CN102405765B - Fungus agent inoculating method for improving nutrients of pinellia tuber and quality of medicinal material - Google Patents
Fungus agent inoculating method for improving nutrients of pinellia tuber and quality of medicinal material Download PDFInfo
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- 241001522129 Pinellia Species 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 38
- 235000015097 nutrients Nutrition 0.000 title claims abstract description 24
- 239000000463 material Substances 0.000 title claims abstract description 5
- 241000233866 Fungi Species 0.000 title claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 16
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 16
- 239000011574 phosphorus Substances 0.000 claims abstract description 16
- 230000033228 biological regulation Effects 0.000 claims abstract description 9
- 239000011159 matrix material Substances 0.000 claims description 33
- 239000002068 microbial inoculum Substances 0.000 claims description 21
- 238000011081 inoculation Methods 0.000 claims description 17
- 238000004659 sterilization and disinfection Methods 0.000 claims description 13
- 230000001954 sterilising effect Effects 0.000 claims description 11
- 241001123597 Funneliformis mosseae Species 0.000 claims description 9
- 239000012567 medical material Substances 0.000 claims description 8
- 238000009331 sowing Methods 0.000 claims description 7
- 239000012286 potassium permanganate Substances 0.000 claims description 6
- 239000004576 sand Substances 0.000 claims description 6
- 235000019354 vermiculite Nutrition 0.000 claims description 6
- 239000010455 vermiculite Substances 0.000 claims description 6
- 229910052902 vermiculite Inorganic materials 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 230000003203 everyday effect Effects 0.000 claims description 3
- 238000010899 nucleation Methods 0.000 claims description 3
- 150000007524 organic acids Chemical class 0.000 claims description 3
- 235000005985 organic acids Nutrition 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 230000003442 weekly effect Effects 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 2
- 230000008030 elimination Effects 0.000 claims description 2
- 238000003379 elimination reaction Methods 0.000 claims description 2
- 239000012535 impurity Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims 3
- 239000000243 solution Substances 0.000 claims 2
- 238000000205 computational method Methods 0.000 claims 1
- 239000012895 dilution Substances 0.000 claims 1
- 238000010790 dilution Methods 0.000 claims 1
- 239000003337 fertilizer Substances 0.000 abstract description 11
- 239000003814 drug Substances 0.000 abstract description 7
- 230000000813 microbial effect Effects 0.000 abstract description 5
- 235000021049 nutrient content Nutrition 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000013459 approach Methods 0.000 abstract description 2
- 241000773655 Ambispora fennica Species 0.000 abstract 3
- 239000002609 medium Substances 0.000 abstract 2
- 230000007850 degeneration Effects 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000010871 livestock manure Substances 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 2
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241001522232 Pinellia ternata Species 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- 240000006064 Urena lobata Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 208000024386 fungal infectious disease Diseases 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 238000013332 literature search Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002686 phosphate fertilizer Substances 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 230000001863 plant nutrition Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 239000001384 succinic acid Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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- Mushroom Cultivation (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses an arbuscular mycorrhiza fungus inoculating method for improving nutrients of pinellia tuber and quality of medicinal material. In the invention, through the manual regulation of factors such as culture medium, quantity of arbuscular mycorrhiza fungus, phosphorus nutrient content and water content in medium, and coaction of the medium and other microbial fertilizers, the establishment of the symbiotic relationship between pinellia and arbuscular mycorrhiza fungi is promoted effectively, and the nutrients and medicinal quality of pinellia tuber are improved obviously. The method represents a new thought and approach for solving the degeneration problem of pinellia by using microbial fertilizers. The method has certain practical significance for improving medicinal quality of Chinese medicinal materials by reasonably utilizing microbial fertilizers in a modern Chinese medicine production system.
Description
Technical field
The invention belongs to traditional Chinese medicine culture technique field.
