CN114231465A - Microbial preparation for improving iron deficiency stress resistance of crops and application thereof - Google Patents

Microbial preparation for improving iron deficiency stress resistance of crops and application thereof Download PDF

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CN114231465A
CN114231465A CN202111680982.XA CN202111680982A CN114231465A CN 114231465 A CN114231465 A CN 114231465A CN 202111680982 A CN202111680982 A CN 202111680982A CN 114231465 A CN114231465 A CN 114231465A
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李华一
罗钰彬
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Guangdong Lihao Biological Agriculture Co ltd
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Abstract

The invention belongs to the field of microbial agents, and particularly relates to a microbial agent for improving the iron deficiency stress resistance of crops and an application thereof.

Description

Microbial preparation for improving iron deficiency stress resistance of crops and application thereof
Technical Field
The invention belongs to the field of microbial agents, and particularly relates to a microbial agent for improving iron deficiency stress resistance of crops and application thereof.
Background
In recent years, Chinese scholars generally study the bacillus to perform crop experiments. The research on the microorganisms of bacillus is more, such as bacillus subtilis, bacillus megaterium, bacillus licheniformis and the like. Meanwhile, the bacillus bacterial manure is only applied to a low-yield field or a new growth area, and the effect is obvious. The research and test on the relationship between bacteria are not complete, the microbial inoculum is single, the bacteria lack cooperation and are easy to repel each other, and finally the bacteria are disintegrated and isolated. It is also easy to damage the environment and bring irreversible damage to crops, with the following consequences: excess deficiency of elemental iron and stress of bicarbonate. Leading to reduced income for farmers and consequently reduced agricultural production levels.
The existing microbial bacillus agents in the market are similar in pesticide effect, lack of supplement to carbon sources, little and little supplement to nitrogen elements, short in pesticide effect duration, lack of stress of iron elements excessively, and have the probability of bringing irreversible damage to plants, so that farmers lose income and income is reduced. The existing bacillus inocula are flooded in the market, so that corresponding environment-friendly inocula are needed to be made, and sustainable development agricultural inocula are brought.
Disclosure of Invention
The invention aims to provide a microbial preparation for improving the iron deficiency stress resistance of crops and application thereof, which can effectively improve the yellowing phenomenon of iron deficiency stress of crops, promote the germination of crop seeds and the synthesis and accumulation of chlorophyll and nitrogen elements, and further promote the growth of crops.
Based on the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides a microbial preparation for improving the capacity of crops to resist iron deficiency stress, wherein the microbial preparation is a compound microbial preparation formed by mixed culture of arbuscular mycorrhizal fungi and azospirillum brasilense.
Arbuscular mycorrhizal fungi (Glomus intraradics) have a common symbiosis phenomenon with plants in nature, and the so-called mycorrhiza is a union formed by mycorrhizal fungi hypha in soil and a higher plant nutrient root system. The symbiotic fungi obtain necessary carbohydrates and other nutrients from the plant body, and the plant also obtains the required nutrients and moisture from the fungi, so that the high unification of mutual benefit and mutual assistance and communication is achieved, the capability of resisting adverse environment of the plant is improved, the plant growth is promoted, and the survival of mycorrhizal fungi is facilitated. This relationship sometimes develops to the extent that both parties are difficult to distinguish, plants lack mycorrhiza and cannot survive, and mycorrhiza lacks essential plant root system symbiosis, cannot complete life history, and cannot continue to reproduce. It has the characteristics of common plant root system and the characteristics of obligate fungi. Arbuscular mycorrhizal fungi (Glomus intraradics) are root endophytic fungi capable of establishing reciprocal symbiotic relationship with plants, and can fix nitrogen, promote plant growth and increase crop yield.
Azospirillum brasilense (Azospirillum brasilense) is a microorganism capable of fixing nitrogen by an autonomous organism, can be symbiotically combined with roots of various crops and important gramineous crops such as rice, corn, wheat, sugarcane and the like, and has strong adaptability. It can remove N in air2Into NH effective for crops4 +、NH3 +The nitrogen is combined with metal ions in the soil, so that the root system of the plant can absorb sufficient nitrogen elements by crops, and the plant height, the stem thickness, the quality of the above-ground parts and the quality of the below-ground parts of the crops are obviously improved.
