CN104774106A - Preparation method and application of chaetomium globosum biological bacterial fertilizer granules - Google Patents
Preparation method and application of chaetomium globosum biological bacterial fertilizer granules Download PDFInfo
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Abstract
The invention discloses a preparation method and application of chaetomium globosum biological bacterial fertilizer granules. The preparation method of the chaetomium globosum biological bacterial fertilizer granules of the invention comprises the following steps: 1) uniformly mixing chaetomium globosum ND35 CGMCC No. 4136 spores, humic acid, chitosan and auxiliaries to obtain raw materials; and 2) granulating the raw materials to obtain a biological bacterial fertilizer, wherein the auxiliaries are xanthan gum, sodium carboxymethylcellulose and calcium chloride. Tests prove that the biological bacterial fertilizer granules can be used for not only preventing and treating a variety of plant diseases, but also realizing a good effect of promoting growth, significantly improving the biomass and the yield of crops, and realizing good economic benefits; and furthermore, the biological bacterial fertilizer granules are simple to use and can avoid flying of micro-powder and reduce environmental pollution. The preparation method has very great feasibility in the aspects of theory and market demands.
Description
Technical field
The present invention relates to a kind of preparation method and application of chaetomium globosum bio-bacterial manure granule, particularly relating to the thecaspore of chaetomium globosum (Chaetomium globosum) ND35 bacterial strain is the preparation method of the bio-feritlizer granule of activeconstituents, belongs to field of agricultural microorganism.
Background technology
Biocontrol fungi for Biological Control Test mainly contains: Trichoderma (Trichoderma), chaetomium (Chaetomium), mycorrhizal fungi (Mycorrhiza fungi), sticky mould (Gliocladium), Paecilomyces lilacinus (Paecilomyces lilacinus), coniothyrium minitans (Coniothyrium minitans) and ferment (king advocate constitution, 2005) such as female bacterium (Yeasts).But commercial prod can be made into be applied to and only have a little part in agriculture production.At present, being Trichoderma agriculturally applying the most successfully biocontrol strain, now having made various biocontrol fungicide and being applied in plant pest management; Coniothyrium minitans, by the commercialization of minority European countries, is made into wetting properties particle (Paulitz T C, 2001; McQuilken M P, 1995).
Chaetomium globosum (C.globosum) ND35 bacterial strain is the plant endogenesis epiphyte that a strain is separated from healthy Cortex Populi Tomentosae, belongs to Ascomycota, gang pyrenomycetes, Sphaerials, Hei Bao shell section, Chaetomium.The antimicrobial spectrum of chaetomium globosum ND35 bacterial strain is very wide, obvious antagonistic action can be had to the mycelial growth of cucumber fusarium axysporum, bitter gourd wilt bacterium, withered germ of water-melon, bacterium of downy mildew of cucumber, Botryosphaeria berengeriana f. sp, apple rot pathogen, Cytospora chrysosperma and Dothiorella gregaria and dry thread Pyrenomycetes, the antibiotics with bacteriostatic activity can be produced in metabolic process, and the spore germination of cucumber fusarium axysporum, bitter gourd wilt bacterium, Botryosphaeria berengeriana f. sp and Botryosphaeria dothidea bacterium etc. can be suppressed.Observe under transmission electron microscope, find that chaetomium globosum ND35 contacts with dry thread Pyrenomycetes and after there is hyperparasitism, after chaetomium globosum ND35 mycelia parasitism to pathogenic bacteria mycelia inside, chitinase and β-1 is created in Interaction, 3-dextranase, can make that its cellularstructure gets muddled, cell wall degradation.
