Some standard content:
GB17716—1999
This standard is formulated to identify black carp, protect and preserve the excellent traits and germplasm of black carp original species, avoid germplasm mixing and degradation during seed production, and carry out effective monitoring of black carp germplasm. This standard is mainly the result of the "Eighth Five-Year Plan" scientific and technological research, and absorbs the relevant results of the "Sixth Five-Year Plan" and "Seventh Five-Year Plan" scientific and technological research and predecessors. Appendix A and Appendix B of this standard are the appendices of the standard. Appendix C of this standard is a reminder appendix.
This standard is proposed by the Ministry of Agriculture of the People's Republic of China. This standard is under the jurisdiction of the Freshwater Aquaculture Sub-Technical Committee of the National Technical Committee for Aquatic Standardization. The drafting units of this standard are: Shanghai Fisheries University, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences. The main drafters of this standard are: Li Sifa, Zhao Jinliang, Cai Wanqi, Zhou Biyun, Lv Guoqing, Fan Zhaoting, Xu Zhongfa. This standard shall be implemented from July 1, 1999. 83
1 Scope
National Standard of the People's Republic of China
Black carp
GB17716—1999
This standard gives the main morphological and structural characteristics, growth and reproduction, genetic characteristics, and detection methods of black carp (Mylopharyngodonpiceus).
This standard applies to the identification of black carp germplasm. Name and classificationwwW.bzxz.Net
2.1 Scientific name
Black carp (Mylopharyngodonpiceus). 2.2 Classification position
Belongs to the order Cypriniformes, the family Cyprinidae, the subfamily Leuciscinae, and the genus Mylophuryngodom.
3 Main morphological and structural characteristics
3.1 External morphological characteristics
3.1.1 Appearance
The body is elongated and fusiform. The abdomen is round and has no ventral ridges. The mouth is terminal and the upper jaw is arc-shaped. The snout is pointed and the eyes are lateral. The body is covered with large round scales and the lateral line is complete. The gill rakers are sparse and short. The tail fin is deeply forked, and the upper and lower lobes are equal in length. The body is blue-black, the back is dark black, the abdomen is grayish white, and all fins are grayish black. The external morphology of the black carp is shown in Figure 1.
Figure 1 Appearance of the black carp
Approved by the State Administration of Quality and Technical Supervision on April 5, 1999, 84
Implemented on July 1, 1999
3.1.2 Countable traits
3.1.2.1 Dorsal fin style: D3.7.
3.1.2.2 Anal fin style: A3.7~9.
3.1.2.3 Lateral line scale type: 40
3.1.2.4 Number of gill stripes on the outer side of the first gill arch: 15-21. 3.1.3 Measurable traits
GB 17716—1999
For individuals with a body length of 17.2-53.4 cm and a body weight of 110.0-2250.0 g, the proportion values of the measured traits are shown in Table 1. Table 1 Proportion of measured measurable traits
Total length/body length
Body length/body height
Body length/head length
Head length/snout length
Head length/eye diameter
Head length/distance between eyes
1.159±0.0321.050±0.3504.318±0.2953.318±0.5806.619±0.7982.290±0.3963.2 Internal structural characteristics
Ventricle, posterior chamber is relatively slender.
3.2.2 Hypopharynx
1 row, sun-toothed, tooth formula 4/5.
3.2.3 Vertebrae
Total number of vertebrae: 39~~42.
3.2.4 Peritoneum
Black.
