CN100417719C - Pseudomonas RRj228 helpful for crop growth and microbe preparation containing the same - Google Patents
Pseudomonas RRj228 helpful for crop growth and microbe preparation containing the same Download PDFInfo
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- CN100417719C CN100417719C CNB2005100750367A CN200510075036A CN100417719C CN 100417719 C CN100417719 C CN 100417719C CN B2005100750367 A CNB2005100750367 A CN B2005100750367A CN 200510075036 A CN200510075036 A CN 200510075036A CN 100417719 C CN100417719 C CN 100417719C
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- 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 present invention relates to a pseudomonas RRj228 represented by sequence 1. The strain replaces fertilizer to promote plant growth by establishing in a plant root system to promote the plant growth in a growth process of crops. The present invention also relates to a microbe preparation with environment affinity, and the microbe preparation with the environment affinity contains the pseudomonas RRj228 or a secondary metabolite generated by the pseudomonas RRj228.
Description
Technical field
The present invention relates to that (this bacterial strain is deposited in Korea S agricultural culture collection center (KACC) on June 11st, 2003, and deposit number is KACC 91052 by the pseudomonas RRj228 (Pseudomonas sp.RRj228) of No. 1 sequence representative; Again be preserved in Korea S typical case culture collection center (KCTC) on May 27th, 2005, deposit number is KCTC 10812BP), in the crop planting process, this bacterial strain replaces chemical fertilizer to promote plant-growth inside and outside root system of plant by growing surely.The invention still further relates to and contain pseudomonas RRj228 or by the environment affinity microbial preparation of the secondary metabolites of its generation.
Environment affinity microbial preparation according to the present invention by contain pseudomonas RRj228 or can reduce by the secondary metabolites of its generation chemical fertilizer usage quantity, stablize crop yield, protection water quality and edatope and improve the health of soil ecosystem.
Background technology
Fertilizer is a kind of important agriculture goods and materials of decision crop yield, but it as the factor inferior of crop growth can cause crops quality degenerations, river eutrophication, groundwater pollution, soil nutrient is unbalance and the output reduction.Therefore, the soil microorganisms for promoting the protection of sustainable development of agriculture and agricultural environment, must rationally use to have the function that promotes plant-growth and helping the surging growth of farm crop substitutes chemical fertilizer.
Do not form symbiotic relationship and can promote the short endophytic bacteria (Plant growth-promoting rhizobacteria) of plant rhizosphere of plant-growth (PGPR) to name by people such as Kloepper in 1980 with host plant, it comprises Azospirillum (Azospirrilum sp.) (Okon, 1985), Pseudomonas (Kloepper etc., 1980), bacillus (Backman etc., 1994) etc.The plant-growth that is called as PGPR promote microorganism have multiple function as the nitrogen in the fixing atmosphere, produce plant-growth and promote hormone, promotion nutrition absorption (Hallmann etc., 1997) and avoid plant to be subjected to disease invasion and attack (Pleban etc., 1995), therefore help plant-growth.
People such as Ward point out in 1992 annual report roads to classify and this microbe species of identifying account for live in all categories in the soil less than 5%, and people such as Torisvik nineteen ninety report point out in soil, to exist many not by the microorganism of artificial culture.Therefore, also have the more microorganism of finding than in the past with excellent functions in soil, this causes the follow-up study to them thus.
In addition, confirmed to inoculate different plant-growths and promoted microorganism can make rice yield improve 19-60% (Baldani, 2000), and can improve tomato (Kloepper and Schroth, 1981), beans (Polonenko etc., 1987; And Dashti etc., 1998), the output of wheat (Freitas and Germida, 1990) and beet (Suslow and Schrith, 1982).Plant-growth promotes that application of microorganism possesses some special knowledge.
Therefore, inventor of the present invention separates in the research soil microorganisms and has identified the pseudomonas RRj228 that can grow surely inside and outside the rice plants root system, and finds that this bacterial strain can promote crop growth, therefore alternative chemical fertilizer.
Summary of the invention
The purpose of this invention is to provide the pseudomonas RRj228 by No. 1 sequence representative, in the crop planting process, this bacterial strain replaces chemical fertilizer to promote plant-growth inside and outside root system of plant by growing surely.The present invention also aims to provide and contain pseudomonas RRj228 or by the environment affinity microbial preparation of the secondary metabolites of its generation.
