CN104987139B - A kind of composite microbiological fertilizer and its application in southerm yew growth and taxol accumulation is promoted - Google Patents
A kind of composite microbiological fertilizer and its application in southerm yew growth and taxol accumulation is promoted Download PDFInfo
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- CN104987139B CN104987139B CN201510089892.1A CN201510089892A CN104987139B CN 104987139 B CN104987139 B CN 104987139B CN 201510089892 A CN201510089892 A CN 201510089892A CN 104987139 B CN104987139 B CN 104987139B
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- pseudomonas fluorescens
- southerm yew
- glomus versiforme
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Abstract
The invention discloses a kind of composite microbiological fertilizer, and it includes Pseudomonas fluorescens Pseudomonas fluorescens and Glomus versiforme Glomus versiforme；Wherein, Pseudomonas fluorescens Pseudomonas fluorescens are CCTCC M 2010158 in the deposit number of China typical culture collection center；Glomus versiforme Glomus versiforme are BGC NM03C in Arbuscular Mycorrhizal Fungi In China germplasm resource bank BGC deposit number, are 1511C0001BGCAM0032 in National Nature scientific and technological resources platform resource number.The invention also discloses application of the above-mentioned composite microbiological fertilizer in southerm yew growth promotion and taxol accumulation.The present invention can provide excellent strain resource for exploitation in future southerm yew microbial fertilizer special.
Description
Technical field
The invention belongs to the microbial bacterial agent technical field in biological fertilizer field, and in particular to one kind is by being isolated from south
Chinese yew rhizosphere efficiently solves compound made of Pseudomonas fluorescens and AMF Glomus versiforme (G.versiforme)
Fertilizer and its application in southerm yew growth and taxol accumulation is promoted.
Background technology
Southerm yew (Taxus chinensis var.mairei) is Chinese endemic tree, is Tertiary Period few survivors seeds
It is one of main source of new type anticancer natural drug taxol (Taxol) with first-grade state protection rare tree species.Taxol is
What (1971) such as Americanized scholar Wall were extracted from Pacific yew (Taxus brevifolia) bark first has five first
The diterpene-kind compound of the carbene skeleton of base 15.It is wide spectrum generally acknowledged in the world, the cancer therapy drug of high activity.Due to domestic and international
An urgent demand to Chinese yew extract taxol, China is set unprecedented taxus resource exploitation upsurge, various regions occur
Natural taxus resource is even endangered by heavy damage, some places.Therefore, how to protect, develop and utilize it is red
Beans China fir resource, realizes the sustainable development of taxol industry, it has also become the important topic of urgent need to resolve.
Southerm yew is the species that the Taxus kind speed of growth is most fast, distribution is most wide, is Chinese Chinese yew people
The principal item of work cultivation.Artificially-induced bezoar, improving the biosynthesis of taxol and its precursor in plant body is
Solves the important channel of taxol resource bottleneck.To realize the high-yield and high-efficiency operation of the short cycle Medical forests of southerm yew, fertilising
It is an important link.At present, the various microbial manures developed based on microorganism obtain in agriculture and forestry production
To extensive use, such fertilizer has the characteristics that fertilizer efficiency is high, itself is nontoxic, free from environmental pollution, cost is low, saves the energy, being
Learn the maximally effective substitute of fertilizer.
Phosphate-solubilizing bacteria serves as important role in the related biology system of soil phosphorus circulation, and it can be by indissoluble
Phosphorus is converted into the titanium pigment that plant can absorb, and especially in plant rhizosphere part, phosphate-solubilizing bacteria also acts as to a certain degree
Growth-promoting functions.After the phosphate-solubilizing bacteria separated on calacareous soil is manured into soil by EI-Tarabily etc. (2008), add big
The biomass of beans, available phosphorus content in soil is not only increased, and improve soybean plant strain to the nutrition such as N, P, K, Zn, Mg member
Absorption.