According to the invention, the composite microbial inoculum prepared by mixed culture of the arbuscular mycorrhizal fungi and the azospirillum brasilense is found through a crop irrigation test, and compared with the mixed microbial inoculum formed by mixed culture of the arbuscular mycorrhizal fungi, the single microbial inoculum of the azospirillum brasilense and other azospirillum and arbuscular mycorrhizal fungi, the microbial preparation provided by the invention has better effects of promoting germination of crop seeds and promoting growth of crops, and can effectively improve the symptom of iron deficiency and yellowing of crops.
Further, the viable count of the arbuscular mycorrhizal fungi in the microbial preparation is 2.5X 106～1.3×109(ii) a The number of viable bacteria of Azospirillum brasilense is 1.6 × 107～3.1×108。
Further, the preservation number of the arbuscular mycorrhizal fungi is BGC BJ03, and the preservation number of the azospirillum brasilense is CGMCC 1.10379.
Further, the crops include rice, wheat, corn, citrus, soybean, sorghum, sesame, and peanut.
Tests show that the compound microbial inoculum formed by mixed culture of the arbuscular mycorrhizal fungi and the azospirillum brasilense provided by the invention has the effects of promoting seed germination, promoting crop growth and improving the phenomena of iron deficiency stress yellowing of the crops.
In a second aspect, the present invention provides the use of the aforementioned microbial preparation in crops against stress of iron deficiency.
Experiments show that after the microbial inoculum treats crops which are yellowed under the stress of iron deficiency, the content of iron elements in the crops is improved, and the yellowing phenomenon of the crops is obviously improved.
In a third aspect, the present invention provides the use of a microbial preparation as described above for growth promotion in crops.
Tests show that after the fungicide is irrigated, chlorophyll synthesis and accumulation are effectively promoted, the increase of biological properties of crops such as plant height, root length, root number, leaf length, leaf width, root fresh quality, overground fresh quality, root trunk quality and overground dry quality is promoted, and the growth of the crops is promoted.
In a fourth aspect, the invention provides the use of the aforementioned microbial preparation in breeding of low-iron tolerant crops.
Compared with the prior art, the invention has the following beneficial effects:
the mixed microbial inoculum provided by the invention can be used for providing nitrogen sources for crops such as rice and the like by autonomous biological nitrogen fixation, so that the absorption of the nitrogen sources by the crops is enhanced, the secretion of the auxin is induced, and the growth of the crops is promoted.
Compared with the mixed microbial inoculum formed by mixed culture of the arbuscular mycorrhizal fungi, the single microbial inoculum of the azospirillum brasilense and other azospirillum and arbuscular mycorrhizal fungi, the microbial preparation provided by the invention has better effects of promoting the germination of crop seeds and the growth of crops, and can effectively improve the symptom of iron deficiency and yellowing of crops.
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FIG. 1 is a graph showing the variation of chlorophyll content in rice treated by different treatments;
FIG. 2 is a graph showing the variation of chlorophyll content in wheat according to different treatments;
FIG. 3 is a graph showing the variation of chlorophyll content in corn for different treatments;
FIG. 4 is a graph showing the variation of chlorophyll content in citrus fruits according to different treatments;
FIG. 5 is a graph showing the variation of chlorophyll content in soybeans for different treatments;
FIG. 6 is a graph showing the change of chlorophyll content in sorghum according to different treatments;
FIG. 7 is a graph showing the change of chlorophyll content in sesame seeds according to different treatments;
FIG. 8 is a graph showing the variation of chlorophyll content in peanuts according to different treatments.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples. It will be understood by those skilled in the art that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The test methods used in the examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are commercially available unless otherwise specified.
Example 1
The embodiment provides a microbial preparation, which is a composite microbial inoculum formed by mixed culture of arbuscular mycorrhizal fungi and azospirillum brasilense, wherein arbuscular mycorrhizal fungi (Glomus intraradics) strains are purchased from the research center for plant nutrition and resources of agriculture and forestry academy of sciences of Beijing, China arbuscular mycorrhizal fungi (BGC), and the preservation number is BJ 03; the Azospirillum brasilense strain is purchased from China general microbiological culture Collection center with the preservation number of CGMCC 1.10379.