Background technology
The tuber of pinellia (Pinellia ternata Breit.) is Araeceae Pinellia Tenore plant, is the traditional large parts of generic medicinal plants of China.Due to the excessive collection of people to the Wild Rhizoma Pinelliae resource, cause the Wild Rhizoma Pinelliae resource very deficient.And in the artificial cultivation process, the unreasonable of fertilizer applies, and the reasons such as the unreasonable regulation and control of moisture cause the tuber of pinellia kind of a sexual involution problem to occur, cause the stability decreases of Pinellia Ternate quality, output.
Arbuscular mycorrhizal fungi (AM) is a most ancient parasymbiosis fungi, almost occurs on earth simultaneously with terrestrial plant.Form the various physiological metabolism activities of plant corpus acrial part except hereditary capacity of mycorrhizal fungi, all can be subject to the impact of arbuscular mycorrhiza to the growth of plant, nutrition condition, yield and quality etc., and majority is favorable influence.
EM (effective microorganisms) bacterium is CEM, and it is by short living the composition of the microorganisms such as light and bacterium, lactic acid bacteria, saccharomycete, bacillus, acetic acid bacteria, actinomycetes.The EM microbial inoculum can be bought acquisition by routine in market, EM microbial inoculum used herein is bought in Jiangxi Tianyi Biologic Technology Development Co., Ltd., People's Republic of China's fertilizer registration number is accurate No. 90222 of microorganism fertilizer (2005), and product standard number is Q/JTY006-2005.
By literature search, in order to stablize tuber of pinellia output and to be used as medicine quality, large quantity research has been carried out in this laboratory, find that there is the arbuscular mycorrhizal fungi structure in the tuber of pinellia, and arbuscular mycorrhizal fungi is existed stronger dependence, under single condition, after both symbiosis, the growth of the tuber of pinellia, physiologically active and output all increase.But arbuscular mycorrhizal fungi improves effective inoculation method of tuber of pinellia stem tuber nitrogen, phosphorus, potassium nutrition accumulation and the quality of being used as medicine to be published.
Bacterial strain Glomus mosseae (Glomus mosseae used herein, hereinafter referred GM) be a kind of of arbuscular mycorrhizal fungi, can conventional the purchase obtain on market, the GM microbial inoculum that experiment is used is bought in Inst. of Plant Nutrition ﹠. Resource, Beijing City Academy of Agricultural ﹠. Fo, for national fund is subsidized " Arbuscular Mycorrhizal Fungi In China germplasm resource bank (BGC) ", numbering BGC XZ01, National Nature scientific and technological resources platform resource 1511C0001BGCAM0024.
The invention provides a kind of arbuscular mycorrhizal fungi inoculation method that improves tuber of pinellia stem tuber nutrient and medical material quanlity, under this inoculation method, mycorrhizal fungi is infected situation and has obtained increase, has promoted the accumulation of tuber of pinellia stem tuber nitrogen, phosphorus, potassium nutrition, has improved the tuber of pinellia quality of being used as medicine.Has certain realistic meaning for utilizing microbial manure to solve tuber of pinellia degenerate problem.
Summary of the invention
The purpose of this invention is to provide a kind of arbuscular mycorrhizal fungi inoculation method that improves tuber of pinellia stem tuber nutrient and medical material quanlity, the method is by carrying out artificial adjustment to culture matrix, inoculum concentration, matrix phosphorus nutrient content, moisture with the factor such as other short living bacterium acting in conjunction, effectively promote the foundation of the tuber of pinellia and arbuscular mycorrhizal fungi symbiotic relation, and improve the nutrient of tuber of pinellia stem tuber and the quality of being used as medicine.
