was lower than 0.5. The iron carrier activity unit (SU) was used to represent the iron carrier production of the strain. The calculation formula was: SU = [(Ar-As) / Ar] × 100.

[0025] S4. In the high temperature season from early July to mid-August, the yellowing symptoms disappeared and the divalent iron content in the surface soil was less than 0.01 μmol·L -1 When the iron carrier is high, apply the high-yield iron carrier agent, apply the fluorescent Pseudomonas agent near the drip irrigation pipe, and apply 10g / m 2 , apply Bacillus compound bacterial agent on the far side of the drip irrigation pipe, 5-10cm deep soil in the rhizosphere, 30g / m 2 .

[0026] In a possible implementation, the composite bacterial agent added in step S22 is Bacillus subtilis and Aspergillus niger, and the bacterial count ratio of Bacillus subtilis to Aspergillus niger is 60:2.

[0027] In a possible implementation, the degree of maturity in step S22 is measured by the humic acid E4 / E6 value, and the specific determination method is: distilled water is used to extract the sample (the mass ratio of distilled water to sample is 20:1), and the filtrate is filtered and the absorbance is measured by a spectrophotometer at wavelengths of 465nm and 665nm, and the ratio is calculated as E4 / E6. Maturity requires C / N to be less than 6.50, protein N degradation rate to be higher than 22%, humic acid E4 / E6 value to be less than 3.50, and total amino acid content to be greater than 27.3mg / g.

[0028] In a possible implementation, the LB culture medium in step S312 is prepared by sterilizing 10 g of tryptone, 5 g of yeast powder, 10 g of sodium chloride and 1 L of distilled water at 121° C. for 20 min.

[0029] In a possible implementation, the liquid seed KB medium (1 L) in step S321 comprises: 15 mL of glycerol, 20 g of peptone, 0.3 g of K2HPO4, 1.5 g of MgSO4·7H20, and a pH of 7.0; solid seed KB medium: 20 g of agar is added to Kings′B medium.

[0030] In a possible implementation, the components of the NB medium (1 L) in step S33 are: 3.0 g beef extract powder, 10.0 g peptone, 5.0 g NaCl, and 18.0 g agar; the components of the MSA medium (1 L) are: 20.0 g sucrose, 2.0 g L-asparagine, 1.0 g K2HPO4, and 0.5 g MgSO4·7 H2O.

[0031] In a possible implementation, the composition of the CAS culture medium in step S33 is: 1 mL of 20% sucrose solution, 3 mL of 10% acid hydrolyzed casein, 100 μL of 1 mmol / L CaCl2, 2 mL of 1 mmol / L MgSO4, and 1.8 g of agar per 100 mL, and 5 mL of phosphate buffer and CAS dye solution are slowly added at about 60° C. to obtain CAS blue culture medium;

[0032] The composition of CAS detection solution is as follows: Solution A: 500mL of 1mmol / L CAS (chrome azure), 100mL of 0.1mmol / L FeCl3·6H2O (prepared with 12mmol / L hydrochloric acid); Solution B: 400mL of 2mmol / L HDTMA (hexadecyltrimethylammonium bromide); slowly pour Solution A into Solution B to prepare 1L of blue CAS dye solution.

[0033] Beneficial effects: The comprehensive prevention and control method for iron deficiency in Polygonatum cyrtonema under the forest provided by the present invention regulates the pH of the soil and supplies effective iron elements, combines a composite microbial agent of Bacillus subtilis, Bacillus pumilus, Bacillus tequila, Bacillus mycoides and Bacillus amyloliquefaciens, and a Pseudomonas fluorescens agent to release iron carriers, fundamentally solving the symptoms of young leaves of Polygonatum cyrtonema under the forest losing their green color and becoming yellow-white, oily yellow-brown spots appearing on the leaves, wilting of young and tender tips, elongated internodes, and easy lodging; for the physiological diseases caused by iron deficiency that occur in Polygonatum cyrtonema when it is ecologically cultivated in the forest, a three-step comprehensive measure of pH value regulation-effective iron supply-iron carrier agent application is proposed to eliminate the deficiency symptoms, so that the chloroplast thylakoid membrane and chlorophyll synthesis of Polygonatum cyrtonema plants under the forest can be restored, photosynthesis returns to a normal state, and the chlorophyll fluorescence kinetic parameters show a healthy state. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 Schematic diagram of the structure of biochar prepared by pyrolysis of cow dung.

[0035] Figure 2 This is an indication map of the locations for applying rhizosphere fungal agents to Polygonatum cyrtonema under the forest.

[0036] Figure 3 These are the chlorosis symptoms caused by iron deficiency in Polygonatum cyrtonema under the forest.

[0037] Figure 4 This is the leaf yellowing symptom of Polygonatum cyrtonema in Case 1.

[0038] Figure 5 In Case 1, the iron deficiency of Polygonatum cyrtonema showed severe chlorosis of young leaves, loss of green between the veins of semi-mature leaves, and mild chlorosis of old leaves.

[0039] Figure 6 This is a detailed observation of the interveinal chlorosis symptoms caused by iron deficiency in Polygonatum cyrtonema in Case 1.

