CN103907481A - Process for manufacturing high-quality morchella strains - Google Patents
Process for manufacturing high-quality morchella strains Download PDFInfo
- Publication number
- CN103907481A CN103907481A CN201410157198.4A CN201410157198A CN103907481A CN 103907481 A CN103907481 A CN 103907481A CN 201410157198 A CN201410157198 A CN 201410157198A CN 103907481 A CN103907481 A CN 103907481A
- Authority
- CN
- China
- Prior art keywords
- spore
- hickory chick
- level
- suspension
- morchella
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a process for manufacturing high-quality morchella strains. The process includes selecting acquisition time of morchella; screening high-quality strains; cultivating mycelia; manufacturing and cultivating mother strains; manufacturing and cultivating cultivated strains. The morchella is acquired in the seventh-tenth days when first or second crops of morchella are fruited in February or March of the second year of morchella planting under the conditions that morchella individuals are longer than or equal to 10cm and are shorter than or equal to 15cm, pilei of the morchella individuals are plump, ridges and pits on the pilei are completely spread and stipites of the morchella individuals are milky; the high-quality strains are ejected and screened step by step according to forms and colors of the pilei, colors and forms of the stipites, colors and quantities of spores and sclerotium growing ability of the strains. The process has the advantages that the yield of fresh morchella which grows from the high-quality morchella strains manufactured by the aid of the process is stable for three continuous years and is stabilized at about 80kg, the fruiting time is stable, the growing speeds of the individuals are similar to one another, and the technical problem of instable yields in current morchella industrial development can be solved.
Description
Technical field
The present invention relates to a kind of production technology of hickory chick good quality strain.
Background technology
Hickory chick (
morchella) have another name called sheep tripe dish, delicious hickory chick, sheep mushroom, gain the name because its appearance exactly likes sheep tripe, be Ascomycotina (
ascomy-cotina), discomycete (
discomycetes), Pezizale (
pezizales) Morchellaceae (
morchellaceae), morchella (
morchel-la).Hickory chick distributes more extensive in China, but the hickory chick species resource richness difference in each area, and also have very big-difference.Up to now, the hickory chick of having reported has 29 kinds, there are 15 kinds in the hickory chick kind of distribution in China, Yunnan has 4 kinds: balck vein hickory chick, Morchellaconica, high sheep liver bacterium and sheep liver bacterium, being distributed in Kunming, Dali, Lijing, Zhong Dian, Zhaotong, height above sea level 1000-3000 meter territory, Deng Dizhou city, Qujing, is one of Chinese hickory chick major production areas.Traditional medicine proves: hickory chick property is flat, and taste is sweet cold, nontoxic; Useful stomach, aid digestion, phlegm-eliminiating and qi-regulating, tonifying kidney and strengthening yang, benefit brain effect such as refresh oneself.In addition, its delicious flavour of hickory chick, therefore enjoys domestic and international favor.
Wild toadstool is more extensive in the distribution in Lijiang County In Yunnan Province city, is mainly distributed in stone township, Jiu He township and Ludian township.But constantly soaring due to hickory chick price in recent years, causes the continuous collection of hickory chick wild resource, and grown place, hickory chick open country has been difficult to collect hickory chick again now, and therefore, hickory chick bionic cultivation is extremely urgent.From 2006, Alpine Economic Plants Inst., Yunnan Prov. Academy of Agricultural Sciences has carried out collection and the experiment in cultivation of hickory chick bacterial classification, test has obtained success, per mu yield fresh goods hickory chick 60kg left and right, but we find that the annual output of hickory chick is unsettled subsequently, we have carried out output assessment to the M001 hickory chick of 2006-2008 plantation, hickory chick per mu yield in 2006 is 62.57kg, hickory chick per mu yield in 2007 is 54.32kg, hickory chick per mu yield in 2008 is 49.51kg, 2007 and per mu yield in 2008 have declined 13.19% and 20.87% than per mu yield in 2006 respectively, we have done correlative study to this, research is found, the stable yields of hickory chick is except outside the Pass having with natural conditions and management, there is direct relation with the quality of bacterial classification, the acquisition time of for example bacterial strain and the mode of appearance of hickory chick, the quality of female kind and cultivated species etc.Therefore, keep the stable high yield of hickory chick must be using the bacterial classification of high-quality as prerequisite.The invention provides a kind of production technology of hickory chick good quality strain, be intended to specification hickory chick good quality strain production method, support for the unstable important technology that provides of output in hickory chick production process is provided.
Summary of the invention
The object of the invention is to provide a kind of production technology of hickory chick good quality strain, to have solved the unsettled technical problem of current hickory chick output.
