Some standard content:
Appendix A, Appendix B and Appendix C of this standard are all normative appendices. This standard is proposed and managed by the Ministry of Agriculture of the People's Republic of China. GB19172-2003
The drafting units of this standard are: Institute of Soil and Fertilizer, Chinese Academy of Agricultural Sciences, Agricultural Microbiology Center of China Microbiological Culture Collection Administration Committee, and Edible Fungi Engineering Technology Center of Henan Academy of Sciences. The main drafters of this standard are: Zhang Jinxia, Jia Shenmao, Zuo Xuemei, Shen Jinwen1
GB19172-2003
The strains used for cultivating Pleurotus ostreatus are a mixture of artificially cultivated pure mycelium and its culture medium. my country adopts a three-level expansion breeding procedure (i.e., mother strain, original strain, and cultivated strain) to cultivate Pleurotus ostreatus strains. In order to standardize the production, distribution and use of Pleurotus ostreatus strains in my country and ensure the sustainable and healthy development of Pleurotus ostreatus production in my country, this standard is specially formulated. 1 Scope
Pleurotus ostreatus strains
GB19172—2003
This standard specifies the quality requirements, test methods, inspection rules, labels, signs, packaging, storage and transportation of Pleurotus ostreatus strains.
This standard applies to Pleurotus ostreatus strains of the genus Pleurotus, and also applies to the production, circulation and use of Pleurotus sapidus, Pleurotus cornucopiae, Pleurotus pulmonariuss and Pleurotus florida of the genus Pleurotus. 2 Normative references
The clauses in the following documents become the clauses of this standard through reference in this standard. For all referenced documents in the note period, all subsequent amendments (excluding errata) or revisions are not applicable to this standard. However, parties to an agreement based on this standard are encouraged to study whether the latest versions of these documents can be used. For any undated referenced document, the latest version shall apply to this standard. GB/T191 Pictorial marking for packaging, storage and transportation (GB/T191-2000, eqvISO780:1997) GB/T4789.28 Food hygiene microbiological examination staining methods, culture media and reagents GB/T12728-1991 Edible fungi terminology
NY/T528-2002 Technical regulations for the production of edible fungi strains 3 Terms and definitions
The following terms and definitions apply to this standard. 3.1
Stock culture
The pure culture of mycelium and its subculture obtained by various methods with strong properties, using glass test tubes as culture containers and use units, is also called grade species and test tube species. [NY/T528--2002, definition 3.3]
Pre-culture spawn
Pure culture of mycelium obtained by transplanting and expanding the mother culture. Glass culture bottles, plastic culture bottles or 15 cm×28 cm polyolefin plastic bags are usually used as containers.
[NY/T528--2002, definition 3.4]
Cultivated spawn
Pure culture of mycelium obtained by transplanting and expanding the mother culture. Glass bottles or plastic bags are usually used as containers. Cultivated spawn can only be used for cultivation and cannot be expanded for reproduction.
[NY/T 528--2002, definition 3.5]
Antagonism
The phenomenon of no-growth zones or different forms of linear edges between colonies with different genetic genes. 3.5
Angular variation sector
The phenomenon that the mycelium becomes thinner, grows slowly, and the mycelium surface features become abnormally angular due to local variation of the mycelium or infection with viruses. 3.6
high temperatured line
High temperature inhibition line
GB 19172—2003
The phenomenon that the culture of edible fungi becomes yellowish, dark, or the mycelium becomes sparse and weak due to the adverse effects of high temperature during the production process.
Biological efficiencybiological efficiency The ratio between the dry matter of a unit amount of culture medium and the dry weight of the fruiting body or mycelium produced by the culture. [GB/T 12728—1991, definition 2. 1.20] 3.8
characters of variety
The variety characteristics of edible fungi are one of the important criteria for distinguishing the quality of edible fungi species or varieties. Generally, it includes the requirements for temperature, humidity, pH, light and oxygen, stress resistance, high yield, early or late fruiting, fruiting tide number, cultivation cycle, commodity quality and cultivation habits and other agronomic traits.
[NY/T 528—2002, definition 3.8]
4 Quality requirements
4.1 Mother stock
4.1.1 Container specifications shall comply with the provisions of 4.7.1.1 in NY/T528--2002. 4.1.2 Sensory requirements shall comply with the provisions of Table 1. Qiu 1
Sensory requirements for mother culture
Cotton plug or cotton-free plastic cover
Amount of culture medium filled
Length of slope
Size of inoculation block (inoculum amount)
Growth of mycelium
Characteristics of mycelium
Appearance of strain
Surface of myceliumWww.bzxZ.net
Mycelial secretion
Edge of colony
Colony of other bacteria
Appearance of the back of slope
4.1.3 Microbiological requirements shall comply with the requirements of Table 2. Requirements
Complete, intact
Dry, clean, moderately tight, able to meet the requirements of ventilation and bacteria filtration. One-fourth to one-fifth of the total volume of the test tube. 40mm~～50mm from the top of the cotton plug.
(3~5)mm×(3~5)mm
Growing all over the slope
White, dense, vigorous, cotton-like
Uniform, stretched, flat, no angular changes
The culture medium does not shrink, the color is uniform, without dark spots and pigments. It has a unique fragrance of Pleurotus ostreatus. Taste, no sour, smelly, moldy and other peculiar smells 2 Mother strain microbiological requirements
Mycelial growth state
Lock-shaped joint
Requirements
Thick, plump, even
4.1.4 Mycelial growth rate: On PDA medium, at a suitable temperature (25℃ ± 2℃), it will cover the slope in 6 to 8 days. 4.1.5 Mother strain cultivation characteristics: The mother strain provided by the seed supply unit must be confirmed by mushroom fruiting test for agronomic and commercial characteristics before it can be used for expanded reproduction or sale. The biological efficiency of yield characteristics under normal conditions should not be less than 10%. 2
4.2 Original strain
The container specifications should comply with the provisions of 4.7.1.2 of NY/T528-2002. 4.2. 1
4.2.2 Sensory requirements should comply with the provisions of Table 3. Sensory requirements for original strain
Cotton plug or cotton-free plastic cover
Distance between the upper surface of culture medium and the mouth of the bottle (bag)Inoculation amount (number of original strains inoculated with each mother strain, inoculum size)Mycelial growth
Mycelial characteristics
Mycelium on the culture surface
Appearance of strains
Culture medium and mycelium
Secretion on the culture surface
Miscellaneous bacterial colonies
Color phenomenon
Fruiting body primordium
Microbiological requirements should comply with the requirements of Table 2.
Requirements
Complete, intact
GB 19172-—2003
Dry, clean, moderate tightness, can meet the requirements of air permeability and bacteria filtration 50mm±5mm
(4~6) bottles (bags), ≥12 mm×15 mm
Full the container
White and dense, vigorous growth
Grow evenly, no angular change, no high temperature inhibition line Close to the bottle wall, no shrinkage
No, a small amount of colorless or light yellow water drops is allowed None
With the unique fragrance of Pleurotus ostreatus, no sour, smelly, moldy and other odors Mycelium growth rate: On a suitable culture medium, at a suitable temperature (25℃ ± 2℃), it will take 25 to 30 days to fill the container. Cultivated species
Container specifications should comply with 4.7.1 of NY/T528-2002.3. The sensory requirements shall comply with the provisions of Table 4.
Sensory requirements for cultivated species
Cotton plug or cotton-free plastic cover
Distance between the upper surface of the culture medium and the mouth of the bottle (bag)Inoculation amount [Number of cultivated species inoculated with original seed per bottle (bag)]Mycelial growth
Mycelial characteristics
Mycelium in different parts
Appearance of strains
Culture medium and mycelium
Secretion on the culture surface
Miscellaneous bacterial colonies
Face color phenomenon
Fruiting body primordium
Microbiological requirements shall comply with the provisions of Table 2.
Complete, intact
Dry, clean, moderate tightness, meet the requirements of air permeability and bacteria filtration 50 mm±5 mm
(30~～50) bottles (bags)
Full container
White and dense, vigorous and full
Growing evenly, uniform color, no angular change, no high temperature inhibition line, tightly adhere to the bottle (bag) wall, no shrinkage
No, a small amount of colorless or light yellow water droplets is allowedNo
A small amount is allowed, the total amount of primordium is ≤5%
Has the unique fragrance of Pleurotus ostreatus, no sour, smelly, moldy and other odors3
GB 19172--2003
4.3.4 Mycelium growth rate: Under the appropriate temperature (25℃±2℃), it takes (15±2) days for mycelium to fill the bottle and (20±2) days for mycelium to fill the bag on the grain culture medium; it takes 20-25 days for mycelium to fill the bottle and 30-35 days for mycelium to fill the bag on other culture media. 5 Sampling
5.1 The sampling of the quality inspection department should be representative. 5.2 The mother strains shall be numbered in batches according to the variety, culture conditions and inoculation time, and the original strains and cultivated strains shall be numbered in batches according to the strain source, seed production method and inoculation time. The samples to be inspected shall be randomly selected by batch.
5.3 The sampling quantity of the mother strains, original strains and cultivated strains shall be 10%, 5% and 1% of the strain quantity of the batch respectively. However, the sampling quantity of each batch shall not be less than 10 (bottles, bags); if it exceeds 100 (bottles, bags), two-level sampling may be carried out. 6 Test methods
6.1 Sensory test
Perform each test item by item in Table 5.
Table 5 Sensory requirements Test methods
Test items
Tampons, cotton-free plastic covers
Amount of mother culture medium filled
Length of mother culture slope
Appearance of the back of mother culture slope
6.2 Microbiological test
Test methods
Visual observation
Visual observation
Visual observation
Visual observation
Visual observation
Visual observation
Test items
Inoculation amount
Mother culture, original culture
Cultivated culture
Surface of culture medium Distance from the bottle (bag) mouth Various aspects of bacterial appearance (except for colonies of other bacteria) Colonies of other bacteria Inspection methods Visual observation and measurement Inspection of production records Visual observation Visual observation, and use a 5× magnifying glass for observation when necessary 6.2.1 The mycelial growth status and lock-shaped joints in Table 2 should be observed using an optical microscope with a magnification of not less than 10×40 on the water seal of the culture. Each inspection sample should be observed for not less than 50 fields of view. 6.2.2 Bacterial inspection: Take a small amount of culture suspected of bacterial contamination and inoculate it into the nutrient broth culture solution specified in 4.8 of GB/T4789.28 according to aseptic operation. Culture at 25℃～28℃ for 1 to 2 days with shaking, and observe whether the culture solution is turbid. If the culture fluid is turbid, it is contaminated by bacteria. If the culture fluid is clear, it is not contaminated by bacteria.
6.2.3 Fungal test: Take a small amount of culture suspected of fungal contamination and inoculate it into PDA culture medium (see A.1 in Appendix A) according to aseptic operation. Cultivate it at 25℃～28℃ for 3～4 days. If colonies with colors other than white or colonies with non-oyster mushroom hyphae morphology or peculiar smell appear, it is a fungal contaminant. If necessary, conduct water-sealed microscopic examination. 6.3 Mycelial growth rate
6.3.1 Mother stock: PDA culture medium, culture at 25℃ and soil 2℃, and calculate the number of days required for full growth. 6.3.2 Original stock and cultivar: Choose one of the formulas specified in Chapters B.1, B.2, B.3, and B.4, and culture at 25℃ and soil 2℃, and calculate the number of days required for full growth.
6.4 Agronomic and commercial traits in the cultivation of mother stock The mother stock to be tested is made into original stock. Use the culture medium formula specified in Appendix C to make 45 bags. After inoculation, divide into three groups (15 bags in each group) for routine management. According to the items listed in Table 6, make cultivation records and statistically test results. At the same time, set the starting strain of the mother strain as the control and do the same treatment. Compare the test results of the two. Among the test items measured by time, if any time of the mother strain tested is delayed by more than five days (including five days) compared with the control strain, it is unqualified; if the yield is significantly lower than the control strain, it is unqualified; if the appearance of the mushroom body is significantly different from that of the control strain or is deformed, it is unqualified.