[0040] Figure 7 This is a diagram showing the growth status of Polygonatum cyrtonema in Case 1 after being controlled by the system of the present invention.

[0041] Figure 8 This is the mild iron deficiency symptom of Polygonatum sibiricum in Case 2.

[0042] Fig. 9 This is a graph showing the growth status of Polygonatum cyrtonema after treatment in treatment group 2 in Case 2.

[0043] Fig.10 This is a graph showing the growth status of Polygonatum cyrtonema in Case 2 after treatment in Group 3.

[0044] Fig.11 These are the symptoms of iron deficiency in Polygonatum cyrtonema in Case 3.

[0045] Fig.12 These are the symptoms of iron deficiency in case 3 when Polygonatum cyrtonema and Polygonatum cyrtonema were mixed together.

[0046] Fig.13 This is a graph showing the growth status of Polygonatum cyrtonema and Polygonatum cyrtonema after treatment in Group 2 in Case 3.

[0047] In the figure: 1. nitrogen source; 2. gas flow meter; 3. air inlet; 4. pyrolysis carbonization furnace; 5. air outlet; 6. alkali solution; 7. operating table. DETAILED DESCRIPTION

[0048] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described below, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0049] A comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forests comprises the following steps:

[0050] S0, iron deficiency evaluation;

[0051] S01. Determination of chlorophyll content

[0052] Accurately weigh 100 mg of Polygonatum cyrtonema leaves, transfer to a mortar, add a pre-prepared mixed extract of anhydrous ethanol, acetone and distilled water in a ratio of 4.5:4.5:1 and a small amount of quartz sand, grind to a homogenous slurry; after dark treatment for 2 hours, use the extract as a control, use an ultraviolet spectrophotometer to measure the absorbance of the sample at 645nm and 663nm, respectively, and use Arnon's formula to calculate the chlorophyll content in the leaves.

[0053] S02, Chlorophyll fluorescence measurement

[0054] The fluorescence parameters were measured from 8:00 to 12:00 using the MINI-PAM (ultra-portable modulated chlorophyll fluorescence instrument) from the German WALZ company. After 20 minutes of dark reaction, saturated light intensity treatment was performed to measure the initial fluorescence F0, maximum fluorescence Fm, and PS II maximum light quantum conversion efficiency Fv / Fm.

[0055] S03. Leaf color determination

[0056] The leaf color parameters were measured using a fully automatic colorimeter CR-400 (Konica Minolta, Japan), and the L*, a*, and b* values ​​were recorded. Each index was repeated 3 times. Among them, L* is the gloss brightness, and the larger the value, the higher the brightness; a* is the red and green saturation of the sample, with a positive value for red and a negative value for green. The larger the a* value, the darker the red, and vice versa. The b* is the yellow and blue saturation of the sample, with a positive value for yellow and a negative value for blue. The larger the b* value, the darker the yellow, and vice versa.

[0057] S04. Determination of effective iron content in leaves

[0058] The leaves were washed with 0.1% sodium dodecyl sulfate solution, then rinsed with tap water and then rinsed with deionized water three times. The total Fe content in the leaves was determined by inductively coupled plasma atomic emission spectrometry. 2+ The content was determined by dilute hydrochloric acid extraction method and o-phenanthroline colorimetry.

[0059] S05. Available iron content in soil

[0060] The effective iron was determined by diethylenetriaminepentaacetic acid (DTPA) extraction method, using DTPA-CaCl2-TEA buffer at pH 7.3 as the extractant to extract the effective iron in the soil, and the iron content in the extract was determined by atomic absorption spectrophotometer.

[0061] When the soil pH is higher than 7.2 and the effective iron content is lower than 1.93-4.38 mg / kg, Polygonatum cyrtonema under the forest is prone to iron deficiency. Symptom identification is shown in Table 1.

[0062] Table 1 Symptoms and characteristics of iron deficiency in Polygonatum cyrtonema under forest

[0063]

[0064] The higher the disease index, the more serious the iron deficiency in the leaves.

[0065] S1. pH regulation of rhizosphere soil;

[0066] S11, crushing the tree waste obtained from forest cultivation into particles with a particle size of 2 mm, placing them in a pyrolysis carbonization furnace 4 and anaerobic pyrolysis them at 520° C. for 2 h under nitrogen conditions to obtain garden biochar C0;

[0067] S12, soak the biochar C0 in 0.5 mol·L -1 FeCl3 and 0.5 mol·L -1 In the FeSO4 solution, the Fe:C mass ratio is 0.56, the mass volume ratio of biochar C0:solution is 1:8, after stirring for 30 min until uniform, it is placed in a furnace and calcined at 430 °C for 2 h to obtain modified biochar C1;

[0068] S13. Wood vinegar was provided by the Wood Laboratory of Beijing Forestry University, with a pH of 3.1, EC (electrical conductivity) of 3.48 mS / cm, and an organic carbon content of 32.87 g / L. The wood vinegar stock solution was diluted 10 times, and the modified biochar C1 was mixed with the diluted wood vinegar stock solution at a ratio of 1 g:10 ml. After sufficient stirring, the mixture was allowed to stand for 2 days for acidification to obtain acidified modified biochar with a pH of about 4.51, a carbon content of more than 75%, and an iron content of more than 40 mg / kg. In early spring, a 10 cm shallow ditch was dug and applied to the rhizosphere of Polygonatum cyrtonema under the forest. 5 L of the mixed solution was irrigated per square meter, and 1500 L was used per mu of Polygonatum cyrtonema under the forest.