Inoculating arbuscular mycorrhizal fungi (Glomus intraradics) and Azospirillum brasilense (Azospirillum brasilense) into 2L LB liquid culture medium, and performing mixed culture for 7d in a light-proof constant temperature oscillator at 30 ℃ and 180r/min to obtain the microbial preparation, namely the mixed microbial preparation, wherein the viable count of the arbuscular mycorrhizal fungi in the mixed microbial preparation is 3.2 multiplied by 108(ii) a The number of viable bacteria of Azospirillum brasilense is 2.7 × 107。
The effects of the mixed microbial inoculum of the invention on crop breeding and crop growth promotion are analyzed by taking a single arbuscular mycorrhizal fungi microbial inoculum, a nitrogen fixing spirillum brasilense microbial inoculum and sterile water as references as follows. The specific test method is as follows:
1. test inoculum or blank control
(1) The mixed microbial inoculum is obtained by the method described above.
(2) Arbuscular mycorrhizal fungi agent
Inoculating arbuscular mycorrhizal fungi (Glomus intraradics) on sterilized LB liquid culture medium, culturing in dark constant temperature oscillator at 30 deg.C and 180r/min for 7d to obtain single arbuscular mycorrhizal fungi liquid with viable count of 1.3 × 109。
(3) Azospirillum brasilense microbial inoculum
Inoculating Azospirillum brasilense (Azospirillum brasilense) on sterilized LB liquid culture medium, culturing in dark constant temperature oscillator at 30 deg.C and 180r/min for 7d to obtain single Azospirillum brasilense bacterial liquid with viable count of 3.1 × 108。
(4) Sterile water
2. Test seeds and treatment thereof
The tested seeds are selected from rice (Gudun No. 18), wheat (Zhoumai No. 16), corn (dry Zhen 958), orange (Aiyun No. 38), soybean (Zhonghuang 37), sorghum (Jia dwarf 60), sesame (Aitian black giant sesame) and peanut (golden crown 308). Respectively selecting healthy and plump 20 seeds, placing the seeds in a 70% ethanol solution for 8min, washing the seeds with sterile water for 3 times, placing the seeds in a 0.5% sodium hypochlorite solution for 10min, washing the seeds with sterile water for a plurality of times, spreading the seeds on wet filter paper, accelerating germination in a constant-temperature incubator at 28-30 ℃ in a dark environment, pouring 200ml of sterile water once every 2 days, and respectively selecting seedlings with consistent growth vigor for later use after 14 days.
3. Test method and test results
(1) Test for improving iron deficiency stress of crops
After 14 days, selecting seedlings with consistent growth vigor, setting 10 pots of seedlings and 80 pots of seedlings in total, placing the seedlings on an MS culture medium plate (without adding cane sugar) and culturing the seedlings in a light-dark alternating incubator with the temperature of 28 ℃ and the illumination of 16 h/the darkness of 8 h. After one month of culture, 200ml of iron-removing Hoagland's nutrient solution is applied to each pot, and the solution is applied every 2 days. And respectively irrigating the mixed microbial inoculum, the arbuscular mycorrhizal fungal microbial inoculum, the azospirillum braziliani microbial inoculum and sterile water (contrast) for treatment until the seedlings have iron deficiency and yellowing symptoms, and irrigating once every 6 days, wherein the mixed microbial inoculum, the arbuscular mycorrhizal fungal microbial inoculum and the azospirillum braziliani microbial inoculum are diluted by 50 times by the sterile water during irrigation and irrigated.
And (3) testing the content of the iron element in each crop by adopting a hydrochloric acid leaching method 20 days from the beginning of irrigating the mixed microbial inoculum, wherein the testing method comprises the following steps:
washing each crop leaf with distilled water, deactivating enzymes at 105 ℃, then placing the crop leaves in a room for airing, grinding the leaves through a 0.5mm sieve by using a porcelain mortar or a stainless steel pulverizer, drying the leaves for 4-6 h in a drying box at 70 ℃, weighing 0.5-1.0 g of a dried sample, and adding 0.24mol/L hydrochloric acid solution into the dried sample, wherein the weight ratio of the sample to the hydrochloric acid solution is 1: 50. Stirring, precipitating and filtering to obtain clear filtrate, namely the plant sample solution to be detected.
