CN104496574A - Novel soybean or leguminous forage rhizosphere biological-bacterial fertilizer as well as preparation method and application thereof - Google Patents
Novel soybean or leguminous forage rhizosphere biological-bacterial fertilizer as well as preparation method and application thereof Download PDFInfo
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- 244000068988 Glycine max Species 0.000 title claims abstract description 57
- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 57
- 239000004459 forage Substances 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000003337 fertilizer Substances 0.000 title abstract description 24
- 230000001580 bacterial effect Effects 0.000 claims abstract description 79
- 239000002689 soil Substances 0.000 claims abstract description 23
- 239000007788 liquid Substances 0.000 claims abstract description 20
- 235000015097 nutrients Nutrition 0.000 claims abstract description 11
- 230000012010 growth Effects 0.000 claims abstract description 10
- 210000003608 fece Anatomy 0.000 claims description 72
- 239000010871 livestock manure Substances 0.000 claims description 72
- 241000894006 Bacteria Species 0.000 claims description 51
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- 235000008898 Morinda citrifolia Nutrition 0.000 claims description 20
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- 241000219793 Trifolium Species 0.000 claims description 20
- 235000017524 noni Nutrition 0.000 claims description 20
- 230000028327 secretion Effects 0.000 claims description 16
- 241000589180 Rhizobium Species 0.000 claims description 13
- 239000003375 plant hormone Substances 0.000 claims description 12
- 229910019142 PO4 Inorganic materials 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 9
- 230000000050 nutritive effect Effects 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 9
- 239000010452 phosphate Substances 0.000 claims description 9
- 230000001737 promoting effect Effects 0.000 claims description 9
- 238000005063 solubilization Methods 0.000 claims description 9
- 230000007928 solubilization Effects 0.000 claims description 9
- 238000009472 formulation Methods 0.000 claims description 7
- 239000004615 ingredient Substances 0.000 claims description 7
- 239000002054 inoculum Substances 0.000 claims description 7
- 238000010521 absorption reaction Methods 0.000 claims description 6
- 241000588914 Enterobacter Species 0.000 claims description 5
- 241000588748 Klebsiella Species 0.000 claims description 4
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 4
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- 230000008859 change Effects 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 3
- 238000012856 packing Methods 0.000 claims description 3
- 241000588697 Enterobacter cloacae Species 0.000 claims description 2
- 241000305071 Enterobacterales Species 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 45
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 38
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 abstract description 24
- 229910052698 phosphorus Inorganic materials 0.000 abstract description 24
- 239000011574 phosphorus Substances 0.000 abstract description 24
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 19
- 230000001965 increasing effect Effects 0.000 abstract description 8
- 235000016709 nutrition Nutrition 0.000 abstract description 7
- 230000008635 plant growth Effects 0.000 abstract description 6
- 239000000122 growth hormone Substances 0.000 abstract description 4
- 230000035764 nutrition Effects 0.000 abstract description 3
- 102000018997 Growth Hormone Human genes 0.000 abstract description 2
- 108010051696 Growth Hormone Proteins 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 23
- 230000008569 process Effects 0.000 description 19
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- 239000000126 substance Substances 0.000 description 11
- 238000004519 manufacturing process Methods 0.000 description 10
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- 235000019750 Crude protein Nutrition 0.000 description 7
- 241001647841 Leclercia adecarboxylata Species 0.000 description 7
- 241000589194 Rhizobium leguminosarum Species 0.000 description 7
- 235000019784 crude fat Nutrition 0.000 description 7
- 238000002156 mixing Methods 0.000 description 7
- 241001014264 Klebsiella variicola Species 0.000 description 6
- 241000881812 Kluyvera intermedia Species 0.000 description 6
- 241001353184 Rhizobium soli Species 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 6
- 235000013339 cereals Nutrition 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 150000007524 organic acids Chemical class 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 240000007594 Oryza sativa Species 0.000 description 4
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- 238000003756 stirring Methods 0.000 description 3
- 241000589941 Azospirillum Species 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 239000006035 Tryptophane Substances 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 230000035558 fertility Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- 239000002686 phosphate fertilizer Substances 0.000 description 2
- 235000014786 phosphorus Nutrition 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 229960004799 tryptophan Drugs 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000579722 Kocuria Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000589157 Rhizobiales Species 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 244000042324 Trifolium repens Species 0.000 description 1
- 235000010729 Trifolium repens Nutrition 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- YUWBVKYVJWNVLE-UHFFFAOYSA-N [N].[P] Chemical compound [N].[P] YUWBVKYVJWNVLE-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
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- 238000012271 agricultural production Methods 0.000 description 1
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- 230000008485 antagonism Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 244000037666 field crops Species 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
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- 238000011835 investigation Methods 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000024121 nodulation Effects 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 230000001863 plant nutrition Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 239000002688 soil aggregate Substances 0.000 description 1
- 239000004016 soil organic matter Substances 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 230000001228 trophic effect Effects 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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Abstract
The invention discloses a novel soybean or leguminous forage rhizosphere biological-bacterial fertilizer as well as a preparation method and an application thereof. The biological-bacterial fertilizer is prepared from the following strains: strain DR19, strain DR9, strain DR24, strain DR4, strain D2 and strain D4 at a bacterial liquid volume ratio of 1: 1: 1: 2: 1: 1. By applying the biological-bacterial fertilizer disclosed by the invention, insoluble phosphorus in the soil can be activated, nitrogen in the air can be fixed, the effective nitrogen and phosphorus nutrients in the soil for soybean and leguminous forage can be increased, plant growth hormone can be secreted, plants are promoted to absorb other nutrients and thus the growth of the plants is promoted, the nutrition structure of the plants is improved, the yield of soybean is increased and the nutritional quality of the forage is improved.