[0069] S2. Effective supply of iron element: applying chelated iron organic fertilizer to the rhizosphere soil of Polygonatum cyrtonema under the forest improved by step S1, and spraying thin chelated iron organic fertilizer on the leaves for multiple times.

[0070] S21, crush the horseshoe slices, pass through a 40-mesh sieve, take 12 kg and put it into a corrosion-resistant container, add 4.6 kg of water, the pH of the water is 7.2, and add Actinomadura keratinilytica (EU637009, strain WCC-2265 T ) 48g of bacterial agent, of which the content of live bacteria is 2×10 10 CFU / g, stir evenly, spray water once every 12 hours at 42-45°C, keep it moist, and treat the stubborn keratin by biodegradation for 5 days; then add 30L of 1.0mol / L sulfuric acid, stir evenly with a glass rod, and react for 12h; evaporate and deacidify in a water bath (≤90°C) and concentrate to 10L, slowly inject it into another anti-corrosion container containing 20L water, evaporate and deacidify again, and do this operation 3 times to obtain the hydrolyzed amino acid solution of horseshoe.

[0071] S22. Purchase sesame cake with a crude protein content of more than 40% and an iron content of more than 900 mg / kg. Crush 260 kg of sesame cake into pieces with a diameter of less than 1 cm. Put 60 kg of millet bran into a fermentation tank with a length of 300 cm, a width of 200 cm and a depth of 90 cm. Add 240 L of water and stir evenly. Slowly inject the water chestnut hydrolyzed amino acid solution and stir evenly. Add the composite bacterial agent, mix and stir evenly so that the bacterial count of the total material after mixing is not less than 700,000 / gram, wherein the composite bacterial agent is Bacillus subtilis and Aspergillus niger, and the bacterial count ratio of Bacillus subtilis and Aspergillus niger is 60:2. Record the central temperature and ambient temperature of the fertilizer pile at 10:00 every day, turn the pile over every 2 days, and add water to keep the moisture content of the whole composting process at 62%. The fertilizer composting time is 30 days.

[0072] After the fertilizer is composted, accurately weigh it and uniformly sample more than 5 kg to determine the moisture content, protein N, total N, mineral nitrogen (NH4+ +NO3 - ), total amino acids, and organic C content, and calculated the maturity and protein N degradation rate of cake fertilizer under different bacterial agent treatments.

[0073] The degree of maturity is measured by the humic acid E4 / E6 value, and the determination method is as follows: extract the sample with distilled water, filter and measure the absorbance of the filtrate with a spectrophotometer at wavelengths of 465nm and 665nm, and calculate the ratio of the absorbance at the two wavelengths, i.e. E4 / E6; maturity requires C / N less than 6.50, protein N degradation rate higher than 22%, humic acid E4 / E6 value less than 3.50, and total amino acid content greater than 27.3mg / g, wherein the mass ratio of distilled water to sample is 20:1.

[0074] S23, add 22kg of ferrous sulfate, add a large amount of water to completely dissolve, finally the liquid level reaches 50cm, stir evenly, chelate reaction for one week, and obtain chelated iron organic fertilizer;

[0075] S24. From late April to early May, take 100L of fermented fertilizer solution and dilute it 10 times, and spray it on the leaves 4 times, with an interval of 7-15 days between two times; take 900L of fermented fertilizer solution and dilute it 5 times, and use it as drip irrigation to the roots of Polygonatum cyrtonema under the forest.

[0076] S3, high-yield iron carrier bacterial agent secretes iron carrier

[0077] S31, preparation of Bacillus composite bacterial agent;

[0078] S311. Preparation of cow dung biochar: After collecting fresh cow dung, clean out the stones and other debris, spread the cow dung on a clean large core board, the lower layer is a flat cement floor, and air-dry it under full light and natural ventilation conditions to a moisture content of less than 18%. Spread the dried cow dung at 105°C and dry it for 3 hours for preheating. Load the preheated cow dung into a pyrolysis carbonization furnace 4, control the heating rate to 100°C·h-1, and when the pyrolysis temperature reaches 380°C, a nitrogen source 1 is passed through the air inlet. 3. Blow nitrogen into the pyrolysis carbonization furnace 4. The gas flowmeter 2 can observe the amount of nitrogen introduced. The pyrolysis time is 3 hours under oxygen-deficient conditions. After closing the pyrolysis carbonization furnace 4, cool it to below 80° C. The cooked cow dung is cooled for standby use. The cow dung biochar prepared by pyrolysis is put into a pulverizer and crushed. The 40-mesh sieve is passed to obtain cow dung biochar. The nitrogen is finally discharged from the gas outlet 5 and absorbed by the alkali solution 6. The nitrogen source 1, the gas flowmeter 2, the air inlet 3, the pyrolysis carbonization furnace 4, the gas outlet 5, and the alkali solution 6 are all located on the operating table 7.