The invention provides the pseudomonas RRj228 by No. 1 sequence representative, in the crop planting process, this bacterial strain replaces chemical fertilizer to promote plant-growth inside and outside root system of plant by growing surely.
Pseudomonas RRj228 is a kind of soil bacteria of being found by the present inventor, under field conditions (factors), this bacterium grows surely inside and outside root system of plant, and the secretion plant-growth promotes material, and suppresses the growth of multiple pathogenic agent by the antagonistic action to plant pathogenic fungi and bacterium.
The Gram-negative shaft-like thermophilic bacteria of pseudomonas RRj228 of the present invention in 28 ± 2 ℃ PDA (potato dextrose agar) substratum, cultivating.
The effect of using pseudomonas RRj228 of the present invention to obtain is as follows:
When 1) microbial preparation that contains this bacterial strain when use is as the farm crop inoculum,,, also can improve crop yield although therefore use a spot of chemical fertilizer because pseudomonas RRj228 of the present invention can provide outstanding nutrient supply and promote plant-growth.
2) the pseudomonas RRj228 of the present invention substitute that is chemical fertilizer has use range widely to farm crop, as can be used for paddy rice, capsicum and cucumber.
3) when pseudomonas RRj228 of the present invention is inoculated in the farm crop, this bacterial strain can be by suppressing the harm that plant pathogenic microorganisms in the soil is helped crop growth and avoided pathogenic agent.
4) pseudomonas RRj228 of the present invention can be by great expression in the farm crop that inoculate by growing surely in farm crop root system inside.
5) according to the present invention, except the effect of alternative chemical fertilizer, by will also producing more value from the plant-growth promotion of pseudomonas RRj228 nutrient solution or the secondary metabolites that the cause of disease growth-inhibiting substance separates and purifying obtains.
6) owing to the impossible contaminate environment of pseudomonas RRj228 of the present invention, thus water quality and edatope can be protected, and improve the health of soil ecosystem.
The present invention also provides the environment affinity microbial preparation of the secondary metabolites that contains pseudomonas RRj228 or its generation.
Can use method well known in the art that microbial preparation of the present invention is mixed with plant growth promoter or pathogenic growth inhibitor, but the present invention is not limited thereto.For substituting chemical fertilizer, microbial preparation of the present invention preferably is mixed with the organic fertilizer that is used to promote plant-growth.
When microbial preparation of the present invention was the plant growth promoter of dry powder or liquid fertilizer shape, the pseudomonas RRj228 content in per 1 gram dry powder or 1 milliliters of liquid was 10
5-10
7
When microbial preparation of the present invention was the cause of disease growth inhibitor of dry powder or liquid fertilizer shape, the pseudomonas RRj228 content in per 1 gram dry powder or 1 milliliters of liquid should surpass 10
7In this case, it is characterized in that this bacterial strain content is than the bacterial strain content height in the plant growth promoter.
Environment affinity microbial preparation according to the present invention can reduce by containing pseudomonas RRj228 chemical fertilizer consumption, stablize crop yield, protection water quality and edatope and improve the health of soil ecosystem.
Pseudomonas RRj228 can produce becomes suitable microbial preparation, and said preparation can be inoculated in farm crop such as paddy rice, capsicum and the cucumber.
Biological preservation
RRj228 bacterial strain of the present invention is named as pseudomonas RRj228, and being deposited in the Korea S agricultural culture collection center (KACC) that is attached to Developing Rural Office, ,Korea/Agricultural biotechnologies institute of country on June 11st, 2003, deposit number is KACC 91052.
On May 27th, 2005, preservation is to Korea S typical case culture collection center (KCTC) again for the RRj228 quilt, and deposit number is KCTC10812BP.
Description of drawings
Fig. 1 represents the flat board of pseudomonas RRj228 culture;
Fig. 2 a and 2b are the electromicroscopic photographs of pseudomonas RRj228;
Fig. 3 is the photo of growing surely in the pseudomonas RRj228 of roots of plants inside.
