CN1943323B - Pleurotus eryngii cross breeding new strain of fruiting body in fine stick shape and its selective breeding method - Google Patents
Pleurotus eryngii cross breeding new strain of fruiting body in fine stick shape and its selective breeding method Download PDFInfo
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- CN1943323B CN1943323B CN200610114135A CN200610114135A CN1943323B CN 1943323 B CN1943323 B CN 1943323B CN 200610114135 A CN200610114135 A CN 200610114135A CN 200610114135 A CN200610114135 A CN 200610114135A CN 1943323 B CN1943323 B CN 1943323B
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Abstract
A new hybrid strain of Pleurotus eryngii (eryngii mushroom)has its fruiting body in the form of fine stick shape, with a conservative number of CGMCC NO.1820, a selective breeding method of said strain is also provided, which comprises the steps: selecting the Pleurotus eryngii No. 9 and Pleurotus eryngii No. 11 as its parent strains for hybridization; separating monospore from either of the parent strains respectively; picking up the hyphae of monokaryon and cultivating them and matching pairs for hybridization; and choosing and cultivating the new hybrid strain, etc. The fruiting body of thepresent new hybrid strain of Pleurotus eryngii has a distinct shape unlike its parent strains.
Description
Technical field
The present invention relates to a kind of Xingbao mushroom cross new strain, and selection.
Background technology
The edible mushroom crossbreeding is a kind of breeding technique commonly used, and is the most accurate with the monospore hybrid method in the crossbreeding technology, so use more.Xingbao mushroom (Pleurotus eryngii) has another name called eryngo picks up the ears, and is a kind of market sale prospect high-grade edible bacterium preferably.The sexual reproduction mode of Xingbao mushroom is different ancestor combination, utilizes the uninucleate hyphae pairing hybridization of the different mating types that produce between different strains to form the principle of nucleated mycelium, the hybrid strain that can seed selection makes new advances.
Summary of the invention
According to above-mentioned principle, the present invention makes parent strain with No. 9 bacterial strains of Xingbao mushroom and No. 11 bacterial strains of Xingbao mushroom, carries out the monospore crossbreeding, selects the club-shaped Xingbao mushroom cross new strain of fruiting body in fine.
Technical scheme provided by the present invention is: the selection of a kind of Xingbao mushroom cross new strain (Pleurotus eryngii) is characterized in that may further comprise the steps:
1) selects the hybrid strain bacterial strain: choose No. 9 bacterial strains of Xingbao mushroom (Pleurotus eryngii) and No. 11 bacterial strains of Xingbao mushroom (Pleurotus eryngii) as the hybrid strain bacterial strain;
2) described parent strain is carried out monospore respectively and separate, and the single spore that separation obtains is seeded on the PDA medium;
3) select uninucleate hyphae, cultivate pairing: the uninucleate hyphae that will not have described two parent strains of clamp connection is seeded on the PDA culture medium flat plate of same ware;
4) selection and cultivation cross new strain: will have the mycelium inoculation of clamp connection on the PDA medium, to cultivate in the step 3), from hybridization hybrid combination, select the hybrid strain that mycelial growth rate is fast and mycelial growth potential is strong, transfer and on the PDA test tube slant, cultivate, obtain cross new strain.
For the cross new strain of above-mentioned acquisition, by antagonistic effect, the relation between esterase isozyme electrophoresis verification experimental verification cross new strain and parent strain.
5) the conventional cultivation carried out antagonistic effect: adopt 3 inoculation methods respectively described cross new strain and described two parent strains to be seeded in the same PDA medium plate and cultivate antagonism situation between each bacterial strain of observed and recorded;
6) carry out the test of esterase isozyme electrophoresis: the mycelia of described cross new strain and described two parent strains is carried out the test of esterase isozyme electrophoresis, observed and recorded esterase isozyme electrophoresis pattern;
7) randomly, carry out the fruiting comparative trial, measure the fruiting body yield of described cross new strain and described two parent strains
,Single mushroom fresh weight and sporophore shape feature.
Two above-mentioned parent strains are all open, can buy from the commercial sources of public offering to obtain, and wherein, No. 9 bacterium sources of Xingbao mushroom are in Beijing City Agriculture and Forestry Institute plant protection environmental protection research institute, and No. 11 bacterium sources of Xingbao mushroom are in Sanming City, Fujian fungal studies institute.