[0079] The biochar sample was placed at room temperature for degassing, cooled down after degassing, and transferred to a physical adsorption instrument for analysis and treatment to obtain the specific surface area of ​​the biochar, which should be ≥130m 2 / g.

[0080] S312. Preparation of fermentation broth: Pick single colonies of Bacillus tekila, Bacillus mycoides, Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus brevis and inoculate them into a conical flask filled with LB medium, and culture them at 30°C and 180 r / min for 12 h as seed liquid, inoculate them into a fermenter filled with LB medium at an inoculum size of 2%, and continue to culture at 30°C and 180 r / min for 24 h to obtain fermentation broth; LB medium is prepared by sterilizing 10 g of tryptone, 5 g of yeast powder, 10 g of sodium chloride and 1 L of distilled water at 121°C for 20 min.

[0081] S313, preparation of bacterial powder: the fermentation liquid cultured in the fermentation tank was taken out, centrifuged at 5000rpm for 10min, the supernatant was removed, the protective agent maltodextrin was added at a ratio of 20:3, oscillated for 30s, and placed on a shaker at a speed of 150rpm for incubation at 30°C for 30min, spray dried to obtain bacterial powder; the bacterial powder and cow dung biochar were uniformly mixed at a volume ratio of 23:77 to obtain a Bacillus composite bacterial agent; the bacterial agent prepared by the formula had a spore content of 3.9×10 8 CFU·g -1 , moisture content (w) ≤ 0.9%, fineness ≥ 99%, wetting time is 33.0s, suspension rate reaches 81.0%, and all indicators meet national standards.

[0082] S32, Preparation of Pseudomonas fluorescens Agent

[0083] S321. Activate Pseudomonas fluorescens on a KB plate at 28°C for 24h. After activation, pick a single colony with an inoculation loop, inoculate it into 5mL of liquid seed KB culture medium, culture it in a shaker at 28°C and 150r / min for 24h, then inoculate it into solid seed KB culture medium, culture it at 28°C and 24h, pick one loop of the cultured first-level seeds with an inoculation loop, inoculate it into 50mL of liquid seed KB culture medium, culture it in a shaker at 28°C and 150r / min for 24h, and obtain second-level seeds; the components of liquid seed KB culture medium (1L) are: 15mL of glycerol, 20g of peptone, 0.3g of K2HPO4, 1.5g of MgSO4·7H2O, and pH 7.0; solid seed KB culture medium: add 20g of agar to Kings′B culture medium.

[0084] S322, the fermentation medium was first treated with 1g 8-hydroxyquinoline to remove trace iron, the fermentation medium composition was: 1.5g glycerol, 1.0g hydrolyzed casein, 2.5g magnesium sulfate, 2.5g dipotassium hydrogen phosphate and 7.0g glucose, the pH was 6.5, ammonia water and phosphoric acid were used to control the pH of the fermentation medium at 6.5, the culture conditions were bottling volume 200 / 500 (mL), inoculation size 2%, the optimal culture time was 48h, Fe 3+ The concentration is 0.8 mg / L, and the final viable count is 5.0×10 9 CFU·g -1 .

[0085] S323. Under sterile conditions, collect the fluorescent Pseudomonas cells in the logarithmic growth phase by centrifugation and suspend them in fresh liquid seed KB medium to make the number of viable cells reach 5.0×10 9 CFU·g -1 The above, then add sodium carboxymethyl cellulose according to the volume ratio of bacterial liquid: stabilizer = 10:1, mix thoroughly and mix with a wetting agent, a dispersant, a protective agent and a carrier, wherein the wetting agent is sodium dodecyl sulfate (9%), the dispersant is sodium tripolyphosphate (9%), the protective agent is humic acid (1.2%), and the carrier is diatomaceous earth with a particle size of 4 mm (52.8%). After standing at room temperature for 48 hours, encapsulate and store at room temperature to obtain a fluorescent Pseudomonas agent.

[0086] S33. Detection of the ability of strains to produce iron carriers. The strains (Bacillus composite bacteria and Pseudomonas fluorescens) were activated and cultured on NB plates for 24 hours. Single colonies were picked up with toothpicks and spotted on CAS plates. After culturing at 28°C for 2 days, single colonies were picked up and inoculated into MSA culture medium. The culture was incubated at 28°C and 150 r / min for 2 days. The bacterial suspension was centrifuged at 10,000 r / min for 10 min. An equal volume of bacterial culture supernatant was mixed with CAS detection solution. The mixture was reacted in the dark for 1 hour. The absorbance As was detected at a wavelength of 680 nm. At the same time, the absorbance Ar of the reaction between the blank culture medium and the equal volume of CAS detection solution was determined. The iron carrier production ability of the strain was analyzed by As / Ar. As / Ar was between 0-1. The As / Ar of bacteria with higher iron carrier production ability was lower than 0.5. The iron carrier activity unit (SU) was used to represent the iron carrier production of the strain, and the calculation formula was: SU=[(Ar-As) / Ar]×100.