The present invention is achieved by the following technical solutions:
A kind of arbuscular mycorrhizal fungi inoculation method that improves tuber of pinellia stem tuber nutrient and medical material quanlity comprises the following step:
The half summer sowing stem that A. will collect soaks 20min with 1% potassium permanganate through 75% ethanol disinfection 3～5min after cleaning, and cleans and puts into the 24h that soaks seed under 26 ℃ of dark conditions, dries in the shade, and is standby;
B. culture vessel and matrix all need sterilization to process, and are specially: cultivate plastic basin and soak 1h with 5% potassium permanganate, clean and dry in the shade; Cultivation matrix is sand: and vermiculite (v: v)=1: 1, sand, vermiculite are dried, cross 20 mesh sieves, be placed on 121 ℃ of lower moist heat sterilization 1h, repeat sterilization once after placement 2d, standby;
C. the concrete mode of artificial adjustment inoculation is as follows:
A. moisture regulation method: moisture was weighed to execute at dusk and is watered not contain P element Hoagland nutrient solution according to matrix mass water content (moisture weight/cultivation matrix dry weight) every day in 5 o'clock, and keeping the matrix mass water content is 30%～50%;
B. phosphorus nutrient applying method: it is converted into KH in the Hoagland nutrient solution prescription
2PO
4Content dissolves in weekly moisture and applies in processing, and phosphorus content is 30～90mgP/kg;
C. microbial inoculum applying method: Mixed Microbes inoculation method.Arbuscular mycorrhizal fungus strain Glomus mosseae (Glomus mosseae), arbuscular mycorrhizal fungi germplasm resource bank numbering BGC BZ01A, National Nature scientific and technological resources platform numbering 1511C0001BGCAM0012, microbial inoculum and matrix ratio are 1: 10 (v: v); The former dew of EM is bought in Jiangxi Tianyi Biologic Technology Development Co., Ltd., People's Republic of China's fertilizer registration number is accurate No. 90222 of microorganism fertilizer (2005), product standard number is Q/JTY006-2005, execute after diluting 500 times and water, volume with the matrix weight ratio with v: m counts 1: 10;
D. first half summer sowing stem being placed in the plastic seeding culturing basin cultivates, conventional water management during this time, take seedling and move to the cultivation plastic basin that fills sterilization matrix when seedling grows, an initial stage Arbuscular Mycorrhizal Fungi microbial inoculum and EM microbial inoculum, regulation and control matrix phosphorus nutrient and moisture after emerging fully.
The invention has the advantages that:
The present invention is directed to that in the actual cultivation of the tuber of pinellia, fertilizer, moisture apply the problems such as unreasonable, amount of application and method of application by phosphate fertilizer, microbial manure and moisture in the growth of the artificial regulatory tuber of pinellia, effectively set up the inoculation method that the tuber of pinellia and arbuscular mycorrhiza form symbiont, have stronger guiding value in tuber of pinellia cultivation process, and be suitable for production application.
Embodiment
For further illustrating the beneficial effect of the invention, the spy will test and the mensuration process is summarized as follows:
A. plant the stem acquisition process:
With the half summer sowing stem impurity elimination that collects, choose the kind stem of diameter 2～3cm through 75% alcohol disinfecting 3～5min, soak 20min with 1% potassium permanganate after cleaning, clean and put into the 24h that soaks seed under 26 ℃ of dark conditions, dry in the shade;
B. culture vessel and matrix treatments:
A. cultivate plastic basin and soak 1h with 5% potassium permanganate, clean dry in the shade standby;
B. cultivation matrix is sand: and vermiculite (v: v)=1: 1, sand, vermiculite are dried, cross 20 mesh sieves, put into moist heat sterilization 1h under 121 ℃ of high temperature, repeat sterilization once after placing 2d, place a couple of days stand-by;
C. sowing
A. moisture regulation method: moisture was weighed to execute at dusk and is watered not contain P element Hoagland nutrient solution according to matrix mass water content (moisture weight/cultivation matrix dry weight) every day in 5 o'clock, and keeping the matrix mass water content is 30%～50%;
B. phosphorus nutrient applying method: it is converted into KH in the Hoagland nutrient solution prescription
2PO
4Content dissolves in weekly moisture and applies in processing, and phosphorus content is 30～90mg/kg;
C. microbial inoculum applying method: Mixed Microbes inoculation method.Arbuscular mycorrhizal fungus strain Glomus mosseae (Glomus mosseae), arbuscular mycorrhizal fungi germplasm resource bank numbering BGC BZ01A, National Nature scientific and technological resources platform numbering 1511C0001BGCAM0012, microbial inoculum and matrix ratio are 1: 10 (v: v); The former dew of EM is bought in Jiangxi Tianyi Biologic Technology Development Co., Ltd., People's Republic of China's fertilizer registration number is accurate No. 90222 of microorganism fertilizer (2005), product standard number is Q/JTY006-2005, execute after diluting 500 times and water, volume with the matrix weight ratio with v: m counts 1: 10;
D. first half summer sowing stem is placed in the plastic seeding culturing basin and cultivates (culture vessel and matrix treatments method are as shown in B), conventional water management during this time, when seedling grows, select the consistent seedling of growing way (plant height 3～5cm, blade 2～3cm), take and move to the cultivation plastic basin that fills sterilization matrix, an initial stage Arbuscular Mycorrhizal Fungi microbial inoculum and EM microbial inoculum, approximately emerged in 1 week fully rear regulation and control matrix phosphorus nutrient and moisture.
The tuber of pinellia maturing stage mycorrhizal fungi infection rate that obtains according to the embodiment cultivation reaches more than 73%, its fresh weight of general measure, stem tuber nutrient content and be used as medicine quality index value such as table 1 after the results stem tuber:
Table 1 improves tuber of pinellia stem tuber nutrient and medical material quanlity desired value that new method obtains
Matrix mass water content 30%～50%, matrix phosphorus content are 30～90mg/kg, and GM and EM combined inoculation be in tuber of pinellia stem tuber, can be so that the tuber of pinellia and endogenetic fungus symbiosis rate reach more than 73%, thus improved tuber of pinellia stem tuber nitrogen, phosphorus, three nutrient contents of potassium.2010 editions pharmacopeia are made standard to the quality of Pinellia Ternate, and moisture must not cross 14.0%, and water-soluble extractives content must not be less than 9.0%, must not be less than 0.25% in the total acid content of succinic acid.This method obtains Pinellia Ternate moisture, water-soluble extractives content and total organic acids content and all is significantly higher than standards of pharmacopoeia.
Repeat above experiment 5 times, the tuber of pinellia stem tuber medicinal material water content that measures all not higher than 2010 editions Chinese Pharmacopoeias regulations 14%, water-soluble extractives content all is not less than 16%, total organic acids content all is not less than 0.3%.
Wherein, EM microbial inoculum and AM microbial inoculum play to work in coordination and improve the effect of Pinellia Ternate quality.
The Pinellia Ternate index of correlation numerical value that provides can illustrate more effectively that this inoculation method can be used as a kind of new method that improves tuber of pinellia stem tuber nutrient and medical material quanlity, has stronger guiding value in tuber of pinellia cultivation process, and being suitable for production application, is also a kind of useful approach that solves tuber of pinellia cultivation degenerate problem in prior art.In the present invention, all experiments and embodiment are only to the enumerating of beneficial effect of the present invention, and are not the restrictions to invention, and the simple change that all those skilled in the art can make according to general knowledge all belongs in the protection domain of invention.