Table 6 Inspection records of agronomic and commercial traits in mother seed cultivation Inspection items
Time required for mother seed to grow fully/day
Time required for original seed to grow fully/day
Time required to fill the mushroom bag/day
Time required for the first wave of mushrooms/day
Yield of the first wave of mushrooms/kg
Biological efficiency of the first wave of mushrooms/(%)
6.5 Sample retention
Inspection results
Inspection items
Total yield/kg
Average yield per unit area/kg
Biological efficiency/(%)
Color, texture
Cap diameter, stem length/mm
GB 19172—2003
Test results
All levels of strains must be kept for inspection. The number of samples should be 3 to 5 bottles (bottles, bags) of each batch of mother strains, stored at 4℃ to 6℃, mother strains for 5 months, original strains for 4 months, and culture strains for 2 months. 7 Inspection rules
Judgment rules are carried out according to quality requirements. When all the inspection items meet the quality requirements, it is a qualified strain. If any one of them does not meet the requirements, it is an unqualified strain.
8 Labels, signs, packaging, transportation, storage 8.1 Labels, signs
8.1.1 Product labels
Each bottle (bottle, bag) of fungus must be affixed with a label clearly indicating the following elements: a)
Product name (e.g.: Pleurotus ostreatus mother culture);
Variety name (e.g.: Zhongshu No. 10);
Production unit (×× fungus plant);
Inoculation date (e.g.: 2002.××.××); e)
Implementation standards.
Packaging labels
Each box of fungus must be affixed with a packaging label clearly indicating the following elements: a)
Product name, variety name;
Factory name, address, contact number;
Factory date;
Shelf life, storage conditions;
Quantity;
Implementation standards.
Packing, storage and transportation diagram
According to GB/T191, the following diagrams and signs should be marked: a)
Handle with care sign;
Waterproof, moisture-proof, anti-freeze sign;
Sun protection, high temperature protection sign;
Prevent inversion sign;
Prevent heavy pressure sign.
GB 19172-—2003
8.2 Packaging
8.2.1 The outer packaging of mother stocks shall be made of wooden boxes or cartons made of paper with sufficient strength, and the inside shall be filled with light materials with cushioning effect such as cotton, shredded paper, newspapers, etc.
8.2.2 Original stocks and cutting stocks: The outer packaging shall be made of cartons made of paper with sufficient strength, and the spaces between the strains shall be filled with light materials with cushioning effect such as shredded paper, newspapers, etc. The top and bottom of the carton are sealed with 8cm wide tape and tied with packing tape twice. The box contains the product certificate and instructions for use (including strain type, culture medium formula and applicable scope, etc.). 8.3 Transportation
8.3.1 It shall not be mixed with toxic substances.
8.3.2 When the temperature reaches above 30℃, it shall be transported in a refrigerated truck with a temperature of 2℃~20℃. 8.3.3 During transportation, measures must be taken to prevent shock, sun exposure, dust, rain, frost and contamination by miscellaneous bacteria. 8.4 Purchase and storage
8.4.1 The mother seed shall be stored in a refrigerator at 5℃±1℃, and the storage period shall not exceed 90 days. 8.4.2 The original seed shall be used as soon as possible. The grain seed shall be stored in a clean, dry and ventilated room (relative air humidity 50%~~70%) and light-proof for no more than 7 days, and the original seed of other culture media shall not exceed 14 days. Store at 5℃±1℃, the storage period shall not exceed 45 days. 8.4.3 The seedlings should be used as soon as possible. Grain seeds should be stored in a clean, ventilated, dry (relative humidity 50%~70%), light-proof room at a temperature not exceeding 25℃ for no more than 10 days, and seedlings of other culture media should be stored for no more than 20 days. When stored at 1℃～6℃, the storage period shall not exceed 45 days.
A.1PDA culture medium
Appendix A
(Normative Appendix)
Common mother culture medium and its formula
200g potato, 20g glucose, 20g agar. 2CPDA culture medium
200g potato, 20g glucose, 2g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 20g agar. Appendix B
(Normative Appendix)
Common culture media for original species and cultivated species and their formulas B.1 Grain culture media
98% wheat, millet, corn or sorghum, 2% gypsum, 50% water content and 1% soil. Cottonseed hull and wheat bran culture media
84% cottonseed hull, 15% wheat bran, 1% gypsum, 60% water content and 2% soil. B.3 Cottonseed hull culture media
100% cottonseed hull, 62%±2% water content.
B.4 Wood culture media
79% broadleaf tree chips, 20% wheat bran, 1% gypsum, 60% water content and 2% soil. Appendix C
(Normative Appendix)
Common culture media for testing cultivation traits
98% cottonseed hull, 2% lime, 60%±2% water content. GB 19172—2003
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