CN103828617A - Biological method for inducing heartwood formation of dalbergia odorifera - Google Patents
Biological method for inducing heartwood formation of dalbergia odorifera Download PDFInfo
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- CN103828617A CN103828617A CN201410104193.5A CN201410104193A CN103828617A CN 103828617 A CN103828617 A CN 103828617A CN 201410104193 A CN201410104193 A CN 201410104193A CN 103828617 A CN103828617 A CN 103828617A
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Abstract
The invention belongs to the field of biotechnology and relates to a biological method for inducing heartwood formation of the dalbergia odorifera, heartwoods of adult dalbergia odorifera trees with formed heartwoods are collected in a drilling mode, smashed and then added into an ISP2 liquid medium for being cultivated, and an endophyte mycelium culture solution of rosewood heartwoods is prepared; the dalbergia odorifera trees of 6-7 years without the formed heartwoods are selected, drilling is performed on trunks, the depths of holes reach xylems, the endophyte mycelium culture solution is then injected into the holes, hole openings are sealed, and after 12 months, the claret-colored xylems dug from the processed parts are the rosewood heartwoods. The biological method is simple in technology and easy and convenient to operate, the endophyte mycelia of the rosewood heartwoods are used for inducing the rosewood heartwoods, the production period of heartwood formation of the dalbergia odorifera is effectively shortened, and the biological method has the advantages of being natural and free of hazard and is one of the safest and most promising methods. The biological method is suitable for large-scale standard and commercial production of the rosewood heartwoods, facilitates protection, development and utilization of dalbergia odorifera resources and has important economic, social and ecological benefits.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of method that stimulates dalbergia odorifera Edgeworthia chrysantha, specifically a kind of method that adopts biological method induction dalbergia odorifera Edgeworthia chrysantha.
Background technology
Chinese medicine dalbergia wood is the trunk of leguminous plant dalbergia odorifera (Dalbergia odorifera T.Chen) and the dry duramen of root, dalbergia wood mainly contains promoting qi circulation and relieving pain, invigorate blood circulation, stop blooding, stop the effects such as dysentery, preventing or arresting vomiting, that clinical common dosage forms is as the primary raw material of the Chinese patent drugs such as compound dalbergia wood capsule, GUANXIN DANSHEN PIAN breast knot dissipation sheet, can be used for treating traumatic injury, wound appearance, wrist abdomen pain, obstruction of qi in the chest shouting pain etc., there is higher medical value and economic worth.In recent years, along with the market demand constantly increases, cause wild resource to be fell trees unlawfully seriously, resource faces the exhausted even condition of extinction.In order to alleviate this present situation, at present, all there is artificial commerial growing various places, south China, but under dalbergia odorifera natural growthing condition, heartwood forms very slow, after general growth 7-8, the trunk of dalbergia odorifera just starts to form heartwood, 20-30 just can run up to some with hyoscine, Edgeworthia chrysantha process is very very long, cannot meet social medication demand, therefore, how to make full use of dalbergia odorifera tree and accelerate its Edgeworthia chrysantha process and increase output, become key issue urgently to be resolved hurrily in current production to alleviate dalbergia wood imbalance between supply and demand.
At present, mainly in production form medicinal material by dalbergia odorifera self-sow, bibliographical information environmental stimuli produces that fragrant method mainly contains that physics is chopped wound, chemical stimulation, plant hormone induction and utilize the methods such as fungal induction.But the heartwood endophyte fungus strain induction dalbergia odorifera that utilizes dalbergia odorifera self-assembling formation forms fast the method for heartwood and has no report.
Summary of the invention
The object of the invention is provides a kind of biological method of inducing dalbergia odorifera Edgeworthia chrysantha for the deficiencies in the prior art; utilize the endophyte fungus strain of dalbergia wood itself to induce dalbergia wood; not only the production cycle shortens; and there is natural, non-harmful advantage; it is the safest one of the most promising method; be applicable to dalbergia wood large-scale standardized, commercialization production, be conducive to protection, the development and utilization of dalbergia odorifera resource, there are important economy, society and ecological benefits.
The technical solution adopted in the present invention:
A kind of biological method of inducing dalbergia odorifera Edgeworthia chrysantha, to choose on the adult dalbergia odorifera tree of Edgeworthia chrysantha to drill through heartwood, after surface sterilization, pulverizing, join in ISP2 liquid nutrient medium, 25～30 ℃ of shaken cultivation 5～10 days, making the endophyte fungus strain culture fluid of dalbergia wood, is wherein 1:10～50 by the ratio of mass volume ratio (g/mL) dalbergia odorifera pith material and ISP2 liquid nutrient medium; Choose the dalbergia odorifera tree that does not form heartwood for 6～7 years, overhead holing on trunk in the scope below 50cm height, hole depth is to xylem, then by endophyte fungus strain culture fluid hand-hole, again encapsulation process is carried out in aperture, after 12 months, take mauve xylem from treatment sites and be dalbergia wood.