The production technology of a kind of hickory chick good quality strain provided by the present invention, comprises the following steps:
(1) acquisition time of hickory chick
The February～March of plantation hickory chick Second Year, after first batch or second batch of hickory chick fruiting 7th～10 days, 10cm≤hickory chick individual lengths≤15cm, cap is full, on cap, rib and pit launch completely, gather the hickory chick bacterial strain without damage by disease and insect when stem is milky;
(2) screening of quality strains, eliminate step by step screening by following level Four:
1. cap form and dithering
Width≤the 5cm of cap length >=4 cm, 2 cm≤cap the widest part, cap color are black or filemot for one-level, and all the other be secondary, the superseded second level;
2. stem color and form screening
By step (2) 1. not superseded bacterial strain first carry out the comparison of stem color, stem color be milky be primary election one-level; Stem color is white or filemot for primary election secondary, eliminates primary election secondary; Be that milky primary election one-level bacterial strain carries out the comparison of stem form again by stem color, wherein disease-free, stem the narrowest place width >=1.5cm be one-level, all the other are secondary, eliminate the second level;
3. spore color and quantity screening
A. the collection of spore, by clean A
4paper is converted into envelope shape, take off step (2) 2. not superseded hickory chick cap and remove the earth on cap, then cap put into envelope top down, seal, put and shine in the sun 2～3 days, obtain hickory chick spore print, scraping spore print obtains spore, spore color be yellow be one-level spore, spore color be white be secondary, eliminate the second level;
B. one-level spore is put into the test tube of 10ml sterile water, shake up, be made into the female suspension of spore, getting the female suspension of 1ml spore adds in the sterile water test tube of 100ml, shake up to such an extent that dilute for the first time spore suspension, get and dilute for the first time in the sterile water test tube that spore suspension 1ml adds 100ml, shake up to such an extent that dilute for the second time spore suspension, get and dilute for the second time in the sterile water test tube that spore suspension 1ml adds 100ml, shake up to such an extent that dilute for the third time spore suspension, get and dilute for the third time spore suspension 1ml and be placed on slide, carry out under the microscope spore counting, spore number>=10
2individual is one-level, spore number < 10
2individual is secondary, eliminates the second level;
4. sclerotium generative capacity screening
Under aseptic condition, the bacterial strain spore of a small amount of step of picking (2) hickory chick that 3. B does not eliminate is put into 10ml sterile water, shake up and obtain the female suspension of spore, get the female suspension 1ml of spore and put into 100ml sterile water, shake up and obtain spore suspension, now get the spore suspension of 1ml and put into PDA medium, be coated with the spreading rod of sterilizing, 25 DEG C of dark cultivations, in the time growing bacterium colony, single bacterium colony of picking accesses in PDA medium again, 25 DEG C of dark cultivations, now obtain single spore mycelia, cover with after plate, get in 1.0cm × 1.0cm bacterium piece access PDA medium, under 25 DEG C of conditions, cultivate, observed and recorded mycelial growth rate, sclerotium rise time and sclerotium generate quantity, mycelial growth rate >=6.4mm/d, sclerotium quantity >=40, the bacterial strain that sclerotium rise time≤7d and mycelia are covered with the plate of radius r=45mm is one-level, all the other are secondary, eliminate the second level, the one-level bacterial strain obtaining is hickory chick quality strains,
(3) cultivation of mycelia
By after PDA medium autoclaving, be cooled to 50-60 DEG C, under aseptic condition, get 15ml PDA medium and pour in flat board after cooled and solidified, for subsequent use; The spore of the hickory chick quality strains 4. obtaining by a small amount of step of transfer needle picking (2) is put into 10ml sterile water, shake up, obtain the female suspension of spore, get the female suspension 1ml of spore and put into 100ml sterile water, shake up and obtain spore suspension, now get the spore suspension of 1ml and put into PDA medium, be coated with the spreading rod of sterilizing, 25 DEG C of dark cultivations, in the time growing bacterium colony, single bacterium colony of picking accesses in PDA medium again, and 25 DEG C of dark cultivations are for subsequent use after covering with plate;
(4) female making and cultivation of planting
1. the preparation of mother culture media: pull out after the wheat soaking 48 hours is boiled, in the wheat boiling, add sucrose and wheat bran, the addition of sucrose is 0.1% of the wheat quality that boils, and the addition of wheat bran is 0.2% of the wheat quality that boils, mixes and controls water content thoroughly and be
30%obtain mother culture media;
2. female making and cultivation of planting, the mother culture media of preparation is packed in the polypropylene blake bottle of 300ml, the Mother culture base unit weight packing into is 2/3 of polypropylene blake bottle volume, then by polypropylene blake bottle at 126 DEG C, under the condition of pressure 0.15Mpa, after sterilizing 1h, be chilled to room temperature; Under aseptic condition, after the mycelia that the step (3) of every polypropylene blake bottle access 1cm × 1cm is cultivated, be placed in 25 DEG C of incubators, under 100 lx illumination, cultivate, after mycelia is covered with polypropylene blake bottle, proceed to the dark 10～15d of cultivation in 15 DEG C of incubators;
(5) making of cultivated species and cultivation
1. the preparation of cultivated species medium: in mass fraction, with wood chip be 78%, wheat bran is 20%, sucrose is 1%, lime is 0.1%, and potassium dihydrogen phosphate is 0.1%, and the wheat soaking 48 hours is that to mix and control water content thoroughly be 30% to obtain cultivated species medium in 0.8% mixing;
2. the making of cultivated species and cultivation: the 1kg cultivated species medium of preparation is packed in the Polypropylene Bag that specification is 17cm × 33cm, limit rim compacting, making cultivated species culture volume is 2/3 of Polypropylene Bag volume, with glass fiber tying and stay gap, again by Polypropylene Bag at 121～126 DEG C, under the condition of pressure 0.12～0.15Mpa, after sterilizing 1.5h, be cooled to room temperature, for subsequent use; The mother that step (4) is 2. cultivated plants described cultivated species medium being housed and being cooled in the Polypropylene Bag of room temperature through sterilizing of access, at 25 DEG C, cultivate 20-30d, mycelia is covered with after bacterium bag, proceed under room temperature and continue and cultivate 10-15d, in every Polypropylene Bag, female kind access amount is 10g, when cultivated species medium surrounding covers with golden yellow sclerotium, obtain hickory chick good quality strain.