Arbuscular mycorrhiza (Arbuscular Mycorrhizae, AM) fungi is one of most important member in soil.AM is true
Bacterium significantly affects the nascent metabolic process of plant, and it can promote root system of plant to the mineral elements such as phosphorus, copper, zinc, cadmium and nutrient
Absorption, adjust the synthesis and distribution of plant hormone, improve the rhizosphere microorganism environment of plant and strengthen the disease resistance of plant, together
The tolerance of Shi Tigao plant pair environment-stress, so as to improve the upgrowth situation of host plant comprehensively.In recent years, it is many to study
It is observed that to AM fungies and the correlation of Secondary Metabolism of Plant, AM fungies can directly or indirectly influence the secondary metabolism of plant
Process.Rojas-Andrade etc. (2003) is inoculated with algarroba with the huge sporangiocyst of rose-red mould (Gigaspora rosea)
Found when (Prosopis laevigata), trigonelline (trigonelline, a kind of pyrrole in the screwbean mesquite tree root of AM fungal infections
Pyridine alkaloid) content add 1.8 times than control.Some existing results of study are also shown that mycorrhizal fungi by infecting shape
Influence into mycorhiza to Secondary Metabolism of Plant has obvious species variation.Zhao Xin etc. (2006) is with 6 kinds of AM fungies to Camptotheca acuminata seedlings
After being inoculated with, it is found that different vaccination processing camptothecine (camptothecin, CPT) content is variant, Glomus etunicatum, printing opacity
Sacculus is mould, nectarine Acaulospora and light wall Acaulospora improve the content of camptothecine in Camptotheca acuminata seedlings, and Glomus versiforme influences not
Substantially, and Glomus manihotis reduces the content of camptothecine.At present, AM fungies technology has been applied in the planting of medicinal materials research
Gradually attract people's attention.Many medicinal plant active components such as alkaloid, terpene etc. belong to secondary metabolite, at present
Influence research of the medicinal plant AM fungies to its host plant secondary metabolite is less, quantity and gos deep into degree and all seems not
It is enough.AM fungies can infect southerm yew root formed typical AM (Wang Sen etc., 2008；Ren Jiahong etc., 2008), but current state
It is inside and outside there is not yet AM fungies on southerm yew grow and its secondary metabolite content of taxol influence relevant report.Cause
This, excellent AM fungies are screened from taxus chinensis var. mairei rhizosphere, are promoted using biological technology of mycorrhiza that southerm yew grows and secondary
The accumulation of raw metabolite taxol and precursor, this will be helpful to the artificial domesticating cultivation of rare tree southerm yew and open country
The recovery of raw southerm yew resource.
Biological technology of mycorrhiza turns into most valued research and development field in current sustainable development technology.It is but single
The microbial manure of function can not meet the development need of modern agriculture and forestry, develop multi-functional compound (or compound) microorganism
Fertilizer turned into needed for the epoch.As the Major Members of edaphon family, many between phosphate-solubilizing bacteria and mycorrhizal fungi
Aspect is interactional.Some scholars have carried out the interaction of phosphate-solubilizing bacteria and mycorrhizal fungi and its host plant have been grown
The research influenceed with Nutrient Absorption, it is indicated that phosphate-solubilizing bacteria is with mycorrhizal fungi interaction in increase P acquisition and the growth-promoting functions of plant
Above it is better than effect during single inoculation.Kim etc. (1998) gathers Glomus etunicatum (Glomus etunicatum) with phosphate-solubilizing bacteria
Group's enterobacteria (Enterobacter agglomerans) co-inoculation after tomato, significantly promotes the growth of tomato and to phosphorus
The absorption of element.At present, the area research have focused largely on crops AM fungies and phosphate-solubilizing bacteria mutually oppose its growth and support
Divide in the relation absorbed.At present, the application study about taxus chinensis var. mairei rhizosphere high-efficient phosphate-solubilizing bacterium and AM fungies has no report.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of southerm yew Special compound microbial fertilizer.
The present invention also technical problems to be solved are to provide above-mentioned composite microbiological fertilizer and are promoting the life of southerm yew seedling
Application in the accumulation of long and taxol.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention is as follows：
A kind of composite microbiological fertilizer, it includes Pseudomonas fluorescens Pseudomonas fluorescens and earth's surface ball
The mould Glomus versiforme of capsule；
Wherein,
Pseudomonas fluorescens Pseudomonas fluorescens (bacterial strain CLW17) are in China typical culture collection
The deposit number at center is CCTCC M 2010158；The bacterial strain is documented in Chinese patent ZL201010221242.5.
AMF Glomus versiforme Glomus rersiforme are in Arbuscular Mycorrhizal Fungi In China germplasm resource bank
BGC deposit number is BGC NM03C, is 1511C0001BGCAM0032 in National Nature scientific and technological resources platform resource number.
Preferably, Pseudomonas fluorescens Pseudomonas fluorescens and Glomus versiforme Glomus
Versiforme proportioning is 50 milliliters：25 grams.Wherein Pseudomonas fluorescens is NB culture mediums fermentation 48h nutrient solution；Earth's surface
The spore that it is Glomus versiforme Glomus versiforme that sacculus is mould is to carry with butch clover (Trifolium Nepens L.)
Body enrichment culture, culture matrix are sterilized soil and husky (mass ratio 2: 1) mixture, after cultivating 2 months, remove aerial part,
Culture is made.
Application of the above-mentioned composite microbiological fertilizer in the accumulation of Chinese yew seedlings taxol is promoted.
Wherein, described Chinese yew seedlings are southerm yew seedling.
Above-mentioned composite microbiological fertilizer applies in Chinese yew seedlings growth is promoted
Wherein, described Chinese yew seedlings are southerm yew seedling.
Beneficial effect：
Composite bacteria agent made of phosphate-solubilizing bacteria Pseudomonas CLW17 and AM fungi Glomus versiforme is inoculated in southern red
Beans China fir, the results showed that, the microbial inoculum is obviously promoted the accumulation grown with taxol of southerm yew seedling.Therefore, it is of the invention
Excellent strain combination is provided for exploitation southerm yew dedicated fertilizer.
Brief description of the drawings
Fig. 1 is the growth-promoting effect of CLW17, G.versiforme to southerm yew seedling.
Embodiment
According to following embodiments, those skilled in the art can be made to more fully understand the present invention.Described by embodiment
The present invention is merely to illustrate, without should be also without limitation on the present invention described in detail in claims.
Pseudomonas fluorescens Pseudomonas fluorescens (bacterial strain CLW17) used in following examples are in
The deposit number of state's Type Tissue Collection is CCTCC M 2010158；The bacterial strain is documented in Chinese patent
In ZL201010221242.5.
AMF Glomus versiforme Glomus versiforme used in following examples are in Chinese arbuscular mycorrhiza
Fungi germplasm resource bank BGC deposit number is BGC NM03C, is in National Nature scientific and technological resources platform resource number
1511C0001BGCAM0032。
Embodiment 1：It is inoculated with the preparation of microbial inoculum
(1) the spore 30%H of AM fungies Glomus versiforme (G.versiforme)2O210min is sterilized, it is then sterile
Water is rinsed for several times, and the spore of rinsed AM fungies is trained with butch clover (Trifolium Nepens L.) for carrier enrichment
Foster, culture matrix is sterilized soil and husky (2: 1) mixture, after cultivating 2 months, removes aerial part, is made containing spore, bacterium
Silk and infect the mixture of root segment as AM fungi Inoculants；
(2) after CLW17 is activated 2~3 times, it is inoculated in a small amount of thalline of oese picking containing 50mL NB culture medium (oxen
Meat extract 3g, peptone 10g, sodium chloride 5g, distilled water 1000mL, pH 7.2~7.4) in 100mL triangular flasks, 30 DEG C, 200r/
Inoculant is used as after min shaken cultivations 48h.
Embodiment 2：Microbial inoculum inoculation greenhouse pot culture experiment
The Inoculant prepared in embodiment 1 is inoculated in 1 year raw seedling root of southerm yew respectively, trained with blank NB
Support base and AM microbial inoculums expand numerous matrix as control, set 4 processing altogether.4 processing respectively (1) G.versiforme, inoculum concentration
For 25 grams/plant；(2) CLW17, inoculum concentration are 50mL/ strains；(3) CLW17+G.versiforme, AM inoculum concentration are 25 grams/plant, solution
Phosphobacteria inoculum concentration is 50mL/ strains；(4) blank control CK.15 repetitions are often handled, are placed in greenhouse unified management, illumination is
12h/ days, watering in good time.
360 days growing states (being shown in Table 1) of southerm yew seedling.As can be seen from Table 1, inoculation microbial inoculum has been obviously promoted south
The growth of square Chinese yew seedling.Height of seedling, ground diameter, the biomass of inoculation processing southerm yew seedling have not compared with CK
With the growth of degree.Wherein, best with double effect of inoculation, height of seedling and ground diameter ratio are not inoculated with control (CK) and increased respectively
76.79% and 16.43%；Biomass increases 85.50% respectively than CK.
The influence that table 1 phosphorus bacteria fertilizer CLW17, AM fungi grows to southerm yew
Note：P ＜ 0.05, same column lowercase of not going together differ and represent significant difference.