Existing test-results shows, chaetomium globosum ND35 bacterial strain can be grown effectively surely in vegetable plant if balsam pear, cucumber, tomato, Kidney bean and commodity trees are if willow, walnut, Chinese chestnut and field crop are as in the plant of sweet potato, wheat, potato, tobacco, ginger, and has obvious promoter action to the growth of cucumber, balsam pear, eggplant and tomato; The generation of cucumber at seedling stage samping off effectively can be prevented and treated under condition of pot.After inoculation chaetomium globosum ND35 bacterial strain, the side root of willow and breast diameter growth are had to certain promoter action and induce it to produce resistance to rotten pathogenic bacteria and leaf rust bacterium, tomato and Kidney bean also can be induced to produce systemic disease resistance to ash arrhizus bacteria; Also can effectively prevent and treat Kidney bean damping-off.The growth of chaetomium globosum ND35 bacterial strain on Chinese chestnut side root and capillary root has positive impact, can increase the quantity of Chinese chestnut inflorescence, side root and capillary radical amount, and in the later stage, Stem nematode and one-level branch amount also produce obvious growth-promoting functions over the ground; Growth-promoting functions is revealed to the growth table of willow cuttage seeding, Kidney bean, cucumber, tobacco, ginger.By scanning electron microscopic observation chaetomium globosum ND35 bacterial strain, surely discovery is grown to infecting of willow tissue cultured seedling: after thecaspore is sprouted, the mycelia produced focuses mostly in root hair, but do not infect root hair, invade mainly through the gap between root, stem's superficial cell, or form appressorium or mycelia foot in root table face, directly invade from epidermic cell by being formed to infect to follow closely, mainly blade interior is invaded from pore in leaf portion, the mycelia invading root mainly grows surely in epidermic cell, exodermis cell and intercellular substance, does not enter endodermis and vascular tissue.
The environmental compatibility of chaetomium globosum ND35 bacterial strain is very strong, can adapt to severe saline and alkaline condition.In indoor, substratum based on PDA substratum, research potential of hydrogen and the mycelial growth of salt concentration on chaetomium globosum ND35 and the impact of spore germination, found that: within the scope of pH value 3-12, the mycelia of chaetomium globosum ND35 can normal growth, at the Ca containing 1%-10%
2+, K
+, N and P substratum in, the mycelia of chaetomium globosum ND35 still can normal growth, and growth velocity is not subject to the impact of potential of hydrogen and salinity; The thecaspore of chaetomium globosum ND35 is in the PDB nutrient solution within the scope of 3-12 in pH value, and still can normally sprout, the sprouting of spore is not affected.
Granule as one tradition bacterial manure preparation, have easy to use, safe, reduce the advantages such as environmental pollution.When granule can be avoided using, particulate flies upward, contaminate environment, personnel suck the problems such as micro mist; The release rate of effective constituent can be controlled.Delay the lasting period; The place of needs can be arrived during use exactly, be not attached on the cauline leaf of plant.
Summary of the invention
An object of the present invention is to provide a kind of method preparing bio-bacterial manure.
The method preparing bio-bacterial manure provided by the invention comprises the steps: the spore of chaetomium globosum (Chaetomiumglobosum) ND35CGMCC No.4136, humic acids, chitosan and auxiliary agent to mix, and obtains bio-bacterial manure.
In aforesaid method, described auxiliary agent is xanthan gum, Xylo-Mucine and calcium chloride.
In aforesaid method, the proportioning of the spore of described chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136, described humic acids, described chitosan, described xanthan gum, described Xylo-Mucine and described calcium chloride is 3 × 10
11cfu:(265-285) g:(5-20) g:(5-20) g:(15-35) g:(30-50) g.
In aforesaid method, the spore of described chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136 is for being less than 200 object spore powder.
In aforesaid method, the spore powder of described chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136 is the product obtained that fermented in culture material by chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136 spore.
In aforesaid method, described fermentation is with (1-3) × 10 by described spore
8the amount of cfu/ml is seeded in the culture material containing edible fungus bran; Described spore is specifically with 2 × 10
8the amount of cfu/ml is seeded in described containing in the culture material of edible fungus bran.
In aforesaid method, the described culture material containing edible fungus bran is edible fungus bran and nutrient solution are mixed; The mixing of potato, glucose and water obtains by described nutrient solution; Described potato, described glucose and described water proportioning are 70-100) g ︰ (7-10) g ︰ 1000ml; The proportioning of described potato, described glucose and described water is specially 70g ︰ 7g ︰ 1000ml.