4 Growth and reproduction
4.1 Growth
Body length/tail peduncle length
7.657±0.624
Caudal peduncle length/tail peduncle height
1.059±0.100
The measured values of body length and weight of fish of different age groups are shown in Table 2. The average calculated values of body length and weight of fish of different age groups corresponding to Table 2, growth equation, and body length and weight relationship equation are shown in Appendix C (suggestive appendix). Table 2 Measured values of body length and weight of fish of different age groups Age
Measured body length
Measured weight
Measured body length
Measured weight
3100～
1) Age is mainly identified based on the age number on the scales. 4.2 Reproduction
4.2.1 Sexual maturity age: Female fish 5~7 years old, male fish 4~~6 years old. Sexually mature individuals have their gonads mature once a year and spawn once. 4.2.2
3 Egg carrying capacity: The egg carrying capacity of individuals in different age groups is shown in Table 3.5+
11 100~
12900~
15900~
15900~
22900~
17 400~
20 250~
20250~
Age, age
Weight g
Absolute egg carrying capacity, granules
Relative egg carrying capacity, granules/g
Genetic characteristics
5.1 Cytogenetic characteristics
5.1.1 Chromosome number
Somatic cell chromosome 2-fold, 2n-48
5.1.2 Karyotype
GB 17716 --1999
Egg carrying capacity of individuals in different age groups,
Medium centromere chromosomes (m group) 14 pairs; sub-metacentric chromosomes (sm group) 4 pairs; sub-telocentric chromosomes (st group) 6 pairs, number of chromosome arms (NF) 84 (see Figure 2). x X xX Xx xx r
XA ax n K xu x
其场其北x
x xxnAx 1nr
Figure 2 Chromosome karyotype of black carp
5.2 Biochemical genetic characteristics
5.2.1 Lactate dehydrogenase (LDH) isozymes in muscle 5.2.1.1 The zymogram of lactate dehydrogenase (LDH) isozymes in muscle is shown in Figure 3. 2 The enzyme band scanning diagram of lactate dehydrogenase (LDH) isozymes in muscle is shown in Figure 4. 5. 2. 1. 2
LDH4LDHs
3 Electrophoretic zymogram of LDH isozymes in black carp muscle
Figure 4 Enzyme band scanning diagram of ILDH isozymes in black carp muscle Figure 5. 2. 1. 3
The activity intensity of the lactate dehydrogenase (LDH) isozyme bands in muscle is shown in Table 4. Table 4 Activity intensity of LDH isozyme bands in black carp muscle Enzyme band
Activity intensity
Population variation range: The proportion of polymorphic loci in the black carp population in the middle and lower reaches of the Yangtze River is 5.9%~～11.8%, and the average heterozygosity is 0.001 0~0.035 0.
6 Detection method
6.1 Identification of age
6.1.1 Scale selection site
GB 17716—1999
Between the lower part of the dorsal fin base and the upper part of the lateral line scales on the side of the body. 6.1.2 Operation steps
a) Remove the dorsal scales and store them in a scale bag; b) Take out the scales stored in the scale bag, put them in culture blood, and wash them clean; c) Press the cleaned scales on the fish body and clamp them between two glass slides, label them, and seal the two ends with transparent tape; d) Observe the growth rings of the scales under a microscope or projector to determine the age of the fish. Note: Regenerated scales cannot be used for age identification. 6.1.3 Take a grinded slice (thickness 0.20~～0.25mm) from the base of the first fin ray of the pectoral fin as a control. 6.2 Determination of fecundity
The fecundity of black carp can be estimated by counting the actual number of eggs laid by the broodstock to avoid killing the broodstock. During the breeding season, weigh the weight of the broodstock before and after spawning (select the broodstock that has spawned once) (accurate to 50g), and the difference between the two weighings is the weight of the eggs laid. Weigh 1.0g of eggs with an electronic balance, and count the average number of eggs per gram of eggs under a dissecting microscope. The total number of eggs contained in the ovary is the absolute egg-bearing capacity, and the number of eggs contained per unit body weight (g) is the relative egg-bearing capacity. 6.3 Chromosome detection
6.3.1 Specimen preparation
Use in vivo injection of phytohemagglutinin (PHA), take a drop of kidney cell suspension, air dry and prepare a slide, and stain with Giemsa. The preparation of Giemsa staining solution is shown in Appendix A (Standard Appendix). 6.3.2 Determination of chromosome number
Observe under the oil microscope, count the number of chromosomes in more than 100 clear and well-dispersed metaphases, and determine the chromosome number. 6.3.3 Karyotype analysis
Select some of the best metaphases, take microphotographs, cut and paste the magnified photos, and perform chromosome karyotype analysis. The morphological categories of chromosomes are divided according to the following regulations. That is, arm ratios of 1.0 to 1.7 are mid-centromere chromosomes (group m); 1.7 to 3.0 is the subcentral centromere chromosome (sm group); 3.0~7.0 is the subtelomeric centromere chromosome (st group); 7.0~~cc is the telomeric centromere chromosome (t group). The number of chromosome arms in the m and sm groups is 2; the number of chromosome arms in the st and t groups is 1.6.4 Biochemical genetic analysis
6.4.1 Collection and preservation of samples
First, cut the ventral aorta to bleed, dissect the body alive, take the back white muscle, put it in a numbered small plastic bag, and store it in liquid nitrogen. After transporting it back to the laboratory, store it in a 25C refrigerator.
6.4.2 Sample preparation
Add 3% coenzyme 1 solution to the sample and centrifuge it at low temperature until the supernatant is clear. The entire preparation process is operated at low temperature. 6.4.3 Electrophoresis separation
Use horizontal plate electrophoresis instrument and 4.0% polyacrylamide gel electrophoresis for separation. Before adding the sample, pre-electrophoresis: 50mA, 30min. After adding the sample, pre-electrophoresis: 25mA, 10min; formal electrophoresis: 275V.100min. After the electrophoresis, put it into the isozyme staining solution that has been prepared in advance and kept warm in a 37C constant temperature box for staining. The preparation of the isozyme staining solution is shown in Appendix B (Standard Appendix). 6.4.4 Scanning determination
Scan the electrophoresis pattern with a laser scanner. 87
A1 Preparation of Giemsa staining solution
GB17716 -1999
Appendix A
(Appendix of the standard)
Preparation of Giemsa staining solution
Weigh 0.5g Giemsa powder, measure 33mL glycerol, mix a small amount of glycerol with Giemsa powder in a mortar, grind until there are no particles, then add the remaining glycerol, keep warm at 56°C for 2h, then add 33mL methanol, and store in a brown bottle. A2 Preparation of phosphate buffer (0.2mol/L·pH7.2) Phosphate buffer is prepared by mixing liquid A and liquid B in proportion. A2.1A solution (0.2mol/L, NazHPO.) preparation: Take 36.61g of disodium hydrogen phosphate (NazHPO4·H,O) containing 1 crystal water and dilute it to 1000mL. distilled water, or take 71.64g of disodium hydrogen phosphate (NazHPO·H,O) containing 2 crystal waters and dilute it to 1000mL distilled water.
A2.2B solution (0.2mol/LNaH,PO.) preparation: Take 27.6g of sodium dihydrogen phosphate (NaH,PO4·H,O) containing 1 crystal water and dilute it to 1000mL distilled water, or take 31.21g of sodium dihydrogen phosphate (NaHzPO.·2H,O) containing 2 crystal waters and dilute it to 1.000mL distilled water.
A2.3 Take 720 mL of solution A, add 28.0 mL of solution B, and mix to form the required phosphate buffer. A3 Preparation of Giemsa working staining solution
Take 100 mL of phosphate buffer and add 3 mL of Giemsa staining stock solution. Appendix B
(Standard Appendix)
Preparation of isoenzyme staining solution
B1 Preparation of Tris-HCl staining buffer (1. 5 mol/L) Take 181.71 g of Tris and dissolve it in 900 mL of distilled water, adjust the pH to 9.5 with hydrochloric acid, and then dilute it to 1000 mL with distilled water. B2 Preparation of fluorinated nitro tetrazolium blue (NBT) solution (mg/mL) Take 250 mg of chlorinated nitro tetrazolium blue and dissolve it in 250 mL of distilled water. B3 Preparation of sodium lactate solution (1 mol/L)
Take 203.76 mL of lactic acid, add 700 mL of distilled water, mix, adjust to pH 7.0 with 50% sodium hydroxide solution, and then dilute to 1000 ml with distilled water.
B4 Preparation of isozyme staining solution
Prepare the required isozyme staining solution according to Table B1. 88
Tris-HCl staining buffer
Coenzyme 1
GB 17716-1999
Formula of isozyme staining solution
Phenazine methyl ester sulfate
Appendix C
(Suggested Appendix)
Sodium lactate solution
Calculated values of body length and weight of black carp in different age groups, growth equation, body length and weight relationship formula C1
Calculated values of body length and weight
The average calculated values of body length and weight of fish in different age groups corresponding to Table 2 are shown in Table C1. Table C1
Age, age
Deferred body length
Deferred weight
Deferred body length
Deferred weight
Growth equation
Female fish:
Male fish:
Average deferred values of body length and weight of fish in different age groups2+
L, - 220. 4E1 -- e-0. 078 9(r+0. 298 2)7W, =157 384. 0[1 - e-0. 078 9(++0. 298 2)73L, 198. 1[1 - e-0. 086 2(+0. 343 7)]W.122466.4[1e0.086 2+0.3437# In the formula: L--fish body length at age t, cm; W.-fish body weight at age t, g;
e ---natural logarithm;
t——-fish age.
C3 Body length and weight relationship formula
C3.1 Fish stage
In the formula.·Fish body weight.
L…fish body length, cm.
Edible commercial fish stage
C3.3Broodstock stage
W = 0. 022 2 X L2 953 4
Distilled water
15 123. 1
·(C1）
.c0.rc .cg..( C2 )
·(C3)
+++**+...+*++*.+****o++++....+*.-( C5 )W 0. 026 5 × L2. 895 5
（C6)
Female fish:
Male fish:
17716-1999
W = 0. 017 8 × L2. 965 3
W 0.001 4×L3.5191
..........( C7 )
centercenter
centercenter#center2mol/LNaH,PO.) Preparation: Take 27.6g of sodium dihydrogen phosphate (NaH,PO4·H,O) containing 1 crystal water and dilute it to 1000mL distilled water, or take 31.21g of sodium dihydrogen phosphate (NaHzPO.·2H,O) containing 2 crystal waters and dilute it to 1.000mL distilled water.
A2.3 Take 720mL of solution A, add 28.0ml of solution B, mix to get the required phosphate buffer. A3 Preparation of Giemsa working staining solution
Take 100ml of phosphate buffer and add 3mL of Giemsa staining mother solution. Appendix B
(Standard Appendix)
Preparation of isoenzyme staining solution
B1 Preparation of Tris-HCl staining buffer (1.5 mol/L) Take 181.71 g of Tris and dissolve it in 900 ml of distilled water, adjust the pH to 9.5 with hydrochloric acid, and dilute it to 1000 ml with distilled water. B2 Preparation of fluorinated nitro blue tetrazolium (NBT) solution (mg/mL) Take 250 mg of chlorinated nitro blue tetrazolium and dissolve it in 250 ml of distilled water. B3 Preparation of sodium lactate solution (1 mol/L)
Take 203.76 ml of lactic acid, add 700 ml of distilled water, mix, adjust the pH to 7.0 with 50% sodium hydroxide solution, and dilute it to 1000 ml with distilled water.
B4 Preparation of isozyme staining solution
The required isozyme staining solution is prepared according to Table B1. 88
Tris-HCl staining buffer
Coenzyme 1
GB 17716-1999
Formula of isozyme staining solution
Phenazine methyl ester sulfate
Appendix C
(Suggested Appendix)
Sodium lactate solution
Body length and weight decalcified values of black carp in different age groups, growth equation, body length and weight relationship C1
Body length and weight decalcified values
The average decalcified values of body length and weight of fish in different age groups corresponding to Table 2 are shown in Table C1. Table C1
Age, age
Deferred body length
Deferred weight
Deferred body length
Deferred weight
Growth equation
Female fish:
Male fish:
Average deferred values of body length and weight of fish in different age groups2+
L, - 220. 4E1 -- e-0. 078 9(r+0. 298 2)7W, =157 384. 0[1 - e-0. 078 9(++0. 298 2)73L, 198. 1[1 - e-0. 086 2(+0. 343 7)]W.122466.4[1e0.086 2+0.3437# In the formula: L--fish body length at age t, cm; W.-fish body weight at age t, g;
e ---natural logarithm;
t——-fish age.
C3 Body length and weight relationship formula
C3.1 Fish stage
In the formula.·Fish body weight.
L…fish body length, cm.
Edible commercial fish stage
C3.3Broodstock stage
W = 0. 022 2 X L2 953 4
Distilled water
15 123. 1
·(C1）
.c0.rc .cg..( C2 )
·(C3)
+++**+...+*++*.+****o++++....+*.-( C5 )W 0. 026 5 × L2. 895 5
（C6)
Female fish:
Male fish:
17716-1999
W = 0. 017 8 × L2. 965 3
W 0.001 4×L3.5191
..........( C7 )
centercenter
centercenter#center2mol/LNaH,PO.) Preparation: Take 27.6g of sodium dihydrogen phosphate (NaH,PO4·H,O) containing 1 crystal water and dilute it to 1000mL distilled water, or take 31.21g of sodium dihydrogen phosphate (NaHzPO.·2H,O) containing 2 crystal waters and dilute it to 1.000mL distilled water.
A2.3 Take 720mL of solution A, add 28.0ml of solution B, mix to get the required phosphate buffer. A3 Preparation of Giemsa working staining solution
Take 100ml of phosphate buffer and add 3mL of Giemsa staining mother solution. Appendix B
(Standard Appendix)
Preparation of isoenzyme staining solution
B1 Preparation of Tris-HCl staining buffer (1.5 mol/L) Take 181.71 g of Tris and dissolve it in 900 ml of distilled water, adjust the pH to 9.5 with hydrochloric acid, and dilute it to 1000 ml with distilled water. B2 Preparation of fluorinated nitro blue tetrazolium (NBT) solution (mg/mL) Take 250 mg of chlorinated nitro blue tetrazolium and dissolve it in 250 ml of distilled water. B3 Preparation of sodium lactate solution (1 mol/L)
Take 203.76 ml of lactic acid, add 700 ml of distilled water, mix, adjust the pH to 7.0 with 50% sodium hydroxide solution, and dilute it to 1000 ml with distilled water.
B4 Preparation of isozyme staining solution
The required isozyme staining solution is prepared according to Table B1. 88
Tris-HCl staining buffer
Coenzyme 1
GB 17716-1999
Formula of isozyme staining solution
Phenazine methyl ester sulfate
Appendix C
(Suggested Appendix)
Sodium lactate solution
Body length and weight decalcified values of black carp in different age groups, growth equation, body length and weight relationship C1
Body length and weight decalcified values
The average decalcified values of body length and weight of fish in different age groups corresponding to Table 2 are shown in Table C1. Table C1
Age, age
Deferred body length
Deferred weight
Deferred body length
Deferred weight
Growth equation
Female fish:
Male fish:
Average deferred values of body length and weight of fish in different age groups2+
L, - 220. 4E1 -- e-0. 078 9(r+0. 298 2)7W, =157 384. 0[1 - e-0. 078 9(++0. 298 2)73L, 198. 1[1 - e-0. 086 2(+0. 343 7)]W.122466.4[1e0.086 2+0.3437# In the formula: L--fish body length at age t, cm; W.-fish body weight at age t, g;
e ---natural logarithm;
t——-fish age.
C3 Body length and weight relationship formula
C3.1 Fish stage
In the formula.·Fish body weight.
L…fish body length, cm.
Edible commercial fish stage
C3.3Broodstock stage
W = 0. 022 2 X L2 953 4
Distilled water
15 123. 1
·(C1）
.c0.rc .cg..( C2 )
·(C3)
+++**+...+*++*.+****o++++....+*.-( C5 )W 0. 026 5 × L2. 895 5
（C6)
Female fish:
Male fish:
17716-1999
W = 0. 017 8 × L2. 965 3
W 0.001 4×L3.5191
..........( C7 )
centercenter
centercenter#center
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