Description
Technical field
The present invention relates to fertilizer field, particularly relate to the application of a kind of novel soybean or the bio-bacterial manure of leguminous forage rhizosphere and preparation method thereof and this bio-bacterial manure.
Background technology
Nitrogen, phosphorus are one of necessary macroelement of growth and development of plants, and Nitrogen In Soils, phosphoric lack the common issue having become world many countries and faced.With regard to China, investigation result show, the arable land of 74% lacks phosphorus, and in agricultural land soil, phosphorus exists with phosphatic combined form, the phosphorus in chemical combination state due to the Fe in soil
3+, Ca
2+ and Al
3+ wait combining and in water-insoluble, being difficult to be absorbed by plants, so a large amount of soluble phosphoric manures must be applied to soil every year in agriculture production.For applied phosphate fertilizer, the utilization ratio of this season crop generally only has 5% ~ 10%, adds the aftereffect of crop, is also no more than 25%, and the most of phosphate fertilizer be manured into soil is actually and accumulates in soil with invalid state.In agricultural production process, in order to maintain and improve crop yield, chemical fertilizer is considered to the main or even unique channel solving soil nitrogen and phosphorus element shortage problem always.Special in China, the annual production of chemical fertilizer, ultimate production and unit surface usage quantity all rank first in the world.But chemical fertilizers production cost is high, along with the increase of amount of application, also with having occurred the phenomenon such as fertilizer efficiency decline, utilization ratio reduction, and along with NO in soil and plant
2, inevitably there is contaminate environment, jeopardized the ecological problem such as people and animals and food safety in-isoionic accumulation.Meanwhile, produce chemical fertilizer comparatively large to non-renewable energy consumption, use chemical fertilizer and can cause the negative effects such as soil aggregate and microbial diversity are damaged again.Therefore, seek other fertilizer originate particularly bio-feritlizer with the effect of Some substitute chemical fertilizer be produce at present in problem demanding prompt solution.
Domestic and international research shows, ploughs and does not open up wasteland the place that soil is the most enrichment of occurring in nature microorganism, there is a large amount of beneficial microorganism in plant rhizosphere soil.Microorganism is the Living Organism in soil, is indispensable activeconstituents of increasing soil fertility, as the vinelandii in soil can N in fixed air
2, by plant is utilized, Promoting plant growth; The phosphorus element be difficult in soil by plant directly absorbs can be converted into plant available state by phosphorus-solubilizing bacteria, and in addition, the molten phosphorus microorganism had has fixed nitrogen concurrently and secretes plant hormone as the characteristic of growth hormone etc.With the complex inoculant (microbial-bacterial fertilizer) being separated the efficient Promoting bacteria that obtains from plant rhizosphere and being developed into, not only can activating soil Insoluble phosphates, nitrogen in fixed air, nitrogen, phosphorus nourishing are provided, also contribute to the absorption of farm crop to other nutritive element simultaneously, improve plant nutrition structure, regulate and Promoting plant growth, strengthen plant disease-resistant ability.The bio-bacterial manure utilizing beneficial microbe strain to make has compared with chemical fertilizer that cost is low, use safety, lasting effect are good, volume increase is stable, non-renewable energy consumption less, high financial profit, without the advantage such as environment and food contamination, meanwhile, also can improve Soil structure, improve soil organic matter content and improvement saltings.Therefore, the research and development of the research and bio-bacterial manure of carrying out plant growth-promoting rhizobacteria resource have extremely important realistic meaning.
At present, developed country (as the U.S.) and some developing countries (as India, Pakistan, Brazil, Egypt etc.) all attach great importance to the research of bio-feritlizer abroad, have oneself achievement and product.As India to there has been the bio-bacterial manure of wheat, paddy rice etc. in the 1980s; The national biotechnology of Pakistan and genetically engineered institute utilized in 1996 and promote that strains A zoarcusK-1, Azospirillum N-4, Azospirillum ER-20, Psedomonas 96-51 etc. develop into BIOPOWER bio-bacterial manure from isolated growth of grass rhizosphere, with cheap price, significantly produce effects, outstanding environmental protection characteristic and being favored.But about the research of leguminous crop rhizosphere bio-bacterial manure has no report.
A large amount of research work has been done in root nodule bacterium by China, and achieves some gratifying results, but about leguminous plants rhizosphere phosphorus-solubilizing bacteria, secretion plant hormone bacterium and report seldom with the research of the aspect such as root nodule bacterium do mutually.Although the Promoting bacteria fertilizer product of external existing mass-producing, the plant rhizosphere in different ecological environment is survived different growth-promoting bacterial strains, and the adaptability of different strains to environment is different.We can the successful experience of foreign, constantly be separated from the specific weather of China, habitat, upgrade active bacterial strain, the microbial preparation of Study and Development to one's name intellecture property, produce the bio-bacterial manure product innovation of the specific weather of applicable China, specific environment, this plays irreplaceable effect by the sustainable development of China's Modern Green agricultural.
In leguminous crop, soybean is the important crops originating in a kind of grain of China, oil plant, feed and industrial raw material dual-purpose, people, stock keeping can be supported, increase soil fertility, can export goods and earn foreign currency again, in agriculture production, planting structure and distribution and the national economic development, occupy critical role.In history, China's soybean acreage and ultimate production occupy the first in the world for a long time, but generally speaking, soybean in China per unit area yield is not high, always produces shakiness, year rich apology change is large, area output is uneven, and especially compared with the soybean main product state such as U.S., per unit area yield has a long way to go, its reason, except kind and planting patterns, also has direct relation with fertilizer type.On the other hand, in leguminous plants, leguminous forage is a kind of Important Economic crop connecting plant husbandry and aquaculture.In the good forage of China's plantation, the proportion of leguminous forage in occupation of more than 50%, meadow turf establishment, improve regional agriculture produce structure, promotion " ternary structural agricultural " and save-grain animal husbandry development in play an important role.But the level of the productive forces of leguminous forage is widely different under different cultivation management condition.Leguminous plants all has stronger dependency to phosphorus element, because phosphorus element is the necessary element of rhizobium leguminosarum noclulation, directly have influence on quantity and the nitrogen-fixing efficiency of nodule formation, simultaneously, phosphorus element can also affect the absorption of crop to potassium element, and then affects the output of crop, quality and anti-adversity ability.In the prior art, relevant leguminous crop Inoculant is all based on single root nodule bacterium both at home and abroad, there is no the complex inoculant gathered Rhizobium strains, phosphorus-solubilizing bacteria strain and secrete plant hormone bacterial strain, is badly in need of solving the practical problems that Inoculant kind is single, growgh promoting effects is low.Therefore, the leguminous crop rhizosphere screening efficient phosphorus-dissolution bacterial strain in matter never of the same race, habitat, Rhizobium strains, secretion plant hormone bacterial strain, the bio-bacterial manure of research and development leguminous crop rhizosphere, produces for leguminous crop and provides novel " environmental friendliness " fertilizer, be significant.
Summary of the invention
The object of the invention is in prior art, chemical fertilizers production cost is high, fertilizer efficiency is low, nonrenewable resource consumption is large, harm people and animals, the feature of environmental protection is poor, of less types in leguminous crop bio-bacterial manure, bacterial strain function singleness, growth-promoting effect is lower, and the soybean crops that China extensively plants lacks the problems such as the strong bio-bacterial manure of specificity, propose a kind of bio-bacterial manure for soybean and leguminous forage to fill a prescription and its preparation method and application, low to realize fertiliser production cost, nonrenewable resource consumption is little, have molten phosphorus concurrently simultaneously, fixed nitrogen, secretion plant hormone function and fertilizer efficiency persistence is good, security is high, environmental protection, the target that bio-bacterial manure is with strong points, and then reach object soybean and leguminous forage to good growth-promoting increasing of quality and production effect.
The soybean rhizosphere of the present invention's matter never of the same race, ecotope filters out phosphorus-solubilizing bacteria and root nodule bacterium, by phosphate solubilization, nitrogen fixation effect, secretion plant growth hormones (IAA), organic acid secretion amount, produce acid produce alkalescence can, the indices such as antagonism interaction characteristics between each bacterial strain measures and land for growing field crops uses test determination to soybean, leguminous forage Trifolium repense and Root or stem of Littleleaf Indianmulberry growth-promoting effect and qualitative effects, filter out efficient phosphorus-dissolution bacterial strain, plant hormone secretion bacterial strain and Rhizobium strains, have developed soybean and leguminous forage rhizosphere growth-promoting complex inoculant---bio-bacterial manure.
Technical scheme of the present invention is:
A kind of novel soybean or the bio-bacterial manure of leguminous forage rhizosphere: it is prepared from from soybean or leguminous forage rhizosphere biological bacteria by being separated, described rhizosphere biological bacteria comprises plant hormone secretion bacterial strain and at least two kinds of Rhizobium strains that the phosphorus-solubilizing bacteria strain having IAA secretion capacity concurrently of at least three kinds and at least one have phosphate solubilization concurrently.
Further, the phosphorus-solubilizing bacteria strain of the described IAA of having concurrently secretion capacity comprises one or more the combination in bacterial strain DR19, DR9, DR24, the described plant hormone secretion bacterial strain having phosphate solubilization concurrently comprises bacterial strain DR4, and described Rhizobium strains comprises one or more the combination in bacterial strain D2, D4.Bio-bacterial manure of the present invention is made up of the excellent growth-promoting bacterium of 6 strain, and bacterial strain code name is respectively: DR19, DR9, DR24, DR4, D2, D4.In classification, bacterial strain DR19 is the Leclercia adecarboxylata (Leclercia adecarboxylata) strangled in Kocuria, the DR9 change belonged in Klebsiella is dwelt klebsiella (Klebsiella variicola), DR24 belongs to the enterobacter cloacae (Enterobacter cloacea) in enterobacter, DR4 belongs to the osculant enterobacteria (Enterobacter intermedium) in enterobacter, D2 belongs to the beans root nodule bacterium (Rhizobiumleguminosarum) in rhizobium, D4 belongs to the soil root nodule bacterium (Rhizobium soli) in rhizobium.
Wherein, the D in bacterial strain code name represents and is separated from the excellent Rhizobium strains of soybean (Glycine max) rhizosphere, and DR represents the excellent phosphorus-solubilizing bacteria strain be separated from soybean rhizosphere, and the Arabic numerals such as 19,9,24,4,2,4 are strain number when screening in a large number.Wherein, bacterial strain DR19, DR9, DR24 have stronger phosphate solubilization, have IAA secretion capacity concurrently simultaneously; Bacterial strain DR4 has stronger plant hormone IAA secretion capacity, has phosphate solubilization concurrently; Bacterial strain D2, D4 have stronger nitrogen fixing capacity.The strain name of bacterial strain DR19, DR9, DR24, DR4, D2, D4 of the excellent growth-promoting bacterium of 6 strain filtered out is respectively: L.adecarboxylata strain DR19, K.variicola strain DR9, E.cloacea strain DR24, E.intermedium strain DR4, R.leguminosarum strain D2, R.soli strain D4.In title, first capitalization adds the abbreviation that lower-right most point is generic name, after be kind of a name, the bacterial strain code name used when strain DR19 etc. is Screening and Identification.These six kinds of bacterial strains are existing bacterial strain.
Three of the component of bio-bacterial manure of the present invention are provided specifically to fill a prescription below:
basic recipe:
The long-pending ratio of L.adecarboxylata strain DR19:K.variicola strain DR9:E.cloacea strain DR24:E.intermedium strain DR4:R.leguminosarum strain D2:R.soli strain D4=1:1:1:1:1:1(bacteria liquid).
screening formulation:
The long-pending ratio of L.adecarboxylata strain DR19:K.variicola strain DR9:E.cloacea strain DR24:E.intermedium strain DR4:R.leguminosarum strain D2:R.soli strain D4=1:1:1:2:1:1(bacteria liquid).Its technique effect than basic components is slightly good.
optimization formula:
The long-pending ratio of L.adecarboxylata strain DR19:K.variicola strain DR9:E.cloacea strain DR24:E.intermedium strain DR4:R.leguminosarum strain D2:R.soli strain D4=2:2:1:3:1:1(bacteria liquid).Under optimization formula, the technique effect of bio-bacterial manure is compared basic components and is significantly improved.
According to above-mentioned formula, the preparation method of soybean of the present invention and leguminous forage bio-bacterial manure is as follows:
By the bacterial strain L.adecarboxylata strain DR19 after activated, K.variicola strain DR9, E.cloacea strain DR24, E.intermedium strain DR4, R.leguminosarum strain D2, R.soli strain D4 is inoculated in SOC liquid nutrient medium by the bacterium liquid inoculum size of 1:1:1:1:1:1 or 1:1:1:2:1:1 or 2:2:1:3:1:1 respectively, at 26 ~ 28 DEG C, 180 ~ 200rpm(rpm) condition under shaking culture, to the light absorption value of bacterium liquid OD600(at wavelength 600nm place) stop fermentation obtaining liquid active bacteria formulation when reaching 0.5 ~ 0.8, with vegetable mould as carrier, active bacteria formulation is cultivated through sterile carrier absorption, weigh packing and make solid live bacteria preparation.
Further, the present invention proposes this novel soybean or the application of leguminous forage rhizosphere bio-bacterial manure on promotion soybean is born pods.
Further, the present invention proposes this novel soybean or the bio-bacterial manure of leguminous forage rhizosphere promoting Trifolium repense growth and improving the application of Trifolium repense nutritive ingredient.
Further, the present invention proposes this novel soybean or the bio-bacterial manure of leguminous forage rhizosphere promoting Root or stem of Littleleaf Indianmulberry growth and improving the application of Root or stem of Littleleaf Indianmulberry nutritive ingredient.
Determination experiment result for the property indices of the soybean in the present invention and leguminous forage bio-bacterial manure is as follows respectively:
1, the Antagonistic reaction of hybrid bacterial strain system measures
By bacterial strain
l.adecarboxylatastrain DR19,
k.variicolastrain DR9,
e.cloaceastrain DR24,
e.intermediumstrain DR4,
r.leguminosarumstrain D2,
r.solistrain D4 is the ratio mixing of 1:1:1:1:1:1,1:1:1:2:1:1 and 2:2:1:3:1:1 respectively by volume, carries out Antagonistic reaction mensuration, find bacterial strain to hybrid bacterial strain system
l.adecarboxylatastrain DR19,
k.variicolastrain DR9,
e.cloaceastrain DR24,
e.intermediumstrain DR4,
r.leguminosarumstrain D2,
r.soliall without Antagonistic reaction between strain D4.
2, the phosphate solubilization of hybrid bacterial strain system measures
By bacterial strain
l.adecarboxylatastrain DR19,
k.variicolastrain DR9,
e.cloaceastrain DR24,
e.intermediumstrain DR4,
r.leguminosarumstrain D2,
r.solistrain D4 is the ratio mixing of 1:1:1:2:1:1 and 2:2:1:3:1:1 respectively by volume, carry out phosphate solubilization mensuration, find that mixed bacterial strain system is respectively 189.27 μ g/mL and 193.65 μ g/mL to the meltage of phosphorus in calcium phosphate, the available phosphorus increment compared when the first six bacterial strain of mixing is individually cultivated improves 25% ~ 80%.
3, the IAA of hybrid bacterial strain system and organic acid secretion ability measure
By bacterial strain
l.adecarboxylatastrain DR19,
k.variicolastrain DR9,
e.cloaceastrain DR24,
e.intermediumstrain DR4,
r.leguminosarumstrain D2,
r.solistrain D4 is the ratio mixed culture of 1:1:1:2:1:1 and 2:2:1:3:1:1 respectively by volume, carry out IAA and organic acid secretion capacity mensuration, find when adding precursor substance tryptophane in hybrid bacterial strain system, in nutrient solution, the content of IAA is 19.96 μ g/mL and 21.43 μ g/mL, and the IAA content compared when the first six bacterial strain of mixing is individually cultivated improves 65% ~ 98%; When not adding precursor substance tryptophane in hybrid bacterial strain system, in nutrient solution, IAA content is 18.07 μ g/ml and 18.76 μ g/mL, and the IAA content compared when the first six bacterial strain of mixing is individually cultivated improves 70% ~ 100%; In mixed bacterial strain system, total organic acids content is 14.09mmol/L and 14.58mmol/L, and the total organic acids content compared when the first six bacterial strain of mixing is individually cultivated improves 121%.
4, the living bacteria count of hybrid bacterial strain system measures
Bacterial strain
l.adecarboxylatastrain DR19,
k.variicolastrain DR9,
e.cloaceastrain DR24,
e.intermediumstrain DR4,
r.leguminosarumstrain D2,
r.solistrain D4 is the ratio mixing of 1:1:1:2:1:1 and 2:2:1:3:1:1 by volume, hybrid bacterial strain is cultivated in the matrix being carrier with aseptic vegetable mould, and 180 days are stored under room temperature condition, record wherein living bacteria count content and be 4.63 × 10
8cfu/g, requirement preserved by the bacterial manure met in " microbial fertilizer " NY227-94.
For the growth-promoting effect of increasing production that checking the present invention brings leguminous crop, the present invention has carried out field experiment for soybean, leguminous forage Trifolium repense and Root or stem of Littleleaf Indianmulberry respectively.Respectively choose the three identical edatopes in place and the two pieces of experimental plots be connected together, select the seed of same batch of same kind of soybean, Trifolium repense and Root or stem of Littleleaf Indianmulberry, equivalent sowing is in respective two pieces of experimental plots respectively, single administration bio-bacterial manure of the present invention in first piece of experimental plot, any fertilizer is not used in second piece of experimental plot in contrast, carry out identical cultivation management, carry out the growing state of trial crops, output or nutrient component determining, result is as follows simultaneously simultaneously.
Test one: bio-bacterial manure is to the growth-promoting growth effect of soybean
Field experiment result shows, uses the experimental plot after bio-bacterial manure process of the present invention
in soybean, be 69.93cm at the average plant height of branching stage, comparatively controlled trial field
in average plant height 55.53cm exceed 25.93%; Soybean after process is slightly 2.87cm at the average stem of branching stage, comparatively contrasts the thick 2.11cm of average stem and exceeds 35.69%.Using the soybean after bio-bacterial manure process of the present invention, is 100.40cm at the average plant height in flowering period, comparatively contrasts average plant height 82.80cm and exceeds 21.26%; Soybean after process is slightly 3.04cm at the average stem in flowering period, comparatively contrasts the thick 2.16cm of average stem and exceeds 40.79%.
Test two: the impact that bio-bacterial manure is born pods on soybean and grain is heavy
In harvesting time respectively to experimental plot
,
in bear pods number, heavy, the every pod simple grain number of pod and Single seed weight of the every strain of soybean measure.Result shows, use the soybean after bio-bacterial manure process of the present invention, every strain on average bear pods number, every pod weight in average, every strain average single number and every strain average single be heavily respectively 6.33,7.4g, 11.47 and 1.71g, exceed 55.74%, 20.21%, 56.36% and 57.20% respectively than 4.07 in contrast, 6.2g, 7.33 and 1.09g.
Test three: the growth of bio-bacterial manure on Trifolium repense and the impact of nutritive ingredient
At first batch of cut issue respectively to experimental plot
,
in the plant height of Trifolium repense and ground biomass measure.Result shows, uses the Trifolium repense after bio-bacterial manure process of the present invention, and average plant height is 27.45cm, and the average ground biomass of individual plant is 4.479g, exceeds 20.82% and 55.20% respectively than 24.21cm, 2.886g in contrast; The total nitrogen content of the Trifolium repense after bio-bacterial manure process is 3.02%, content of tatal phosphorus 0.31%, crude protein content 18.88%, crude fat content 7.12%, exceeds 9.82%, 19.23%, 9.71% and 7.07% respectively than 2.75%, 0.26%, 17.21%, 6.65% in contrast.
Test four: the growth of bio-bacterial manure on Root or stem of Littleleaf Indianmulberry and the impact of nutritive ingredient
At first batch of cut issue respectively to experimental plot
,
in the plant height of Root or stem of Littleleaf Indianmulberry, ground biomass and nutritive ingredient measure.Result shows, uses the Root or stem of Littleleaf Indianmulberry after bio-bacterial manure process of the present invention, and average plant height is 25.49cm, and the average ground biomass of individual plant is 9.26g, exceeds 20.82% and 106.14% respectively than 21.10cm, 4.49g in contrast; Root or stem of Littleleaf Indianmulberry total nitrogen content after bio-bacterial manure process is 2.18%, content of tatal phosphorus 0.77%, crude protein content 13.63%, crude fat content 6.20%, exceeds 19.34%, 61.88%, 19.34% and 25.08% respectively than 1.83%, 0.47%, 11.42%, 4.96% in contrast.
Result shows, the soybean after bio-bacterial manure process of the present invention, compares untreated same kind soybean, and its average plant height, stem are thick, the number that bears pods, pod weight in average, often strain simple grain number and Single seed weight are all significantly increased; Leguminous forage Trifolium repense after bio-bacterial manure process of the present invention and Root or stem of Littleleaf Indianmulberry, compare Trifolium repense and the Root or stem of Littleleaf Indianmulberry of untreated same kind, its average plant height, ground biomass, total nitrogen content, content of tatal phosphorus, crude protein content and crude fat content are all significantly increased.Visible, use bio-bacterial manure of the present invention, can the insoluble phosphorus element in activating soil, the nitrogen in fixed air, increase the nutrient such as soybean and the available nitrogen phosphorus of leguminous forage in soil, simultaneously can also secrete plant growth hormones, promote that plant is to the absorption of other nutritive element, and then promote the growth of plant and improve the trophic structure of plant, increase the output of soybean, improve the nutritional quality of herbage.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
Embodiment 1: the preparation of bio-bacterial manure
By the excellent phosphorus-solubilizing bacteria strain DR19 filtered out, DR9, DR24, good plant hormone secretion bacterial strain DR4, excellent Rhizobium strains D2, D4, the bacterial classification difference streak inoculation of totally 6 bacterial strains is on LB solid medium, 26 DEG C of inversion activation culture of spending the night, in time growing apparent single bacterium colony, picking list bacterium colony is inoculated in the SOC liquid nutrient medium of same volume respectively, 26 DEG C shake cultivation simultaneously, when nutrient solution starts to become muddiness, DR19, DR9, DR24, DR4, D2, these 6 bacterial strains of D4 are inoculated in same SOC liquid nutrient medium by the bacterium liquid inoculum size of 1:1:1:2:1:1 respectively, at 26 ~ 28 DEG C, 180 ~ 200rpm(rpm) condition under mix shaking culture, to the absorbancy of bacterium liquid OD600(in wavelength 600 nanometers) stop fermentation when reaching 0.5 ~ 0.8, obtain liquid active bacteria formulation, with vegetable mould as carrier, again active bacteria formulation is cultivated through sterile carrier absorption, weigh packing, every bag of 500g, obtain solid biologic bacterial manure.
Embodiment 2: bio-bacterial manure is to the growth-promoting yield increasing effect of soybean
In early June, 2012, choose the seed 20kg of same batch of soybean varieties " No. 1, Guizhou Province beans ", be divided into 2 parts, every part of 10kg.Get obtained bio-bacterial manure 2 bags, add compared with the appropriate furnishing pasty state of the thin rice gruel of thickness, then add the seed of wherein 1 part of 10kg, fully stir and mix well, the soya bean of another 1 part of 10kg without bio-bacterial manure seed dressing in contrast, sows two pieces of connected experimental plots in the identical edatope of equal area on the same day respectively
,
in, carry out identical cultivation management simultaneously.Mid-July in two pieces of experimental plots respectively random selecting 100 strain bean seedlings measure the plant height of branching stage and stem thick, the plant height and the stem that measure flowering period are by the end of July thick, measure the number that bears pods, heavy, the every pod simple grain number of pod and Single seed weight by the end of August.Result proves, uses the soybean after bio-bacterial manure process of the present invention, slightly comparatively contrasts exceed 25.93% and 35.69% respectively at the average plant height of branching stage and stem, slightly comparatively contrasts exceed 21.26% and 40.79% respectively at the average plant height in flowering period and stem; On average bear pods number, every pod weight in average, every strain average single number and average Single seed weight of every strain exceeds 55.74%, 20.21%, 56.36% and 57.20% than contrast respectively.Ultimate capacity measures display, is 187.36kg with the soya bean per mu yield after bio-bacterial manure seed dressing of the present invention, than contrast 152.05kg volume increase 23.22%.
Embodiment 3: bio-bacterial manure increases matter effect to the growth-promoting of Trifolium repense
In mid-September, 2012, choose the seed 2.0kg of same batch of Trifolium repens cultivar " Haifa ", be divided into 2 parts, every part of 1.0kg.Get obtained bio-bacterial manure 1 bag, add compared with the appropriate furnishing pasty state of the thin rice gruel of thickness, then the seed of wherein 1 part of 1.0kg is added, abundant stirring is mixed well, the Trifolium repense of another 1 part of 1.0kg without bio-bacterial manure seed dressing in contrast, sows two pieces of connected experimental plots in the identical edatope of equal area on the same day respectively
,
in, carry out identical cultivation management simultaneously.The difference random selecting 100 strain Trifolium repense plant in two pieces of experimental plots in flowering period that buddings in April, 2013, measures plant height and ground biomass.Biomass estimation method is, 100 strain Trifolium repense are cut off root system part, stays cauline leaf part on the ground to complete at 105 DEG C 15 minutes, then dries to constant weight at 65 DEG C, obtain average biomass of individual tree after weighing.Respectively adopt 100 strains in addition, measure the full nitrogen of above-ground plant parts, full phosphorus, crude protein and crude fat content respectively, Comparative nutrition level.Result shows, uses the Trifolium repense after bio-bacterial manure process of the present invention, comparatively contrasts exceed 20.82% and 55.20% respectively at the average plant height in flowering period of buddingging and ground biomass; The total nitrogen content of the Trifolium repense after bio-bacterial manure process of the present invention, content of tatal phosphorus, crude protein content and crude fat content are than exceeding 9.82%, 19.23%, 9.71% and 7.07% respectively in contrast.Prove the Trifolium repense after using bio-bacterial manure process of the present invention, output and nutritional quality are all significantly improved.
Embodiment 4: bio-bacterial manure increases matter effect to the growth-promoting of Root or stem of Littleleaf Indianmulberry
In early March, 2013, chooses the seed 2.0kg of same batch of Root or stem of Littleleaf Indianmulberry, is divided into 2 parts, every part of 1.0kg.Get obtained bio-bacterial manure 1 bag, add compared with the appropriate furnishing pasty state of the thin rice gruel of thickness, then the seed of wherein 1 part of 1.0kg is added, abundant stirring is mixed well, the Root or stem of Littleleaf Indianmulberry of another 1 part of 1.0kg without bio-bacterial manure seed dressing in contrast, sows two pieces of connected experimental plots in the identical edatope of equal area on the same day respectively
,
in, carry out identical cultivation management simultaneously.First time in late May, 2013 cut issue respectively random selecting 100 strain Root or stem of Littleleaf Indianmulberry plant in two pieces of experimental plots, measures plant height and ground biomass, identical with embodiment 3 of measuring method.Respectively adopt 100 strains in addition, measure the full nitrogen of above-ground plant parts, full phosphorus, crude protein and crude fat content respectively, Comparative nutrition level.Result shows, and uses biology of the present invention
Root or stem of Littleleaf Indianmulberry after bacterial manure process, first time cut issue average plant height and ground biomass comparatively contrast and exceed 20.82% and 106.14% respectively; The total nitrogen content of the Root or stem of Littleleaf Indianmulberry after bio-bacterial manure process of the present invention, content of tatal phosphorus, crude protein content and crude fat content are than exceeding 19.34%, 61.88%, 19.34% and 25.08% respectively in contrast.Prove to use bio-bacterial manure process Root or stem of Littleleaf Indianmulberry of the present invention, effectively can improve output and the nutritional quality of Root or stem of Littleleaf Indianmulberry.
Certainly, the embody rule example of more than just invention, the technical scheme that the present invention also has other embodiment, all employings to be equal to replacement or equivalent transformation to be formed, all drops within protection domain of the presently claimed invention.
Claims (9)
1. a novel soybean or the bio-bacterial manure of leguminous forage rhizosphere, it is characterized in that: it is prepared from by the rhizosphere biological bacteria be separated from soybean, described rhizosphere biological bacteria comprises plant hormone secretion bacterial strain and at least two kinds of Rhizobium strains that the phosphorus-solubilizing bacteria strain having IAA secretion capacity concurrently of at least three kinds and at least one have phosphate solubilization concurrently.
2. novel soybean according to claim 1 or the bio-bacterial manure of leguminous forage rhizosphere, it is characterized in that: described in have IAA secretion capacity concurrently phosphorus-solubilizing bacteria strain comprise one or more combination in bacterial strain DR19, DR9, DR24, the described plant hormone secretion bacterial strain having phosphate solubilization concurrently comprises bacterial strain DR4, and described Rhizobium strains comprises one or more the combination in bacterial strain D2, D4; Wherein in classification, the DR9 change belonged in Klebsiella is dwelt klebsiella, DR24 belongs to the enterobacter cloacae in enterobacter, DR4 belongs to the osculant enterobacteria in enterobacter, and D2 belongs to the beans root nodule bacterium in rhizobium, and D4 belongs to the soil root nodule bacterium in rhizobium.
3. novel soybean according to claim 2 or the bio-bacterial manure of leguminous forage rhizosphere, is characterized in that:
The bacterial strain that this bio-bacterial manure amasss ratio by following bacteria liquid is made: bacterial strain DR19: bacterial strain DR9: bacterial strain DR24: bacterial strain DR4: bacterial strain D2: bacterial strain D4:=1:1:1:1:1:1.
4. novel soybean according to claim 2 or the bio-bacterial manure of leguminous forage rhizosphere, is characterized in that: the bacterial strain that this bio-bacterial manure amasss ratio by following bacteria liquid is made: bacterial strain DR19: bacterial strain DR9: bacterial strain DR24: bacterial strain DR4: bacterial strain D2: bacterial strain D4:=1:1:1:2:1:1.
5. novel soybean according to claim 2 or the bio-bacterial manure of leguminous forage rhizosphere, is characterized in that: the bacterial strain that this bio-bacterial manure amasss ratio by following bacteria liquid is made: bacterial strain DR19: bacterial strain DR9: bacterial strain DR24: bacterial strain DR4: bacterial strain D2: bacterial strain D4:=2:2:1:3:1:1.
6. the preparation method of a novel soybean or the bio-bacterial manure of leguminous forage rhizosphere, it is characterized in that: bacterial strain DR19, DR9, DR24, DR4, D2, D4 after activated are inoculated in SOC liquid nutrient medium by the bacterium liquid inoculum size of 1:1:1:2:1:1,26 ~ 28 DEG C, shaking culture under the condition of 180 ~ 200rpm, when bacterium liquid OD600 reaches 0.5 ~ 0.8, stop fermentation obtaining liquid active bacteria formulation, with vegetable mould as carrier, active bacteria formulation is cultivated through sterile carrier absorption, weighs packing and make solid live bacteria preparation.
7. claim 3 or the novel soybean described in 4 or 5 or the bio-bacterial manure of leguminous forage rhizosphere promote soybean bear pods on application.
8. claim 3 or the novel soybean described in 4 or 5 or the bio-bacterial manure of leguminous forage rhizosphere are promoting Trifolium repense growth and are improving the application of Trifolium repense nutritive ingredient.
9. claim 3 or the novel soybean described in 4 or 5 or the bio-bacterial manure of leguminous forage rhizosphere are promoting Root or stem of Littleleaf Indianmulberry growth and are improving the application of Root or stem of Littleleaf Indianmulberry nutritive ingredient.
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