Compared with prior art, the invention has the beneficial effects as follows:
?by conventional farmland cultivation mode, (conventional farmland cultivation mode refers in farmland the hickory chick good quality strain that production technology of the present invention is produced, the method of utilizing roundleaf poplar to plant hickory chick for bacterium assortment) plant, carry out continuous three annual production assessments, 2011, 2012, within 2013 continuous 3 years, fresh mushroom production reaches respectively 83.52kg, 79.95kg, 81.74kg, the fruiting time of continuous 3 years is respectively on February 26th, 2011, on February 29th, 2012, on February 28th, 2013, within continuous 3 years, individual lengths is respectively 8～12cm, 6～13cm, 5～10cm, therefore, the continuous 3 years fresh mushroom productions of hickory chick good quality strain that the inventive method is produced are steady, within continuous 3 years, fresh mushroom production is stabilized in 80kg left and right, it is maximum less than 5% that each annual production differs, the fruiting time is stable, individual growth speed is close, solve the unsettled technical problem of output in current hickory chick industrialized development.
Brief description of the drawings
Fig. 1 is the individual each spot size instrumentation plan of strain of steepletop hickory chick.In figure, each mark represents successively: 1-Morchellaconica cap, and 2-Morchellaconica stem, L represents bacterial strain individual lengths, and LC represents cap length, and WC represents the width of cap the widest part, and WS represents the stem width at narrow place.
Embodiment
Referring to Fig. 1, bacterial strain individual lengths: be bacterial strain individual lengths from top outer rim to the length L of the bottom outer rim of bacterial strain stem of bacterial strain cap.
Cap length: be cap length from the top outer rim of bacterial strain cap to the length L C of the bottom outer rim of bacterial strain cap.
embodiment 1 the inventive method
1,the acquisition time of hickory chick
The acquisition time of the hickory chick of cultivation is generally between annual February-June, during this period, hickory chick is fruiting constantly, the hickory chick in first batch and second batch (February～March) is individual large, form is good, micro-Microscopic observation, the spore shape ellipse of hickory chick, energetic, therefore, the 7-10 days of the Best Times that gathers hickory chick after first batch or second batch of hickory chick fruiting, 10cm≤hickory chick individual lengths≤15cm, cap is full, without damage by disease and insect, stem milky, cap rib and pit launch completely, now spore comes to the ripening period, and also do not launch spore, it is now the best period that gathers hickory chick, plantation hickory chick is generally annual August-October.Therefore, February～the March of the Second Year after plantation hickory chick, after first batch or second batch of hickory chick fruiting 7th～10 days, 10cm≤hickory chick individual lengths≤15cm, cap is full, on cap, rib and pit launch completely, and stem is the best period that gathers hickory chick while being milky, need gather the hickory chick bacterial strain of complete stool without damage by disease and insect when collection.
The present embodiment is in February, 2010, the Morchellaconica of Yulong county of Lijiang County In Yunnan Province city stone township random acquisition (
m.conica) 20 strains, gathering first batch is after fruiting in February the 8th day, be numbered M001, M002 ... .M010, second batch is within after fruiting in February the 18th day, to be numbered M011, M012 ... 10cm≤hickory chick individual lengths L≤15cm of the Morchellaconica .M020, gathering, cap are full, rib and pit launches completely, stem is that milky, complete stool are without damage by disease and insect on cap.