[0087] The components of NB medium (1L) are: 3.0g beef extract powder, 10.0g peptone, 5.0g NaCI, and 18.0g agar; the components of MSA medium (1L) are: 20.0g sucrose, 2.0g L-asparagine, 1.0g K2HPO4, and 0.5g MgSO4·7H20.

[0088] The composition of CAS culture medium is: 1 mL of 20% sucrose solution, 3 mL of 10% acid hydrolyzed casein, 100 μL of 1 mmol / L CaCl2, 2 mL of 1 mmol / L MgSO4, and 1.8 g of agar per 100 mL. Slowly add 5 mL of phosphate buffer and CAS dye solution at about 60°C to obtain CAS blue culture medium.

[0089] The composition of CAS detection solution is as follows: Solution A: 500 mL of 1 mmol / L CAS (chrome azure), 100 mL of 0.1 mmol / L FeCl3·6H20; Solution B: 400 mL of 2 mmol / L HDTMA (hexadecyltrimethylammonium bromide); slowly pour Solution A into Solution B to prepare 1 L of blue CAS dye solution.

[0090] S4. In the high temperature and high humidity season from early July to mid-August, the yellowing symptoms disappeared and the divalent iron content in the surface soil was less than 0.01 μmol·L -1 When the iron carrier is high, apply the high-yield iron carrier agent, apply the fluorescent Pseudomonas agent near the drip irrigation pipe, and apply 10g / m 2 , apply Bacillus compound bacterial agent on the far side of the drip irrigation pipe, 5-10cm deep soil in the rhizosphere, 30g / m 2 .

[0091] Case 1: Iron deficiency chlorosis of Polygonatum cyrtonema under the Sophora japonica forest in Tongzhou District, Beijing

[0092] The two-year-old Polygonatum cyrtonema under the Sophora japonica forest in Tongzhou District, Beijing, has a brown tidal soil with a pH value of 8.05, an effective iron content of less than 2.17 mg / kg, and a bulk density of 1.372 g / cm3, which is an alkaline iron-deficient soil. The forest stand is a plain ecological forest, the dominant tree species Sophora japonica is 12 years old, and the forest stand canopy density is 0.7.

[0093] The comparative test found that the Polygonatum cyrtonema under the Sophora japonica forest, which did not implement the comprehensive technology of pH value control, effective iron supply and iron carrier agent application, began to show iron deficiency symptoms in mid-May, and the symptoms became more obvious in early June. Figure 4 , Figure 5 and Figure 6 .

[0094] In the test plots that were treated with pH value control, effective iron supply, and iron carrier bacteria application (ternary system treatment method), Polygonatum cyrtonema grew normally.

[0095] Data were measured in mid-July. Plant samples were randomly selected according to the grid method. 50 plants were selected from the control group and the treatment group, and the average value was calculated after measurement. The measurement methods were as follows: using an ultraviolet spectrophotometer to measure the urease in the rhizosphere 10cm soil according to the sodium phenol-sodium hypochlorite colorimetric method, using an ultraviolet spectrophotometer to measure the absorbance of the sample at 645nm and 663nm to calculate the chlorophyll content; using the German WALZ company MINI-PAM to measure the chlorophyll fluorescence parameter Fv / Fm (PSII maximum photon conversion efficiency), using the automatic colorimeter CR-400 (Konica Minolta, Japan) to measure the red-green saturation parameter a* of the leaf color, using the dilute hydrochloric acid extraction method and the o-phenanthroline colorimetric method to determine the effective iron content of the leaves. The results are shown in Table 2.

[0096] Table 2 The effect of the three-element system treatment method on the prevention and control of iron deficiency of Polygonatum cyrtonema under the forest

[0097]

[0098] As can be seen from Table 2, in the test plots treated by the method of the present invention, there was no iron deficiency in the Polygonatum cyrtonema under the forest. The Fv / Fm in the chlorophyll fluorescence kinetic parameter represents the health status of the plant. The value obtained in this test is greater than 0.8, indicating that the Polygonatum cyrtonema plants treated by the ternary system are in a healthy growth state, and the urease activity of the rhizosphere soil is 23% higher than that of the control, indicating that the soil activity is higher and can more effectively supply mineral nutrients. The application effect of the present invention in Case 1 is shown in FIG. Figure 7 .

[0099] Case 2: Ecological planting of Polygonatum cyrtonema under chestnut forest in Chengde City, Hebei Province

[0100] The soil of the ecological planting plot of Polygonatum cyrtonema under the chestnut forest in Chengde City, Hebei Province is mountain brown soil with a pH value of 7.4 and a surface soil bulk density of 1.302g / cm 3 , the effective iron content is 4.61 mg / kg, which is nearly neutral soil, and the canopy density of chestnut economic forest is 0.5.

[0101] The experiment was designed with three treatment groups: treatment group one was a blank control, treatment group two was iron-loaded biochar acidified with wood vinegar + foliar spraying of ferrous sulfate, and treatment group three was iron-loaded biochar acidified with wood vinegar + foliar spraying of chelated iron organic fertilizer + high-yield iron carrier agent.

[0102] like Figure 8 As shown, the iron deficiency symptoms of Polygonatum cyrtonema in the blank control plot were yellow-white leaves at the top, lighter leaf color between the middle leaf veins, and basically normal leaf color of the lower leaves.