Putting 5-10 ml of clarified plant sample solution to be detected into a 25ml volumetric flask, adding 2 drops of 2, 4-dinitrophenol into the volumetric flask, adjusting the solution to be yellow by using 1mol/L NaOH solution, and adding 0.1mol/L HCl until the yellow color is just faded; 2ml of 10% hydroxylamine hydrochloride solution was added to the flask, shaken well and left for several minutes. Then 1ml of phenanthroline is added, and the volume is determined by water. After 30min of incubation, the colorimetric assay was carried out, 1cm cuvette, 510nm wavelength. The Fe concentration was found from the standard curve. Standard curve: respectively taking 0, 1, 2, 3, 4 and 5ml of 10ppm Fe standard solution, placing the solution into a 25ml volumetric flask, carrying out color development measurement by the same operation steps, and respectively obtaining standard series containing 0, 0.4, 0.8, 1.2, 1.6 and 2.0ppm Fe, and drawing a standard curve of the concentration of Fe to the absorbance A. The iron content in the plant sample to be detected is detected based on the standard curve, the detection result is shown in table 1, and the iron content units in table 1 are ppm/g.
TABLE 1 iron content in various crops treated differently
Species of Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Rice (Gudun No. 18) 21.4 240.3 30.4 251.2
Wheat (Zhoumai 16) 22.5 230.5 31.6 261.5
Corn (Liandan 958) 42.3 298.8 53.6 302.2
Orange (pretty girl 38) 35.6 246.3 45.1 256.3
Soybean (middle yellow 37) 32.5 261.6 45.3 269.8
Jowar (Jia low 60) 39.8 301.6 45.6 311.2
Sesame (airline black giant sesame) 29.8 301.3 35.6 312.1
Peanut (golden crown 308) 25.8 220.6 35.6 226.3
On the appearance of each seedling, it can be seen that after the mixed microbial inoculum is irrigated for 20d, each crop grows obviously and the symptom of iron deficiency yellowing is obviously relieved, and the results in table 1 show that compared with a control group, the method relieves the symptom of iron deficiency yellowing to a certain extent after the microbial inoculum is used for treatment, and particularly obviously improves the content of iron element in each test seedling after the arbuscular mycorrhizal fungi and the mixed microbial inoculum are used for treatment. And compared with arbuscular mycorrhizal fungi, the iron element content in each test seedling is further improved after the mixed fungicide is treated. Compared with a single microbial inoculum, the mixed microbial inoculum consisting of the arbuscular mycorrhizal fungi and the azospirillum brasilense has the advantages that the symptom of iron deficiency and yellowing of crops can be better improved, and the mixed microbial inoculum is expected to be applied to breeding of low-iron-resistant crops.
(2) Test for growth promoting effect on crops
Taking 14d seedlings with consistent growth vigor of sprouts, respectively taking arbuscular mycorrhizal fungi liquid, Azospirillum brasilense liquid and mixed microbial inoculum liquid which are diluted by 50 times of sterile water, irrigating the seedlings by using the diluted bacterial liquid every 7d, taking sterile water with the same irrigation quantity as a control group, and measuring the average value of the plant height, the root length, the root number, the maximum leaf length, the maximum leaf width, the fresh root quality, the fresh overground part quality, the dry root quality and the overground part quality of the seedlings in each experimental group after the irrigation treatment day 40; oven-drying in 70 deg.C oven to constant weight, and weighing aerial parts. After 6 weeks, it was found that each of the seedlings significantly grew vigorously in the test group to which the arbuscular mycorrhizal fungi (Glomus intraradics) bacterial solution, Azospirillum brasilense (Azospirillum brasilense) bacterial solution, and mixed bacterial solution was applied, as compared with the control group, and the results are shown in tables 2 to 9. As can be seen from the results in tables 2 to 9, the growth promoting effect on crops is better after the mixed microbial inoculum treatment compared with the single microbial inoculum treatment.