Embodiment
The present invention can promote active relevant embodiment to embody with plant-growth by the separating of following and pseudomonas RRj228, evaluation, pseudomonas RRj228 separates from the soil of St. Petersburg, Russia (St.Petersburg), and it also is one of isolated 172 kinds of bacterial strains from Korea S and Russian soil.Yet, provide these embodiment just to be used for explaining, can not be considered to limitation of the scope of the invention, appended claim has been done appropriate description to scope of the present invention.
Embodiment 1
The separation of soil microorganisms RRj228
The separation method of soil microorganisms RRj228 is as follows.
As shown in Figure 1, a rice paddy seed of sprouting in aseptic culture medium is transferred in the 0.6% agarose substratum.0.5 gram pedotheque (from St. Petersburg, Russia) is seeded in the centimeters apart from rice root 1-2, then substratum was cultivated 48 hours at 28 ℃.With root pulverize make it to form suspension after, this suspension is coated on the LC substratum (this substratum contains microbial culture with Tryptones 10 grams, yeast extract 5 grams, MgSO
47H
21 liter of O 8 gram, 1 milliliter of 1M Tris and distilled water), with isolated strains.
Embodiment 2
The evaluation of soil microorganisms Rj228
A. physio-biochemical characteristics
The physio-biochemical characteristics of RRj228 bacterial strain are presented in the following table 1.
Described RRj228 bacterial strain is shaft-like, and size is 2.0-3.0 * 0.5-0.7 micron, and it has flagellum shown in Fig. 2 a and 2b.When this bacterial strain is cultivated 24 hours in the PDB substratum after, can form yellow reddish bacterium colony.Described RRj228 bacterial strain is a Gram-negative bacteria, carries out aerobic metabolism and has catalase and oxidase activity.When the carbon source that adds substratum is pectinose, fructose, semi-lactosi, glucose, N.F,USP MANNITOL, sucrose, glycerine or wood sugar, this bacterial strain can produce acid, and when the carbon source that adds substratum was lactose, dextrin, ramnose, Sorbitol Powder, maltose or melampyrum, this bacterial strain can produce alkali.In addition, as shown in Figure 3, the feature of RRj228 bacterial strain of the present invention is to be present in the root system of plant outside also to grow surely in root system inside.
[table 1]
B. sequential analysis
In order to analyze the RRj228DNA sequence, adopt 16S rDNA sequence analysis method.At first,, product is inserted in the pGEMT-T carrier, use automatic dna sequencer ABI3100 (Applied Biosystem, the U.S.) to carry out dna sequence analysis then with PCR (polymerase chain reaction) amplification 16S rDNA part.The fD1 (5 '-agagtttgatggctcag-3 ') of No. 2 sequences and the rP2 of No. 4 sequences (5 '-acggctaccttgttacgactt-3 ') are used in PCR reaction.In addition, use pUC/M 13 forward primer (5 '-gttttcccagtcacgac-3 ': No. 4 sequences) and reverse primer (5 '-gcggataacaatttcacacagg-3 ': No. 5 sequences) react two end regions of analyzing above-mentioned primer, and use 4 primers to react to analyze the dna sequence dna of inside.
According to intestinal bacteria (Escherichia coli) 16S rDNA sequence (GenBank J01695 number), the 311-330 zone of each dna sequence dna use 16SP-1 (5 '-gccacactggaactgagacac-3 '), the 684-703 zone of 16SP-2 (5 '-tgtagcg gtgaaatgcgtg-3 '), the 1228-1247 zone of the 944-963 zone of 16SP-3 (5 '-ggagcatgtgt tattcg-3 ') and 16SP-4 (5 '-ctacacacgtgctacaatgg-3 ').
No. 1 sequence is represented the 16S rDNA sequence of RRj228 bacterial strain.Homology between the dna sequence dna in this bacterial strain and the GenBank database is discovered, the homology of this bacterial strain and many Pseudomonas members such as Pseudomonas chlororaphis (Pseudomonas chlororaphis), gauffer pseudomonas (P.Corrugata) and P.tolaasii reaches 99%, thus this bacterial strain is classified as Pseudomonas.
Therefore, RRj228 bacterial strain of the present invention is named as pseudomonas RRj228.
