CN102934586A - Low-carbon and high-yield poria culture method - Google Patents
Low-carbon and high-yield poria culture method Download PDFInfo
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Abstract
The invention relates to a low-carbon and high-yield poria culture method and belongs to the technical field of edible (medicinal) fungus culture. According to the method, poria is cultured naturally by utilizing underwood waste pine root and stem, and a specific poria strain, which is Wolfiporia extents Chuanjie-1-A5 with preservation number of CGMCC No.6660, suitable to be cultured by using pine root and stem is selected and is cultured in combination with a taproot inoculation method. The culture method comprises the following steps of: producing a strain; pre-treating the pine root and stem; inoculating; and conventionally managing and harvesting. According to the method, waste pine root and stem of forestry is effectively utilized to naturally culture poria in a low-carbon manner, 'conflict between fungus and forest' in poria production can be effectively solved, and harm of termite and other insect pests caused by forest waste discharge can be reduced, so that the production cost can be reduced, labor intensity of poria planting can be greatly reduced, time and labor can be saved, and management is facilitated. The culture method is beneficial to sustainable development of the poria industry and has remarkable economical benefit.
Description
Technical field
The invention belongs to food (medicine) and use the bacteria cultivation technical field, more specifically relate to a kind of method of low-carbon (LC) high-yield culturing Poria cocos
Background technology
Poria cocos [
Wolfiporia extensa(Peck) Ginns] have another name called Fu Tu, Yun Ling, Fu spirit, loose potato, Song Ling etc.Be subordinate to Basidiomycota, agaric guiding principle, Aphyllophorales, Polyporaceae, Fu a kind of reed mentioned in ancient books genus on taxonomy.Poria cocos typically refers to the sclerotium colonized on the pinaceae plant tree root.Poria cocos, at China's medicinal history of existing more than 2000 year, is one of kind of current domestic medicinal fungus artificial cultivation output maximum.Although the artificial cultivation Poria cocos history of existing more than 1000 year is still prolonged traditional kind Siberian cocklebur pattern of attacking in cultivation.According to annual about more than the 20000 tons of dry Poria cocos that need in the pharmaceuticals industry incomplete statistics whole nation; traditional conventional cultivation mode is to adopt pine tree section wood to produce; 1 ton of dry Poria cocos of every production approximately needs the pine of 20 cubic metres; therefore the annual pine for the production of Poria cocos just reaches more than 40 ten thousand steres; therefore Poria cocos and forestry is particularly thorny; production cost pressure is large; cultivation output is generally (1.5～2kg/ pine section wooden 10kg) on the low side again; lack market competitive power, greatly restricted the segment wood cultivated large-scale production of Poria cocos.National Poria cocos output in 2009 is 29.3314 ten thousand tons (fresh goods), and Poria cocos output in 2010 falls sharply and only has 6.9055 ten thousand tons.And China's pine tree root and stem of certain plants resource is extremely abundant, annual forest department all wants a large amount of pine tree of intermediate cutting to stay many pine tree root and stem of certain plants, allows a large amount of pine tree root and stem of certain plants be placed on mountain and rots, and not only wastes resource, life area is provided also can to white ant, endangers other forests.Utilize pine tree root and stem of certain plants cultivation Poria cocos can not only effectively utilize the forest resources, keep ecological, the development low-carbon economy, turn waste into wealth, meeting the Important Action of the development conservation-minded society that country advocates, meet the requirement developed eco-agriculture, is the Eco-agriculture project of a sustainable development.The method about pine tree root and stem of certain plants cultivation Poria cocos of having reported at present existing document, adopt Poria cocos be inoculated in to root and stem of certain plants top, root and stem of certain plants side and lateral root, and bacterial strain output is not remarkable, and the bacterial strain genetic stability is not high.
Summary of the invention
The purpose of this invention is to provide that a kind of inoculation survival rate is high, the knot Siberian cocklebur early, the method for low-carbon (LC) high-yield culturing Poria cocos that output is high.
At first the present invention provides a strain Poria cocos bacterial strain, described Poria cocos bacterial strain be Poria cocos (
Wolfiporia extens) the outstanding No. 1-A5 in river, on October 29th, 2012, being deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, it is called for short CGMCC, and address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preserving number is CGMCC NO. 6660.