Get part aubergine xylem, dry, after comminuted powder with appropriate high-purity distilled water immersion, with volatile oil extractor routinely the boiling of steam circumfluence method until volatile oil no longer increase, after anhydrous sodium sulfate drying volatile oil.Volatile oil is water white transparency oily thing, has special fragrant.By with reference to 2010 Pharmacopoeia of the People's Republic of China determination of volatile oil method (appendix XD first method) measure the content of its volatile oil, by gas chromatography-mass spectrum (GC-MS), chemical composition is identified, determine relative percentage composition by normalization method.
Technique of the present invention is simple; easy and simple to handle; utilize the endophyte fungus strain of dalbergia wood itself to induce dalbergia wood; effectively shorten the production cycle of dalbergia odorifera Edgeworthia chrysantha; having natural, non-harmful advantage, is one of the safest the most promising method, is applicable to dalbergia wood large-scale standardized, commercialization is produced; be conducive to protection, the development and utilization of dalbergia odorifera resource, there are important economy, society and ecological benefits.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, conventionally according to normal condition, or the condition of advising according to manufacturer.
Embodiment mono-
From 60 years adult dalbergia odorifera tree, drill through 5g heartwood with vegetative cone, after 70% alcohol surface sterilization, the pulverizing of milling under aseptic condition, put into 200mL ISP2 liquid nutrient medium, 28 ℃, 180rpm shaken cultivation, after 1 week, makes the endophyte fungus strain culture fluid of dalbergia wood.Choose the dalbergia odorifera that 6-7 do not form heartwood and set 30 strains, the drill bit that is 1cm with diameter in the scope below 50cm height overhead bores from top to bottom 3 holes (tilting 45 °) on trunk, hole depth is to xylem, each hole between the upper and lower be spaced apart 10～15cm, in each hole, inject 1mL culture fluid, by preservative film and adhesive tape sealed envelope, process after 12 months, cut down trees, take mauve xylem from treatment sites.By the aubergine material obtaining, to dry, accurately weigh, weight is 6090g, average strain output: 203g.
Embodiment bis-
From 40 years adult dalbergia odorifera tree, drill through 3g heartwood with vegetative cone, after 70% alcohol surface sterilization, the pulverizing of milling under aseptic condition, put into 100mL ISP2 liquid nutrient medium, 28 ℃, 180rpm shaken cultivation, after 1 week, makes the endophyte fungus strain culture fluid of dalbergia wood.Choose the dalbergia odorifera that 6-7 do not form heartwood and set 30 strains, the drill bit that is 1cm with diameter in the scope below 50cm height overhead bores from top to bottom 3 holes (tilting 45 °) on trunk, hole depth is to xylem, each hole between the upper and lower be spaced apart 10～15cm, in each hole, inject 1mL culture fluid, by preservative film and adhesive tape sealed envelope, process after 12 months, cut down trees, take mauve xylem from treatment sites.By the aubergine material obtaining, to dry, accurately weigh, weight is 6420g, average strain output: 214g.
Embodiment tri-
From 70 years adult dalbergia odorifera tree, drill through 2g heartwood with vegetative cone, after 70% alcohol surface sterilization, the pulverizing of milling under aseptic condition, put into 50mL ISP2 liquid nutrient medium, 28 ℃, 180rpm shaken cultivation, after 1 week, makes the endophyte fungus strain culture fluid of dalbergia wood.Choose the dalbergia odorifera that 6-7 do not form heartwood and set 30 strains, the drill bit that is 1cm with diameter in the scope below 50cm height overhead bores from top to bottom 3 holes (tilting 45 °) on trunk, hole depth is to xylem, each hole between the upper and lower be spaced apart 10～15cm, in each hole, inject 1mL culture fluid, by preservative film and adhesive tape sealed envelope, process after 12 months, cut down trees, take mauve xylem from treatment sites.By the aubergine material obtaining, to dry, accurately weigh, weight is 6270g, average strain output: 209g.
Check:
1, volatile oil content in product
Accurately take respectively the product 50g of embodiment mono-, two, three, after pulverizing with appropriate high-purity distilled water immersion 5h, with volatile oil extractor routinely 100 ℃ of boiling 16h of steam circumfluence method until volatile oil no longer increase, must volatile oil after anhydrous sodium sulfate drying.Volatile oil is water white transparency oily thing, has special fragrant.By with reference to 2010 Pharmacopoeia of the People's Republic of China determination of volatile oil method (appendix XD first method) measure the content of its volatile oil, the results are shown in Table 1.
Table 1 volatile oil content index
| Project | Aubergine product (g) | Volatile oil (mL) | Recovery rate (mL/g) |
| Embodiment mono- | 50 | 0.68 | 1.36% |
| Embodiment bis- | 50 | 0.73 | 1.47% |
| Embodiment tri- | 50 | 0.64 | 1.28% |
The standard that must not be less than 1.0% (mL/g) according to the dalbergia wood of Pharmacopoeia of the People's Republic of China regulation in 2010 containing volatile oil, the volatile oil content of the preparation-obtained aubergine product of the present invention meets standard.