Described WA medium is the single spore isolation medium, and its prescription is: agar 20g, water 1000ml.
Described PDA medium is a uninucleate hyphae pairing hybridization medium, and its prescription is: potato 200g, glucose 20g, agar 20g, water 1000ml.
The invention provides the cross new strain that obtains by said method.Particularly, the club-shaped Xingbao mushroom of a kind of fruiting body in fine (Pleurotus eryngii) cross new strain is provided, this bacterial strain sample has been the preservation of common micro-organisms center C GMCC institute of China Committee for Culture Collection of Microorganisms on September 21st, 2006, the preservation address is: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City Institute of Microorganism, Academia Sinica, its preserving number is CGMCC No.1820, bacterial strain is survived after testing, classification called after: Agaricales, Pleurotaceae, the Xingbao mushroom hybridization novel bacterial of Pleurotus.
Further, the club-shaped Xingbao mushroom cross new strain of described fruiting body in fine, its average biology efficient is 84.64%, the single mushroom fresh weight of average fruit body 70.10g, sporophore shape is thin club-like, and average bacteria cover diameter is 4.81cm, and average bacterial context thickness is 1.63cm, average stem length is 12.36cm, and average stem diameter is 2.56cm.
The present invention has following advantage: the fruit body shape of Xingbao mushroom cross new strain of the present invention is all different with two parent strains, be typical thin club-like, the present domestic bacterial strain that does not also have this kind shape, this bacterial strain are fit to the supermarket inner wrapping very much to be sold, and has good market prospects.
Description of drawings
Fig. 1 is the antagonistic effect result of Xingbao mushroom cross new strain of the present invention and two parent strain.
Fig. 2 is the esterase isozyme zymogram of Xingbao mushroom cross new strain of the present invention and two parent strain.
Fig. 3 is the sporophore shape figure of Xingbao mushroom cross new strain of the present invention and two parent strain.
Embodiment
The selection of Xingbao mushroom of the present invention (Pleurotus eryngii) cross new strain is as follows:
1) selects the hybrid strain bacterial strain
Choose No. 9 bacterial strains of Xingbao mushroom and No. 11 bacterial strains of Xingbao mushroom as the hybrid strain bacterial strain; these two parent strains all can be bought from the commercial sources of public offering and obtain; for example; No. 9 bacterium sources of Xingbao mushroom are in Beijing City Agriculture and Forestry Institute plant protection environmental protection research institute, and No. 11 bacterium sources of Xingbao mushroom are in Sanming City, Fujian fungal studies institute.
2) make medium
The single spore isolation medium, i.e. WA medium, its prescription is: agar 20g, water 1000ml makes according to a conventional method.
Uninucleate hyphae pairing hybridization medium, i.e. PDA medium, its prescription is: potato 200g, glucose 20g, agar 20g, water 1000ml makes according to a conventional method.
3) preparation composts or fertilisers of cultivating
The composts or fertilisers of cultivating prescription is: cotton seed hulls 80%, wheat bran 18%, sucrose 1%, gypsum 1%, water content 60%, preparation according to a conventional method.
4) two parent strains being carried out monospore respectively separates
Choose each one of No. 9 bacterial strains of Xingbao mushroom of robust growth and No. 11 bacterial strain fruit body of Xingbao mushroom respectively, sterilize, on aseptic superclean bench, have the 1cm of lamella with the sterile scissors clip with 75% cotton ball soaked in alcohol wipe surfaces
3Fruit body, hang in filling the triangular flask of sterile water.Hang 20h at normal temperatures, collect basidiospore.
The stoste of collecting basidiospore is diluted, and extension rate is 10
-1, 10
-2, 10
-3Use the glass rod method of scoring then, in WA medium plate, inoculate the basidiospore suspension of different extension rates respectively.After cultivating 2～3 days in 25 ℃ of incubators, at microscopically, the single spore with aseptic inoculation pin picking has just been sprouted is seeded in the plate of PDA medium thereupon.
5) select uninucleate hyphae, cultivate pairing
The uninucleate hyphae that the microscopy single spore is sprouted, observation has or not clamp connection.The uninucleate hyphae of no clamp connection is transferred on the PDA test tube slant, cultivates down, treat that mycelia covers with that to put into refrigerator behind the slant medium standby in 25 ℃.
The uninucleate hyphae of No. 9 bacterial strains of Xingbao mushroom and No. 11 bacterial strains of Xingbao mushroom is seeded on the PDA culture medium flat plate of same ware, and the inoculation point is cultivated down at 25 ℃ at a distance of 3cm.At two kinds of uninucleate hyphae contact position microscopies, find that the explanation pairing of clamp connection is hybridized successfully.
6) select and cultivate cross new strain
The mycelium inoculation that clamp connection will be arranged is on PDA plate medium, cultivated 5 days down in 25 ℃, from hybrid combination, select the hybrid strain that mycelial growth rate is fast and mycelial growth potential is strong, transfer on the PDA test tube slant, it is standby to cultivate the back, promptly obtain cross new strain, the inventor is called No. 14 hybrid strains of Xingbao mushroom to a concrete bacterial strain that wherein obtains, and being preserved in the common micro-organisms center C GMCC of China Committee for Culture Collection of Microorganisms on September 21st, 2006, its preserving number is CGMCC No.1820.
7) carry out antagonistic effect
In PDA medium plate, adopt 3 inoculation methods inoculate Xingbao mushroom respectively No. 9, No. 11 two parent strains of Xingbao mushroom and No. 14 hybrid strains of Xingbao mushroom.Every point to point is at a distance of 2cm, and antagonism situation between each bacterial strain of observed and recorded is cultivated in the inoculation back in 25 ℃ of incubators.
Result of the test sees also Fig. 1, and the antagonistic effect effect is obvious between No. 9, No. 14 hybrid strains of Xingbao mushroom and Xingbao mushroom and No. 11 two parent strains, between per two bacterial strains tangible antagonism line is arranged all.
8) carry out the test of esterase isozyme electrophoresis
Each sample is got the 20mg mycelia, adds 60 μ L zyme extracts, and after grinding in ice bath, the centrifuging and taking supernatant is standby.Adopt the polyacrylamide vertical slab gel electrophoresis, concentrated gum concentration is 2.5% (the pH value is 6.8), and resolving gel concentration is 7% (the pH value is 8.8), and at 4 ℃ of following electrophoresis, concentrating gel electrophoresis voltage is 150V, and the separation gel electrophoretic voltage is 300V.Gel behind the electrophoresis is immersed in the esterase dyeing liquid, and 30 ℃ were dyeed 1 hour.The esterase isozyme electrophoresis pattern of No. 9, observed and recorded Xingbao mushroom, No. 11 two parent strains and No. 14 hybrid strains.
Result of the test sees also Fig. 2, and wherein, No. 1 and No. 2 is No. 9 bacterial strain enzymes of Xingbao mushroom band, and No. 3 and No. 4 is No. 14 bacterial strain enzymes of Xingbao mushroom band, and No. 5, No. 6 and No. 7 is No. 11 bacterial strain enzymes of Xingbao mushroom band.
The esterase isozyme zymogram as shown in Figure 2 between No. 9, No. 14 hybrid strains of Xingbao mushroom and Xingbao mushroom and No. 11 two parent strains, in the Rf value is between the 0.0488-0.919, No. 14 hybrid strains of Xingbao mushroom and No. 9 and No. 11 two parent strains have 10 common enzyme bands, the Rf value is respectively 0.0488,0.0851,0.173,0.199,0.275,0.422,0.485,0.696,0.741,0.919, but at width, slightly different on deep or light.In the Rf value is 0.341 and 0.586 place, has only No. 14 hybrid strains that the enzyme band is arranged.In the Rf value is 0.234 place, and No. 9 bacterial strains of Xingbao mushroom and No. 14 hybrid strains have the enzyme band, but slightly different on deep or light.In the Rf value is 0.389,0.542,0.631 and 0.867 place, and No. 9 bacterial strains of Xingbao mushroom and No. 11 bacterial strains have the enzyme band, but at width, slightly different on deep or light.In the Rf value is 0.946 place, and No. 11, Xingbao mushroom and No. 14 hybrid strains have the enzyme band, but slightly different on deep or light.