The present invention also provides a kind of method of low-carbon (LC) high-yield culturing Poria cocos, utilizes the sylvan life discarded object pine tree root and stem of certain plants to carry out Poria cocos and naturally cultivates, and selects the specificity Poria cocos bacterial strain of suitable pine tree root and stem of certain plants cultivation, preserving number be CGMCC NO. 6660 Poria cocos (
Wolfiporia extens) the outstanding No. 1-A5 in river, simultaneously in conjunction with main root inoculation method cultivation Poria cocos; The cultivation step is: the pretreatment of bacterial classification production → pine tree root and stem of certain plants → inoculation → Routine Management and gathering.
The step that described bacterial classification is produced is as follows: (1) at first carries out the rejuvenation of bacterial strain, is vigorous " wavy " while growing forward on the PDA slant tube with mycelia and is arranged as the production that target is carried out the female kind of Poria cocos one-level; (2) Poria cocos seed production: the pine sawdust of take is produced the Poria cocos original seed according to a conventional method as major ingredient; (3) tuckahoe cultivated species production: take pine sheet as major ingredient, the cultivation of access Poria cocos original seed.
The pretreatment of the described pine tree root and stem of certain plants comprises: by pine tree root and stem of certain plants periphery eliminating impurities, and carry out termite-proof processing, find out the main root place of the pine tree root and stem of certain plants, the position dry near the root and stem of certain plants wide 5～8 cm that prune to main root, the root skin of long 15 cm, the large butt that diameter surpasses 25cm needs to select the processing of being peeled of 2～3 thicker lateral roots again.
Management of the present invention with gather: focus on the control water of Siberian cocklebur field with ventilative.Meeting the rainy day prevents from around the pine tree root and stem of certain plants becoming silted up water, meets humidification of drought; The cultivation process remains the loose, ventilative of soil layer.Inoculate 3～4 months by normal cruising Siberian cocklebur field, without blindly finding, as long as whether the table soil in concern pine tree root and stem of certain plants 1m diameter range ftractures, timely earthing just can be guaranteed the normal growth of poria cocos sclerotium.Be full of cracks no longer appears in the table soil after inoculation in 10～12 months butt 1m diameter ranges, and white slight crack, no longer appear in the poria cocos sclerotium crust sepia that darkens, and just can gather.Because of the characteristic of pine tree root and stem of certain plants 1m diameter range knot Siberian cocklebur, the intensity that has greatly reduced the Poria cocos cultivation management and gathered, saving of work and time.
By application the present invention, can make the abundant useless pine tree root and stem of certain plants resource in each province and city be fully utilized, this invention has not only reduced the labour intensity of pine tree root and stem of certain plants kind Siberian cocklebur, improve work efficiency simultaneously, make pine tree root and stem of certain plants kind Siberian cocklebur be easy to management, average single root and stem of certain plants Poria cocos output can reach 15kg/ pine tree root and stem of certain plants D25 ± 5cm, makes the forestry waste material bring into play maximum economic benefit, turn waste into wealth, the development low-carbon economy.The method of applying this low-carbon (LC) high-yield culturing Poria cocos has not only solved " the bacterium woods contradiction " existed in the Edible Fungi, has dissolved the insect pests such as termite simultaneously and has grown the harm to forest, and economy, society and ecological benefits are remarkable.
Remarkable advantage of the present invention is:
1) the present invention utilizes forestry waste-pine tree root and stem of certain plants to carry out the inventive method of Poria cocos low-carbon (LC) ecosystem Efficient Cultivation; solved the culture technique problem of pine tree root and stem of certain plants cultivation large-scale production; not only reduce production costs; and reduced the labour intensity of pine tree root and stem of certain plants kind Siberian cocklebur; improved pine tree root and stem of certain plants availability; be conducive to the sustainable development of Poria cocos industry; improve the productivity effect of edible mushroom; and realize the recycling of forestry waste-pine tree root and stem of certain plants; improve the value of the pine tree root and stem of certain plants; reduced the waste discharge of forestry, to being significant of the recycling economy that develops forestry.
2) the present invention is by the selection of suitable pine tree root and stem of certain plants cultivation specific strains, select " the outstanding No. 1-A5 in river " Poria cocos kind of this institute seed selection to guarantee that in cultivation, poria cocos sclerotium concentrates the knot Siberian cocklebur in pine tree root and stem of certain plants 1m diameter range, and output is high, average single root and stem of certain plants output can reach 15kg/ root and stem of certain plants diameter 25 ± 5cm.
3) the present invention, by the production of corresponding cultivation bacterial classification, is the feature that vigorous " wavy " arranged while growing forward on the mycelia in the female PDA of kind of one-level slant tube must possess.The original seed mycelial growth of secondary high-quality is vigorous, pure white, even, fine and close, has rhizomorph to distribute; The cultivated species mycelia of three grades of high-qualitys should pure white densification evenly be covered with wood chip, and section is faint yellow, and has the peculiar fragrance of strong Poria cocos, and without the jaundice mycelia, the bacterium bag is complete.Guarantee that cultivation strain is not only in the specificity of the 1m knot Siberian cocklebur that keeps its bacterial strain, vigorous mycelia vigor can improve the survival rate of pine tree root and stem of certain plants inoculation simultaneously.The Poria cocos strain excellent of suitable pine tree root and stem of certain plants cultivation is that high yield is had laid a good foundation.
4) pretreatment of the pine tree root and stem of certain plants of the present invention, without the processing of being completely cured of the pine stump to all, greatly reduce the labour intensity of the kind Siberian cocklebur of the pine tree root and stem of certain plants.
5) inoculation of the present invention: main root is not dialled a bag inoculation method: pine sheet cultivation strain (0.5kg/ bag): suitable cell age is 30～40 d, and it is healthy and strong fine and close that mycelia evenly is covered with wood chip, the special aromatic flavour of tool Poria cocos, and the bacterium bag is complete, without living contaminants; Only cultivated species bacterium bag is cut open to a slit that 2cm is wide during inoculation, do not dial the survival rate that can guarantee the rainy day inoculation from the bacterium bag; The position of pine tree root and stem of certain plants cultivation inoculation is very crucial, what the opening part of bacterium bag should be close to pine tree root and stem of certain plants main root and the dry infall of the root and stem of certain plants newly splits mouth, main root for the pine tree root and stem of certain plants especially as human body arteries, the mouth of newly splitting that is connected on pine tree root and stem of certain plants main root and the dry infall of the root and stem of certain plants is located to improve the survival rate that the pine tree root and stem of certain plants is inoculated, can improve again the material feeding speed of Poria cocos mycelia in the pine tree root and stem of certain plants, guarantee the supply of Poria cocos later stage sclerotium Fast Growth to nutrition.Inoculum concentration (0.5kg/ bag): 1 bag/pine tree root and stem of certain plants diameter, 20 ± 5cm cm, 2 bags/pine tree root and stem of certain plants diameter, 30 ± 5cm, the above pine tree root and stem of certain plants of 3～4 bags/pine tree root and stem of certain plants diameter 40cm.Inoculum concentration is too much, depleted in the nutrition meeting of the vegetative stage pine tree root and stem of certain plants, knot oligotrophy during Siberian cocklebur, and sclerotium can't Fast Growth causes the degradation of Poria cocos output; Inoculum concentration is very few, and mycelia can't rapid spread, and material feeding speed is slow, and vegetative growth phase extends the formation of having postponed sclerotium, can cause equally the decline of output.Suitable pine tree root and stem of certain plants inoculation method is also the crux place of high-yield culture.
6) average single root and stem of certain plants (pine tree root and stem of certain plants diameter 25 ± 5cm) output reaches 15kg, poria cocos sclerotium diameter 10cm～30cm, monokaryon weighs 2～5kg, have the distinctive tissue morphology of poria cocos sclerotium, the sclerotium surface all is pitchy or sepia, and crust is thin and coarse, the warty shrinkage is arranged, matter is soft, easy to crack, and bacterial context is pure white solid, is suitable for the processing of traditional Chinese medicine.Product meets 2010<<Chinese Pharmacopoeia and China's agricultural industry criteria agricultural industry criteria [pollution-free food edible mushroom] sense organ (NY5095-2006) require and safety requirements.
Embodiment
Embodiment 1
1 strain breeding thereof
Take " Fujian Siberian cocklebur " carry out the Poria cocos breeding research as the parent applies the Mutagenesis technology: the preparation " Fujian Siberian cocklebur " Mycelial Protoplasts, protoplast is diluted to 1.25 * 10
6Individual/ml, respectively get 0.1 ml and coat on the CYM regeneration culture medium, with the ultra violet lamp mutagenesis of 15w.Culture dish, apart from fluorescent tube 30 cm, after preheating 20 min that turn on light, is opened the ware lid and is irradiated, and irradiation time is 150s.After ultraviolet mutagenesis finishes, dull and stereotyped tight with black paper bag, put under 28 ℃ and cultivate 5～7 days.After protoplast regeneration, select at first and grow, 120 of healthy and strong, pure bacterium colonies (being numbered W1～W120), move into the PDA medium culture.The mycelial growth rate on the PDA flat board of usining, as the primary dcreening operation standard, filters out the regeneration strain that 30 (being numbered P1～P30) growth rate rank forefront.With mycelial growth rate on the pine sawdust medium for sieving again standard, sift out again 10 (numbering A1～A10) regeneration strains that mycelial growth rate ranks forefront on the pine sawdust medium, through the genetic stability analytical test and in 2002, in Qingliu County, high mouthful logging headquarter carries out the knot Siberian cocklebur comparative trial of 10 protoplasms regeneration isolated strains (A1～A10) and Fujian Siberian cocklebur (A), the output that Siberian cocklebur is tied in the Poria cocos pine root and stem of certain plants cultivation of take is the screening target, the result of the test statistical analysis shows that between 10 test strains, volume variance reaches utmost point significance level, the A5 bacterial strain is than contrast " Fujian Siberian cocklebur " output high by 25.93% (in Table 1), finally select the new strains A 5 of high yield of the suitable loose root and stem of certain plants cultivation of 1 strain.
Within 2008, at Changting County South Mountain town hill, carry out the knot Siberian cocklebur comparative trial (in Table 2) of new bacterial strain and 5 Poria cocos main breeds.Knot Siberian cocklebur comparative trial shows that new bacterial strain mycelial growth is vigorous, the survival rate of inoculation is high, the knot Siberian cocklebur early, output is high, average single root and stem of certain plants output is 16.40kg/ root and stem of certain plants diameter 25 ± 5cm, compare output with the main breed of domestic 5 Poria cocos and improve a lot, higher by 57.69% than current Poria cocos govern-house-variety " 5.78 " output, statistical analysis shows that between test strain, volume variance reaches utmost point significance level.Test has also further verified that new bacterial strain knot Siberian cocklebur is all in 1 rice diameter scope of the loose root and stem of certain plants--the stability of " not running Siberian cocklebur " characteristic, during Poria cocos kind Siberian cocklebur, without the processing of being completely cured that is tired of rope, it is the good Poria cocos bacterial strain of the suitable loose root and stem of certain plants cultivation of a strain.
The cultivation comparative test result of 1:10 protoplast regeneration isolated strains of table
Annotate: the mean value that in table, the data of each repetition are random 10 root and stem of certain plants determination datas
Table 2: the knot Siberian cocklebur comparing result of new bacterial strain and domestic Poria cocos main breed
Annotate: the mean value that in table, the data of each repetition are random 10 root and stem of certain plants determination datas
Table 3: new bacterial strain and Poria cocos govern-house-variety " 5.78 " knot Siberian cocklebur comparing result
2 features
2.1, morphological feature:
The colony morphology characteristic that the new bacterial strain of Poria cocos " river outstanding No. 1-A5 " is grown on the PDA plating medium is obvious, just is radial growth, is close to stromal surface, after be sparse alternate wavy " concentric ring " feature (seeing Fig. 1) while growing forward with mycelia; Mycelia density: medium; Aerial hyphae development degree: medium; Bacterium colony surface color: white; Bacterium colony back side color: white, there is secretion yellowish-brown pigment in the later stage; The irregular shape of sclerotium profile, diameter 10cm～30cm, monokaryon weigh 2～5kg, sometimes can reach 10～30 kg.The sclerotium appearance: surperficial pitchy or sepia, crust is thin and coarse, and the warty shrinkage is arranged, during the sclerotium children, cot is bronzing, matter is soft, easy to crack, sclerotium inside: be white powder graininess bacterial context, matter density is medium, easily cut open, the special fragrance of strong Poria cocos is arranged.
2.2 biological property
2.2.1 thermal adaptability
Will " river outstanding No. 1-A5 " be inoculated in moisture 60% pine sawdust medium and (take pine sawdust as main matrix, add the auxiliary material such as wheat bran), under 10,15,20,25,30,35,38 ℃, cultivate respectively, every kind 3 is repeated, and observes bacterial strain and walk bacterium speed under low temperature and high temperature.The humid test result shows, the mycelial growth temperature scope of new bacterial strain " the outstanding No. 1-A5 in river " is 10 ℃～35 ℃, and suitable growth temperature is 25 ℃, and killing temperature is 38 ℃ (table 4).
Table 4: " the outstanding No. 1-A5 in river " mycelial growth temperature result of the test (mm)
2.2.2 moisture content in medium adaptability
Will " river outstanding No. 1-A5 " bacterial strain be inoculated in respectively in moisture 50,55,60,65 and 70% pine sawdust medium and (take pine sawdust as main matrix, add the auxiliary material such as wheat bran), under 25 ℃, cultivate, every kind 3 is repeated, observes this bacterial strain and walk bacterium speed in the medium of different moisture content.Medium different moisture content result of the test shows, the applicable scope of the mycelial growth of new bacterial strain " the outstanding No. 1-A5 in river " is 50%～70%, and the scope of suitable growth is 55%～65%(table 5).
Table 5: " the outstanding No. 1-A5 in river " mycelial growth humid test result (mm)
2.2.3 acid-base value adaptability
" river outstanding No. 1-A5 " bacterial strain is inoculated in respectively to acid-base value detection range 2,3,4, in 5,6,7 PDA flat board, 3 repetitions of each pH value, the observation bacterial strain is walked bacterium speed under different pH values.Dull and stereotyped acid-base value result of the test shows, the pH scope that the mycelia of new bacterial strain " the outstanding No. 1-A5 in river " grows on the PDA flat board is 2～6, and the pH of its this bacterial strain suitable growth of ligative hyphae growing way is 5(table 6).
Table 6: " the outstanding No. 1-A5 in river " dull and stereotyped pH result of the test (mm) of mycelial growth
3 cultivation characteristics
The cultivation of " the outstanding No. 1-A5 in river " suitable Poria cocos pine of new bacterial strain root and stem of certain plants, 10～35 ℃ of mycelia suitable growth temperatures, optimum temperature is 24～28 ℃.Higher than 35 ℃, mycelium is easily aging, and the duration is length or excess Temperature, causes death; Below 20 ℃, poor growth; Humidity: the scope that mycelia is 50～75 ﹪ in water content all can be grown, and the optimum moisture content of original seed and cultivated species medium is 55～60%.Cultivation process soil moisture content should remain on 20%～25%.Acid-base value: the mycelial growth slant acidity, mycelia all can normal growth under the acid soil environment of pH3～7, take the soil of pH4～6 as good.Illumination: mycelial growth is without illumination, and it is good that sunshine abundant tailo should be selected in cultivation Siberian cocklebur field.Air: circulation of air, well-drained sandy soil (conglomeratic sand approximately 70%) should be selected in the Siberian cocklebur field, and earthing can not be blocked up, and soil compaction will loosen the soil in time.Be " the outstanding No. 1-A5 in river " new inoculation optimum period annual 5～September, within 3～4 months, starts to tie Siberian cocklebur after inoculation, and the knot Siberian cocklebur concentrates in the 1m diameter range of the loose root and stem of certain plants, within 10～12 months, gathers.
4 output performances
Poria cocos " river outstanding No. 1-A5 " bacterial strain from June, 2008, respectively in Liancheng of our province, 10 points such as Changting, Songxi, Youxi, land for growing field crops, Shaowu carry out experiment in cultivation and industrial experimentation.Adopt conventional loose root and stem of certain plants kind Siberian cocklebur pattern, the conventional method management.Result shows: this bacterial strain strong stress resistance, inoculation survival rate are 95%
More than exceed 3～5 percentage points of check varieties; The knot Siberian cocklebur was than Zao 20～30 days of contrast, and knot Siberian cocklebur scope is all in the 1m of loose root and stem of certain plants diameter range; In 6 demonstration pilot project experiments in cultivation, average single root and stem of certain plants output is 147.30～157.69% (in Table 7) for contrast " 5.78 " kind at 14.60～16.47kg/ root and stem of certain plants diameter, 25 ± 5cm, obvious effect of increasing production.
Table 7: Poria cocos " the outstanding No. 1-A5 in river " bacterial strain output performance
Annotate: in table, data are random 10 root and stem of certain plants determination datas, the mean value of 3 repetitions (its integer is got in round off)
Quality analysis: " the outstanding No. 1-A5 in river " bacterial strain sclerotium form is irregular type, and sclerotium size 10cm～30cm, monokaryon are heavily 2～5kg, sometimes can reach 10～40 kg; The sclerotium surface all is pitchy or sepia, crust is thin and coarse, the warty shrinkage is arranged, matter is soft, easy to crack, the bacterial context compactedness is suitable, its main component detects through Fujian Province Institute of Analysis, and " outstanding No. 1 of river--A5 " (Poria cocos piece dry product) crude protein content is that 1.36 %, crude fat content are 0.6%, triterpenes content is 0.059%, total reducing sugar is 0.50%, ash content 1.42%, moisture are 14.4%, the processing of crude drugs of suitable Poria cocos.
Embodiment 2
A kind of method of low-carbon (LC) high-yield culturing Poria cocos, concrete operations are as follows:
1, the root and stem of certain plants is selected in fixed field
The Siberian cocklebur field should be away from pollution source, air quality reaches GB3095-96 atmosphere quality secondary, soil quality reaches GB15618-95 standard of soil environment quality secondary, water source reaches GB5084-92 field irrigation standards of water quality, the Siberian cocklebur field should be selected and sits in the north facing the south or sit Xi Chaodong, height above sea level 300-1500M, the gradient 10-50 degree, full sun, the not ponding that ventilates, soil is loose sand hill soil, is subacidity (pH5.5-6).
Select the root and stem of certain plants: the healthy pine tree root and stem of certain plants of selecting tree-walk to cut down 1-2 years more than 20CM.
2, bacterial classification production
2.1 bacterial classification requirement
Select the Poria cocos kind " the outstanding No. 1-A5 in river " of high-quality, high yield, strong stress resistance, the cultivation of the suitable pine tree root and stem of certain plants.According to the requirement of NY/T528-2002 bacterial classification production technology regulation, make mother's kind, original seed, the cultivated species of Poria cocos.
Plant (one-level kind) and original seed (secondary kind) 2.2 female
One-level kind, secondary kind must by the national correlation department regulation have qualification for the supply of arranging production of kind of a unit.The I and II kind how expand numerous by generation.The one-level kind is teat glass agar medium bacterial classification (PDA medium: potato 200g, glucose 20g, agar 20, water 1000ml, pH nature; CYM medium: sucrose 200g, peptone 4g, KH
2PO
43g, MgSO
41.5g, agar 20g, water 1000ml, pH nature).The secondary kind is 750ml standard wide-mouth seed bottle bacterial classification (original seed recommended formula: take pine sawdust as major ingredient 82%, wheat bran 15%, sucrose 2%, gypsum 1%, water content 55-60%).The one-level kind must be through the rejuvenation of CYM medium, and keeping " the outstanding No. 1-A5 in river " distinctive growth characteristics of bacterial strain with its mycelia is that mycelia is vigorous " wavy " collarium while growing forward on the PDA slant tube and is arranged as target and carries out the female production of planting of Poria cocos one-level; The Poria cocos seed production is produced according to a conventional method: pine sawdust and other auxiliary materials are puddled evenly, and the 750ml standard of packing into wide-mouth seed bottle is by autoclaving (1.4 * 10
5Pa) 2～2.5h, cooling latter 25 ± 3 ℃, under sterile working, the access Poria cocos is female plants, and puts under 25 ± 3 ℃ and cultivates 20-25 days, regular sense organ inspection.
2.3 cultivated species (three grades of kinds)
Cultivated species must, by after having the examination of relevant authorities of country qualified, be issued the professional field of bacterial of bacterial classification production licence and make.Tuckahoe cultivated species is produced recommended formula: pine sheet ((1 * 2 * 12cm) 68%, wheat bran 15%, pine sawdust 15%, gypsum 2% etc. 20%.Before preparation, the pine sheet that dries is soaked to 10～12h, after draining and other auxiliary materials evenly mix and stir rear pack (15 * 30 * 0.4cm), every sacked material 0.5kg, autoclaving (1.4 * 10
5Pa) 2～2.5h, after bacterium bag cooling (25 ± 3 ℃), access Poria cocos original seed under sterile working, put under 25 ± 3 ℃ and cultivate 25-30 days, regularly sense organ inspection.
2.4 keep sample
Bacterial classifications at different levels should keep sample for future reference.The quantity kept sample is propped up with the female 3-5 that plants of each lot number, and original seed and cultivated species 5-7 bottle (bag) are stored under 4-6 ℃, for future reference.
2.5 Poria cocos strain quality standard
2.5.1 the female quality standard of planting of Poria cocos
The mycelia look white is, even, fine and close, sturdy, and the tool Poria cocos is special to give off a strong fragrance, cell age 7～10 d; The visible sparkling and crystal-clear dew of mycelium surface drips shape secretion, and the bacterial classification test tube is intact, and tampon is tight, without living contaminants.2.5.2 Poria cocos original seed quality standard
Mycelial growth is vigorous, closely questions whitely, even, fine and close, climbs the wall phenomenon obvious, visible rhizomorph, cell age 15～30 d; The most advanced and sophisticated visible milky of mycelium is revealed a shape secretion, and Poria cocos is special to give off a strong fragrance, and seed bottle is intact, without living contaminants.2.5.3 Poria cocos cultivation strain quality standard
The pure white densification of mycelia evenly is covered with wood chip, and the bacterium sheet slightly is rotten wooden shape but soft corruption within reason, and section is faint yellow, the special aromatic flavour of tool Poria cocos, and cell age 30～45 d, the bacterium bag is complete, without the jaundice mycelia, without fruit body, grows, without living contaminants.2.5.4 the first-selected bacterial classification cell age that checks of Poria cocos strain quality standard test rules, be 7～10 d as mother plants, original seed 20～30 d, cultivation strain 30～45 d.Check bacterial classification culture vessel, as test tube, seed bottle, bacterial classification culture bag etc., must be complete, without damaged, tampon or bottle cap, sack are tight, sealing.Mycelial growth is vigorous, and look pure white is, even, fine and close, is covered with in culture vessel, without living contaminants.The most advanced and sophisticated visible milky of mycelium is revealed a shape secretion.Open culture vessel, can smell the special fragrance of strong Poria cocos.
3, the pretreatment of the pine tree root and stem of certain plants
By pine tree root and stem of certain plants periphery eliminating impurities, the dry crust of being cut of the root and stem of certain plants is processed, plane pine tree root and stem of certain plants limit soil, find out the main root place of the pine tree root and stem of certain plants, the position dry near the root and stem of certain plants wide 5～8cm that prunes to the one main root, the root skin of long 15 cm, the large root and stem of certain plants needs to select the processing of being peeled of pine stump that the 2-3 root is sturdy again, and according to Siberian cocklebur field actual landform, drainage trenching.
4 inoculations
4.1 bacterial classification: pine sheet bacterial classification
4.2 inoculum concentration: 1 bag of bacterial classification/pine tree root and stem of certain plants diameter 20 ± 5cm, 2 bags of bacterial classification/pine tree root and stem of certain plants diameter 30 ± 5cm, 3～4 bags of pine tree root and stem of certain plants that bacterial classification/pine tree root and stem of certain plants diameter 40cm is above.
4.3 seed stage: be the inoculation optimum period 5-August, and during inoculation, the suitableeest temperature is 25 ± 3 ℃, meet the high temperature fine day should early morning or the dusk inoculated.
4.4 inoculation method: main root does not remove a bag inoculation method.
During inoculation without cultivating bacteria bag is dialled from, as long as cultivated species bacterium bag is cut open to a slit that 2cm is wide, the mouth of newly splitting that the opening slit of bacterium bag is close to pine tree root and stem of certain plants main root and the dry infall of the root and stem of certain plants gets final product.
5 manage and gather
5.1 note control water and ventilation
Meet the rainy day after inoculation and want duty inspections of going up a hill, the ditch cleaning draining, guarantee that not ponding prevents from around the pine tree root and stem of certain plants becoming silted up water, as Continuous Drought or dry and cold season, should thicken the overburden layer humidification, but will keep overburden layer to loosen, ventilate.
5.2 carry out earthing in good time
Unify earthing on pine tree root and stem of certain plants limit after inoculation, thickness 6-10CM.If the chance heavy rain, soil layer runs off, and after fine day, should mend in time soil.Inoculate after 60-120 days and want frequent cruising Siberian cocklebur field, once find that pine tree root and stem of certain plants periphery table soil cracking has the outer earthing in time of sclerotium, guarantees the continued growth of poria cocos sclerotium.
5.3 the extermination of disease and insect pest
5.3.1 insect pest
Termite is the primary pest in the Poria cocos cultivation, executes one time the low-toxin farm chemicals control termites such as chlorine valerate when the pretreatment of the pine tree root and stem of certain plants.The cultivation process is used agricultural chemicals in strict accordance with NY/T393-2000 standards for safe use and the rationally regulation execution of Application standard.Do not use the agricultural chemicals of high poison, high residue, allow to use low-toxin farm chemicals, dispenser is apart from more than picking time interval 30d.Must not spray any chemical agent picking time.
5.3.3.2 disease
The Major Diseases of Poria cocos is that " rotten maize ear rot " is physiological disturbance.Cardinal symptom: large tracts of land atrophy ulcer appears in the sclerotium epidermis, the recessed deformity of collapsing of sclerotium.Control principle should be followed the method for relying mainly on prevention.The knot Siberian cocklebur initial stage is suitably thickeied overburden layer according to actual conditions, makes the not fusion with temperature of soil temperature; Hold drain ditch during rainy season successfully, the water and airtight that prevents from becoming silted up, hot and humid and cause rotten Siberian cocklebur; Should gather in time when Poria cocos is ripe, the Siberian cocklebur rope of immature young Siberian cocklebur not broken while gathering.
5.4 gather
5.4.1 gather period
After introducing a fine variety, table soil cracking no longer appears in 10-12 months Siberian cocklebur ground, and the poria cocos sclerotium skin darkens, and white slight crack no longer occurs, is Harvesting Date.The principle of gathering: the young pines root and stem of certain plants is first adopted, and after the large root and stem of certain plants, adopts, and ripe sclerotium is first adopted, and immature young Siberian cocklebur does not injure through earthing and can again gather after 2-4 months while adopting Siberian cocklebur.Especially big butt can be gathered 3-5 time, lasts 2-3, until the change of whole pine tree root and stem of certain plants approach exhaustion is rotten.
The knot Siberian cocklebur comparative trial of embodiment 3 " the outstanding No. 1-A5 in outstanding No. 1 ”Yu“ river, river "
The beginning of the nineties I select " river outstanding No. 1 " kind by the inducing and acclimating to wild Poria cocos bacterial strain, the early stage result of the test of this kind shows it is also the kind of the more suitable Poria cocos pine of strain root and stem of certain plants cultivation, but the cultivation application through the several years shows that its genetic stability is slightly poor, and bacterial strain has degradation phenomena to produce.So 2000 we to take Poria cocos wild strain " Fujian Siberian cocklebur " be the parent, application Mutagenesis technology selects Poria cocos new varieties " the outstanding No. 1-A5 in river ", this kind genetic stability is good, on the knot Siberian cocklebur output of loose root and stem of certain plants cultivation, performance is particularly outstanding, more than average single root and stem of certain plants output can reach 15kg/ pine root and stem of certain plants diameter 25 ± 5cm.Within 2010, carried out the knot Siberian cocklebur comparative trial of two kinds in forest farm, Xian Shui Beizhen City, Shaowu, the result of the test statistical analysis is in Table 8." the outstanding No. 1-A5 in river " ties Siberian cocklebur than " outstanding No. 1 of river " early as known from Table 1, inoculates to survive also to exceed 5 percentage points, and average single root and stem of certain plants output exceeds 25.61%.
Table 8: " the outstanding No. 1-A5 in outstanding No. 1 ”Yu“ river, river " knot Siberian cocklebur comparing result
Claims (4)
1. a strain Poria cocos bacterial strain is characterized in that: described Poria cocos bacterial strain be Poria cocos (
Wolfiporia extens) the outstanding No. 1-A5 in river, on October 29th, 2012, being deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, it is called for short CGMCC, and preserving number is CGMCC NO. 6660.
2. the method for a low-carbon (LC) high-yield culturing Poria cocos is characterized in that: utilize the sylvan life discarded object pine tree root and stem of certain plants to carry out Poria cocos and naturally cultivate, select the specificity Poria cocos bacterial strain of suitable pine tree root and stem of certain plants cultivation, preserving number be CGMCC NO. 6660 Poria cocos (
Wolfiporia extens) the outstanding No. 1-A5 in river, simultaneously in conjunction with main root inoculation method cultivation Poria cocos; The cultivation step is: the pretreatment of bacterial classification production → pine tree root and stem of certain plants → inoculation → Routine Management and gathering.
3. the method for low-carbon (LC) high-yield culturing Poria cocos according to claim 2, it is characterized in that: the step that described bacterial classification is produced is as follows: (1) at first carries out the rejuvenation of bacterial strain, is vigorous " wavy " while growing forward on the PDA slant tube with mycelia and is arranged as the production that target is carried out the female kind of Poria cocos one-level; (2) Poria cocos seed production: the pine sawdust of take is produced the Poria cocos original seed according to a conventional method as major ingredient; (3) tuckahoe cultivated species production: take pine sheet as major ingredient, the cultivation of access Poria cocos original seed.
4. the method for low-carbon (LC) high-yield culturing Poria cocos according to claim 1, it is characterized in that: the pretreatment of the described pine tree root and stem of certain plants comprises: by pine tree root and stem of certain plants periphery eliminating impurities, and carry out termite-proof processing, find out the main root place of the pine tree root and stem of certain plants, the position dry near the root and stem of certain plants wide 5～8 cm that prune to main root, the root skin of long 15 cm, the large butt that diameter surpasses 25cm needs to select the processing of being peeled of 2～3 thicker lateral roots again.
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| CN109588210A (en) * | 2019-01-07 | 2019-04-09 | 重庆市药物种植研究所 | Artificial induction's method of stone-like omphalia fructification |
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| CN110651663A (en) * | 2019-11-12 | 2020-01-07 | 吴丽娟 | Production method of green selenium-rich poria cocos |
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