Embodiment 3：The detection experiment of greenhouse pot culture seedling content of taxol
After being inoculated with 1 year to the southerm yew seedling in embodiment 2, the root, stem, leaf of different disposal are dried at 56 DEG C respectively
It is standby that dry grinding is broken into powder.5 grams of powders are weighed respectively to be placed in 500ml conical flask with stopper, measure the extraction of 250ml methanol
1h, then ultrasonic extraction 20min, is filtered.Repeat above step 3 times.Filtered fluid is incorporated in vacuum rotating at 37 DEG C and is evaporated, will
To dry be dissolved in 10ml methanol, obtain taxol crude extract after crossing 0.45 μm of filter membrane, be placed in -20 DEG C it is standby.With Japanese yew
Alcohol (Sigma) is reference material, and the measure of different disposal content of taxol is carried out using HPLC.
As can be seen from Table 2, single inoculation AM fungies Glomus versiforme (G.versiforme) and combined inoculation phosphorus decomposing are thin
Bacterium Pseudomonas fluorescens CLW17 and G.versiforme has certain to southerm yew seedling root, stem, leaf medicinal ingredient taxol
Summation；Wherein, best with double effect of inoculation, root, the yew alcohol content of leaf are respectively 824.61,143.83,88.26 μ g
g-1Dry weight, it is only 161.61,20.95,55.62 μ gg without being inoculated with processing-1Dry weight.But single inoculation CLW17 is to taxol
Accumulation does not almost have effect, does not have difference with compareing CK.
The influence that phosphate-solubilizing bacteria CLW17, AM fungus G .versiforme of table 2 accumulates to southerm yew seedling taxol
Note：P ＜ 0.05, same column lowercase of not going together differ and represent significant difference.
Claims (1)
1. a kind of application of composite microbiological fertilizer in the accumulation of southerm yew seedling taxol is promoted, wherein, it is described
Composite microbiological fertilizer includes Pseudomonas fluorescens Pseudomonas fluorescens and Glomus versiforme Glomus
versiforme；
Wherein,
Pseudomonas fluorescens Pseudomonas fluorescens are in the deposit number of China typical culture collection center
CCTCC M 2010158；
Glomus versiforme Glomus versiforme are BGC in Arbuscular Mycorrhizal Fungi In China germplasm resource bank BGC deposit number
NM03C, it is 1511C0001BGCAM0032 in National Nature scientific and technological resources platform resource number.
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| CN106258057A (en) * | 2016-08-09 | 2017-01-04 | 中国矿业大学(北京) | Arbuscular mycorrhiza and phosphate solubilizing bacteria work in coordination with promotion plant growing and the method for soil improvement |
| CN108605706B (en) * | 2018-03-20 | 2021-04-27 | 中山大学 | Method for improving cadmium resistance of tomatoes |
| CN108781979A (en) * | 2018-07-03 | 2018-11-13 | 东北林业大学 | A method of promoting southerm yew growth |
| CN115354003B (en) * | 2022-08-25 | 2023-12-19 | 安徽省农业科学院园艺研究所 | Strain for promoting high-density growth of cucurbits and reducing cadmium absorption and application thereof |
| CN116199538A (en) * | 2022-11-30 | 2023-06-02 | 乐施康农业科技有限公司 | Microbial agent for promoting peanut growth and preparation method thereof |
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| CN101693641A (en) * | 2009-10-22 | 2010-04-14 | 南京信息工程大学 | Compound bio-fertilizer containing ardealite and application thereof |
| CN101899409A (en) * | 2010-07-07 | 2010-12-01 | 长治学院 | Taxus chinensis var. mairei rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens and application thereof |
| CN104016795A (en) * | 2014-06-06 | 2014-09-03 | 成都新柯力化工科技有限公司 | Biological phosphatic fertilizer as well as preparation method thereof |
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Patent Citations (3)
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|---|---|---|---|---|
| CN101693641A (en) * | 2009-10-22 | 2010-04-14 | 南京信息工程大学 | Compound bio-fertilizer containing ardealite and application thereof |
| CN101899409A (en) * | 2010-07-07 | 2010-12-01 | 长治学院 | Taxus chinensis var. mairei rhizosphere high-efficient phosphate-solubilizing pseudomonas fluorescens and application thereof |
| CN104016795A (en) * | 2014-06-06 | 2014-09-03 | 成都新柯力化工科技有限公司 | Biological phosphatic fertilizer as well as preparation method thereof |
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| 南方红豆杉根际溶无机磷细菌的筛选、鉴定及其促生效果;任嘉红，刘辉，吴晓蕙等;《微生物学报》;20120304;第52卷(第3期);295-303 * |
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