EXPERIMENTAL EXAMPLE 1
The plant-growth facilitation effect of pseudomonas RRj228
In order to detect the plant-growth facilitation effect of pseudomonas RRj228, corn seedling is experimentized.Corn root was immersed in these strain culture suspension 1.5 hours for terminal 3 millimeters, and other corn root immerses in aseptic culture fluid in contrast.Corn seedling is placed on the wet filter paper in the plastic box, with filter paper the root end at start line place is built then, cultivated 24 hours for 28 ℃.Measure the length of the root of new elongation, repeat 10 times, the corn growth facilitation effect of described RRj228 bacterial strain is presented in the following table 2.
[table 2]
The corn growth facilitation effect of RRj228 bacterial strain
As shown in table 2, with stoste (4.5 * 10
9The c.f.u/ milliliter) with dilution in 1: 50 and when inoculating, the growth level of pseudomonas RRj228 of the present invention has reduced by 26%, and with stoste with dilution in 1: 100 and when inoculating, its growth level has improved 13%.Therefore, bacterial strain of the present invention is more effective when inoculating with low concentration.
EXPERIMENTAL EXAMPLE 2
The soil disease antagonistic action of pseudomonas RRj 228
Use following method to detect the soil disease antagonistic action of pseudomonas RRj228.At first, in PDA, inoculate pathogenic fungi and bacterium respectively, then nutrient solution is transferred in the petri diss and made it and solidify.In each flat board, do a radius and be 4 millimeters well, with the pseudomonas RRj228 of 100 microlitres in the PDA liquid nutrient medium 28 ℃ cultivated 5 days, be inoculated into then in this well.Flat board was placed 5 days down at 28 ℃, detected the growth-inhibiting scope of PGPR then, this result is presented in the following table 3.With 5 milliliter (1 * 10
6Individual spore/milliliter) fungi and 5 milliliter (1 * 10
8The cfu/ milliliter) bacterium mixes with 250 milliliters of PDA substratum respectively and is incorporated in 45 ℃ of vibrations, and each petri diss is respectively got 25 milliliters of coated plates of solution, triplicate.
[table 3]
The RRj228 bacterial strain suppresses the growth scope * (suppressing growth scope Ф mm) of plant pathogenic fungi and bacterium
* inoculate the pathogenic growth of this bacterial strain after 5 days and suppress scope
The result shows that RRj228 bacterial strain of the present invention shows very strong growth inhibitory effect to Phytophthora capsici, pythium spp, carrot soft rot erwinia and Pseudomonas syringae.
EXPERIMENTAL EXAMPLE 3
Pseudomonas RRj228 competition enters the relative capacity of plant root
For studying the relative capacity that pseudomonas RRj228 and fluorescent pseudomonas (Pseudomonasfluorescens) WCS365 competition enters plant root, two kinds of bacterial strains are inoculated into simultaneously after the root of paddy rice (Kumo paddy rice) and radish (Saxa nova), detect pseudomonas RRj228 and account for the per-cent of growing all bacterial strains in root surely, the result shows in following table 4.
[table 4]
Pseudomonas RRj228 and fluorescent pseudomonas WCS365 competition enter the relative capacity of plant root
EXPERIMENTAL EXAMPLE 4
The effect of inoculation pseudomonas RRj228 in farm crop
The nutrient solution of the pseudomonas RRj228 of the present invention that routine is cultivated is inoculated on paddy rice, capsicum, cucumber and the tomato, and the result is presented in the following table 5.
[table 5]
As shown in table 5, confirm that pseudomonas RRj228 makes that paddy growth has increased by 33.2%, chili growth has increased by 8.5%, cucumber growth has increased by 21.4%, tomato growth has increased by 17.5%, and pseudomonas RRj228 of the present invention can be used as microbial fertilizer and substitutes chemical fertilizer.
Because pseudomonas RRj228 of the present invention directly separates from plant (paddy rice) root, rather than the bacterial strain before resembling is isolating from soil, so this bacterial strain can suitably be grown surely in the root system of farm crop such as paddy rice and cucumber, therefore when with this inoculation to farm crop the time, it can be by stably growing self duplication in the farm crop root system and inside and outside root system surely, between the farm crop survival time, because rainwater or arid are difficult to make its density to reduce, so its effect of inoculation can keep for a long time.