9) cultivate comparative trial
Adopt randomised block design, each bacterial strain is established 3 repetitions, each repeats 20 bags. set by step 3) in the prescription weighing, composts or fertilisers of cultivating is stirred, water content transfers to about 65%, adopt 17cm * 40cm * 0.05cm polypropylene plastics sacked material, every packed siccative 300g, autoclaving 2h, when being cooled to below 30 ℃, under aseptic condition, insert No. 9 bacterial strains of Xingbao mushroom respectively, the cultivated species of No. 11 bacterial strains and No. 14 bacterial strains, putting 22 ℃ of-26 ℃ of culturing room cultivates, move into the mushroom room after the mycelia purseful and carry out management of producing mushroom. in the fruiting stage, regulate every environmental condition, make it to satisfy the requirement that the Xingbao mushroom sporophore growth is grown. measure the fruiting body yield of each bacterial strain when gathering, calculate its biology efficient (biology efficient=bright mushroom weight/composts or fertilisers of cultivating dry weight), to the fruit body random sampling of each bacterial strain, measure the single mushroom fresh weight and the sporophore shape feature of each bacterial strain.
The fruiting body yield of Xingbao mushroom different strains and biology efficient see Table 1:
The fruiting body yield of table 1 Xingbao mushroom different strains and biology efficient
As can be seen from Table 1, the fruiting body yield of No. 14 hybrid strains of Xingbao mushroom is between No. 9 bacterial strains of Xingbao mushroom and No. 11 bacterial strains of Xingbao mushroom, and average biology efficient is 84.64%.
The single mushroom fresh weight of the fruit body of Xingbao mushroom different strains sees Table 2:
The single mushroom fresh weight of the fruit body of table 2 Xingbao mushroom different strains
As can be seen from Table 2, the single mushroom fresh weight of No. 14 hybrid strain fruit bodys of Xingbao mushroom minimum, average out to 70.10g, the single mushroom fresh weight of No. 9 bacterial strain fruit bodys of Xingbao mushroom maximum, average out to 95.03g.
The sporophore shape feature of Xingbao mushroom different strains sees Table 3 and Fig. 3:
The sporophore shape feature of table 3 Xingbao mushroom different strains
From table 3 and Fig. 3 as can be seen, the sporophore shape of No. 14 hybrid strains of Xingbao mushroom is thin club-like, and bacteria cover diameter is 4.81cm, the numerical value minimum, bacterial context thickness is 1.63cm, and numerical value is less, and stem length is 12.36cm, numerical value maximum, stem thickness are 2.56cm, the numerical value minimum.
As can be seen from Figure 3, the fruit body shape of No. 14 hybrid strains of Xingbao mushroom is all different with two parent strains, is typical thin club-like, at present the domestic bacterial strain that does not also have this kind shape, this bacterial strain is fit to the supermarket inner wrapping very much to be sold, and has good market prospects.
Claims (2)
1. an Xingbao mushroom (Pleurotus eryngii) cross new strain, it is that preserving number is the Xingbao mushroom cross new strain of CGMCC No.1820.
2. cross new strain according to claim 1, it is characterized in that: its average biology efficient is 84.64%, the single mushroom fresh weight of average fruit body 70.10g, sporophore shape is thin club-like, average bacteria cover diameter is 4.81cm, average bacterial context thickness is 1.63cm, and average stem length is 12.36cm, and average stem diameter is 2.56cm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN200610114135A CN1943323B (en) | 2006-10-30 | 2006-10-30 | Pleurotus eryngii cross breeding new strain of fruiting body in fine stick shape and its selective breeding method |
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| CN200610114135A CN1943323B (en) | 2006-10-30 | 2006-10-30 | Pleurotus eryngii cross breeding new strain of fruiting body in fine stick shape and its selective breeding method |
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| CN1943323B true CN1943323B (en) | 2010-05-12 |
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Non-Patent Citations (4)
| Title |
|---|
| 刘宇等.杏鲍菇13号杂交菌株选育研究.食用菌学报11 3.2004,11(3),61-62. |
| 刘宇等.杏鲍菇13号杂交菌株选育研究.食用菌学报11 3.2004,11(3),61-62. * |
| 王波等.10个杏鲍菇菌株生产性能评价.吉林农业大学学报27 4.2005,27(4),363-365. |
| 王波等.10个杏鲍菇菌株生产性能评价.吉林农业大学学报27 4.2005,27(4),363-365. * |
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