2, the screening of quality strains, eliminate step by step screening by following level Four:
1. cap form and dithering
The bacterial strain that step 1 is gathered carries out the comparison of cap form and color, and cap length L C >=4 cm, the width W C≤5cm of 2 cm≤cap the widest part, cap color are black or filemot for one-level, and all the other be secondary, the superseded second level;
Result:
One-level strain of steepletop hickory chick has 13 strains, respectively:
M002、M003、M004、M005、M007、M008、M009、M010、M011、M012、M015、M018、M020。
Secondary strain of steepletop hickory chick has 7 strains, respectively:
M001、M006、M013、M014、M016、M017、M019。
2. stem color and form screening
By step (2) 1. not superseded bacterial strain first carry out the comparison of stem color, stem color be milky be primary election one-level; Stem color is white or filemot for primary election secondary, eliminates primary election secondary; Be that milky primary election one-level bacterial strain carries out the comparison of stem form again by stem color, wherein disease-free, stem the narrowest place width W S >=1.5cm be one-level, all the other are secondary, eliminate the second level;
Result:
8 strains of primary election one-level strain of steepletop hickory chick, respectively:
M002、M004、M005、M008、M010、M011、M012、M020。
5 strains of primary election secondary strain of steepletop hickory chick, respectively:
M003、M007、M009?、M015、M018。
6 strains of one-level strain of steepletop hickory chick, respectively: M002, M005, M008, M010, M012, M020.
2 strains of secondary strain of steepletop hickory chick, respectively: M004, M011.
3. spore color and quantity screening
A. the collection of spore, by clean A
4paper is converted into envelope shape, take off step (2) 2. not superseded hickory chick cap and remove the earth on cap, then cap put into envelope top down, seal, put and shine in the sun 2～3 days, obtain hickory chick spore print, scraping spore print obtains spore, spore color be yellow be one-level spore, spore color be white be secondary, eliminate the second level;
B. one-level spore is put into the test tube of 10ml sterile water, shake up, be made into the female suspension of spore, getting the female suspension of 1ml spore adds in the sterile water test tube of 100ml, shake up to such an extent that dilute for the first time spore suspension, get and dilute for the first time in the sterile water test tube that spore suspension 1ml adds 100ml, shake up to such an extent that dilute for the second time spore suspension, get and dilute for the second time in the sterile water test tube that spore suspension 1ml adds 100ml, shake up to such an extent that dilute for the third time spore suspension, get and dilute for the third time spore suspension 1ml and be placed on slide, carry out under the microscope spore counting, spore number>=10
2individual is one-level, spore number < 10
2individual is secondary, eliminates the second level;
Result:
One-level bacterial strain has 4 strains: M002, M005, M010, M012.
Secondary bacterial strain has 2 strains: M008, M020, eliminates the second level.
4. sclerotium generative capacity screening
Under aseptic condition, picking is put into respectively 10ml sterile water through M002, M005, M010, the M012 bacterial strain spore of step 2 Morchellaconica that 3. B does not eliminate on a small quantity respectively, shakes up, and obtains respectively the female suspension of four kinds of spores, and every kind of female suspension of spore operates by following:
Get the female suspension 1ml of spore and put into 100ml sterile water, shake up and obtain spore suspension, now get the spore suspension of 1ml and put into PDA medium, be coated with the spreading rod of sterilizing, 25 DEG C of dark cultivations, in the time growing bacterium colony, single bacterium colony of picking accesses in the PDA medium of newly joining again, 25 DEG C of dark cultivations, now obtain single spore mycelia, cover with after plate, getting 1.0cm × 1.0cm bacterium piece accesses in the PDA medium of newly joining, under 25 DEG C of conditions, cultivate, observed and recorded mycelial growth rate, sclerotium rise time and sclerotium generate quantity, mycelial growth rate >=6.4mm/d, sclerotium quantity >=40, the bacterial strain that sclerotium rise time≤7d and mycelia are covered with the plate of radius r=45mm is one-level, all the other are secondary, eliminate the second level, the one-level bacterial strain obtaining is hickory chick quality strains.
Result:
One-level bacterial strain has 1 strain: M012.
Secondary bacterial strain has 3 strains: M002, M005, M010, eliminates the second level.
3, the cultivation of mycelia
By after PDA medium autoclaving, be cooled to 50-60 DEG C, under aseptic condition, get 15ml PDA
Medium is poured in flat board after cooled and solidified, for subsequent use; 4. the spore of the hickory chick quality strains M012 obtaining by transfer needle picking step 2 is put into 10ml sterile water, shake up, obtain the female suspension of spore, get the female suspension 1ml of spore and put into 100ml sterile water, shake up and obtain spore suspension, now get the spore suspension of 1ml and put into PDA medium, be coated with the spreading rod of sterilizing, 25 DEG C of dark cultivations, in the time growing bacterium colony, single bacterium colony of picking accesses in the PDA medium of newly joining again, 25 DEG C of dark cultivations, the mycelia now obtaining is single spore mycelia, for subsequent use after covering with plate;
4, female making and cultivation of planting
1. the preparation of mother culture media: pull out after the wheat soaking 48 hours is boiled, in the wheat boiling, add sucrose and wheat bran, the addition of sucrose is 0.1% of the wheat quality that boils, and the addition of wheat bran is 0.2% of the wheat quality that boils, mixes and controls water content thoroughly and be
30%obtain mother culture media;
2. female making and cultivation of planting, the mother culture media of preparation is packed in the polypropylene blake bottle of 300ml, the Mother culture base unit weight packing into is 2/3 of polypropylene blake bottle volume, then by polypropylene blake bottle at 126 DEG C, under the condition of pressure 0.15Mpa, after sterilizing 1h, be chilled to room temperature; Under aseptic condition, after the mycelia that the step 3 of every polypropylene blake bottle access 1cm × 1cm is cultivated, be placed in 25 DEG C of incubators, under 100 lx illumination, cultivate, after mycelia is covered with polypropylene blake bottle (after 10 days, mycelia is covered with bacterium bottle), proceed to the dark 10d of cultivation in 15 DEG C of incubators; Sclerotium of morchella esculenta generates in a large number, and is covered with whole bottle, and sclerotium quality is hard, golden yellow.
