CN108409411B - Biological seedling strengthening agent for enhancing stress resistance of rice and preparation method thereof - Google Patents

Biological seedling strengthening agent for enhancing stress resistance of rice and preparation method thereof Download PDF

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CN108409411B
CN108409411B CN201810275666.6A CN201810275666A CN108409411B CN 108409411 B CN108409411 B CN 108409411B CN 201810275666 A CN201810275666 A CN 201810275666A CN 108409411 B CN108409411 B CN 108409411B
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rice
biological
seedling
strengthening agent
vinasse
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CN108409411A (en
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李学辉
李亚华
周天惠
王灵敏
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Shaanxi Fengdan Baili Biotechnology Co ltd
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    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05GMIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05BPHOSPHATIC FERTILISERS
    • C05B1/00Superphosphates, i.e. fertilisers produced by reacting rock or bone phosphates with sulfuric or phosphoric acid in such amounts and concentrations as to yield solid products directly
    • C05B1/02Superphosphates

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Abstract

The invention discloses a biological seedling strengthening agent capable of enhancing stress resistance of rice and a preparation method of the seedling strengthening agent. Belongs to the technical field of agricultural production. The seedling strengthening agent comprises the following raw materials in parts by weight: 5-20 parts of biological agent, 5-40 parts of vinasse, 10-30 parts of urea, 2-10 parts of magnesium sulfate potassium, 6-15 parts of calcium superphosphate, 0.1-1 part of paclobutrazol and 0.1-1 part of naphthylacetic acid. The biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117. The biological seedling-strengthening agent is safer to use and does not generate phytotoxicity. Can enhance the low temperature resistance of the rice and avoid the influence of low temperature on the normal growth of the rice. Changing the vinasse into valuable, regulating the soil by utilizing the acidity of the vinasse, and providing nutrition for the rice by utilizing medium and trace elements contained in the vinasse.

Description

Biological seedling strengthening agent for enhancing stress resistance of rice and preparation method thereof
Technical Field
The invention discloses a biological seedling strengthening agent capable of enhancing stress resistance of rice and a preparation method of the seedling strengthening agent. Belongs to the technical field of agricultural production.
Background
Rice is a traditional crop in China and also a main variety for grain production in China. The growth condition of rice on the seedbed directly influences the growth and quality yield of the rice in the later period. The rice is an acid-loving crop, the pH value is 4.5-5.5, and the acid environment can effectively promote the growth of plants in the growth stage of seedlings, promote the root system growth and strengthen and protect the seedlings. The rice has various pathogenic bacteria in the soil in the seedling stage, and diseases such as damping-off, bacterial wilt and the like are easily induced by the rice. The seedling strengthening agent is generally used in the early growth stage of rice, the current rice seedling strengthening agent is a preparation for effectively promoting the growth of rice seedlings, and the preparation integrates sterilization, acid regulation, nutrition and chemical control into a whole according to the physiological and ecological characteristics of rice, and the bactericide, the chemical control agent, the acid regulating agent and the nutrient interact with each other to achieve complementary effects, so that a proper growth environment for the rice seedlings is created, and the aim of strengthening the seedlings is fulfilled.
The invention discloses a novel rice seedling strengthening agent with publication (bulletin) number CN101723746A, which comprises 1.5-2.5 parts by weight of humic acid, 1.5-2.5 parts by weight of wood vinegar and 0.5-1.5 parts by weight of organic fertilizer, wherein during implementation, the humic acid is 40g/L, the wood vinegar is 40g/L and the organic fertilizer is 20g/L, the organic fertilizer comprises nitrogen fertilizer, phosphate fertilizer and potash fertilizer, the content of the microbial preparation is 20-30ml/L, and the pH value is 3.5-4.5.
The invention of publication (announcement) No. CN105175159A, discloses a method for producing a biological matrix for strengthening rice seedlings in dry breeding and an application thereof, which comprises the following steps: firstly, mixing rice hulls and cow dung, adding mixed bacteria liquid, stacking and decomposing outdoors, adding lactic acid bacteria liquid, mixing uniformly, and stacking for 7-10 days; secondly, adding pure nitrogen, pure phosphorus, pure potassium and pure zinc and then uniformly mixing; thirdly, adding a binder, compressing into a sheet and drying. The biological matrix for dry-raised and strong seedling of rice is characterized by that every biological matrix is flatly spread in the rice seedling-raising disk, after the rice seedling is watered thoroughly, the rice seeds whose germination is hasten are uniformly spread in the rice seedling-raising disk, and placed in a plastic greenhouse to raise seedling. The invention can achieve the same effect of strengthening the seedlings as the currently applied rice seedling strengthening agent, and is safe to the environment and pollution-free.
The invention discloses a rice seedling-raising seedling-strengthening agent and a use method thereof with publication (announcement) number CN101113122A, and the agent comprises nutrient, fulvic acid, acid regulating agent, chelating agent, water-retaining agent, disinfectant, plant growth regulator, thermal element, mildew preventive and additive.
The invention of publication (announcement) No. CN107382503A, namely a rice seedling strengthening agent, discloses a rice seedling strengthening agent, which consists of the following raw materials: 50-60% of powder obtained by carrying out harmless treatment on the livestock and poultry manure; 20-22% of concentrated sulfuric acid; 8-10% of urea; 8-10% of calcium superphosphate; 4-8% of potassium sulfate; zinc sulfate in 5/1000 weight ratio of the matrix; 2/1000 parts by weight of the substrate; 1/1000 parts by weight of the matrix. The invention has the advantages that: compared with the conventional rice seedling strengthening agent, the rice seedling strengthening agent has the characteristics of integrating waste resource utilization, nutrition, acid regulation, disinfection, rooting and chemical control functions, can improve soil air permeability and improve the tillering rate of rice seedlings, so that the rice seedlings have long roots, multiple roots, heavy roots, thick stems, stiff stems and leaves, strong stress resistance, quick green return after rice transplanting, early maturity and high yield, and the rice seedling strengthening agent has no pollution to soil environment, and is a preferred production data for green rice production.