In aforesaid method, the mass ratio of described edible fungus bran and nutrient solution is 1 ︰ (2-4); The mass ratio of described edible fungus bran and nutrient solution is specially 1 ︰ 2.
In aforesaid method, described fermentation time is 25 ~ 30 days; Described leavening temperature is 20-30 DEG C.
In aforesaid method, described fermentation time is specially 28 days; Described leavening temperature is specially 25 DEG C.
In aforesaid method, when being also included in mixing after the spore of described chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136, described humic acids, described chitosan, described xanthan gum, described Xylo-Mucine and described calcium chloride being mixed, add the step of filler.
In aforesaid method, after the spore of described chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136, described humic acids, described chitosan, described xanthan gum, described Xylo-Mucine and described calcium chloride being mixed, also comprise the step of described raw material being carried out granulation, obtain bio-bacterial manure.
In aforesaid method, described filler is diatomite.
Another object of the present invention is to provide the bio-bacterial manure prepared by aforesaid method.
Last object of the present invention is to provide the application of above-mentioned bio-bacterial manure in following (1)-(14) at least one:
(1) plant height of plant is increased;
(2) Chlorophyll-a Content of plant is improved;
(3) content of chlorophyll b of plant is improved;
(4) total chlorophyll content of plant is improved;
(5) carotenoid content of plant is improved;
(6) reducing sugar content of plant leaf is improved;
(7) proline content of plant leaf is improved;
(8) yield per plant is improved;
(9) leaf increasing plant is wide;
(10) leaf increasing plant is long;
(11) leading thread of plant is increased;
(12) growth in plant seedling stage is promoted;
(13) plant Blooming result is impelled;
(14) plant seedling sudden fall ill is prevented and treated.
In above-mentioned application, described plant is specially walnut and/or cucumber and/or purple potato and/or willow.
Above-mentioned chaetomium globosum (Chaetomium globosum) ND35CGMCC No.4136 or its spore or its tunning are as bio-bacterial manure additive or the application in the product of preparation bio-bacterial manure additive.
Shown in following (1)-(4) of advantage of the present invention:
(1) sexual spore that the spore that the present invention uses is chaetomium, relative to the asexual spore of the fungi such as Trichoderma and non-pathogenic sickle-like bacteria, sexual spore has more advantage in resistance.
(2) the ferment starting material of chaetomium globosum ND35 of the present invention are waste mushroom leftovers after needle mushroom fruiting, the advantages such as this raw material has low, the easy acquisition of cost, cultural colony is excellent, physical behavior is good.
(3) with the addition of humic acids and chitosan in Granular formulations of the present invention.Humic acids not only has fertilizer synergistic, improves soil, stimulates plant growth, improves the functions such as agricultural product quality, can also provide a buffer environment for chaetomium globosum spore, improves the viability of spore; Chitosan provides nutrition can not only to the sprouting of spore, and chaetomium globosum can be induced to produce chitinase, strengthens chaetomium globosum to the antagonistic activity of various plants pathogenic bacteria.
(4) granule adds thickening material, binding agent and helps the auxiliary agents such as disintegrating agent in making processes, make granule in a wetted condition can disintegration rapidly, promote spore rapid dispersion in soil, can spore germination be made as early as possible, plant be infected and surely grows and set up symbiotic relationship with plant.
The present invention adopts the spore powder of simple production technology and production unit fermentative production chaetomium globosum ND35; then after pulverizing, sieving, add thickening material, the binding agent of different ratios, help disintegrating agent etc.; and through disc granulation or extruder grain, produce cheap bio-bacterial manure granule.Show by experiment: this granule can not only prevent and treat plurality of plant diseases, but also have good growth-promoting functions to crop, growth-promoting functions is mainly manifested in the raising of biomass and the raising of output aspect, has good economic benefit; And this granule uses simple, avoids the micro mist flown upward, decreases environmental pollution, have good ecological benefits.Chaetomium globosum bio-bacterial manure granule of the present invention has very large feasibility in theory with in the market requirement.
Accompanying drawing explanation
Fig. 1 is chaetomium globosum bio-bacterial manure granule.
Fig. 2 is that chaetomium globosum bio-bacterial manure granule is on the impact of willow growth of cuttage seedlings.
Fig. 3 is the impact that chaetomium globosum bio-bacterial manure granule grows purple potato.
Fig. 4 is that chaetomium globosum bio-bacterial manure granule is on the impact of the growth of cucumber seedling.
Fig. 5 is that chaetomium globosum bio-bacterial manure granule is to the prevention effect of cucumber at seedling stage samping off disease.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
The Classification And Nomenclature of chaetomium globosum of the present invention (Chaetomium globosum) ND35 bacterial strain is chaetomium globosum, and preserving number is CGMCC No.4136, is be disclosed in CN 102311925B at publication number.
PDA culture medium prescription: potato 200g, glucose 20g, agar 17g, tap water 1000ml.
The preparation method of PDA substratum: take peeled potatoes 200g, puts into after 1000ml tap water boils 30min, filters out potato residue with double gauze, put into agar 17g in filtrate, heating makes it melt, then adds glucose 20g, after dissolving, add tap water and be settled to 1000ml.Then in packing triangular flask, after 121 DEG C of sterilizing 25min, for subsequent use.
PDB culture medium prescription: potato 200g, glucose 20g, tap water 1000ml.The preparation method of PDB substratum is basic identical with the preparation method of PDA substratum, does not just add agar, and preparation becomes liquid nutrient medium.
Humic acids is the product of Shandong Henkel biotechnology company limited;
Chitosan is the product of taste bio tech ltd, Hebei hundred;
Xanthan gum is the product that enrichment chemical product company limited is added in Zhengzhou;
Xylo-Mucine (CMC) is the product of Tianjin Kai Tong chemical reagent company limited;
Calcium chloride is the product of Tianjin Kai Tong chemical reagent company limited.
biological organic fertilizer take S. ahygroscopicus as main active ingredient, and living bacteria count is 0.2 hundred million cfu/g, is the product of Liaoning Wkioc Bioengineering Co., Ltd..
The preparation method of embodiment 1, chaetomium globosum bio-bacterial manure granule
One, the preparation of chaetomium globosum bio-bacterial manure granule raw materials for production
1, raw materials
With the waste mushroom leftover after the needle mushroom fruiting such as (flat mushroom, mushroom, needle mushroom edible mushrooms) for starting material, being pulverized, being dried the rear raw material as cultivating chaetomium globosum.
2, nutritive medium is prepared
The compound method of nutritive medium boils maintenance certain hour (40min) for a certain amount of potato, glucose and water (potato 70g, glucose 7g, tap water 1000ml) being put into pot, is mixed with nutritive medium.
3, the preparation of culture material
The pulverizing that step 1 is prepared dry after bacterium chaff and the nutritive medium prepared of step 2 be the ratio stirring and evenly mixing (humid control about 55%, natural pH) of 1:2 according to mass ratio, obtain culture material; Culture material is filled Polypropylene Bag.
4, the sterilizing of culture material
Culture material step 3 prepared, at 121 DEG C of autoclave sterilization 100min, obtains the culture material after sterilizing for subsequent use.
5, the preparation of inoculation liquid
Chaetomium globosum ND35 inoculation cultivated on PDA substratum, after product spore, scraping spore adds in PDB substratum, and (concentration is 2 × 10 to obtain spore suspension
8cfu/ml) i.e. inoculation liquid.
6, inoculation and fermentation culture
By in the culture material after sterilizing in the spore suspension inoculation step 4 for preparing in step 5, spore suspension is with 2 × 10
8the amount of cfu/ml is seeded in described containing in the culture material of edible fungus bran.Under 25 DEG C of conditions, postvaccinal Compost fermentation being cultivated 28 days (when cultivating by 25 days, sack being untied), obtaining containing spore powder tunning.
7, the acquisition of thick spore powder
After chaetomium globosum ND35 bacterial strain produces spore completely, to take out containing spore powder tunning and dry, supper micron mill is used to be pulverized, successively by 80 orders and 200 object screen clothes, obtain being less than 200 objects and contain spore powder tunning (the thick spore powder of called after), adopt gradient dilution method to detect spore quantity in thick spore powder again, concrete detection method is as follows:
(1) thick spore powder 0.02-0.05g is taken;
(2) the thick spore powder taken is put into the beaker containing 5ml sterilized water, stir, place 5-10min;
(3) getting suspension drips on ready blood counting chamber, carries out microscopy;
(4) measure according to the service regeulations of blood counting chamber;
(5) total count (total count=A/5 × 25 × 10000 in 1ml bacterium liquid × B, A is the total count in 5 middle lattice, and B is bacterium liquid extension rate) in 1ml bacterium liquid is calculated according to the calculation formula of blood counting chamber;
(6) spore count in 1g sample is calculated according to the total count in 1ml bacterium liquid;
(7) repeat above-mentioned steps 2-3 time, calculate mean number.
After testing, the spore amount in thick spore powder is 2 × 10
8cfu/g, it can be used as the raw materials for production of chaetomium globosum bio-bacterial manure granule for subsequent use.
Two, the preparation of chaetomium globosum bio-bacterial manure granule
1, take thick spore powder 1kg, humic acids 273g, chitosan 13.7g, xanthan gum 13.7g, Xylo-Mucine (CMC) 27.4g, calcium chloride 41.1g fully mix, obtain the raw material after mixing;
2, the raw material after mixing step 1 obtained is put into pelletizing disks and to be sprayed water granulation, obtains the particle made;
3, the particle made step 2 obtained dries under field conditions (factors), obtains chaetomium globosum bio-bacterial manure granule (as shown in Figure 1).
4, according to living bacteria count, mould miscellaneous bacteria number, the index such as moisture and miscellaneous bacteria rate of agricultural microbial agent (GB20287-2006) standard detection chaetomium globosum bio-bacterial manure granule.
Detected result is as follows: living bacteria count is 2.1 × 10
8cfu/g; Mould miscellaneous bacteria number is 2.35 × 10
6individual/g (mL); Miscellaneous bacteria rate is 25.0%; Moisture is 20.0%; Fineness is 85%; PH value is 7.5.
The impact that the granule of embodiment 2, different components grows cucumber at seedling stage
1, test site and method
Test site is in Shandong Agricultural University greenhouse.Divide 4 groups of process:
1st group is blank CK (being denoted as CK);
2nd group is granule A (component is humic acids (20%), chitosan (3%), xanthan gum (1%), CMC (2%), calcium chloride (3%) and thick spore powder (71%));
3rd group is that (component is humic acids (20%), xanthan gum (1%), CMC (2%), calcium chloride (3%) and thick spore powder (74%) to granule B;
4th group is that (component is chitosan (3.8%), xanthan gum (1.3%), CMC (2.5%), calcium chloride (3.8%) and thick spore powder (88.6%) to granule C;
When planting, every cucumber adds 1g granule.
Vegetative period its plant height and ground stem are measured at cucumber at seedling stage, and with SPSS software, statistical study is carried out to data.
2, test-results
Test-results is as shown in table 1: all there were significant differences with blank CK for the cucumber growth of three process (granule A, granule B, granule C) of applying granule.The plant height of the cucumber seedling of granule A, B, C is used to improve 29.22%, 23.13%, 15.18% than the plant height of the cucumber seedling of blank CK respectively; The leading thread of the cucumber seedling of granule A, B, C is used to improve 36.32%, 25.11%, 19.58% than the leading thread of the cucumber seedling of blank CK respectively.And the plant height of the cucumber seedling of granule A treatment group (4-5 leaf phase) and leading thread are all higher than other groups (CK, granule B, granule C).
The impact that table 1, different components granule grow cucumber at seedling stage
Note: in same plant same column, different lowercase represents significant difference in 0.05 level.
Embodiment 3, chaetomium globosum bio-bacterial manure granule are to the growth-promoting functions of walnut Collar diameter growth
1, test site and method
Test site is in Liang Zhuan town, Daiyue District, Tai'an, Shandong Province, in mid-April, 2012, Walnut Seed (Walnut Seed is provided by forest zone, brilliance temple, forest farm, Culai Mountain, Tai'an, Shandong Province) is sowed after vernalization shows money or valuables one carries unintentionally, chaetomium globosum bio-bacterial manure granule embodiment 1 prepared is sprinkling upon in kind of plant hole, overlay film insulation, moisturizing, divide 3 process, each process repetition 3 times.
Granule (treatment group): apply chaetomium globosum bio-bacterial manure granule (below all referred to as granule), 5 grams every during sowing Walnut Seed;
Formulation 1 (positive control): apply the bio-bacterial manure (formulations 1 with in following examples) in the patent " strain of a kind of plant endogenesis epiphyte chaetomium globosum, microbiobacterial agent and application thereof " (ZL201110232477.9) of having authorized during sowing Walnut Seed, 5 grams every;
CK (negative control): sowing Walnut Seed does not apply any bacterial manure formulation.
Measure walnut leading thread and biomass at the beginning of 2013 6 months, and with SPSS software, statistical study is carried out to data.
2, test-results
Result is as shown in table 2: the walnut leading thread of granule treatment improves 26.98% than negative control group, and the walnut leading thread of granule treatment improves 6.59% than positive controls, wherein between treatment group and negative control group in P ﹤ 0.05 level significant difference.
Table 2, the fertile impact on walnut Collar diameter growth of different dosage form chaetomium globosum
Embodiment 4, chaetomium globosum bio-bacterial manure granule are on the impact of willow growth of cuttage seedlings
1, test site and method
Test site is in school district Plant Protection institute experimental plot, Shandong Agricultural University south, and on March 20th, 2013, it is for subsequent use that the cuttings branch of willow with bud being cut into about 25cm does cuttage.Divide three process:
Granule (treatment group): apply chaetomium globosum bio-bacterial manure granule prepared by embodiment 1 during cuttage in jack, each cuttings consumption 5 grams;
Formulation 1 (positive control): apply formulation 1 in jack during cuttage, each cuttings consumption 5 grams;
CK (negative control): do not add any formulation in jack during cuttage.
Cuttage line-spacing 40cm, spacing in the rows 30cm, often organize 300 strains, earthing after cuttage, and spray acetochlor, overlay film, whole process is not watered.Divide and investigate seedling rate, plant height, leading thread, on the ground fresh weight, underground fresh weight for 3 times respectively, enquiry data SPSS software carries out statistical study, compares the significance of difference of different treatment.
2, test-results
In June, (1) 2012 investigation result as shown in Table 3 and Figure 2, the willow cuttage seeding seedling rate of granule treatment improves 117.69%, 8.08% respectively than negative control group and positive controls, between each group in P ﹤ 0.05 level significant difference.
The willow cuttage seeding plant height of the granule treatment (+ND35 as in Fig. 2) improves 54.90%, 17.72% respectively than the negative control group (-ND35 as in Fig. 2) and positive controls respectively, between each group in P ﹤ 0.05 level significant difference.The willow cuttage seeding leading thread of granule treatment improves 32.89%, 9.27% respectively than negative control group and positive controls, between each group in P ﹤ 0.05 level significant difference.
In October, (2) 2012, investigation result was as shown in table 3, and the willow cuttage seeding plant height of granule treatment improves 21.56%, 4.29% respectively than negative control group and positive controls, between each group in P ﹤ 0.05 level equal significant difference.The willow cuttage seeding leading thread of granule treatment improves 24.10%, 6.40% respectively than negative control group and positive controls, between each group in P ﹤ 0.05 level significant difference.
In March, (3) 2013, investigation result was as shown in table 3, and the willow cuttage seeding ground fresh weight of granule treatment improves 88.84%, 26.65% respectively than negative control group and positive controls.The willow cuttage seeding underground fresh weight of granule treatment improves 123.73%, 28.16% respectively than negative control group and positive controls, between each group in P ﹤ 0.05 level significant difference.
The impact that table 3, different dosage form chaetomium fertilizer is emerged on willow cuttage seeding and grown
Embodiment 5, chaetomium globosum bio-bacterial manure granule are on the impact of purple potato single plant yield
1, test site and method
Test site is in Liang Zhuan town, Daiyue District, Tai'an, Shandong Province: chaetomium globosum bio-bacterial manure granule (treatment group prepared by point three groups: CK (negative control group), formulation 1 (positive controls), embodiment 1, hereinafter referred to as granule), each process 200.
CK (negative control): do not apply any formulation during purple potato transplantation of seedlings;
Formulation 1 (positive control): apply formulation 1 during purple potato transplantation of seedlings, every seedling consumption 2 grams;
Granule (treatment group): apply granule during potato transplantation of seedlings, every seedling consumption 2 grams.
2, test-results
Test-results is as shown in table 4 and Fig. 3.As can be seen from Table 3: the purple potato single plant yield of granule treatment, the longest climing length, leading thread, on the ground fresh weight improve 57.89%, 74.62%, 4.66%, 190.48% than negative control group respectively; The single plant yield of the purple potato of granule treatment, the longest climing length, leading thread, on the ground fresh weight, branch amount improve 31.87%, 66.11%, 20.44%, 56.41%, 35.71% than formulation 1 (positive control) respectively.As can be seen from Fig. 3 also: the single plant yield of the purple potato that the granule (+ND35 as in Fig. 3) processes, the longest climing length, leading thread, on the ground fresh weight, branch amount are also significantly higher than the negative control group (-ND35 as in Fig. 3).
The fertile impact on purple potato growth and output of table 4, different dosage form chaetomium globosum
Embodiment 6, chaetomium globosum bio-bacterial manure granule are to the growth-promoting functions of potted plant cucumber seedling
1, test method
Choose the cucumber seeds that full seed is consistent, 75% alcohol disinfecting 30s, clear water is cleaned, and 28 DEG C of vernalization, are seeded into 72 hole seedling pans, using seedling medium as Nutrition Soil after showing money or valuables one carries unintentionally.When height of seedling grows to about 5cm transplant to flowerpot, seedling medium and field soil by volume 2:1 as Nutrition Soil.Every pot transplanting cucumber seedling 3.
CK (negative control): do not add any bacterial manure formulation during transplanting;
Deactivation matrix (positive control): as the deactivation matrix (preparation method of deactivation matrix herein: the culture material that step 1-3 in embodiment 1 is prepared after the chaetomium globosum ND35 high-temperature inactivation that bacterium chaff is cultivated, carry out high pressure steam sterilization 121 DEG C, 30min obtains after cooling completely), during transplanting, every basin applies 2 grams;
Granule (treatment group): get 2 grams of chaetomium globosum bio-bacterial manure granules and add suitable quantity of water and be made into spore suspension, when cucumber is transplanted, pouring is to root.
2, test-results
Test-results is as shown in table 5 and Fig. 4: the cucumber plant height of the granule treatment (+ND35 as in Fig. 4), root length, leading thread, on the ground fresh weight, underground fresh weight improve 23.83%, 11.72%, 1.73%, 17.60%, 39.53% than the negative control group (-ND35 as in Fig. 4) respectively, wherein significant difference in plant height, on the ground fresh weight, underground fresh weight P ﹤ 0.05 level.The cucumber plant height of granule treatment, root length, leading thread, on the ground fresh weight, underground fresh weight improve 16.34%, 9.50%, 0.86%, 22.38%, 36.36% than positive controls respectively, wherein plant height, on the ground fresh weight, underground fresh weight significant difference in P ﹤ 0.05 level.
Table 5, different treatment are on the impact of potted plant cucumber seedling growth
Embodiment 7, chaetomium globosum bio-bacterial manure granule are to the preventive and therapeutic effect of cucumber at seedling stage samping off
1, test method
Choose full seed consistent Tianjin spring No. 4 cucumber seeds (being the product of Xintai City auspicious cloud Zhong Ye company limited of Shandong Province), 28 DEG C of vernalization, be seeded into after showing money or valuables one carries unintentionally in flowerpot, soil is by with causing, the land for growing field crops of cucumber at seedling stage samping off pathogenic bacteria is native to be mixed to get by 1:1 equal-volume with seedling medium.
If three process:
Negative control: do not add any bacterial manure formulation;
biological organic fertilizer (positive control): add 2 grams during sowing.
Chaetomium globosum bio-bacterial manure granule (granule): add 2 grams during sowing.
Often three repetitions are established in process, often repeat 20 young plants.Be placed in greenhouse, normal water management, the protection effect of 8 weeks different bacterial manure groups of " Invest, Then Investigate ".
2, test-results
Test-results is as shown in table 5 and Fig. 5: chaetomium globosum bio-bacterial manure granule of the present invention is 78.33% to the prevention effect of cucumber at seedling stage samping off, improves 18 percentage points than carrying out the prevention effect of fertile biological organic fertilizer to cucumber at seedling stage samping off.Can not only promote that cucumber at seedling stage grows, and certain prevention effect can also be had to cucumber at seedling stage samping off.
Table 5, different treatment are to the prevention effect of cucumber at seedling stage samping off
Claims (10)
1. prepare a method for bio-bacterial manure, comprise the steps: the spore of chaetomium globosum (Chaetomium globosum) ND35 CGMCC No.4136, humic acids, chitosan and auxiliary agent to mix, obtain bio-bacterial manure.
2. bio-bacterial manure according to claim 1, is characterized in that: described auxiliary agent is xanthan gum, Xylo-Mucine and calcium chloride.
3. method according to claim 1 and 2, is characterized in that: the proportioning of the spore of described chaetomium globosum (Chaetomiumglobosum) ND35 CGMCC No.4136, described humic acids, described chitosan, described xanthan gum, described Xylo-Mucine and described calcium chloride is 3 × 10
11cfu:(265-285) g:(5-20) g:(5-20) g:(15-35) g:(30-50) g.
4., according to described method arbitrary in claim 1-3, it is characterized in that: the spore of described chaetomium globosum (Chaetomiumglobosum) ND35 CGMCC No.4136 is for being less than 200 object spore powder.
5. according to described method arbitrary in claim 1-4, it is characterized in that: in described method, when being also included in mixing after the spore of described chaetomium globosum (Chaetomium globosum) ND35 CGMCC No.4136, described humic acids, described chitosan, described xanthan gum, described Xylo-Mucine and described calcium chloride being mixed, add the step of filler.
6. according to described method arbitrary in claim 1-5, it is characterized in that: in described method, after the spore of described chaetomium globosum (Chaetomium globosum) ND35 CGMCC No.4136, described humic acids, described chitosan, described xanthan gum, described Xylo-Mucine and described calcium chloride being mixed, also comprise the step of described raw material being carried out granulation, obtain bio-bacterial manure.
7., according to described method arbitrary in claim 1-6, it is characterized in that: described filler is diatomite.
8. the bio-bacterial manure prepared by described method arbitrary in claim 1-7.
9. the application of bio-bacterial manure according to claim 8 in following (1)-(14) at least one:
(1) plant height of plant is increased;
(2) Chlorophyll-a Content of plant is improved;
(3) content of chlorophyll b of plant is improved;
(4) total chlorophyll content of plant is improved;
(5) carotenoid content of plant is improved;
(6) reducing sugar content of plant leaf is improved;
(7) proline content of plant leaf is improved;
(8) yield per plant is improved;
(9) leaf increasing plant is wide;
(10) leaf increasing plant is long;
(11) leading thread of plant is increased;
(12) growth in plant seedling stage is promoted;
(13) plant Blooming result is impelled;
(14) plant seedling sudden fall ill is prevented and treated;
Described plant is specially walnut and/or cucumber and/or purple potato and/or willow.
10. in claim 1-7 arbitrary described chaetomium globosum (Chaetomium globosum) ND35 CGMCC No.4136 or its spore or its tunning as bio-bacterial manure additive or the application in the product of preparation bio-bacterial manure additive.
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