5, the making of cultivated species and cultivation
1. the preparation of cultivated species medium: in mass fraction, with wood chip be 78%, wheat bran is 20%, sucrose is 1%, lime is 0.1%, potassium dihydrogen phosphate is 0.1%, the wheat soaking 48 hours is that to mix and control water content thoroughly be 30% to obtain cultivated species medium in 0.8% mixing;
2. the making of cultivated species and cultivation: the 1kg cultivated species medium of preparation is packed in the Polypropylene Bag that specification is 17cm × 33cm, limit rim compacting, making cultivated species culture volume is 2/3 of Polypropylene Bag volume, with glass fiber tying and stay gap, to prevent from rising under high pressure bag, then by Polypropylene Bag at 121～126 DEG C, under the condition of pressure 0.12～0.15Mpa after sterilizing 1.5h, be cooled to room temperature, for subsequent use; The mother that 2. step 4 is cultivated plants described cultivated species medium being housed and being cooled in the Polypropylene Bag of room temperature through sterilizing of access, at 25 DEG C, cultivate 20d, mycelia is covered with after bacterium bag, proceed under room temperature and continue and cultivate 10d, in every Polypropylene Bag, female kind access amount is 10g, when cultivated species medium surrounding covers with golden yellow sclerotium, obtain hickory chick good quality strain.
2 kinds of embodiment plant application test
Experiment place: 3 mu of experimental field that Yong Hong villagers' committee of stone township of Yulong county of Lijiang County In Yunnan Province city is adjacent.
The Morchellaconica good quality strain M012 that embodiment 1 is produced is preserved in high mountain economic plants research institute of agricultural science institute of Yunnan Province.This is carried out to the output assessment of continuous 3 years.With before this bacterial strain plantation, first bacterial strain is proceeded under room temperature and continue and cultivate 10-15d, after a large amount of generations of sclerotium, plant by conventional farmland cultivation mode as cultivated species.In 2010,2011 with respectively M012 bacterial strain is activated in June, 2012, in access mother culture media, cultivate, obtain female kind, again mother is planted in access cultivated species medium and obtain cultivated species, 2010,2011 and adopt conventional farmland cultivation mode to plant in August, 2012, application result:
On February 26th, 2011, Morchellaconica M012 started fruiting, and 7 days time, first batch of mushroom Individual Size reaches 8～12cm, and the per mu yield of M012 fresh goods can reach 83.52kg.
On February 29th, 2012, Morchellaconica M012 started fruiting, and 7 days time, first batch of mushroom Individual Size reaches 6～13cm, and the per mu yield of M012 fresh goods can reach 79.95kg.
On February 28th, 2013, Morchellaconica M012 started fruiting, and 7 days time, first batch of mushroom Individual Size reaches 5～10cm, and the per mu yield of M012 fresh goods can reach 81.74kg.
From above result of the test, can find out, the fresh mushroom production of continuous 3 years of M012 steadily, stable, individual growth speed of fruiting time is close, and solved the unsettled technical problem of output in current hickory chick industrialized development.
Described conventional farmland cultivation mode refers in farmland, the method for utilizing roundleaf poplar to plant hickory chick for bacterium assortment.Main Cultivation Measures is as follows: (1) farmland ploughing and roundleaf poplar bacterium material are prepared; Cultivate front 7～10 d, choose the fresh roundleaf poplar of diameter 15～20 cm and remove branches and leaves, be sawn into the long tree section of 1m and the long tree section of 1.5 m some, place at shady and cool place, plant front 1～2 d and every section of bacterium material is indulged to split 4-6 part for subsequent use.(2) utilize lime, wheat bran, wood chip, white sugar cultivated species to be sent out to bacterium for raw material; (3) ditching-put into bacterium material-sprinkle bacterial classification and a small amount of lime and wood chip-blinding, presents different shapes according to different bacterium material accumulation method cultivation heaps, as ground floor 3 root fungus materials, the second layer 2 root fungus materials, the 3rd layer of 1 root fungus material, piles a taper, can improve like this land utilization rate.(4) labor management, the sunshade net that covering light transmittance is 25%, installs equipment of sprinkler irrigation, and control soil moisture content is 55-65%.
Described PDA medium (being potato culture) formula is: potato 200g, glucose 20g, agar 20g, 1000ml water.