Claims (2)
1. microbial inoculum inoculation method that improves tuber of pinellia stem tuber nutrient and medical material quanlity comprises the following step:
The half summer sowing stem impurity elimination that A. will collect through 75% ethanol disinfection 3～5min, is soaked 20min with 1% potassium permanganate after cleaning, and cleans and puts into the 24h that soaks seed under 26 ℃ of dark conditions, dries in the shade, and is standby;
B. culture vessel and matrix sterilization are processed: cultivate plastic basin and soak 1h with 5% potassium permanganate, clean and dry in the shade; Cultivation matrix is with v: v counts sand: vermiculite=1: 1, sand, vermiculite are dried, and cross 20 mesh sieves, be placed on 121 ℃ of lower moist heat sterilization 1h, repeat sterilization once after placing 2d, standby;
C. artificial adjustment inoculation is specially:
A. moisture regulation: weighed to execute in 5 o'clock at dusk every day with the Hoagland nutrient solution that does not contain the P element and water, keeping the matrix mass water content is 30%～50%; Matrix mass water content computational methods are: moisture weight/cultivation matrix dry weight;
B. the phosphorus nutrient applies: phosphorus is converted into KH in the Hoagland nutrient solution prescription
2PO
4Content dissolves in weekly moisture and applies in processing, and making the matrix phosphorus content is 30～90mg/kg;
C. microbial inoculum inoculation: the Mixed Microbes inoculation method is the method that bush mycorrhizal fungi preparation and EM microbial inoculum are inoculated simultaneously; The bacterial strain of bush mycorrhizal fungi preparation wherein is Glomus mosseae (Glomus mosseae), Glomus mosseae is numbered BGC BZ01A the arbuscular mycorrhizal fungi germplasm resource bank, National Nature scientific and technological resources platform numbering 1511C0001BGCAM0012, bush mycorrhizal fungi preparation and matrix ratio are with v: v counts 1: 10; After 500 times of EM microbial inoculum dilutions, count 1: 10 and execute with v: m according to volume and matrix weight ratio and water;
D. first half summer sowing stem being placed in the plastic seeding culturing basin cultivates, conventional water management during this time, take seedling and move to the cultivation plastic basin that fills sterilization matrix when seedling grows, an initial stage Arbuscular Mycorrhizal Fungi microbial inoculum and EM microbial inoculum, emerge fully rear regulation and control matrix phosphorus nutrient and moisture.
2. a kind of microbial inoculum inoculation method that improves tuber of pinellia stem tuber nutrient and medical material quanlity according to claim 1, it is characterized in that, the tuber of pinellia stem tuber medicinal material water content that obtains by the cultivation of described method all not higher than 14%, water-soluble extractives content all is not less than 16%, total organic acids content all is not less than 0.3%.
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| CN110862929B (en) * | 2019-12-09 | 2020-11-06 | 江西农业大学 | Gloeostereum incarnatum, microbial inoculum comprising same and application thereof |
| CN116508599B (en) * | 2023-03-27 | 2024-07-19 | 毕节医学高等专科学校(毕节市卫生学校) | Breeding method for promoting pinellia ternate seeds to emerge and application of breeding method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1988004136A1 (en) * | 1986-12-02 | 1988-06-16 | Kyowa Hakko Kogyo Co., Ltd. | Method of multiplying tubers |
| CN1843073A (en) * | 2006-05-15 | 2006-10-11 | 中国农业科学院蔬菜花卉研究所 | Facility vegetable mycorrhiza production method |
| CN101699960A (en) * | 2009-11-16 | 2010-05-05 | 新疆农业科学院核技术生物技术研究所 | Application method of AM mycorrhizal fungi in virus-free potato test tube plantlet transplanting |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1988004136A1 (en) * | 1986-12-02 | 1988-06-16 | Kyowa Hakko Kogyo Co., Ltd. | Method of multiplying tubers |
| CN1843073A (en) * | 2006-05-15 | 2006-10-11 | 中国农业科学院蔬菜花卉研究所 | Facility vegetable mycorrhiza production method |
| CN101699960A (en) * | 2009-11-16 | 2010-05-05 | 新疆农业科学院核技术生物技术研究所 | Application method of AM mycorrhizal fungi in virus-free potato test tube plantlet transplanting |
Non-Patent Citations (4)
| Title |
|---|
| 丛枝菌根真菌对半夏产量及化学成分的影响;郭巧生等;《中国中药杂志》;20100228;第35卷(第3期);第333-337页 * |
| 半夏丛枝菌根真菌侵染模式及侵染率动态变化;程俐陶等;《贵州农业科学》;20091231;第37卷(第02期);第37-39页 * |
| 程俐陶等.半夏丛枝菌根真菌侵染模式及侵染率动态变化.《贵州农业科学》.2009,第37卷(第02期),第37-39页. |
| 郭巧生等.丛枝菌根真菌对半夏产量及化学成分的影响.《中国中药杂志》.2010,第35卷(第3期), |
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Granted publication date: 20131106 |