The invention discloses a rice seedling strengthening agent and a production process thereof with publication (bulletin) number CN1081433A, the rice seedling strengthening agent and the production process thereof use dry rice hulls as carriers instead of brown coal and the like, and the mixture ratio of various raw materials is as follows: 22% of dry rice husk, 40% of concentrated sulfuric acid, 4.894% of furfural residue, 28% of ammonium sulfate, 3.6% of diammonium phosphate, 1% of potassium sulfate, 0.4% of zinc sulfate, 0.07% of dicarboxylic acid and 0.036% of MET, which are mixed and stirred uniformly in sequence. Can be widely used for rice seedling raising beds and has excellent seedling strengthening effect.
The invention 'DSK Rice seedling strengthening agent' with publication (announcement) number CN1385406A discloses a DSK Rice seedling strengthening agent, a medicine and fertilizer mixed compound preparation, which has the characteristics of obvious disease resistance effect, long-lasting acid adjusting time and obvious fertility increasing effect. The effective components of the fertilizer are medicine, organic matter and fertilizer which are 1: 7. The traditional Chinese medicine consists of 20 percent of 3.3 percent of thiram and metalaxyl compound preparation and 80 percent of argil additive; the organic matter consists of wastes such as plant stalks, plants and the like, and the pH value is 5.0-6.0; the fertilizer consists of 70 percent of macroelements, 20 percent of secondary elements and 10 percent of trace elements, and the content of total available nutrients is more than or equal to 20 percent. The rice seedling strengthening agent can enhance the stress resistance of rice seedlings, has special effect on preventing and treating seedling blight at the seedling stage, has obvious effect on preventing and treating the bacterial wilt, and can be used for breeding the rice seedlings with large seedling age, good quality and strong yield in dry seedling culture of the rice. The invention is packaged in bags, so that the components of the invention keep unique characteristics and functions, and the invention is mixed at the moment when in use and is convenient to use.
The invention discloses a rice seedling strengthening agent with publication (announcement) number CN1176053A, belonging to a seedling strengthening, acid regulating and sterilizing preparation for fertilizing on a rice seedling bed and preventing and treating damping off. The fly ash is used as a carrier, contains N, P, K, Si, Ca, Zn, Fe and a bactericide, and is suitable for dry seedling or tray seedling in the field. The seedling strengthening agent has the advantages of strong cold resistance and drought resistance, good effects of preventing and controlling rhizoctonia rot and bacterial wilt, contribution to cultivating strong seedlings, quick rooting and seedling reviving and early tillering after transplanting, and creation of favorable conditions for obtaining high yield.
The strong seedlings are the basis for high yield of rice, and the quality of the seedlings is seriously affected if the seedlings encounter low-temperature cold weather during the seedling raising of the rice. Mainly characterized by slow growth of rice seedlings, serious disease occurrence, low seedling rate and the like. The conventional rice seedling strengthening agent has certain defects in the aspect of low temperature resistance of rice, particularly has certain influence on the growth of rice seedlings when the external temperature changes greatly, and can easily cause diseases such as bacterial wilt, damping off and the like due to low temperature. The conventional seedling strengthening agent contains a plurality of chemical agents, and has certain risks to plant phytotoxicity. The acid regulating agent is usually concentrated sulfuric acid, sulfur, etc., and has single function and certain destruction effect on soil.
Disclosure of Invention
The invention aims to provide a biological seedling strengthening agent for enhancing stress resistance of rice. On the basis of bacteriostasis, acid regulation, nutrition increase, chemical control and the like, the added biological bactericide with the function of enhancing the stress resistance (low temperature resistance) of rice can improve the rhizosphere environment of seeds and enhance the capability of resisting the external temperature change, and meanwhile, the strain also has good biocontrol effect and can inhibit the growth of harmful bacteria in the soil of a rice seedbed, thereby replacing a chemical bactericide. The invention uses the white spirit vinasse as the acid regulating agent, overcomes the potential safety hazard caused by adopting chemical agents in the traditional seedling strengthening agent, simultaneously, the vinasse contains nitrogen, phosphorus, potassium and various trace elements required by plant growth, can play a role in promoting the growth of rice in the seedling stage, and simultaneously reduces the addition of trace element fertilizers. Not only can reduce the cost and the damage to the soil, but also fully utilizes the vinasse and can change waste into valuable.
A biological seedling strengthening agent for enhancing stress resistance of rice comprises the following raw materials in parts by weight:
5-20 parts of biological agent, 5-40 parts of vinasse, 10-30 parts of urea, 2-10 parts of magnesium sulfate potassium, 6-15 parts of calcium superphosphate, 0.1-1 part of paclobutrazol and 0.1-1 part of naphthylacetic acid;
preferably, the mass fraction of N in the urea is 46%;
preferably, K in the potassium magnesium sulfate2The mass fraction of O is 50 percent;
preferably, P in said superphosphate2O5Is 18 percent;
preferably, the paclobutrazol is more than or equal to 15 percent;
preferably, the naphthylacetic acid in the naphthylacetic acid is more than or equal to 98 percent;
the invention also provides a biological agent for enhancing the stress resistance of rice, and the biological agent has the effects of preventing and treating common soil-borne diseases such as rice damping-off, rice bacterial wilt, rice seedling emergence diseases and the like; the biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117.
The streptomyces albus is separated from soil of a Shanhaiguan cherry garden in Hebei province. The Streptomyces albus provided by the invention is preserved in China general microbiological culture Collection center (CGMCC) in 12 months and 25 days in 2017, the preservation number is CGMCC No.15117, and the address is Beijing university Hokko No.1, North Cheng Xilu No. 3, China academy of sciences institute of microbiology.
Physiological and biochemical characteristics:
the following physiological and biochemical identifications were made for the strain RJ-36b with reference to Bergey's Manual of identification: streptomycin resistance test, gelatin liquefaction, milk peptonization, starch hydrolysis, nitrate reduction, hydrogen sulfide test, tolerance test to salt, the results are shown in table 1.
Table 1: physiological and biochemical identification results
Test of Results
Resistance to streptomycin
Liquefaction of gelatin +
Milk peptone +
Starch hydrolysis +
Nitrate reduction +
Hydrogen sulfide test
NaCl resistance test ≥10%<13%
Note: 1. positive +, 2, negative-
Carbon source utilization
The strains are respectively inoculated on different carbon source plates containing D-glucose, starch, sucrose, D-xylose, D-fructose, L-arabinose, D-galactose, L-rhamnose, D-mannitol and inositol, after the plates are cultured for 7 days at 32 ℃, the growth condition of the strains is observed, and the results are shown in Table 2.
The following results demonstrate that the carbon source utilization range of the strain is wide and that various sugars can be utilized.
Table 2: carbon source utilization results
Carbon source Results
D-glucose +
Sucrose +
D-fructose +
D-galactose +
D-mannitol +
Starch +
D-xylose +
L-arabinose
L-rhamnose
Inositol
Note: 1. use + 2, do not use-
The Streptomyces albus (Streptomyces albus) LM can grow at the temperature of 12-40 ℃, and the optimal growth temperature is 28 ℃; can produce various degradation enzymes such as protease, lipase, etc.
The Streptomyces albus (Streptomyces albus) LM grows in the Gao's No. one liquid culture medium with different pH values of 3-12, after being cultured for 3d at 50 ℃, the Streptomyces albus (Streptomyces albus) LM can grow well when being coated on a Gao's No. one flat plate and cultured for 3d at 50 ℃ in a test tube of the Gao's No. one liquid culture medium, and the results are shown in table 3. Both tests prove that the strain can tolerate a pH value within a range of 3-12, is wide and has strong acid-base tolerance.
Table 3: characteristics of bacterial colony
Culture medium Condition of the condition Aerial hypha Intrabasal hypha Soluble pigments
Gao's first agar culture medium Good taste White colour Yellow colour Is free of
Oat agar medium Good taste White colour Yellow and brown Is free of
Malt extract yeast extract agar culture medium Good taste White colour White colour Is free of
Asparagus agar culture medium Difference (D) White colour White colour Is free of
The invention adopts the residual white spirit vinasse of the brewing as the acid regulating substance, thereby avoiding the damage of concentrated sulfuric acid, sulfur and the like to the soil. The pH value of the seedbed soil is adjusted by utilizing the acidity of the vinasse, and the pH value of the vinasse is about 3. Meanwhile, the rice hulls of the vinasse contain nitrogen, phosphorus, potassium and various trace elements required by plant growth, and the vinasse is physically processed, so that various components in the vinasse can be fully released and utilized.
The invention adopts magnesium potassium sulfate, urea and calcium superphosphate to provide large amount of elements N, P, K as nutrient substances for seedlings.
The invention adopts paclobutrazol and naphthylacetic acid as chemical control agents.
The invention also provides a preparation method of the biological seedling strengthening agent, which comprises the following steps: the urea, the magnesium sulfate potassium, the calcium superphosphate and the ground vinasse are put into a stirrer to be stirred for 15min, and the biological agent, the paclobutrazol and the naphthylacetic acid are put into the mixed materials to be continuously stirred for 5 min. Mixing the mixture for later use.
The invention also provides a preparation method of the biological agent, which comprises the following steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109cfu/mL。
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 1-5% (v/v), culturing at 28 ℃ for 220r/min in a shake flask for 85-95h to obtain a seed liquid;
solid fermentation culture: inoculating the seed liquid into a solid fermentation culture medium with 50% of water content according to the inoculation volume mass ratio of 5-15% (v/m), and performing fermentation culture for 4-7 days at 28 ℃. Air drying at 20-30 deg.C, grinding with a grinder, collecting the microbial inoculum passing through 120 mesh sieve, and storing at room temperature in the shade to obtainTo streptomyces albus biological agent, the strain activity of which is not less than 8 multiplied by 1010cfu/g。
The Gao's I solid culture medium comprises the following components: soluble starch 20g/L, K2HPO4 0.5g/L,KNO3 1g/L,MgSO4.7H2O 0.5g/L,FeSO4.7H20.01g/L of O, 20g/L of agar and the balance of distilled water, wherein the pH value is 7.2-7.4;
the seed liquid culture medium comprises the following components: 10g/L of corn flour, 10g/L of peptone, 5g/L of yeast powder, 2g/L of dipotassium hydrogen phosphate, 0.3g/L of magnesium sulfate and the balance of distilled water, and the pH is natural.
The solid fermentation medium comprises the following components: 18g/L of rice hull, 45.7g/L of bran, 23g/L of corn flour, 12g/L of bean cake powder, 0.8g/L of calcium carbonate, 0.3g/L of dipotassium hydrogen phosphate, 0.2g/L of magnesium sulfate and the balance of distilled water, and the pH is natural.
The pH value of the white spirit vinasse is 2.8-3.5, and the water content is 60% -65%. Firstly, spreading the vinasse in a tray to a thickness of 1-1.5cm, and drying in an oven at 75 ℃ for 40-48 hours.
Further, the dried vinasse is put into a grinder to be ground, and the grinding fineness is about 200 meshes.
Advantageous effects
The seedling strengthening agent is added with a biological agent, so that the stress resistance of rice in a seedling stage is improved, and the interference of the normal growth of the rice due to the instability of external temperature is avoided. The microbial inoculum also has the function of biocontrol, can well inhibit the growth of harmful bacteria in soil, replaces a chemical agent for sterilization in a conventional seedling strengthening agent, and has safer effect and better effect. And the other point is that the vinasse is changed into valuable things, the acidity of the vinasse is utilized to regulate soil, and nitrogen, phosphorus, potassium and various trace elements contained in the vinasse are utilized to provide nutrition for rice.
The vinasse has the effect of adjusting the pH value of soil, reduces the pH value of the soil, is beneficial to the growth of rice, and can provide medium and trace elements for the growth of the rice. The streptomyces albus biological agent can increase the low temperature resistance of rice, is not interfered by low temperature, and simultaneously plays a role in preventing diseases. The potassium magnesium sulfate, the calcium superphosphate and the urea provide necessary nutrient elements for the growth of rice. Paclobutrazol can shorten internodes of rice and promote tillering of rice. The naphthylacetic acid can improve the ear forming rate, promote the plump seeds and enhance the lodging resistance.
Compared with chemical preparations, the biological seedling strengthening agent has unique characteristics, which are mainly shown in that 1, the biological seedling strengthening agent is safer to use than chemical-component seedling strengthening agents, and no phytotoxicity is generated. 2. The biological seedling strengthening agent can enhance the low temperature resistance of rice and avoid the influence of low temperature on the normal growth of the rice. 3. Changing the vinasse into valuable, regulating the soil by utilizing the acidity of the vinasse, and providing nutrition for the rice by utilizing medium and trace elements contained in the vinasse.
Detailed Description
Example 1:
a biological seedling strengthening agent for enhancing stress resistance of rice is composed of the following raw materials in parts by weight:
biological agent: 8 parts of vinasse: 20 parts of urea: 15 parts, magnesium sulfate potassium: 7 parts, calcium superphosphate: 10 parts, paclobutrazol: 0.1 part, naphthylacetic acid: 0.1 part;
the mass fraction of N in the urea is 46 percent;
k in the potassium magnesium sulfate2The mass fraction of O is 50 percent;
p in said superphosphate2O5Is 18 percent;
the paclobutrazol in the paclobutrazol is more than or equal to 15 percent;
the naphthylacetic acid in the naphthylacetic acid is more than or equal to 98 percent;
the biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117.
The preparation method of the biological seedling strengthening agent for enhancing the stress resistance of the rice comprises the following steps:
the urea, the magnesium sulfate potassium, the calcium superphosphate and the ground vinasse are put into a stirrer to be stirred for 15min, and then the streptomyces albus fungicide, the paclobutrazol and the naphthylacetic acid are put into the mixed materials to be continuously stirred for 5 min. Mixing the mixture for later use.
The preparation method of the biological agent comprises the following steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109cfu/mL。
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 3% (v/v), culturing at 28 ℃ for 180r/min for 90h in a shake flask to obtain seed liquid;
solid fermentation culture: inoculating the seed liquid into a solid fermentation culture medium with 50% of water content according to the inoculation volume mass ratio of 10% (v/m), and performing fermentation culture for 5.5 days in a koji tray at 28 ℃. Air drying at 25 deg.C, grinding with a grinder, collecting microbial inoculum passing through 120 mesh sieve, storing at room temperature in shade to obtain Streptomyces albus biological microbial inoculum with strain activity not less than 8 × 1010cfu/g。
The solid culture medium of Gao's synthetic No. one comprises the following components: soluble starch 20g/L, K2HPO4 0.5g/L,KNO31g/L,MgSO4.7H2O 0.5g/L,FeSO4.7H20.01g/L of O, 20g/L of agar and the balance of distilled water, wherein the pH value is 7.2-7.4;
the seed liquid culture medium comprises the following components: 10g/L of corn flour, 10g/L of peptone, 5g/L of yeast powder, 2g/L of dipotassium hydrogen phosphate, 0.3g/L of magnesium sulfate and the balance of distilled water, and the pH is natural.
The solid fermentation medium comprises the following components in percentage by mass: 18g/L of rice hull, 45.7g/L of bran, 23g/L of corn flour, 12g/L of bean cake powder, 0.8g/L of calcium carbonate, 0.3g/L of dipotassium hydrogen phosphate, 0.2g/L of magnesium sulfate and the balance of distilled water, and the pH is natural.
The pH value of the vinasse is 2.8-3.5, and the water content is 60% -65%. Firstly, spreading the vinasse in a tray to a thickness of 1-1.5cm, and drying in an oven at 75 ℃ for 44 hours.
And (3) putting the dried vinasse into a grinder for grinding, wherein the grinding fineness is about 200 meshes.
Comparative experiment:
laboratory experiments: the total number of treatments was 2, 3 replicates. Fully and uniformly mixing 1kg of rice seedling strengthening agent and 50kg of sieved soil powder, respectively loading the mixture into 3 seedling raising trays, and sowing 100 grains of rice in each tray, wherein the treatment 1 is the rice seedling strengthening agent in the embodiment 1; the rice seedling-strengthening agent sold in the market is fully and uniformly mixed with the screened soil, the mixture is subpackaged into 3 seedling-raising trays, and 100 grains of rice are sowed in each tray, namely treatment 2. The variety is pearl rice. The indoor culture temperature is 25 ℃ in the daytime and 16 ℃ at night.
After 3 days of emergence, both treatments were placed in a light incubator at the same time and a low temperature environment was set. The mixture is placed for 48 hours at the temperature of 22 ℃ in the day and 12 ℃ in the night. After 48h both treatments were removed simultaneously and allowed to continue growing in the chamber. The seedling rate and incidence of rice were examined when the rice grows to three leaves and one heart, and the results are shown in Table 4.
TABLE 4 seedling rate and incidence of rice in three leaves and one heart
Figure BDA0001613583350000071
Note that lower case letters indicate the significance of differences at the 5% level
After low temperature treatment, the survival rate of treatment 1 was still high at 95.1% and the incidence was low at 4.8%. The survival rate of the treatment 2 is much lower than that of the treatment 1, and is averagely 10.5 percent lower, and the morbidity is much higher than that of the treatment 1, and is averagely 10.6 percent higher. It can be shown that the rice seedling strengthening agent of the invention enhances the stress resistance of rice seedlings.
And (3) field test:
the 2016 test was completed in Harbin Fangxin county. The seedling raising test is carried out in a rice seedling raising shed. The field test is carried out in the test field of the farmer. The basic fertility is as follows: 18.7g/kg of organic matter, 1.121g/kg of total nitrogen, 13.3mg/kg of quick-acting phosphorus, 198mg/kg of quick-acting potassium and 6.6 of pH.
Preparing rice seedling soil: mixing 2kg of seedling strengthening agent with 100kg of sieved soil powder, loading into 100 seedling raising trays, and sowing rice seeds on the trays.
The rice variety to be tested is No. 12 rice with stalks.
The experiment was divided into 2 treatments in total, 3 replicates: treatment 1 is the rice seedling strengthening agent obtained in example 1 of the present invention; the treatment 2 is a rice seedling strengthening agent sold in the market.
Test results
The test shows that the quality of the seedlings is detected in the three-leaf one-heart period of the rice, the plant height, the leaf length, the leaf width, the number of roots, the leaf area, the overground fresh weight and the underground fresh weight of the rice are detected, and the results are shown in table 5. Treatment 1 was more effective than treatment 2.
TABLE 5 quality of rice seedlings in the three-leaf one-heart period
Figure BDA0001613583350000081
From the data in the above table, it can be seen that the leaf length, leaf width, number of roots, fresh weight on the ground and fresh weight under the ground of treatment 1 are all better than those of treatment 2, and the plant height of treatment 1 is lower than that of treatment 2, so that the plant is not easy to fall down. The indexes of the process 1 are better than those of the process 2.
The results of the tests on tillering, ear length, ear number, grain weight, etc. of the plants are shown in Table 6.
TABLE 6 influence of Rice seedling-strengthening agent on yield-constituting factors
Treatment of The number of acupoints (acupoints/m)2) Tiller number (number/point) Ear length (cm) Grain number of spike (number) Grain weight (g/acupoint)
Process 1 30 18.2 20.8 148.8 52.0
Treatment 2 30 16.8 20.0 127.6 47.2
From the data in Table 6, it can be seen that the tiller number, ear length, ear number and grain weight of treatment 1 were higher than those of treatment 2. Overall significant treatment 1 was better than treatment 2.
Through the two experiments, the rice seedbed seedling strengthening agent has a certain resistance effect on the stress resistance of the rice in the seedling stage, and the rice seedling strengthening agent can improve the quality of rice seedlings and the yield of the rice to a certain extent.
Example 2
A biological seedling strengthening agent for enhancing stress resistance of rice comprises the following raw materials in parts by weight:
15 parts of biological agent, 20 parts of vinasse, 20 parts of urea, 5 parts of magnesium sulfate potassium, 10 parts of calcium superphosphate, 1 part of paclobutrazol and 1 part of naphthylacetic acid;
the mass fraction of N in the urea is 46 percent;
k in the potassium magnesium sulfate2The mass fraction of O is 50 percent;
p in said superphosphate2O5Is 18 percent;
the paclobutrazol in the paclobutrazol is more than or equal to 15 percent;
the naphthylacetic acid in the naphthylacetic acid is more than or equal to 98 percent.
A biological agent for enhancing the stress resistance of rice seedlings and preventing and treating common soil-borne diseases such as rice damping-off, rice bacterial wilt, rice bakanae disease and the like; the biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117.
The preparation method of the biological seedling strengthening agent comprises the following steps: the urea, the magnesium sulfate potassium, the calcium superphosphate and the ground vinasse are put into a stirrer to be stirred for 15min, and the biological agent, the paclobutrazol and the naphthylacetic acid are put into the mixed materials to be continuously stirred for 5 min. Mixing the mixture for later use.
The preparation method of the biological agent comprises the following specific steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109cfu/mL。
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 1% (v/v), culturing at 28 ℃ for 150r/min in a shake flask for 95h to obtain seed liquid;
solid fermentation culture: inoculating the seed liquid into a solid fermentation culture medium with the water content of 50% according to the inoculation volume mass ratio of 15% (v/m), and performing fermentation culture for 7 days at 28 ℃. Air drying at 30 ℃, grinding by a grinder, collecting the microbial inoculum passing through a 120-mesh sieve, and storing in the shade at room temperature to obtain the streptomyces albus biological microbial inoculum, wherein the strain activity of the streptomyces albus biological microbial inoculum is not less than 8 multiplied by 1010cfu/g。
The composition of the Gao's No. I solid culture medium, the seed liquid culture medium and the solid fermentation culture medium is the same as that of the embodiment 1;
the pH value of the white spirit vinasse is 2.8-3.5, and the water content is 60% -65%. Firstly, spreading the vinasse in a tray to a thickness of 1-1.5cm, and drying in an oven at 75 ℃ for 40 hours.
And (3) putting the dried vinasse into a grinder for grinding, wherein the grinding fineness is about 200 meshes.
The experiment was carried out in a laboratory. The total number of treatments was 2, 3 replicates. Fully and uniformly mixing 1kg of rice seedling strengthening agent and 50kg of sieved soil powder, respectively loading the mixture into 3 seedling raising trays, and sowing 100 grains of rice in each tray, wherein the step 1 is treatment; the rice seedling-strengthening agent sold in the market is fully and uniformly mixed with the screened soil, the mixture is subpackaged into 3 seedling-raising trays, and 100 grains of rice are sowed in each tray, namely treatment 2. The variety is pearl rice. The indoor culture temperature is 25 ℃ in the daytime and 16 ℃ at night.
After 3 days of emergence, both treatments were placed in a light incubator at the same time and a low temperature environment was set. The mixture is placed for 48 hours at the temperature of 22 ℃ in the day and 12 ℃ in the night. After 48h both treatments were removed simultaneously and allowed to continue growing in the chamber. The seedling rate and incidence of rice were examined when the rice grows to three leaves and one heart, and the results are shown in Table 7.
TABLE 7 seedling rate and incidence of rice in three leaves and one heart
Treatment of The seedling rate% Incidence rate (damping off, bacterial wilt)
Process 1 93.6a 7.7
Treatment 2 85.1b 15.7
Note that lower case letters indicate the significance of differences at the 5% level
After low temperature treatment, the survival rate of treatment 1 was still 93.6% high, while the incidence rate was relatively low at 7.7%. The survival rate of the treatment 2 is much lower than that of the treatment 1, and is averagely 8.5 percent lower, and the incidence rate is much higher than that of the treatment 1, and is averagely 8.0 percent higher. It can be shown that the rice seedling strengthening agent of the invention enhances the stress resistance of rice seedlings.
Example 3
A biological seedling strengthening agent for enhancing stress resistance of rice comprises the following raw materials in parts by weight:
5 parts of biological agent, 5 parts of vinasse, 30 parts of urea, 10 parts of magnesium sulfate potassium, 15 parts of calcium superphosphate, 1 part of paclobutrazol and 1 part of naphthylacetic acid;
the mass fraction of N in the urea is 46 percent;
k in the potassium magnesium sulfate2The mass fraction of O is 50 percent;
p in said superphosphate2O5Is 18 percent;
the paclobutrazol in the paclobutrazol is more than or equal to 15 percent;
the naphthylacetic acid in the naphthylacetic acid is more than or equal to 98 percent.
A biological agent for enhancing the stress resistance of rice seedlings and preventing and treating common soil-borne diseases such as rice damping-off, rice bacterial wilt, rice bakanae disease and the like; the biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117.
The preparation method of the biological seedling strengthening agent is the same as that of the example 1.
A preparation method of a biological agent comprises the following specific steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109cfu/mL。
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 5% (v/v), performing shake flask culture at 28 ℃ for 95h at 220r/min to obtain a seed liquid;
solid fermentation culture: inoculating the seed liquid into a solid fermentation culture medium with 50% of water content according to the inoculation volume mass ratio of 5% (v/m), and performing fermentation culture for 7 days at 28 ℃. Air drying at 20 deg.C, grinding with a grinder, collecting microbial inoculum passing through 120 mesh sieve, storing at room temperature in shade to obtain Streptomyces albus biological microbial inoculum with strain activity not less than 8 × 1010cfu/g。
The composition of the solid culture medium, the seed liquid culture medium and the solid fermentation culture medium of Gao's I is the same as that of example 1.
The pH value of the white spirit vinasse is 2.8-3.5, and the water content is 60% -65%. Firstly, spreading the vinasse in a tray to a thickness of 1-1.5cm, and drying in an oven at 75 ℃ for 48 hours.
Further, the dried vinasse is put into a grinder to be ground, and the grinding fineness is about 200 meshes.
Comparative experiment:
the experiment was carried out in a laboratory. The total number of treatments was 2, 3 replicates. Fully and uniformly mixing 1kg of rice seedling strengthening agent and 50kg of sieved soil powder, respectively loading the mixture into 3 seedling raising trays, and sowing 100 grains of rice in each tray, wherein the step 1 is treatment; the rice seedling-strengthening agent sold in the market is fully and uniformly mixed with the screened soil, the mixture is subpackaged into 3 seedling-raising trays, and 100 grains of rice are sowed in each tray, namely treatment 2. The variety is pearl rice. The indoor culture temperature is 25 ℃ in the daytime and 16 ℃ at night.
After 3 days of emergence, both treatments were placed in a light incubator at the same time and a low temperature environment was set. The mixture is placed for 48 hours at the temperature of 22 ℃ in the day and 12 ℃ in the night. After 48h both treatments were removed simultaneously and allowed to continue growing in the chamber. The seedling rate and incidence of rice were examined when the rice grows to three leaves and one heart, and the results are shown in Table 8.
TABLE 8 seedling rate and incidence of rice in three leaves and one heart
Treatment of The seedling rate% Incidence rate (damping off, bacterial wilt)
Process 1 92.7a 7.4
Treatment 2 83.8b 17.2
Note that lower case letters indicate the significance of differences at the 5% level
After low temperature treatment, the survival rate of treatment 1 was still high at 92.7% and the incidence rate was relatively low at 7.4%. The survival rate of the treatment 2 is much lower than that of the treatment 1, and is averagely 8.9 percent lower, and the morbidity is much higher than that of the treatment 1, and is averagely 9.8 percent higher. It can be shown that the rice seedling strengthening agent of the invention enhances the stress resistance of rice seedlings.
Example 4
A biological seedling strengthening agent for enhancing stress resistance of rice comprises the following raw materials in parts by weight:
20 parts of biological agent, 30 parts of vinasse, 20 parts of urea, 6 parts of magnesium sulfate potassium, 6 parts of calcium superphosphate, 0.6 part of paclobutrazol and 1 part of naphthylacetic acid;
the mass fraction of N in the urea is 46 percent;
k in the potassium magnesium sulfate2The mass fraction of O is 50 percent;
p in said superphosphate2O5Is 18 percent;
the paclobutrazol in the paclobutrazol is more than or equal to 15 percent;
the naphthylacetic acid in the naphthylacetic acid is more than or equal to 98 percent.
A biological agent for enhancing the stress resistance of rice seedlings and preventing and treating common soil-borne diseases such as rice damping-off, rice bacterial wilt, rice bakanae disease and the like; the biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117.
The preparation method of the biological seedling strengthening agent is the same as that of the example 1.
A preparation method of a biological agent comprises the following specific steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109cfu/mL。
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 4% (v/v), culturing at 28 ℃ for 160r/min for 88h in a shake flask to obtain seed liquid;
solid fermentation culture: inoculating the seed liquid into a solid fermentation culture medium with 50% of water content according to the inoculation volume mass ratio of 8% (v/m), and performing fermentation culture for 5 days in a koji tray at 28 ℃. Air drying at 22 deg.C, grinding with a grinder, collecting microbial inoculum passing through 120 mesh sieve, storing at room temperature in shade to obtain Streptomyces albus biological microbial inoculum with strain activity not less than 8 × 1010cfu/g。
The pH value of the white spirit vinasse is 2.8-3.5, and the water content is 60% -65%. Firstly, spreading the vinasse in a tray to a thickness of 1-1.5cm, and drying in an oven at 75 ℃ for 42 hours.
Comparative experiments were performed in the laboratory. The total number of treatments was 2, 3 replicates. Fully and uniformly mixing 1kg of rice seedling strengthening agent and 50kg of sieved soil powder, respectively loading the mixture into 3 seedling raising trays, and sowing 100 grains of rice in each tray, wherein the step 1 is treatment; the rice seedling-strengthening agent sold in the market is fully and uniformly mixed with the screened soil, the mixture is subpackaged into 3 seedling-raising trays, and 100 grains of rice are sowed in each tray, namely treatment 2. The variety is pearl rice. The indoor culture temperature is 25 ℃ in the daytime and 16 ℃ at night.
After 3 days of emergence, both treatments were placed in a light incubator at the same time and a low temperature environment was set. The mixture is placed for 48 hours at the temperature of 22 ℃ in the day and 12 ℃ in the night. After 48h both treatments were removed simultaneously and allowed to continue growing in the chamber. The seedling rate and incidence of rice were examined when the rice grows to three leaves and one heart, and the results are shown in Table 9.
TABLE 9 seedling rate and incidence of rice in Trifolium and Heart
Treatment of The seedling rate% Incidence rate (damping off, bacterial wilt)
Process 1 90.1a 10.5
Treatment 2 86.8b 15.6
Note that lower case letters indicate the significance of differences at the 5% level
After low temperature treatment, the survival rate of treatment 1 was still high at 90.1% and the incidence rate was relatively low at 10.5%. The survival rate of the treatment 2 is much lower than that of the treatment 1, and is averagely lower by 3.3 percent and averagely higher by 5.1 percent. It can be shown that the rice seedling strengthening agent of the invention enhances the stress resistance of rice seedlings.
Example 5
A biological seedling strengthening agent for enhancing stress resistance of rice comprises the following raw materials in parts by weight:
5 parts of biological agent, 40 parts of vinasse, 10 parts of urea, 10 parts of magnesium sulfate potassium, 15 parts of calcium superphosphate, 0.5 part of paclobutrazol and 0.5 part of naphthylacetic acid;
the mass fraction of N in the urea is 46 percent;
k in the potassium magnesium sulfate2The mass fraction of O is 50 percent;
p in said superphosphate2O5Is 18 percent;
the paclobutrazol in the paclobutrazol is more than or equal to 15 percent;
the naphthylacetic acid in the naphthylacetic acid is more than or equal to 98 percent.
A biological agent for enhancing the stress resistance of rice seedlings and preventing and treating common soil-borne diseases such as rice damping-off, rice bacterial wilt, rice bakanae disease and the like; the biological agent strain is Streptomyces albus (Streptomyces albus) with the preservation number of CGMCC No. 15117.
The preparation method of the biological seedling strengthening agent is the same as that of the example 1.
A preparation method of a biological agent comprises the following specific steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109cfu/mL。
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 4% (v/v), culturing at 28 ℃ for 210r/min in a shake flask for 92h to obtain a seed liquid;
solid fermentation culture: inoculating the seed liquid into a solid fermentation culture medium with the water content of 50% according to the inoculation volume mass ratio of 12% (v/m), and performing fermentation culture for 6 days at 28 ℃. Air drying at 28 deg.C, grinding with a grinder, collecting microbial inoculum passing through 120 mesh sieve, storing at room temperature in shade to obtain Streptomyces albus biological microbial inoculum with strain activity not less than 8 × 1010cfu/g。
The composition of the solid culture medium, the seed liquid culture medium and the solid fermentation culture medium of Gao's I is the same as that of example 1.
The pH value of the white spirit vinasse is 2.8-3.5, and the water content is 60% -65%. Firstly, spreading the vinasse in a tray to a thickness of 1-1.5cm, and drying in an oven at 75 ℃ for 46 hours.
And (3) putting the dried vinasse into a grinder for grinding, wherein the grinding fineness is about 200 meshes.