?
Claims (1)
1. a production technology for hickory chick good quality strain, comprises the following steps:
(1) acquisition time of hickory chick
The February～March of plantation hickory chick Second Year, after first batch or second batch of hickory chick fruiting 7th～10 days, 10cm≤hickory chick individual lengths≤15cm, cap is full, on cap, rib and pit launch completely, gather the hickory chick bacterial strain without damage by disease and insect when stem is milky;
(2) screening of quality strains, eliminate step by step screening by following level Four:
1. cap form and dithering
Width≤the 5cm of cap length >=4 cm, 2 cm≤cap the widest part, cap color are black or filemot for one-level, and all the other be secondary, the superseded second level;
2. stem color and form screening
By step (2) 1. not superseded bacterial strain first carry out the comparison of stem color, stem color be milky be primary election one-level; Stem color is white or filemot for primary election secondary, eliminates primary election secondary; Be that milky primary election one-level bacterial strain carries out the comparison of stem form again by stem color, wherein disease-free, stem the narrowest place width >=1.5cm be one-level, all the other are secondary, eliminate the second level;
3. spore color and quantity screening
A. the collection of spore, by clean A
4paper is converted into envelope shape, take off step (2) 2. not superseded hickory chick cap and remove the earth on cap, then cap put into envelope top down, seal, put and shine in the sun 2～3 days, obtain hickory chick spore print, scraping spore print obtains spore, spore color be yellow be one-level spore, spore color be white be secondary, eliminate the second level;
B. one-level spore is put into the test tube of 10ml sterile water, shake up, be made into the female suspension of spore, getting the female suspension of 1ml spore adds in the sterile water test tube of 100ml, shake up to such an extent that dilute for the first time spore suspension, get and dilute for the first time in the sterile water test tube that spore suspension 1ml adds 100ml, shake up to such an extent that dilute for the second time spore suspension, get and dilute for the second time in the sterile water test tube that spore suspension 1ml adds 100ml, shake up to such an extent that dilute for the third time spore suspension, get and dilute for the third time spore suspension 1ml and be placed on slide, carry out under the microscope spore counting, spore number>=10
2individual is one-level, spore number < 10
2individual is secondary, eliminates the second level;
4. sclerotium generative capacity screening
Under aseptic condition, the bacterial strain spore of picking step (2) hickory chick that 3. B does not eliminate is put into 10ml sterile water, shake up and obtain the female suspension of spore, get the female suspension 1ml of spore and put into 100ml sterile water, shake up and obtain spore suspension, now get the spore suspension of 1ml and put into PDA medium, be coated with the spreading rod of sterilizing, 25 DEG C of dark cultivations, in the time growing bacterium colony, single bacterium colony of picking accesses in PDA medium again, 25 DEG C of dark cultivations, now obtain single spore mycelia, cover with after plate, get in 1.0cm × 1.0cm bacterium piece access PDA medium, under 25 DEG C of conditions, cultivate, observed and recorded mycelial growth rate, sclerotium rise time and sclerotium generate quantity, mycelial growth rate >=6.4mm/d, sclerotium quantity >=40, the bacterial strain that sclerotium rise time≤7d and mycelia are covered with the plate of radius r=45mm is one-level, all the other are secondary, eliminate the second level, the one-level bacterial strain obtaining is hickory chick quality strains,
(3) cultivation of mycelia
By after PDA medium autoclaving, be cooled to 50-60 DEG C, under aseptic condition, get 15ml PDA medium and pour in flat board after cooled and solidified, for subsequent use; 4. spore corresponding to hickory chick quality strains obtaining by transfer needle picking step (2) put into 10ml sterile water, shake up, obtain the female suspension of spore, get the female suspension 1ml of spore and put into 100ml sterile water, shake up and obtain spore suspension, now get the spore suspension of 1ml and put into PDA medium, be coated with the spreading rod of sterilizing, 25 DEG C of dark cultivations, in the time growing bacterium colony, single bacterium colony of picking accesses in PDA medium again, and 25 DEG C of dark cultivations are for subsequent use after covering with plate;
(4) female making and cultivation of planting
1. the preparation of mother culture media: pull out after the wheat soaking 48 hours is boiled, in the wheat boiling, add sucrose and wheat bran, the addition of sucrose is 0.1% of the wheat quality that boils, and the addition of wheat bran is 0.2% of the wheat quality that boils, mixes and controls water content thoroughly and be
