CN115656169A - Method for making scomberomorus niphonius chromosome karyotype diagram - Google Patents
Method for making scomberomorus niphonius chromosome karyotype diagram Download PDFInfo
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Abstract
The invention relates to a method for making a scomberomorus niphonius chromosome karyotype diagram, which belongs to the field of fish cytogenetics and comprises the following steps: 1) Obtaining scomberomorus niphonius larval and young fish; 2) Preparing larval and juvenile fish chromosomes; 3) Analyzing chromosome karyotype of scomberomorus niphonius; according to the invention, two hypotonic solutions with hypotonic concentrations of 0.05mol/L KCl and 0.075mol/LKCl are adopted, and the results of the two hypotonic concentrations are comprehensively analyzed to obtain the chromosome karyotype chart, so that the problems of chromosome loss and insufficient chromosome dispersion can be avoided, and the determination of the position of the chromosome centromere during karyotype analysis is facilitated.
Description
Technical Field
The invention belongs to the field of fish cytogenetics, and particularly relates to a method for making a scomberomorus niphonius chromosome karyotype chart.
Background
Chromosomes are carriers of genetic material and determine the species status and the system evolution stage. The preparation of metaphase chromosomes and the karyotype analysis can help to understand the genetic composition, genetic variation and development mechanism of organisms, have important reference values for predicting interspecies hybridization and polyploid breeding results, understanding sex genetic mechanism and the identification of genome number, species origin, evolution and ethnic relation, and have important significance for the development and utilization of resources and genetic breeding. In actual production, chromosome analysis is also an important means for diagnosing various animal genetic diseases and researching character variation, and is a technology which is necessary to master in operation and breeding of chromosome groups. The aquaculture industry also often needs to have knowledge of the chromosomal composition and structure of the species being cultivated. For the research on fish chromosomes, although the development of China is fast in recent years, china mainly focuses on the cypriniformes and the weedicides, most of the fishes are still fresh water fishes, and the chromosome composition of many important marine economic fishes is unknown. The fish chromosomes have the characteristics of large number, small form and low division index, and a large number of metaphase division phases are difficult to prepare generally, so that a great deal of inconvenience is brought to fish chromosome research, and the difficulty in fish chromosome research is increased.
In 1966, ojima et al (Ojima Y S, hitotsumachi S, makino. Cytogenic students in lower version brates. Proc Jap Acad,1966,42 (1): 62-66) successfully slice by air drying for the first time, the existing chromosome slice basically adopts the method, but the unmodified preparation method is difficult to prepare the slice with clear chromosome morphology and better dispersion, and cannot meet the requirement of chromosome karyotype analysis. To perform accurate cytogenetic analysis, chromosome specimen preparation and karyotype analysis are two important links.
Spanish mackerel is irritable in nature, and dies after hanging the net for a short time and easily dies after leaving water, so that it is extremely difficult to obtain multi-tail living sexually mature parent fish. The water temperature and bait fish are two main factors influencing the survival of mackerel, when the seawater temperature is 16-25 ℃, mackerel chases the fish group to carry out feeding activity, and when the bait fish is insufficient, the larval and young fishes at the 12 th day after fertilization can be mutually killed. The object for chromosome preparation needs to be an individual with good activity except for cultured cells so as to obtain enough dividing cells.
Therefore, the preparation of the mackerel chromosomes needs to fully consider the biological characteristics of the mackerel chromosomes, and the material taking and the early-stage treatment can only be adjusted according to the actually obtained states of the mackerel to ensure that the mackerel tissue cells still keep certain vitality intensity in the colchicine treatment process, so that enough metaphase chromosomes can be prepared. Karyotyping may also be adjusted accordingly.
The hypotonic step in the chromosome preparation process is to make the histiocyte fully imbibe, and the cell membrane is rapidly broken to release the chromosome or cell nucleus through the double actions of beating and high temperature during the dropping process. The conventional chromosome preparation hypotonic solution is a KCl solution with the concentration of 0.075mol/L, but the osmotic pressures of tissue cells of different species and even different individuals are not completely consistent, the hypotonic solution with one concentration is generally adopted in the prior art, the chromosome preparation effect is good and bad, and sometimes even metaphase chromosomes cannot be prepared. Researchers also consider adjusting the concentration of hypotonic solution to prepare metaphase chromosomes, but the obtained results are not ideal. For example, when sweetfish chromosomes are prepared, hypotonic with 0.0375mol/L KC1 solution for 45-60 min at room temperature, the statistical number of diploid chromosomes is 2n =42 (Zhang Chundan, zhang Xiaobo, liming Yun. Sweetfish chromosome karyotype analysis, water conservancy fishery, 2005,25 (4): 16-17, 20), which is far from the actual diploid chromosome mode of the species of 2n = 56.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for making a scomberomorus niphonius chromosome karyotype chart, and the method steps suitable for scomberomorus niphonius chromosome preparation and karyotype analysis are designed by improving the existing marine animal chromosome preparation method, so that a large amount of chromosomes with better dispersion in metaphase are prepared by the most economical, most convenient and accurate method, and the karyotype is quickly analyzed. The method can quickly prepare a large amount of mackerel metaphase chromosomes and prepare an accurate karyotype chart.
The invention is realized by the following technical scheme:
a method for making a scomberomorus niphonius chromosome karyotype chart, which comprises the following steps: 1) Obtaining scomberomorus niphonius larval and young fish; 2) Preparing larval and juvenile fish chromosomes; 3) Analyzing chromosome karyotype of scomberomorus niphonius;
the preparation of the larvae chromosome: fishing out scomberomorus niphonius larvae, putting the scomberomorus niphonius larvae into 0.04% (g/ml) colchicine solution, and standing at room temperature for 30min;
after the sample of the larval and juvenile fish is treated by colchicine, discarding the colchicine solution, dividing the sample into two parts, respectively putting the two parts into hypotonic solutions with the concentrations of 0.05mol/L KCl and 0.075mol/L KCl, and carrying out hypotonic treatment for 45-60 min;
after the hypotonic reaction of the sample is finished, removing the hypotonic solution, adding a fixing solution, and fixing for 15min at 4 ℃; the fixed liquid amount is such that the sample needs to be completely immersed in the fixed liquid; then, removing the fixing solution, then replacing a new precooled fixing solution, fixing for 15min again, carrying out sample fixing at 4 ℃, repeating the fixing step for 3-4 times, and fully fixing the sample; taking 1 sample of the young fish, putting the sample into a precooled sample tube, adding 0.5-1.5 ml of precooled dissociation liquid into the sample tube, and dissociating at 4 ℃;
in the dissociation process, the dissociation of the sample histiocyte can be effectively accelerated by shaking or vibrating the wall of the sample tube or blowing and beating the dissociation liquid by a disposable dropper; preparing slices by dissociation liquid air drying method (also called hot drop slice method);
the locus mackerel chromosome karyotype analysis comprises the following steps: obtaining chromosome mode from chromosome picture prepared from 0.075mol/L KCl hypotonic solution, after determining the chromosome mode, selecting the picture obtained from the hypotonic solution with concentration of 0.05mol/LKCl that the chromosome mode is correct, and the form contour boundary is clear, and the dispersed picture is used for karyotyping.
Further, the 0.04% (g/ml) colchicine solution contains half volume of sterile filtered seawater.
Further, when scomberomorus niphonius larvae were treated with 0.04% (g/ml) colchicine solution, the container lid was opened.
Further, during hypotonic conditions, the top and bottom are inverted several times to ensure adequate hypotonic conditions.
Further, the fixing solution is prepared by mixing absolute ethyl alcohol and glacial acetic acid in a volume ratio of 3:1; the dissociation liquid is a mixture of glacial acetic acid and distilled water in a volume ratio of 1:1.
further, the method for analyzing the karyotype chart comprises the following steps:
(1) Selecting 10-20 well-dispersed metaphase mitotic phases of chromosomes for amplified printing, randomly numbering the 48 chromosomes on each mitotic phase for 1-48, marking the filament-depositing point and the tail ends of the long arm and the short arm of each chromosome by using a conspicuous pen, measuring the lengths of the long arm and the short arm by using a ruler, reading the two bits (0.00 cm) after the decimal point, and sequentially recording the measured values of the 1-48 short arm and the long arm;
(2) Inputting measured values into an Excel table, inputting 1-5 groups of data into each individual, inputting a short arm and a long arm of each group of data in sequence in a transverse direction, inputting numbers of 1-48 in sequence in a longitudinal direction, and spacing at least one row between each group of data; then, calculating the chromosome length, the arm ratio and the relative length of each chromosome by using the Excel table formula corresponding to transverse data, then determining the chromosome type according to the arm ratio, sequencing all the chromosomes from large to small according to the relative length, and selecting and expanding the selected area according to the sequencing;
(3) Checking whether the types of each group of chromosomes are consistent, based on the most consistent types, if the deviation of certain group of data is too large, discarding the data, and if the deviation of certain group of data of individual chromosomes is too large, checking whether the measured data is input wrongly or measured wrongly or the filament-forming point is positioned inaccurately according to the serial number of the individual chromosome data of the certain group of data to perform fine adjustment or modification;
(4) Carrying out sequencing fine adjustment on each group of data according to types, carrying out statistics on the average value +/-standard error of the relative length and arm ratio one by one from large to small on each group of data as the scomberomorus niphonius 48 chromosomes belong to diploids, and finally determining that the karyotype of the scomberomorus niphonius is 2n =48=2sm +4st +42t;
(5) Selecting a metaphase mitosis with good chromosome morphology and good dispersion, copying a picture of the metaphase mitosis by using Photoshop, and newly building a canvas with the same size as the picture; then, sequentially copying and pasting 1-48 chromosomes on the A4 paper to a newly-built canvas, clicking the change in a pull-down menu of an editing button, selecting a rotating button, and rotating the copied chromosomes to a state that the short arm is in the lower state and the upper long arm is in the lower state; erasing the part outside the chromosome contour by using a rubber tool, and only keeping the chromosome shape; and then sequencing from m to sm to st to t according to the relative length actual value and the arm ratio value (form) in an Excel table, and finishing the chromosome karyotype picture.
Compared with the prior art, the invention has the beneficial effects that:
1. the method for preparing the mackerel metaphase chromosome and analyzing the karyotype is simple and rapid, the prepared chromosome slices are clean and clear, observation and counting are convenient, 3 drops of cell rings are arranged in each slice, observation of one chromosome slice can be completed by one week of rotation, and metaphase chromosomes with good dispersion and good shapes in a sample cannot be missed.
2. Because the tissue cell division index of the larval and juvenile fish is high, the invention adopts two hypotonic solutions with hypotonic concentrations of 0.05mol/L KCl and 0.075mol/L KCl respectively, and both hypotonic concentration treatments can prepare a large amount of metaphase chromosomes, but the metaphase chromosomes prepared by the 0.05mol/L KCl hypotonic solution have good shapes and good dispersion, but the chromosomes are seriously lost and are not suitable for chromosome mode statistics; the chromosome morphological contour boundary prepared by 0.075mol/L KCl hypotonic solution is not clear and dispersed insufficiently, chromosome superposition and cross linking are easy to occur, the chromosome mode can be counted, but the karyotype chart analysis is not suitable, so the chromosome karyotype chart is obtained by combining the results of 0.05mol/L hypotonic concentration and 0.075mol/L hypotonic concentration, the problems of chromosome loss and insufficient chromosome dispersion can be avoided, and the determination of the chromosome centromere position in the karyotype analysis is facilitated.
3. The karyotype is analyzed by adopting the method, the number of individuals and the number of split phases can meet the requirements, the manual calculation and measurement errors can be reduced to the greatest extent, the karyotype formula is more accurate and visual than the traditional shearing and pasting measurement method, the corresponding relation of chromosome numerical values is not changed all the time, and the karyotype is convenient for analysts to search and verify in the future.
Drawings
FIG. 1 shows the morphology, amount and dispersion of metaphase chromosomes under 0.05mol/L KCl hypotonic condition;
FIG. 2 shows the metaphase chromosome morphology, number and dispersion under 0.075mol/L KCl hypotonic condition, wherein the chromosome number of all the pictures is about 48;
FIG. 3 Cnidocamphetamine metaphase chromosomes;
FIG. 4 Spanish mackerel karyotype.
Detailed Description
The invention is described in detail below with reference to specific examples and drawings, which are specific operational procedures of preferred embodiments, and the scope of the invention is not limited in any way by the examples.
Example 1:
in 2022, 13 months, ningbo city oceanic and fishery research institute starts artificial insemination to cultivate mackerel fry, hatching out in 15 months in 4 months, opening in 18 months in 4 months, and feeding large yellow croaker initial hatching fry, mullet initial hatching fry and the like. And (4) daily managing the young fishes and the bait young fishes, changing water by 100 percent every day, and culturing the young fishes to 4 months and 24 days, wherein the length of the scomberomorus niphonius fry is 1-2 cm. Because scomberomorus niphonius is impatient in sexual condition and strong in activity, the scomberomorus niphonius can eat the bait fish fry, so that the scomberomorus niphonius can be prevented from killing each other, the number of the fish fry in unit water is rare, and sufficient bait fish fry can be provided. A chromosome preparation device and various reagent consumables are prepared properly in advance, then, rich fish fries are fished out through bolting silk, the thin end of a disposable suction pipe is cut off through scissors, after bait fish fries are removed, 12 mackerel fries are stirred to enter colchicine action, KCl hypotonicity with different gradients, 3-4 times of Carnoy's solution fixation and slice making and dyeing through a hot drop slice method, metaphase chromosome of scomberomorus niphonius is successfully prepared, and the chromosome karyotype of the mackerel is analyzed.
The specific operations for completing the above operation process are as follows:
the scomberomorus niphonius chromosome preparation and karyotype analysis method comprises the following steps: 1) Temporarily breeding scomberomorus niphonius larvae and preparing reagent consumables; 2) Preparing and observing larvae chromosomes; 3) Analyzing chromosome karyotype of scomberomorus niphonius;
before scomberomorus niphonius fry is fished, 0.04% colchicine working solution (50% seawater containing 0.04% colchicine) is prepared in a 10ml centrifugal tube, and the specific operation is carried out by taking the scale on the tube wall of the centrifugal tube as reference: 0.5ml of 0.4 percent colchicine mother liquor (mass volume ratio) is dropped into the mixture, and then 4.5ml of ultrapure water and 5ml of corresponding sea area are added for sterilization and filtration to make the total volume of the seawater be 10ml. Spanish mackerel fry is picked and put into 0.04 percent colchicine working solution for treatment for 30 to 35 minutes. Diluting the prepared KCl hypotonic solution with 0.075mol/L with ultrapure water to a 1 tube (9 ml) with the concentration of 0.05mol/L during the colchicine action period to form 2 gradients, still taking the wall scale of a 10ml centrifuge tube as the standard, and filling 5-10ml of 0.075mol/L KCl in the 1 tube as a normal-concentration hypotonic solution; 6ml of 0.075mol/L KCl and 3ml of ultrapure water are respectively filled into 1 tube to prepare 0.05mol/L KCl solution as low-concentration hypotonic solution. Picking out the colchicine treated young fishes by a disposable dropper, soaking the young fishes in 2 tubes of hypotonic solution for 50 minutes in two equal parts, determining the optimal hypotonic concentration, and slightly reversing the centrifugal tube up and down for several times in the hypotonic process to ensure that the tissue cells are fully hypotonic. After the start of hypotonic reaction, 100ml of Carnoy fixative (ethanolic alcohol: glacial acetic acid = 3). After the hypotonic reaction is finished, the hypotonic solution is sucked out to a waste liquid bottle by a disposable suction tube. Fully fixing with freshly prepared precooled fixing liquid, wherein the fixing liquid amount is 2-5 ml, completely immersing the tissue sample into the fixing liquid, replacing the fixing liquid once after fixing for 15min, pouring the waste fixing liquid into a waste liquid bottle, and repeating the fixing process for 4 times in a refrigerator at 4 ℃. After the sample is fixed for 1 hour, a dissociation solution (10ml) is prepared according to the proportion of glacial acetic acid to ultrapure water =1, and a sample of the intact larval and juvenile fish with 1 tail is placed into 1 centrifugal tube with 1.5ml, the sample is dissociated by using the dissociation solution with about 0.5ml, in order to accelerate the dissociation of the sample, the tube wall is flicked by using fingers or the centrifugal tube is inverted for several times, and then the dissociated sample is placed in a refrigerator with the temperature of 4 ℃ for precooling.
The slide is prepared by a hot drop method, and 3 drops of dissociation liquid are dropped on each glass slide at intervals of 2cm. The filter paper sheet aspirates off excess dissociation liquid. After the dissociation liquid on the hot drop is completely dried, marking the sample group and the slide making time at the ground glass position of the slide by using a pencil. 12 fish samples were dropped with 2 slices each, and 24 slices total. After dyeing with 10% Giemsa dye liquor for 30min, washing off residual dye liquor on the glass slide by running water trickles for 5-10 s, inclining the sample surface at an angle of 45-85 degrees of the glass slide to the corner edge upwards, and filling toilet paper on the bottom for absorbing water to ensure that the sample surface is completely dried and no water stain impression is left. The giemsa-stained sections were placed under a microscope for microscopic examination and photographed. The samples treated by 0.05mol/L KCl hypotonic solution have a plurality of metaphase, and have good chromosome morphology and dispersity, but a plurality of metaphase have chromosome loss, and the chart is shown in figure 1. There were also many metaphase phases in the 0.075mol/L KCl hypotonic solution treated samples, good chromosome morphology, less sharp profile and less than good dispersion, see FIG. 2, but approximately 48 chromosomes. The results of the two hypotonic solutions were combined and analyzed to determine 2n =48 pieces of scomberomorus niphonius chromosome mode from the sample treated with 0.075mol/L KCl hypotonic solution, and then a picture with good chromosome morphology, uniform dispersion, obvious centromere position and correct mode was obtained from the picture of the sample treated with 0.05mol/L KCl hypotonic solution, as shown in FIG. 3.
Selecting 20 chromosome split phases with clear and well dispersed chromosome forms, amplifying and printing the split phases on A4 paper, randomly numbering the 48 chromosomes on each split phase for 1-48, marking the centromere of each chromosome and the tail ends of the long arm and the short arm by using a visible pen, measuring the lengths of the long arm and the short arm by using a ruler, and sequentially recording the measured values of the 1-48 short arm and the long arm.
Inputting measured values into an Excel table, calculating the chromosome length, arm ratio and relative length of each chromosome by adopting an Excel table formula, inputting 1-5 groups of data into each individual, inputting each group of data into a short arm and a long arm in sequence in a transverse direction, inputting 1-48 serial numbers in sequence in a longitudinal direction, and spacing at least one row between each group of data; determining chromosome types according to the arm ratio, sequencing all chromosomes from large to small according to the relative lengths, and selecting and expanding the selected region according to the sequencing;
checking whether the types of each group of chromosomes are consistent, based on the most consistent types, if the deviation of a certain group of data is too large, discarding the data, and if the deviation of the individual chromosome data of the certain group of data is larger, checking whether the measured data is input wrongly or measured wrongly or the filament-forming point is positioned inaccurately according to the serial number of the individual chromosome data to carry out fine adjustment or modification.
And (3) carrying out sequencing fine adjustment on each group of data according to types, wherein as the scomberomorus niphonius 48 chromosomes belong to a diploid, carrying out statistics on the average value +/-standard error of the relative length and arm ratio of each group of data from large to small one by one, and determining that the scomberomorus niphonius chromosome types are classified into 3 types: 1 to sm, 2 to st, 21 to t. Finally, determining the chromosome karyotype of mackerel as 2n =48=2sm +4st +42t.
Finally, selecting a metaphase chromosome with good form and good dispersion, as shown in figure 3, copying the metaphase chromosome by Photoshop, and newly building a canvas with the same size as the picture; then, sequentially copying and pasting 1-48 chromosomes on the A4 paper to a newly-built canvas, clicking the change in a pull-down menu of an editing button, selecting a rotating button, and rotating the copied chromosomes to a state that the short arm is in the lower state and the upper long arm is in the lower state; erasing the part outside the chromosome contour by using a rubber tool, and only keeping the chromosome shape; then sorting the chromosome karyotype image from m to sm to st to t from small to large in an Excel table according to the relative length actual value and the arm ratio (form), and finishing the chromosome karyotype image, which is shown in FIG. 4.
Claims (6)
1. The method for making the scomberomorus niphonius chromosome karyotype chart is characterized by comprising the following steps of: 1) Obtaining scomberomorus niphonius larval fish; 2) Preparing a larval fish chromosome; 3) Analyzing chromosome karyotype of scomberomorus niphonius;
the preparation of the larvae chromosome: fishing out scomberomorus niphonius larvae, putting the scomberomorus niphonius larvae into 0.04% (g/ml) colchicine solution, and standing at room temperature for 30min;
after the sample of the larval and juvenile fish is treated by colchicine, discarding the colchicine solution, dividing the sample into two parts, respectively putting the two parts into hypotonic solutions with the concentrations of 0.05mol/L KCl and 0.075mol/L KCl, and carrying out hypotonic treatment for 45-60 min;
after the hypotonic reaction of the sample is finished, removing the hypotonic solution, adding a fixing solution, and fixing for 15min at 4 ℃; the fixed liquid amount is such that the sample needs to be completely immersed in the fixed liquid; then, removing the fixing solution, then replacing a new precooled fixing solution, fixing for 15min again, carrying out sample fixing at 4 ℃, repeating the fixing step for 3-4 times, and fully fixing the sample; taking 1 sample of the young fish, putting the sample into a pre-cooled sample tube, adding 0.5-1.5 ml of pre-cooled dissociation liquid into the sample tube, and then dissociating at 4 ℃;
in the dissociation process, the dissociation of the sample histiocyte can be effectively accelerated by shaking or vibrating the wall of the sample tube or blowing and beating the dissociation liquid by a disposable dropper; preparing tablets by a dissociation liquid air drying method;
the locus mackerel chromosome karyotype analysis comprises the following steps: obtaining chromosome mode from chromosome picture prepared from 0.075mol/L KCl hypotonic solution, after determining the chromosome mode, selecting the picture obtained from the KCl with concentration of 0.05mol/L hypotonic solution, wherein the chromosome mode is correct, the shape contour boundary is clear, and the dispersed picture is used for karyotyping.
2. The method for preparing a scomberomorus niphonius chromosome karyotype chart according to claim 1, wherein the 0.04% (g/ml) colchicine solution contains half volume of sterile filtered seawater.
3. The method for preparing a scomberomorus niphonius chromosome karyotype chart according to claim 1, wherein a container lid is opened when scomberomorus niphonius larvae are treated with a 0.04% (g/ml) colchicine solution.
4. The method for preparing a scomberomorus niphonius chromosome karyotype chart according to claim 1, wherein during hypotonic conditions, the upper and lower sides are inverted several times to ensure sufficient hypotonic conditions.
5. The method for preparing a scomberomorus niphonius chromosome karyotype chart according to claim 1, wherein the fixing solution is a mixture of absolute ethyl alcohol and glacial acetic acid in a volume ratio of 3:1; the dissociation liquid is a mixture of glacial acetic acid and distilled water in a volume ratio of 1:1.
6. the method for preparing a scomberomorus niphonius chromosome karyotype chart according to claim 1, comprising the steps of:
(1) Selecting 10-20 metaphase mitotic phases with well dispersed chromosome forms for amplified printing, randomly numbering the 48 chromosomes on each mitotic phase for 1-48, marking the centromere of each chromosome and the tail ends of the long arm and the short arm by using a visible pen, measuring the lengths of the long arm and the short arm by using a ruler, and sequentially recording the measured values of the 1-48 short arms and the long arms;
(2) Inputting measured values into an Excel table, wherein 1-5 groups of data can be input into each individual, each group of data is input with a short arm and a long arm in a transverse direction, 1-48 serial numbers are input in a longitudinal direction in a longitudinal sequence, and at least one row of data is arranged between each group of data; then calculating the chromosome length, the arm ratio and the relative length of each chromosome by using Excel table formula corresponding transverse data, then determining the chromosome type according to the arm ratio, sequencing all the chromosomes from large to small according to the relative length, and selecting and expanding the selected region according to the sequencing;
(3) Checking whether the types of each group of chromosomes are consistent, based on the most consistent types, if the deviation of a certain group of data is too large, discarding the data, and if the individual chromosome data of the certain group of data is in large access, checking whether the measured value is input wrongly or measured wrongly or the filament-forming point is positioned inaccurately according to the serial number of the individual chromosome data to carry out fine adjustment or modification;
(4) Carrying out fine adjustment on the sequencing of each group of data according to the type, carrying out statistics on the average value +/-standard error of the relative length and arm ratio of each group of data from large to small one by one because 48 blue mackerel chromosomes belong to a diploid, and finally determining that the karyotype of the blue mackerel chromosomes is 2n =48=2sm +4st +42t;
(5) Selecting a metaphase mitogen with good chromosome morphology and good dispersion, copying a picture of the metaphase mitogen by using Photoshop, and newly building a canvas with the same size as the picture; then, sequentially copying and pasting 1-48 chromosomes on A4 paper to a newly-built canvas, clicking the change in a pull-down menu of an editing button, selecting a rotating button, and rotating the copied chromosomes to a state that the short arm is in the lower state and the long arm is in the lower state; erasing the part outside the chromosome contour by using a rubber tool, and only keeping the chromosome shape; and then sequencing the sequences from m to sm to st to t from small to large in an Excel table according to the actual value of the relative length and the arm ratio value, and finishing the chromosome karyotype picture.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106289907A (en) * | 2016-08-11 | 2017-01-04 | 中国水产科学研究院黄海水产研究所 | Quickly prepare small-scale marine fishes or the method for larva and juvenile division phases chromosome |
| CN109883797A (en) * | 2019-04-01 | 2019-06-14 | 中国水产科学研究院黄海水产研究所 | A kind of method of easy long-tail anchovy chromosome preparation |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106289907A (en) * | 2016-08-11 | 2017-01-04 | 中国水产科学研究院黄海水产研究所 | Quickly prepare small-scale marine fishes or the method for larva and juvenile division phases chromosome |
| CN109883797A (en) * | 2019-04-01 | 2019-06-14 | 中国水产科学研究院黄海水产研究所 | A kind of method of easy long-tail anchovy chromosome preparation |
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| 吴成宾等: "鳊鲂鱼类的染色体核型及 DNA 含量分析", 上海海洋大学学报, vol. 24, no. 16, pages 801 - 808 * |
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