Examples of the experiments
Comparative experiments were performed in the laboratory. The total number of treatments was 2, 3 replicates. Fully and uniformly mixing 1kg of rice seedling strengthening agent and 50kg of sieved soil powder, respectively loading the mixture into 3 seedling raising trays, and sowing 100 grains of rice in each tray, wherein the step 1 is treatment; the rice seedling-strengthening agent sold in the market is fully and uniformly mixed with the screened soil, the mixture is subpackaged into 3 seedling-raising trays, and 100 grains of rice are sowed in each tray, namely treatment 2. The variety is pearl rice. The indoor culture temperature is 25 ℃ in the daytime and 16 ℃ at night.
After 3 days of emergence, both treatments were placed in a light incubator at the same time and a low temperature environment was set. The mixture is placed for 48 hours at the temperature of 22 ℃ in the day and 12 ℃ in the night. After 48h both treatments were removed simultaneously and allowed to continue growing in the chamber. The seedling rate and incidence of rice were examined when the rice grows to three leaves and one heart, and the results are shown in Table 10.
TABLE 10 seedling and incidence rates of rice in three leaves and one heart
Figure BDA0001613583350000131
Figure BDA0001613583350000141
Note that lower case letters indicate the significance of differences at the 5% level
After low temperature treatment, the survival rate of treatment 1 was still high at 92.7% and the incidence rate was relatively low at 6.8%. The survival rate of the treatment 2 is much lower than that of the treatment 1, and is averagely lower by 9.8 percent and averagely higher by 8.6 percent. It can be shown that the rice seedling strengthening agent of the invention enhances the stress resistance of rice seedlings.

Claims (8)

1. A biological seedling strengthening agent for enhancing stress resistance of rice is characterized by comprising the following raw materials in parts by weight:
5-20 parts of biological agent, 5-40 parts of vinasse, 10-30 parts of urea, 2-10 parts of magnesium sulfate potassium, 6-15 parts of calcium superphosphate, 0.1-1 part of paclobutrazol and 0.1-1 part of naphthylacetic acid;
the biological agent strain is streptomyces albus (S.albus)Streptomyces albs) The preservation number is CGMCC No. 15117;
the biological seedling strengthening agent is prepared by the method comprising the following steps:
firstly, putting urea, magnesium sulfate potassium, calcium superphosphate and ground vinasse into a stirrer for stirring, and after stirring for 15min, putting the biological agent, the paclobutrazol and the naphthylacetic acid into the mixed material for continuously stirring for 5 min; mixing the mixture for later use.
2. The biological seedling-strengthening agent for enhancing stress resistance of rice as claimed in claim 1, wherein the vinasse is white spirit vinasse, the pH is 2.8-3.5, and the water content is 60% -65%.
3. The biological seedling-strengthening agent for enhancing stress resistance of rice as claimed in claim 2, wherein the preparation method of the vinasse comprises: spreading the distiller's grains in a tray with thickness of 1-1.5cm, and drying in oven at 75 deg.C for 40-48 hr.
4. The biological seedling-strengthening agent for enhancing stress resistance of rice as claimed in claim 3, wherein the preparation method of the distiller's grains further comprises: and (4) putting the dried vinasse into a grinder for grinding, wherein the grinding fineness is 200 meshes.
5. The biological seedling-strengthening agent for enhancing stress resistance of rice as claimed in claim 1, wherein the mass fraction of N in the urea is 46%; k in the potassium magnesium sulfate2The mass fraction of O is 50 percent; p in said superphosphate2O5Is 18 percent.
6. The biological seedling-strengthening agent as set forth in claim 1, the preparation method of said biological agent comprises the following steps:
preparing spore suspension: adding sterile water to the inclined plane of a Gao's first solid culture medium in which the white streptomycete is stored, further scraping spores, shaking and uniformly mixing to obtain a spore suspension, wherein the thallus concentration of the spore suspension is 108-109 cfu/mL;
Seed liquid culture: inoculating the spore suspension into a seed liquid culture medium according to the inoculation amount of 1-5% (v/v), culturing at 28 ℃ for 220r/min in a shake flask for 85-95h to obtain a seed liquid;
solid fermentation culture: inoculating the seed solution into a solid fermentation culture medium with 50% of water content according to the inoculation volume mass ratio of 5-15% (v/m), and performing fermentation culture on a koji tray at 28 ℃ for 4-7 days; air drying at 20-30 deg.C, grinding with a grinder, collecting microbial inoculum passing through 120 mesh sieve, storing at room temperature in shade to obtain Streptomyces albus biological microbial inoculum with strain activity not less than 8 × 1010 cfu /g。
7. The seedling-strengthening agent as claimed in claim 6, a method for preparing the biological agent,
the Gao's I solid culture medium comprises the following components: soluble starch 20g/L, K2HPO4 0.5 g/L,KNO3 1 g/L,MgSO4 .7H2O 0.5 g/L,FeSO4 .7H20.01g/L of O, 20g/L of agar and the balance of distilled water, wherein the pH value is 7.2-7.4;
the seed liquid culture medium comprises the following components: 10g/L of corn flour, 10g/L of peptone, 5g/L of yeast powder, 2g/L of dipotassium hydrogen phosphate, 0.3g/L of magnesium sulfate and the balance of distilled water, wherein the pH value is natural;
the solid fermentation medium comprises the following components: 18g/L of rice hull, 45.7g/L of bran, 23g/L of corn flour, 12g/L of bean cake powder, 0.8g/L of calcium carbonate, 0.3g/L of dipotassium hydrogen phosphate, 0.2g/L of magnesium sulfate and the balance of distilled water, and the pH is natural.
8. A process for preparing a biological seedling-strengthening agent as claimed in any one of claims 1 to 7, comprising the steps of:
firstly, putting urea, magnesium sulfate potassium, calcium superphosphate and ground vinasse into a stirrer for stirring, and after stirring for 15min, putting the biological agent, the paclobutrazol and the naphthylacetic acid into the mixed material for continuously stirring for 5 min; mixing the mixture for later use.
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