Industrial usability
, can subtract by using pseudomonas RRj228 according to environment compatibility microorganism formulation of the present invention The use of few chemical fertilizer, stablize crop yield, protect water quality and soil environment and improve soil ecosystem Health.
Sequence table
＜110〉Developing Rural Office, ,Korea (Republic of Korea represented by Rural Development Administration)
＜120〉help the pseudomonas of crop growth and contain the microbial preparation of this bacterial strain
<130>I5042YOL
<150>KR10-2004-0041457
<151>2004-06-07
<160>5
<170>PatentIn?3.1
<210>1
<211>1501
<212>DNA
＜213〉pseudomonas RRj228 (Pseudomonas sp.RRj228)
<400>1
gagtttgatc?ctggctcaga?ttgaacgctg?gcggcaggcc?taacacatgc?aagtcgagcg 60
gtagagagaa?gcttgcttct?cttgagagcg?gcggacgggt?gagtaatgcc?taggaatctg 120
cctggtagtg?ggggataacg?tccggaaacg?gacgctaata?ccgcatacgt?cctacgggag 180
aaagcagggg?accttcgggc?cttgcgctat?cagatgagcc?taggtcggat?tagctagttg 240
gtgaggtaat?ggctcaccaa?ggcgacgatc?cgtaactggt?ctgagaggat?gatcagtcac 300
actggaactg?agacacggtc?cagactccta?cgggaggcag?cagtggggaa?tattggacaa 360
tgggcgaaag?cctgatccag?ccatgccgcg?tgtgtgaaga?aggtcttcgg?attgtaaagc 420
actttaagtt?gggaggaagg?gtacttacct?aatacgtgag?tattttgacg?ttaccgacag 480
aataagcacc?ggctaactct?gtgccagcag?ccgcggtaat?acagagggtg?caagcgttaa 540
tcggaattac?tgggcgtaaa?gcgcgcgtag?gtggttcgtt?aagttggatg?tgaaatcccc 600
gggctcaacc?tgggaactgc?atccaaaact?ggcgagctag?agtatggtag?agggtggtgg 660
aatttcctgt?gtagcggtga?aatgcgtaga?tataggaagg?aacaccagtg?gcgaaggcga 720
ccacctggac?atgatactga?cactgaggtg?cgaaagcgtg?gggagcaaac?aggattagat 780
accctggtag?tccacgccgt?aaacgatgtc?aactagccgt?tgggagcctt?gagctcttag 840
tggcgcagct?aacgcattaa?gttgaccgcc?tggggagtac?ggccgcaagg?ttaaaactca 900
aatgaattga?cgggggcccg?cacaagcggt?ggagcatgtg?gtttaattcg?aagcaacgcg 960
aagaacctta?ccaggccttg?acatccaatg?aactttccag?agatggattg?gtgccttcgg 1020
gaacattgag?acaggtgctg?catggctgtc?gtcagctcgt?gtcgtgagat?gttgggttaa 1080
gtcccgtaac?gagcgcaacc?cttgtcctta?gttaccagca?cgttatggtg?ggcactctaa 1140
ggagactgcc?ggtgacaaac?cggaggaagg?tggggatgac?gtcaagtcat?catggccctt 1200
acggcctggg?ctacacacgt?gctacaatgg?tcggtacaga?gggttgccaa?gccgcgaggt 1260
ggagctaatc?ccataaaacc?gatcgtagtc?cggatcgcag?tctgcaactc?gactgcgtga 1320
agtcggaatc?gctagtaatc?gcgaatcaga?atgtcgcggt?gaatacgttc?ccgggccttg 1380
tacacaccgc?ccgtcacacc?atgggagtgg?gttgcaccag?aagtagctag?tctaaccttc 1440
gggaggacgg?ttaccacggt?gtgattcatg?actggggtga?agtcgtaaca?aggtagccgt 1500
a 1501
<210>2
<211>17
<212>DNA
＜213〉artificial sequence
<220>
＜223〉be used to the to increase pcr amplification primer fD1 of pseudomonas RRj228 gene
<400>2
agagtttgat?ggctcag 17
<210>3
<211>21
<212>DNA
＜213〉artificial sequence
<220>
＜223〉be used to the to increase pcr amplification primer rP2 of pseudomonas RRj228 gene
<400>3
acggctacct?tgttacgact?t 21
<210>4
<211>17
<212>DNA
＜213〉artificial sequence
<220>
＜223〉be used to the to increase pcr amplification forward primer of pUC/M13 of pseudomonas RRj228 gene
<400>4
gttttcccag?tcacgac 17
<210>5
<211>22
<212>DNA
＜213〉artificial sequence
<220>
＜223〉be used to the to increase pcr amplification reverse primer of pUC/M13 of pseudomonas RRj228 gene
<400>5
gcggataaca?atttcacaca?gg 22
Claims (6)
1. a pseudomonas (Pseudomonas), this bacterial strain contains the rDNA sequence by SEQ ID NO:1 representative, and the deposit number of this bacterial strain is KCTC10812BP.
2. microbial preparation, this microbial preparation contains pseudomonas as claimed in claim 1 as effective constituent.
3. soil management or plant cultivation method, this method is by handling soil with microbial preparation as claimed in claim 2 or plant carries out soil management or plant cultivation.
4. method according to claim 3, wherein, described preparation is for substituting the organic fertilizer of chemical fertilizer.
5. method according to claim 3, wherein, described preparation is used to promote plant-growth.
6. method according to claim 3, wherein, described preparation is used to suppress pathogenic growth.
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|---|---|---|---|
| KR1020040041457A KR100646597B1 (en) | 2004-06-07 | 2004-06-07 | 228 Pseudomonas sp. RRj228 helping the growth of crops and the microbial agent containing the same |
| KR1020040041457 | 2004-06-07 |
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| CN100417719C true CN100417719C (en) | 2008-09-10 |
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| KR100973168B1 (en) | 2008-05-08 | 2010-07-30 | 충북대학교 산학협력단 | Pseudomonas corrugata mutants promoting plant growth at low temperature and method for promoting plant growth using the same |
| KR101153462B1 (en) | 2010-01-08 | 2012-06-27 | 대한민국 | A New bacteria to enhance rice production |
| US20190320661A1 (en) * | 2017-01-03 | 2019-10-24 | Monsanto Technology, Llc | Microbial compositions and methods |
| KR101972379B1 (en) * | 2018-11-08 | 2019-04-25 | 김경현 | A method for managing golf lawns at low temperatures |
| KR102324981B1 (en) * | 2020-06-11 | 2021-11-12 | 연세대학교 원주산학협력단 | Novel Pseudomonas Fluorescens Strain that Induces Resistance to Drought Stress in Plant and Uses Thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001015524A2 (en) * | 1999-09-02 | 2001-03-08 | Michigan State University | Method for controlling bacterial and fungal disease |
| CN1519309A (en) * | 2003-05-22 | 2004-08-11 | 郑爱萍 | Strain of pseudomonas for promoting growth of paddy as well as preventing and curing sheath blight of rice |
-
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- 2004-06-07 KR KR1020040041457A patent/KR100646597B1/en active IP Right Grant
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001015524A2 (en) * | 1999-09-02 | 2001-03-08 | Michigan State University | Method for controlling bacterial and fungal disease |
| CN1519309A (en) * | 2003-05-22 | 2004-08-11 | 郑爱萍 | Strain of pseudomonas for promoting growth of paddy as well as preventing and curing sheath blight of rice |
Non-Patent Citations (4)
| Title |
|---|
| 国外微生物肥料的发展概况. 赵子定，常玉海.中国农业信息快讯，第7期. 2001 |
| 国外微生物肥料的发展概况. 赵子定，常玉海.中国农业信息快讯，第7期. 2001 * |
| 微生物肥料及其在农业生产中的应用. 李南，李涛.四川农业科技，第4期. 2001 |
| 微生物肥料及其在农业生产中的应用. 李南，李涛.四川农业科技，第4期. 2001 * |
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| KR20050116297A (en) | 2005-12-12 |
| CN1876805A (en) | 2006-12-13 |
| KR100646597B1 (en) | 2006-11-23 |
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