2, the content of nerolidol in volatile oil
Get respectively embodiment mono-, two, three products volatile oil a little, utilize gas chromatography-mass spectrum (GC-MS) to identify chemical composition, determine relative percentage composition by normalization method.Gas chromatography-mass spectrometry analysis condition: chromatographic column: quartz capillary column HP-5MS (30mm × 0.25mm × 0.25mm), chromatographic column temperature programming condition: 50 ℃ of initial temperatures, initial temperature keeps after 3min, is warming up to 200 ℃ and keep 7min, carrier gas He with the speed of 5 ℃/min; Split ratio 1:50; Before post, press 7.6522psi; Injector temperature: 250 ℃; Ion source temperature: 230 ℃; Ionization voltage: 70eV; Quality of scanning scope: 50～400AMU; Scan mode: EI source; Sweep spacing: 0.5s.By above-mentioned GC-MS condition, volatile oil is carried out to constituent analysis, measure the relative percentage composition of nerolidol with area normalization method, the results are shown in Table 2.
Constituent analysis in table 2 volatile oil
| Project | Component kind | The relative percentage composition of nerolidol |
| Embodiment mono- | 14 | 29.25% |
| Embodiment bis- | 14 | 30.71% |
| Embodiment tri- | 14 | 28.12% |
Can be found out by result, from volatile oil, isolate 14 kinds of components, the relative percentage composition that records the nerolidol of volatilization main body of oil with area normalization method reaches high value, illustrates that adopting the aubergine heartwood material that the present invention makes is dalbergia wood really, meets the medicinal material standard of dalbergia wood.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, do not departing under the prerequisite of the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (1)
1. induce the biological method of dalbergia odorifera Edgeworthia chrysantha for one kind, it is characterized in that: choose on the adult dalbergia odorifera tree of Edgeworthia chrysantha and drill through heartwood, after surface sterilization, pulverizing, join in ISP2 liquid nutrient medium, 25～30 ℃ of shaken cultivation 5～10 days, making the endophyte fungus strain culture fluid of dalbergia wood, is wherein 1:10～50 by the ratio of mass volume ratio dalbergia odorifera pith material and ISP2 liquid nutrient medium; Choose the dalbergia odorifera tree that does not form heartwood for 6～7 years, overhead holing on trunk in the scope below 50cm height, hole depth is to xylem, then by endophyte fungus strain culture fluid hand-hole, again encapsulation process is carried out in aperture, after 12 months, take mauve xylem from treatment sites and be dalbergia wood.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104621186A (en) * | 2015-02-10 | 2015-05-20 | 林励 | Composition, method for preparing inducing liquid from composition, application of inducing liquid in exciting heartwood formation of lignum dalbergiae odoriferae |
| CN106508557A (en) * | 2016-10-19 | 2017-03-22 | 宜宾云辰乔木园林有限责任公司 | Method for early forming heartwood by dalbergia odorifera |
| CN110605761A (en) * | 2019-08-20 | 2019-12-24 | 昆明科晔生物技术有限公司 | Hand-held drill bit device capable of promoting trees to generate agilawood |
| CN111657042A (en) * | 2020-06-30 | 2020-09-15 | 中国林业科学研究院资源昆虫研究所 | Planting method for promoting formation of pterocarpus indicus heartwood |
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| CN103168634A (en) * | 2013-03-26 | 2013-06-26 | 中山市沉香黄农林发展有限公司 | Method for promoting aquilaria sinensis to carry out secondary edgeworthia chrysantha |
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| CN102550311A (en) * | 2011-12-29 | 2012-07-11 | 中国林业科学研究院热带林业研究所 | Method for rapid formation of agilawood of agallochum through induction by using fungus fermentation liquid |
| CN102726224A (en) * | 2012-04-17 | 2012-10-17 | 唐文 | Agilawood producing on aquilaria senensis gilg trees by using stick inserting method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104621186A (en) * | 2015-02-10 | 2015-05-20 | 林励 | Composition, method for preparing inducing liquid from composition, application of inducing liquid in exciting heartwood formation of lignum dalbergiae odoriferae |
| CN104621186B (en) * | 2015-02-10 | 2017-11-21 | 林励 | A kind of composition, the application of the method for liquid, the induction liquid on stimulation dalbergia wood Edgeworthia chrysantha is induced by its preparation |
| CN106508557A (en) * | 2016-10-19 | 2017-03-22 | 宜宾云辰乔木园林有限责任公司 | Method for early forming heartwood by dalbergia odorifera |
| CN110605761A (en) * | 2019-08-20 | 2019-12-24 | 昆明科晔生物技术有限公司 | Hand-held drill bit device capable of promoting trees to generate agilawood |
| CN110605761B (en) * | 2019-08-20 | 2022-04-15 | 海南海香园投资有限公司 | Hand-held drill bit device capable of promoting trees to generate agilawood |
| CN111657042A (en) * | 2020-06-30 | 2020-09-15 | 中国林业科学研究院资源昆虫研究所 | Planting method for promoting formation of pterocarpus indicus heartwood |
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| CN103828617B (en) | 2016-09-07 |
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