[0103] like Fig. 9As shown in the figure, the leaf characteristics of Polygonatum cyrtonema in the plot where iron-loaded biochar was acidified with wood vinegar and ferrous sulfate was sprayed on the leaves showed that although the iron deficiency symptoms were alleviated, part of the sprayed ferrous sulfate was oxidized into trivalent iron and retained on the leaf surface, which was invalid iron. The young leaves at the top still showed symptoms of interveinal chlorosis.

[0104] like Fig.10 As shown in the figure, the Polygonatum cyrtonema cultivated in treatment group 3 did not show obvious symptoms of iron deficiency during the vigorous growth season. The specific cultivation process is as follows:

[0105] a. Before the buds of Polygonatum odoratum emerge in early spring, dig shallow trenches at the rhizosphere and apply iron-loaded biochar acidified with wood vinegar. One week later, measure the pH value of the rhizosphere soil, which decreases by 0.3-0.5. The effective iron in the soil can be increased by 30-80% on the basis of the original background of 2.0-6.0 mg / kg, and the maintenance time is about 6-7 weeks.

[0106] b. By May, when the stem segments of Polygonatum cyrtonema were about 6-9 nodes, the pH value of the rhizosphere soil basically recovered to the original 7.0-7.5, and the effective iron in the root soil of Polygonatum cyrtonema was reduced to a level with no significant difference from the base content, that is, an average of about 4.0 mg / kg.

[0107] c. Before Polygonatum cyrtonema faces iron deficiency stress again, spray chelated iron organic fertilizer on the leaves in time. After one week, the iron content of the three leaves below the top can increase from 46.13mg / kg to 52.49mg / kg, an average increase of 13.78%.

[0108] d. Spray once every 10 days. By mid-June, the effective iron content in the leaves will be maintained at a normal level of about 50 mg / kg, the soil pH value will drop slightly by 0.1-0.2, and the average effective iron content in the soil will be about 4.2 mg / kg.

[0109] e. With the coming of the rainy season, stop applying foliar fertilizer, the soil pH value will slowly rise, and the effective iron content in the soil will gradually decrease. At this time, apply fluorescent Pseudomonas agent and high-yield iron carrier Bacillus complex agent respectively according to the moisture gradient, which can keep the effective iron chelated by iron carriers enriched in the roots of Polygonatum cyrtonema for a long time and ensure the full absorption of effective iron.

[0110] Case 3: Ecological planting of Polygonatum cyrtonema under forest in Jizhou District, Tianjin

[0111] The experiment was designed with two treatment groups: treatment group one was a blank control, and treatment group two was the same as treatment group three in case two.

[0112] Mix Polygonatum cyrtonema and Polygonatum cyrtonema under the forest, such as Fig.11 As shown in the blank control test, the three-year-old Polygonatum cyrtonema showed interveinal chlorosis, such as Fig.12As shown in the blank control test, Polygonatum cyrtonema showed interveinal chlorosis, while Polygonatum cyrtonema and Polygonatum cyrtonema cultured in treatment group 2 did not show iron deficiency symptoms. Fig.13 .

[0113] The embodiments of this specific implementation method are all preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention. Therefore, all equivalent changes made based on the structure, shape, and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A comprehensive prevention and treatment method for iron deficiency in Polygonatum cyrtonema under forest, characterized in that: The method comprises the following steps: S1, adjusting the pH value of rhizosphere soil; S11, crushing the tree waste obtained from forest cultivation into particles with a particle size of 2 mm, placing them in a pyrolysis carbonization furnace (4) and performing anaerobically pyrolysis at 520° C. for 2 h under nitrogen conditions to produce garden biochar C0; S12, soak the biochar C0 in 0.5 mol·L -1 FeCl3 and 0.5 mol·L -1 In the FeSO4 solution, the mass ratio of Fe:C is 0.56, the mass volume ratio of biochar C0:solution is 1:8, after stirring for 30 minutes until uniform, it is placed in a pyrolysis carbonization furnace (4) and calcined at 430°C for 2 hours to obtain modified biochar C1; S13, dilute the wood vinegar stock solution 10 times, mix the modified biochar C1 with the diluted wood vinegar stock solution at a ratio of 1g:10ml, stir thoroughly, let stand for 2 days for acidification, and obtain the acidified modified biochar. In early spring, dig a 10cm shallow ditch and apply it to the rhizosphere of Polygonatum cyrtonema under the forest. Water 5L of the mixed solution per square meter, and use 1500L per mu of Polygonatum cyrtonema under the forest. S2, effective supply of iron element, applying chelated iron organic fertilizer to the rhizosphere soil of Polygonatum cyrtonema under the forest improved by step S1, and spraying thin chelated iron organic fertilizer on the leaves multiple times; S3, high-yield iron carrier bacterial agent secretes iron carriers.

2. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 1, characterized in that: The step S2 comprises: S21, crush the horseshoe slices, pass through a 40-mesh sieve, take 12 kg and put it into a corrosion-resistant container, add 4.6 kg of water, and mix with Actinomadura keratinilytica (EU637009, strain WCC-2265 T ) 48g of bacterial agent, of which the content of live bacteria is 2×10 10 CFU / g, stir evenly, spray water once every 12 hours at 42-45°C, keep it moist, and treat the stubborn keratin by biodegradation for 5 days; then add 30L of 1.0mol / L sulfuric acid, stir evenly with a glass rod, and react for 12 hours; evaporate and deacidify in a water bath (≤90°C) to concentrate to 10L, slowly inject into another anti-corrosion container containing 20L of water, evaporate and deacidify again, and do this operation 3 times in total to obtain a hydrolyzed amino acid solution of horseshoe; S22, crush 260kg sesame cake into pieces with a diameter less than 1cm, put 60kg millet bran into a retting tank, add 240L of water, and stir evenly; slowly inject the water solution of hydrolyzed amino acids from horseshoe, stir evenly, add the composite bacterial agent, mix and stir evenly; regularly record the central temperature and ambient temperature of the fertilizer pile, turn the pile once every 2 days, and add water to keep the moisture content of the whole composting process at 62%, and the fertilizer composting time is 30 days; S23, add 22kg of ferrous sulfate, add a large amount of water to completely dissolve, finally the liquid level reaches 50cm, stir evenly, chelate reaction for one week, and obtain chelated iron organic fertilizer; S24. From late April to early May, take 100L of fermented fertilizer solution and dilute it 10 times, and spray it on the leaves 4 times, with an interval of 7-15 days between two times; take 900L of fermented fertilizer solution and dilute it 5 times, and use it as drip irrigation to the roots of Polygonatum cyrtonema under the forest.

3. A comprehensive prevention and treatment method for iron deficiency in Polygonatum cyrtonema under forests according to claim 2, characterized in that: The composite bacterial agent added in step S22 is Bacillus subtilis and Aspergillus niger, and the bacterial count ratio of Bacillus subtilis to Aspergillus niger is 60:

2.

4. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 2, characterized in that: After the fertilizer is piled in step S22, it is accurately weighed and uniformly sampled for more than 5 kg to determine the moisture content, protein N, total N, mineral nitrogen (NH4 + +NO3 - ), total amino acids, organic C content, and calculated the maturity and protein N degradation rate of cake fertilizer under different bacterial agent treatments; The degree of maturity is measured by the humic acid E4 / E6 value, and the determination method is as follows: extract the sample with distilled water, filter and measure the absorbance of the filtrate with a spectrophotometer at wavelengths of 465nm and 665nm, and calculate the ratio of the absorbance at the two wavelengths, i.e. E4 / E6; maturity requires C / N less than 6.50, protein N degradation rate higher than 22%, humic acid E4 / E6 value less than 3.50, and total amino acid content greater than 27.3mg / g, wherein the mass ratio of distilled water to sample is 20:

1.

5. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 1, characterized in that: The step S3 comprises: S31, preparation of Bacillus composite bacterial agent; S311. Preparation of cow dung biochar: fresh cow dung is collected and cleaned of stones and other debris. The cow dung is spread on a clean large core board with a flat cement floor underneath. The cow dung is air-dried under full light and natural ventilation until the water content is below 18%. The dried cow dung is spread at 105° C. and dried for 3 h for preheating. The preheated cow dung is placed in a pyrolysis carbonization furnace (4) and the heating rate is controlled to be 100° C. h -1 When the pyrolysis temperature reaches 380° C., the pyrolysis time is 3 hours under anoxic conditions; after closing the pyrolysis carbonization furnace (4), the pyrolysis cow dung is cooled to below 80° C., the pyrolysis cow dung is cooled for standby use, and the cow dung biochar prepared by pyrolysis is put into a pulverizer and crushed, and passed through a 40-mesh sieve to obtain cow dung biochar; S312, preparation of fermentation liquid: single colonies of Bacillus tekila, Bacillus mycoides, Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus pumilus were selected and inoculated into a conical flask filled with LB medium, and cultured for 12 hours at 30° C. and 180 r / min as seed liquid, and inoculated into a fermentation tank filled with LB medium at an inoculum amount of 2%, and continued to culture for 24 hours at 30° C. and 180 r / min to obtain fermentation liquid; S313, preparation of bacterial powder: the fermentation liquid cultured in the fermentation tank was taken out, centrifuged at 5000rpm for 10min, the supernatant was removed, the protective agent maltodextrin was added at a ratio of 20:3, oscillated for 30s, and placed on a shaker at a speed of 150rpm for incubation at 30°C for 30min, spray dried to obtain bacterial powder; the bacterial powder and cow dung biochar were uniformly mixed at a volume ratio of 23:77 to obtain a Bacillus composite bacterial agent; the bacterial agent prepared by the formula had a spore content of 3.9×10 8 CFU·g -1 , water content (w) ≤ 0.9%, fineness ≥ 99%, wetting time is 33.0s, and suspension rate reaches 81.0%.

6. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 5, characterized in that: The step S3 further comprises: S32, Preparation of Pseudomonas fluorescens Agent S321. Activate Pseudomonas fluorescens on a KB plate at 28°C for 24 hours. After activation, pick a single colony with an inoculation loop, inoculate it into 5 mL of liquid seed KB medium, culture it in a shaker at 28°C and 150 r / min for 24 hours, then inoculate it into a solid seed KB medium, culture it at 28°C for 24 hours, pick a ring of the cultured primary seeds with an inoculation loop, inoculate it into 50 mL of liquid seed KB medium, culture it in a shaker at 28°C and 150 r / min for 24 hours, and obtain secondary seeds; S322, the fermentation medium was first treated with 1g 8-hydroxyquinoline to remove trace iron, the fermentation medium composition was: 1.5g glycerol, 1.0g hydrolyzed casein, 2.5g magnesium sulfate, 2.5g dipotassium hydrogen phosphate and 7.0g glucose, pH was 6.5, the culture conditions were bottling volume 200 / 500 (mL), secondary seed inoculation amount 2%, optimal culture time was 48h, Fe 3+ The concentration is 0.8 mg / L, and the final viable count is 5.0×10 9 CFU·g -1 ; S323. Under sterile conditions, collect the fluorescent Pseudomonas cells in the logarithmic growth phase by centrifugation and suspend them in fresh liquid seed KB medium to make the number of viable cells reach 5.0×10 9 CFU·g -1 The above, then add sodium carboxymethyl cellulose according to the volume ratio of bacterial liquid: stabilizer = 10:1, mix thoroughly, mix with a wetting agent, a dispersant, a protective agent and a carrier, wherein the wetting agent is sodium dodecyl sulfate (9%), the dispersant is sodium tripolyphosphate (9%), the protective agent is humic acid (1.2%), and the carrier is diatomaceous earth (52.8%) with a particle size of 4 mm, and after being placed at room temperature for 48 hours, encapsulate and store at room temperature to obtain a fluorescent pseudomonas agent; S4. In the high temperature and high humidity season from early July to mid-August, the yellowing symptoms disappeared and the divalent iron content in the surface soil was less than 0.01 μmol·L -1 When the iron carrier is high, apply the high-yield iron carrier agent, apply the fluorescent Pseudomonas agent near the drip irrigation pipe, and apply 10g / m 2 , apply Bacillus compound bacterial agent on the far side of the drip irrigation pipe, 5-10cm deep soil in the rhizosphere, 30g / m 2 .

7. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 6, characterized in that: The step S3 further comprises: S33. Detection of the ability of the strain to produce iron carriers. The strain was activated and cultured on NB plates for 24 hours. A single colony was picked up with a toothpick and spotted on a CAS plate. After culturing at 28°C for 2 days, a single colony was picked up and inoculated into MSA culture medium. The culture was incubated at 28°C and 150 r / min for 2 days. The bacterial suspension was centrifuged at 10,000 r / min for 10 min. An equal volume of bacterial culture supernatant was mixed with CAS detection solution. The mixture was reacted in the dark for 1 hour. The absorbance As was detected at a wavelength of 680 nm. At the same time, the absorbance Ar of the reaction between the blank culture medium and the equal volume of CAS detection solution was determined. The iron carrier production ability of the strain was analyzed by As / Ar. As / Ar was between 0-1. The As / Ar of bacteria with higher iron carrier production ability was lower than 0.

5. The iron carrier activity unit (SU) was used to represent the iron carrier production of the strain. The calculation formula was: SU = [(Ar-As) / Ar] × 100.

8. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 5, characterized in that: The LB culture medium in step S312 is prepared by sterilizing 10 g of tryptone, 5 g of yeast powder, 10 g of sodium chloride and 1 L of distilled water at 121° C. for 20 min.

9. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 6, characterized in that: The liquid seed KB medium (1 L) in step S321 comprises: 15 mL of glycerol, 20 g of peptone, 0.3 g of K2HPO4, 1.5 g of MgSO4·7H2O, and a pH of 7.0; the solid seed KB medium is prepared by adding 20 g of agar to Kings′B medium.

10. The comprehensive prevention and treatment method for iron deficiency of Polygonatum cyrtonema under forest according to claim 7, characterized in that: The composition of the NB medium (1 L) in step S33 is: 3.0 g beef extract powder, 10.0 g peptone, 5.0 g NaCl, and 18.0 g agar; the composition of the MSA medium (1 L) is: 20.0 g sucrose, 2.0 g L-asparagine, 1.0 g K2HPO4, and 0.5 g MgSO4·7H2O; The composition of the CAS culture medium in step S33 is as follows: 1 mL of 20% sucrose solution, 3 mL of 10% acid hydrolyzed casein, 100 μL of 1 mmol / L CaCl2, 2 mL of 1 mmol / L MgSO4, and 1.8 g of agar per 100 mL, and 5 mL of phosphate buffer and CAS dye solution are slowly added at about 60° C. to obtain CAS blue culture medium; The composition of CAS detection solution is as follows: Solution A: 500mL of 1mmol / L CAS (chrome azure), 100mL of 0.1mmol / L FeCl3·6H2O; Solution B: 400mL of 2mmol / L HDTMA (hexadecyltrimethylammonium bromide); slowly pour Solution A into Solution B to prepare 1L of blue CAS dye solution.

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