TABLE 2 biological traits of the differently treated Rice
Rice (Gudun No. 18) Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Plant height/cm 16.12 18.81 17.21 19.34
Root length/cm 10.23 12.32 11.5 12.37
Root number of 10.20 13.32 11.56 13.46
Maximum leaf length/cm 10.23 12.56 11.63 13.46
Maximum leaf width/cm 0.21 0.31 0.26 0.35
Root fresh mass/g 0.0310 0.0376 0.0279 0.0389
Fresh weight of overground part/g 0.0753 0.0805 0.0756 0.0885
Dry root mass/g 0.0034 0.0040 0.0012 0.0044
Dry mass of aerial parts per g 0.0060 0.0090 0.0087 0.0098
TABLE 3 biological traits of wheat after different treatments
Wheat (Zhoumai 16) Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Plant height/cm 16.45 18.61 17.12 18.39
Root length/cm 10.79 12.23 11.23 13.56
Root number of 10.5 13.45 11.15 13.45
Maximum leaf length/cm 9.87 12.65 11.45 12.39
Maximum leaf width/cm 0.23 0.30 0.23 0.35
Root fresh mass/g 0.032 0.0356 0.0275 0.0382
Fresh weight of overground part/g 0.0762 0.0815 0.0746 0.0885
Dry root mass/g 0.0035 0.0041 0.0015 0.0041
Dry mass of aerial parts per g 0.0081 0.0090 0.0077 0.1108
TABLE 4 biological traits of maize after different treatments
Corn (C)Lian mono 958) Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Plant height/cm 20.12 18.56 17.13 25.49
Root length/cm 15.79 12.12 11.56 18.56
Root number of 16.5 13.45 11.12 20.45
Maximum leaf length/cm 10.87 12.15 11.12 15.39
Maximum leaf width/cm 2.3 0.32 0.26 0.35
Root of fresh gingerMass/g 0.1210 0.1465 0.0975 15.20
Fresh weight of overground part/g 0.0762 0.0815 0.0745 0.0895
Dry root mass/g 0.01235 0.0135 0.0125 0.0141
Dry mass of aerial parts per g 0.0181 0.0291 0.0171 0.0308
TABLE 5 biological traits of Citrus after different treatments
Orange (pretty girl 38) Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Plant height/cm 10.23 12.56 11.13 13.15
Root length/cm 4.79 5.12 4.96 5.56
Root number of 3.5 5.45 11.12 5.5
Maximum leaf length/cm 3.87 4.15 3.12 4.39
Maximum leaf width/cm 2.23 3.22 2.26 3.35
Root fresh mass/g 0.0322 0.0465 0.0375 0.0381
Fresh weight of overground part/g 0.0761 0.0815 0.0745 0.0881
Dry root mass/g 0.0031 0.0040 0.0025 0.0041
Dry mass of aerial parts per g 0.1081 0.1291 0.1171 0.1307
TABLE 6 biological traits of soybeans after different treatments
Soybean (middle yellow 37) Control Arbuscular mycorrhizal fungi Brazil rubberAzospirillum azotofolium Mixed bacterial agent
Plant height/cm 20.45 24.56 19.13 25.39
Root length/cm 9.79 12.12 10.96 13.56
Root number of 11.5 13.45 11.12 14.45
Maximum leaf length/cm 5.87 6.15 5.12 7.39
Maximum leaf width/cm 3.23 3.56 2.26 4.35
Root fresh mass/g 0.042 0.0465 0.0375 0.0482
Fresh weight of overground part/g 0.0862 0.0815 0.0745 0.0985
Dry root mass/g 0.0025 0.0040 0.0025 0.0031
Dry mass of aerial parts per g 0.1031 0.1291 0.1171 0.1308
TABLE 7 biological traits of sorghum after different treatments
Figure BDA0003446068050000071
Figure BDA0003446068050000081
TABLE 8 biological Properties of sesame after different treatments
Sesame (airline black giant sesame) Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Plant height/cm 15.45 18.56 17.13 23.39
Root length/cm 8.79 12.12 9.96 13.56
Root number of 9.5 5.45 11.12 12.45
Maximum leaf length/cm 3.87 5.15 4.15 6.39
Maximum leaf width/cm 1.89 3.22 3.26 4.03
Root fresh mass/g 0.0420 0.0565 0.0495 0.0582
Fresh weight of overground part/g 0.0862 0.0825 0.0745 0.0985
Dry root mass/g 0.0045 0.0066 0.0056 0.0072
Dry mass of aerial parts per g 0.1082 0.1291 0.1171 0.1345
TABLE 9 biological traits of peanuts after different treatments
Peanut (golden crown 308) Control Arbuscular mycorrhizal fungi Azospirillum brasilense Mixed bacterial agent
Plant height/cm 17.45 18.57 17.13 20.39
Root length/cm 9.79 11.12 9.96 12.56
Root number of 12.5 15.45 13.12 16.45
Maximum leaf length/cm 3.87 5.15 4.15 6.39
Maximum leaf width/cm 2.23 3.22 3.26 4.35
Root fresh mass/g 0.0320 0.0565 0.0495 0.0682
Fresh weight of overground part/g 0.0862 0.1025 0.0945 0.1085
Dry root mass/g 0.0045 0.0066 0.0056 0.0071
Dry mass of aerial parts per g 0.1081 0.1291 0.1171 0.1308
Samples were taken at 7 days, 14 days, 21 days, 28 days and 35 days after the treatment with the microbial inoculum, chlorophyll of the leaves of each sample seedling was extracted with a mixed solution of acetone and alcohol in equal volume, the extract was used as a blank control, absorbances were measured at wavelengths of 645nm and 663nm, and the total chlorophyll content was calculated by the acetone method, the results are shown in fig. 1-8. It can be seen that the chlorophyll content of the crop seedlings treated by the mixed microbial inoculum is obviously higher compared with that of the control group and the single microbial inoculum treatment. Further shows that the mixed microbial inoculum has better growth promoting effect on crops than a single microbial inoculum.
(3) Determination of nitrogen fixation activity of mixed bacteria
The method for measuring nitrogen fixation activity is described in Williams et al (1987) and Wilson et al (2012). The acetylene reducer was sealed, and 5% air was evacuated, then an equal volume of acetylene was injected to bring the acetylene concentration in the reactor to 5%. Acetylene gas is generated into a gas storage bag by a self-made acetylene reactor through the reaction of calcium carbide and pure water. After the reactor is kept stand at 25 ℃ for 8h, the gas in the reactor is collected and injected into a gas collecting bottle, and the ethylene yield is analyzed by a gas chromatograph.
The amount of acetylene produced was measured by gas chromatography (Shanghai apparatus electric analysis). The chromatographic column is a GDX-50260/80mesh 3mm diameter packed column. The concentration of acetylene standard gas is 100 mg/L. Respectively sampling 1ml of arbuscular mycorrhizal fungi (Glomus intraradics) cultured by a single strain in a constant temperature oscillator cultured at 28.8-29.8 ℃ for 48h, Azospirillum brasilense (Azospirillum brasilense) cultured by a single strain, a mixed microbial inoculum of the arbuscular mycorrhizal fungi and the Azospirillum brasilense and a blank control (distilled water) to determine the acetylene reduction activity, cleaning a fresh nodule sample, removing excessive water, putting the nodule sample into a serum bottle with a plug, injecting a certain amount of acetylene gas, and oscillating the nodule sample at 28 ℃ for more than half an hour to determine the amount of generated ethylene. And after the measurement, putting the root nodules into an oven at 80 ℃, drying for 16h, and weighing the dried root nodules.
According to the formula (nmol/mg. h) ═ C2H4nmol/[ amount of mycoprotein (mg). times.reaction time (h)]In which C is2H4nmol＝1000×C2H4Volume (. mu.L). times.273 XP/[ 22.4 × (273+ t ℃ C.) times.760]Wherein P is the gas pressure (mm Hg) and t is the reaction temperature.
The results are shown in table 10, and it can be seen from table 10 that, compared with a single microbial inoculum, the nitrogen fixation performance of the mixed microbial inoculum formed by mixed culture of azospirillum brasilense and arbuscular mycorrhizal fungi is significantly better.
TABLE 10 Nitrogen fixation Performance of different inoculants
Test strains Nitrogenase activity (nmol C)2H4*g-1*h-1)
Azospirillum brasilense 41.56
Arbuscular mycorrhizal fungi 45.45
Mixed bacterial agent 81.45
Blank control 0
Example 2
The embodiment is to explore the growth promoting effect of the mixed microbial inoculum formed by mixed culture of different azotobacter and arbuscular mycorrhizal fungi on crops.
The test microbial inoculum is respectively as follows: 1. mixed bacteria agent formed by mixed culture of azospirillum brasilense and arbuscular mycorrhizal fungi; 2. a mixed microbial inoculum formed by mixed culture of azospirillum melitensis and arbuscular mycorrhizal fungi; 3. a mixed microbial inoculum formed by mixed culture of the iraq azospirillum and the arbuscular mycorrhizal fungi; 4. a mixed microbial inoculum formed by mixed culture of azospirillum salina and arbuscular mycorrhizal fungi; 5. a mixed microbial inoculum formed by mixed culture of azospirillum torvum and arbuscular mycorrhizal fungi; 6. the mixed microbial inoculum is formed by mixed culture of amazon azospirillum and arbuscular mycorrhizal fungi.
The mixed microbial inoculum is obtained by the following method:
the arbuscular mycorrhizal fungi (Glomus intraradics) and Azospirillum brasiliensis (Azospirillum brasilense) are inoculated in 2L of LB liquid culture medium, and mixed and cultured for 7d in a shady constant temperature oscillator under the conditions of 30 ℃ and 180r/min to prepare the mixed microbial inoculum.
The mixed microbial inoculum irrigation method comprises the following steps:
selecting rice (Gudun No. 18), wheat (Zhongmai No. 16), corn (Zhen mono 958), orange (Aiyue No. 38), soybean (Zhonghuang 37), sorghum (Jia dwarf 60), sesame (Aitian black giant glossy ganoderma), and peanut (golden crown 308). Respectively selecting healthy and plump 20 seeds, placing the seeds in a 70% ethanol solution for 8min, washing the seeds with sterile water for 3 times, placing the seeds in a 0.5% sodium hypochlorite solution for 10min, washing the seeds with sterile water for a plurality of times, spreading the seeds on wet filter paper, accelerating germination in a constant-temperature incubator at 28-30 ℃ in a dark environment, pouring 200ml of sterile water once every 2 days, and counting the germination rate.
After 14 days, seedlings with consistent growth vigor are selected for standby, the mixed microbial inoculum liquid diluted by 50 times of sterile water is taken respectively, the diluted microbial inoculum liquid is used for irrigating test seedlings every 7 days, meanwhile, the equivalent sterile water for irrigation is used as clear water for comparison, after the 40 th day of irrigation treatment, the dry matter increment of the overground part, the total quantity increment of the underground part and the total dry matter increment of the seedlings in each test group are measured, and the results are shown in table 11.
TABLE 11 growth promoting effect of different mixed inocula on crops
Figure BDA0003446068050000101
Figure BDA0003446068050000111
From the results in table 11, it is clear that the germination percentage of the crop seeds is significantly improved after the mixed microbial inoculum formed by mixed culture of azospirillum brasilense and arbuscular mycorrhizal fungi is treated, compared with the compound microbial inoculum formed by other azospirillum brasilense and arbuscular mycorrhizal fungi. And after the mixed microbial inoculum formed by mixed culture of azospirillum brasilense and arbuscular mycorrhizal fungi is treated, the increment of the dry matter of the overground part, the increment of the total amount of the underground part and the increment of the total dry matter of the crops are higher than the treatment effect of other mixed microbial inoculants.
In summary, compared with single microbial agents of arbuscular mycorrhizal fungi (Glomus intraradics) and Azospirillum brasilense (Azospirillum brasilense), the mixed microbial agent formed by mixed culture of the two has better growth promoting effect on crops and seed germination rate. Compared with a mixed microbial inoculum formed by mixed culture of other Azospirillum and arbuscular mycorrhizal fungi, the mixed microbial inoculum formed by mixed culture of Azospirillum brasilense and arbuscular mycorrhizal fungi has better effects of promoting seed germination and promoting crop growth. The mixed microbial inoculum promotes the growth of the upper part of the crop and is related to the content of chlorophyll, the mixed microbial inoculum promotes the growth of the upper part of the crop by improving the photosynthetic rate, enhancing the absorption and utilization of iron element nutrition by the crop and inducing the secretion of auxin, and the mixed microbial inoculum automatically fixes nitrogen biologically to provide nitrogen sources for the crops such as rice and the like, enhances the absorption of the nitrogen sources by the crop, induces the secretion of the auxin and promotes the growth of the crop.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims (7)

1. A microbial preparation for improving the capacity of resisting iron deficiency stress of crops is characterized in that the microbial preparation is a compound microbial inoculum formed by mixed culture of arbuscular mycorrhizal fungi and azospirillum brasilense.
2. The microbial preparation of claim 1, wherein the microbial preparation has an active count of 2.5 x 10 for arbuscular mycorrhizal fungi6～1.3×109(ii) a The number of viable bacteria of Azospirillum brasilense is 1.6 × 107～3.1×108。
3. The microbial preparation of claim 1, wherein the arbuscular mycorrhizal fungi has a preservation number of BGC BJ03, and the azospirillum brasilense has a preservation number of CGMCC 1.10379.
4. The microbial formulation of claim 1, wherein the crop comprises rice, wheat, corn, citrus, soy, sorghum, sesame, peanut.
5. Use of a microbial preparation according to claim 1 for crops against iron deficiency stress.
6. Use of a microbial preparation according to claim 1 for growth promotion of crops.
7. Use of the microbial preparation of claim 1 for breeding low-iron tolerant crops.
CN202111680982.XA 2021-12-30 2021-12-30 Microbial preparation for improving iron deficiency stress resistance of crops and application thereof Pending CN114231465A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112852671A (en) * 2021-02-04 2021-05-28 兴安盟莱绅生物农业有限公司 Microbial preparation for promoting corn growth

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103320368A (en) * 2013-07-11 2013-09-25 南京林业大学 Mycorrhiza helper bacterium Bacillus spp and its application in populus growth promotion
CN110063160A (en) * 2018-01-20 2019-07-30 杭州绿风生态环境建设集团有限公司 A kind of arbuscular mycorrhizal fungi screening technique conducive to garden plant growth
CN111713360A (en) * 2020-06-29 2020-09-29 重庆三峡学院 Corydalis tuber planting method

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103320368A (en) * 2013-07-11 2013-09-25 南京林业大学 Mycorrhiza helper bacterium Bacillus spp and its application in populus growth promotion
CN110063160A (en) * 2018-01-20 2019-07-30 杭州绿风生态环境建设集团有限公司 A kind of arbuscular mycorrhizal fungi screening technique conducive to garden plant growth
CN111713360A (en) * 2020-06-29 2020-09-29 重庆三峡学院 Corydalis tuber planting method

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
LIRA-SALDIVAR RH ET AL.: "Azospirillum brasilense and Glomus intraradices co-inoculation stimulates growth and yield of cherry tomato under shadehouse conditions", BIOFERTILIZATION IN CHERRY TOMATO, pages 133 - 138 *
MICHEL RUÍZ-SÁNCHEZ ET AL.: "Azospirillum and arbuscular mycorrhizal colonization enhance rice growth and physiological traits under well-watered and drought conditions", JOURNAL OF PLANT PHYSIOLOGY, pages 1 - 3 *
NEGI ET AL.: "Interaction between Glomus versiforme and Azospirillum brasilense in barley", CURRENT TRENDS IN MYCORRHIZAL *
SÁNCHEZ DE LA CRUZ ET AL.: "Glomus intraradices and Azospirillum brasilense in wheat grown under two regimes of soil humidity", UNIVERSIDAD Y CIENCIA TROPICO HUMEDO, pages 239 - 245 *
VEERASWAMY ET AL.: "Interaction effects of Glomus intraradices and Azospirillum lipoferum on Sorghum", INDIAN J. MICROBIOL, pages 305 - 308 *
YOURY PII ET AL.: "Plant-microorganism-soil interactions influence the Fe availability in the rhizosphere of cucumber plants", PLANT PHYSIOLOGY AND BIOCHEMISTRY *
张玉婷等: "丛枝菌根对防治植物缺铁性黄化病的作用机理与应用研究", 林业科技情报, vol. 52, no. 3 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112852671A (en) * 2021-02-04 2021-05-28 兴安盟莱绅生物农业有限公司 Microbial preparation for promoting corn growth
CN112852671B (en) * 2021-02-04 2023-10-20 兴安盟莱绅生物农业有限公司 Microbial preparation for promoting corn growth

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