30%obtain mother culture media;
2. female making and cultivation of planting, the mother culture media of preparation is packed in the polypropylene blake bottle of 300ml, the Mother culture base unit weight packing into is 2/3 of polypropylene blake bottle volume, then by polypropylene blake bottle at 126 DEG C, under the condition of pressure 0.15Mpa, after sterilizing 1h, be chilled to room temperature; Under aseptic condition, after the mycelia that the step (3) of every polypropylene blake bottle access 1cm × 1cm is cultivated, be placed in 25 DEG C of incubators, under 100 lx illumination, cultivate, after mycelia is covered with polypropylene blake bottle, proceed to the dark 10～15d of cultivation in 15 DEG C of incubators;
(5) making of cultivated species and cultivation
1. the preparation of cultivated species medium: in mass fraction, with wood chip be 78%, wheat bran is 20%, sucrose is 1%, lime is 0.1%, potassium dihydrogen phosphate is 0.1%, the wheat soaking 48 hours is that to mix and control water content thoroughly be 30% to obtain cultivated species medium in 0.8% mixing;
2. the making of cultivated species and cultivation: get cultivated species medium 1kg and pack in the Polypropylene Bag that specification is 17cm × 33cm, limit rim compacting, making cultivated species culture volume is 2/3 of Polypropylene Bag volume, with glass fiber tying and stay gap, again by Polypropylene Bag at 121～126 DEG C, under the condition of pressure 0.12～0.15Mpa, after sterilizing 1.5h, be cooled to room temperature, for subsequent use; The mother that step (4) is 2. cultivated plants described cultivated species medium being housed and being cooled in the Polypropylene Bag of room temperature through sterilizing of access, at 25 DEG C, cultivate 20-30d, mycelia is covered with after bacterium bag, proceed under room temperature and continue and cultivate 10-15d, in every Polypropylene Bag, female kind access amount is 10g, when cultivated species medium surrounding covers with golden yellow sclerotium, obtain hickory chick good quality strain.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410157198.4A CN103907481B (en) | 2014-04-20 | 2014-04-20 | Process for manufacturing high-quality morchella strains |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410157198.4A CN103907481B (en) | 2014-04-20 | 2014-04-20 | Process for manufacturing high-quality morchella strains |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103907481A true CN103907481A (en) | 2014-07-09 |
| CN103907481B CN103907481B (en) | 2015-06-17 |
Family
ID=51033757
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410157198.4A Active CN103907481B (en) | 2014-04-20 | 2014-04-20 | Process for manufacturing high-quality morchella strains |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103907481B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104838883A (en) * | 2015-04-22 | 2015-08-19 | 吴中区胥口精益生物医药研究所 | Toadstool strain preparation process |
| CN104928195A (en) * | 2015-07-10 | 2015-09-23 | 青川县智农农业开发有限公司 | Toadstool strain isolation method |
| CN105684729A (en) * | 2016-01-27 | 2016-06-22 | 宁德师范学院 | Method for rapidly identifying fruiting potential of edible fungus strains |
| CN106544279A (en) * | 2016-11-25 | 2017-03-29 | 秦小波 | A kind of Morchella esculenta (L.) Perss cultivation strain and its induction and cultural method |
| CN107043804A (en) * | 2017-04-24 | 2017-08-15 | 四川省农业科学院植物保护研究所 | The authentication method of the anti-white mould performance of hickory chick |
| CN110447460A (en) * | 2019-09-23 | 2019-11-15 | 襄阳职业技术学院 | A kind of cultural method of hickory chick liquid strain |
| CN110591924A (en) * | 2019-09-03 | 2019-12-20 | 西北农林科技大学 | Morchella strain and application thereof |
| CN111548967A (en) * | 2020-05-27 | 2020-08-18 | 湖南省微生物研究院 | Pseudomonas putida X14 and application method thereof |
| CN113647293A (en) * | 2021-07-20 | 2021-11-16 | 云阳县康诺百草农业科技有限公司 | Cultivation method for preventing toadstool strains from forming mycelium pellets |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1095885A (en) * | 1993-10-30 | 1994-12-07 | 朱斗锡 | The cultivation method of hickory chick |
| CN101585732A (en) * | 2008-05-25 | 2009-11-25 | 冯夫平 | Preparation method of morchella culture material |
| CN101743852A (en) * | 2008-12-18 | 2010-06-23 | 陈宝建 | Technique for cultivating high-quality and high-yield toadstool |
| CN102823429A (en) * | 2012-09-03 | 2012-12-19 | 朱斗锡 | Novel morel cultivation method |
| CN103202177A (en) * | 2013-04-15 | 2013-07-17 | 四川川野食品有限公司 | Method for cultivating morchella |
-
2014
- 2014-04-20 CN CN201410157198.4A patent/CN103907481B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1095885A (en) * | 1993-10-30 | 1994-12-07 | 朱斗锡 | The cultivation method of hickory chick |
| CN101585732A (en) * | 2008-05-25 | 2009-11-25 | 冯夫平 | Preparation method of morchella culture material |
| CN101743852A (en) * | 2008-12-18 | 2010-06-23 | 陈宝建 | Technique for cultivating high-quality and high-yield toadstool |
| CN102823429A (en) * | 2012-09-03 | 2012-12-19 | 朱斗锡 | Novel morel cultivation method |
| CN103202177A (en) * | 2013-04-15 | 2013-07-17 | 四川川野食品有限公司 | Method for cultivating morchella |
Non-Patent Citations (3)
| Title |
|---|
| 刘颖; 丰茂飞; 刘丹; 刘再年: ""羊肚菌栽培技术初探"", 《现代园艺》, 31 October 2013 (2013-10-31) * |
| 王晓云: ""羊肚菌的人工驯化与栽培"", 《现代农业》, 30 November 2011 (2011-11-30) * |
| 赵丹丹; 李凌飞; 赵永昌; 钱金良; 田果廷: ""尖顶羊肚菌人工栽培"", 《食用菌学报》, 31 March 2010 (2010-03-31) * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104838883A (en) * | 2015-04-22 | 2015-08-19 | 吴中区胥口精益生物医药研究所 | Toadstool strain preparation process |
| CN104928195A (en) * | 2015-07-10 | 2015-09-23 | 青川县智农农业开发有限公司 | Toadstool strain isolation method |
| CN105684729A (en) * | 2016-01-27 | 2016-06-22 | 宁德师范学院 | Method for rapidly identifying fruiting potential of edible fungus strains |
| CN106544279A (en) * | 2016-11-25 | 2017-03-29 | 秦小波 | A kind of Morchella esculenta (L.) Perss cultivation strain and its induction and cultural method |
| CN106544279B (en) * | 2016-11-25 | 2019-11-08 | 秦小波 | A kind of hickory chick cultivation strain and its induction and cultural method |
| CN107043804A (en) * | 2017-04-24 | 2017-08-15 | 四川省农业科学院植物保护研究所 | The authentication method of the anti-white mould performance of hickory chick |
| CN107043804B (en) * | 2017-04-24 | 2020-05-19 | 四川省农业科学院植物保护研究所 | Identification method for white mold resistance performance of morchella |
| CN110591924B (en) * | 2019-09-03 | 2021-10-29 | 西北农林科技大学 | Morchella strain and application thereof |
| CN110591924A (en) * | 2019-09-03 | 2019-12-20 | 西北农林科技大学 | Morchella strain and application thereof |
| CN110447460A (en) * | 2019-09-23 | 2019-11-15 | 襄阳职业技术学院 | A kind of cultural method of hickory chick liquid strain |
| CN111548967A (en) * | 2020-05-27 | 2020-08-18 | 湖南省微生物研究院 | Pseudomonas putida X14 and application method thereof |
| CN111548967B (en) * | 2020-05-27 | 2022-01-18 | 湖南省微生物研究院 | Pseudomonas putida X14 and application method thereof |
| CN113647293A (en) * | 2021-07-20 | 2021-11-16 | 云阳县康诺百草农业科技有限公司 | Cultivation method for preventing toadstool strains from forming mycelium pellets |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103907481B (en) | 2015-06-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103907481B (en) | Process for manufacturing high-quality morchella strains | |
| CN105493885B (en) | A kind of cultural method of Termitomyces albuminosus with black skin | |
| CN105474976A (en) | High-yield planting method for potatoes | |
| CN105638362A (en) | Cultivation method for interplanting cherry trees in tea garden | |
| CN107396751B (en) | Artificial cultivation method for grassland black mushroom | |
| CN103314785A (en) | Method of cultivating stropharia mushrooms by using ramie stalks | |
| CN103703983B (en) | A kind of breeding method of Wampee tree | |
| CN103430734A (en) | Method for improving maturing rate of konjak seeds | |
| CN105503331A (en) | Soilless culture seedling substrate for angelica and preparation method thereof | |
| CN109258349A (en) | A kind of high-yield high-efficiency cultivation method of sorghum | |
| CN105191625A (en) | Pollution-free high-yield cultivation method for sweet corn | |
| CN104871913A (en) | Cultivation method of big fruit hawthorn in Karst rock-desertification areas | |
| CN108157115A (en) | A kind of Arid Desert Area oil Paeonia papaveracea composite plant method | |
| CN108243832A (en) | The artificial method for planting of paint face mushroom | |
| CN103039356A (en) | Year-round strawberry hybridizing and breeding method | |
| KR101908518B1 (en) | A Method of Cultivation of Stevia Onion Using Stevia Liquid Compost And Stevia Onion Liquid Thereof | |
| CN107624521A (en) | A kind of organic day lily implantation methods | |
| CN107295896A (en) | A kind of organic implantation methods of asparagus | |
| CN105981507A (en) | Wet sand germination accelerating technology for super sweet corn | |
| CN110432077A (en) | A kind of efficient implantation methods of mulberry fruit hayashishita crop rotation mushroom | |
| CN101595820B (en) | Culturing method for nursery stocks of walnut trees with high survival rate | |
| CN106258437A (en) | A kind of flue-cured tobacco cultivation method that can effectively facilitate flue-cured tobacco root growth | |
| CN109997628A (en) | A kind of high yield strawberry cultivation method | |
| CN111386989B (en) | Chenopodium quinoa planting and cultivating method, chenopodium quinoa rice wine and making method thereof | |
| KR20100084080A (en) | Method for cultivating ginseng by organic farming and manufacturing the cultivated ginseng as a extract |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |