CN117958197A - Rapid discrimination method for mature parent of procambarus clarkia - Google Patents
Rapid discrimination method for mature parent of procambarus clarkia Download PDFInfo
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- CN117958197A CN117958197A CN202410306839.1A CN202410306839A CN117958197A CN 117958197 A CN117958197 A CN 117958197A CN 202410306839 A CN202410306839 A CN 202410306839A CN 117958197 A CN117958197 A CN 117958197A
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- 241000238030 Procambarus clarkii Species 0.000 title claims abstract description 176
- 238000012850 discrimination method Methods 0.000 title claims description 6
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- 230000018109 developmental process Effects 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 26
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention relates to the technical field of modern biological breeding, in particular to a rapid distinguishing method of a mature parent of procambarus clarkia, which comprises the following steps: judging the sex of procambarus clarkia; if the procambarus clarkia is female procambarus clarkia, collecting the area of the tail limbs of the procambarus clarkia and the area of the white plaque on the surfaces of the tail limbs, calculating the coverage of the white plaque, and judging the ovarian development stage of the procambarus clarkia based on the coverage of the white plaque; and if the procambarus clarkia is male, collecting morphological characteristics of a first pair of abdominal limbs and morphological characteristics of a third pair of foot seat sections and a fourth pair of foot seat sections of the procambarus clarkia, and judging the development stage of the testis of the procambarus clarkia based on the morphological characteristics of the first pair of abdominal limbs and the morphological characteristics of the third pair of foot seat sections and the fourth pair of foot seat sections. The method can provide a direct basis for rapid and accurate selection of mature parent of the procambarus clarkia, effectively improve the production efficiency of intensive breeding of procambarus clarkia fries, reduce the production cost and promote sustainable development of procambarus clarkia industry.
Description
Technical Field
The invention relates to the technical field of modern biological breeding, in particular to a rapid distinguishing method of a mature parent of procambarus clarkia.
Background
Procambarus clarkia, also known as crayfish, belongs to the family of crawfish of the order Cryptotaenia of the class Cryptotaenia. In recent years, the procambarus clarkia industry has rapidly developed, the cultivation area of procambarus clarkia is 2800 mu in 2022 years, the yield is 289.07 ten thousand tons, the comprehensive yield value of the industry is 4580 hundred million yuan, however, the shortage of excellent seedlings of procambarus clarkia severely restricts the high-quality development of the industry. At present, the demand of the excellent offspring seeds of procambarus clarkii is over 100 hundred million in Hubei province. To solve this problem, some farms have attempted to mixedly breed a large amount of procambarus clarkii (including female and male shrimps) in mating ponds (ponds, paddy fields, etc.) during the breeding season; after the female shrimp is in eggs, the eggs are introduced into a plurality of spawning ponds (easy-to-manage cultivation places such as cement ponds) for separate feeding; and collecting young shrimps 2 months after spawning by net scattering or net fishing. However, in the existing procambarus clarkia offspring breeding technology mode, due to the fact that gonad development degrees of female procambarus clarkia and male procambarus clarkia are asynchronous, after amphiprotic mating, some female procambarus clarkia can often reach final maturity and spawning after a longer time. The asynchronous gonad development of the procambarus clarkia causes the serious problems of long parent cultivation time, high parent mortality, great variation of offspring seed size, high intraspecies competition and low offspring seed yield, and seriously affects the intensive breeding production efficiency of the procambarus clarkia offspring seeds. And the mature female individuals and the mature male individuals of the procambarus clarkia are selected for mating, so that the aims of mating and spawning in a short period of parents and improving the production efficiency can be fulfilled.
In aquatic animals, farmers typically select mature individuals by changing body types. For example, most female fishes have the phenomenon of swelling and reddish reproduction holes in the abdomen during reproduction, and the abdomen of the female fishes is lightly pressed to have oosperms oozed out of the reproduction holes; the male fish has smaller abdomen than female fish in reproduction period, and sperm oozes from the reproduction hole by slightly pressing the abdomen. However, procambarus clarkia has a hard and opaque chitin shell, whose eggs and sperm cannot be obtained by non-invasive means, so that accurate identification of mature procambarus clarkia individuals in production is challenging. In the past, the mature parent selection of procambarus clarkii mainly depends on the individual size and body surface color for experience judgment, and the overall judgment accuracy is low due to lack of visual and quantitative judgment characteristics. Therefore, visual and quantifiable external morphological characteristics capable of representing the gonad development degree of the procambarus clarkia are required to be found, so that the rapid and accurate discrimination of the mature parent of the procambarus clarkia is realized.
Disclosure of Invention
The invention aims to solve the problem of asynchronous parent gonad development in the intensive breeding process of procambarus clarkia fries, provides a rapid distinguishing method of procambarus clarkia mature parents, provides a direct basis for rapid and accurate selection of procambarus clarkia mature parents, improves the production efficiency of the intensive breeding of procambarus clarkia fries, reduces the production cost, and promotes the sustainable development of procambarus clarkia industry.
In order to achieve the above object, the present invention provides the following solutions:
a method for rapidly distinguishing mature parent of procambarus clarkia comprises the following steps:
judging the sex of procambarus clarkia;
If the procambarus clarkia is female procambarus clarkia, collecting the area of the tail limbs of the procambarus clarkia and the area of the white plaque on the surfaces of the tail limbs, calculating the coverage of the white plaque, and judging the ovarian development stage of the procambarus clarkia based on the coverage of the white plaque;
And if the procambarus clarkia is male procambarus clarkia, collecting morphological characteristics of a first pair of abdominal limbs and morphological characteristics of a third pair and a fourth pair of foot seat sections of the procambarus clarkia, and judging the development stage of the testis of the procambarus clarkia based on the morphological characteristics of the first pair of abdominal limbs and the morphological characteristics of the third pair and the fourth pair of foot seat sections.
Optionally, collecting the tail limb area and the white plaque area of the tail limb surface of the female procambarus clarkia comprises:
Checking whether the cleaned female procambarus clarkia is enriched with white plaques, and if the female procambarus clarkia is not enriched with white plaques, directly discarding the judgment; and if the white plaque is enriched, taking a tail limb photo and a white plaque photo of the female procambarus clarkii, and determining the tail limb area and the white plaque area of the tail limb surface of the female procambarus clarkii based on the tail limb photo and the white plaque photo.
Optionally, the method for calculating the white plaque coverage is as follows: white plaque coverage (%) = white plaque area/caudal area x 100%.
Optionally, the ovarian development stage of the female procambarus clarkia comprises: stage 1-egg primordial stage, stage 2-immature stage, stage 3-pre-vitelline formation stage, stage 4-vitelline formation stage, stage 5-mid vitelline formation stage, stage 6-post vitelline formation stage, and stage 7-post partum development stage.
Optionally, determining the ovarian developmental stage of the female procambarus clarkia based on the white plaque coverage comprises:
Judging the ovarian development stage of the female procambarus clarkii according to the white plaque coverage by referring to a preset white plaque change characteristic plate, and when the ovarian development stage is in the later stage of the 6-phase yolk formation stage, the ovary of the female procambarus clarkii is mature; the white plaque change characteristic plate is constructed according to white plaque coverage of tail limbs of female procambarus clarkia at different ovarian development stages.
Optionally, the white plaque coverage of the tail limbs of female procambarus clarkia at the different ovarian development stages comprises:
the tail limbs of the female procambarus clarkia with the ovarian development stage of 1-3 stages have no white plaque;
white plaque exists on the tail limbs of female procambarus clarkii with the ovarian development stage of 4, and the coverage of the white plaque is less than 30%;
The tail limbs of the female procambarus clarkia with the ovarian development stage of 5 stages have white plaques, and the coverage of the white plaques is more than 30% and less than 45%;
white plaque exists on the tail limbs of female procambarus clarkii with the ovarian development stage of 6 phases, and the coverage of the white plaque is more than 45%;
the white plaque of the tail limbs of female procambarus clarkii with the ovarian development stage of 7 stages disappears.
Optionally, the testis development stage of the male procambarus clarkia comprises: stage 1-spermatogenic stage, stage 2-primary spermatocyte stage, stage 3-secondary spermatocyte stage, stage 4-spermatocyte stage, and stage 5-spermatocyte stage.
Optionally, determining the testis development stage of the male procambarus clarkii based on the morphological features of the first pair of abdominal limbs and the morphological features of the third and fourth pair of foot-seat nodes comprises:
Observing morphological characteristics of the first pair of abdominal limbs and morphological characteristics of the third pair and the fourth pair of foot seat joints of the male procambarus clarkii, wherein when the first pair of abdominal limbs of the male procambarus clarkii have head processes and tail processes with preset sizes and the third pair and the fourth pair of foot seat joints have barbs with preset sizes, the testis development stage of the male procambarus clarkii is in the 5-stage sperm stage, and the testis of the male procambarus clarkii is mature.
The beneficial effects of the invention are as follows:
The rapid distinguishing method for the mature parents of the procambarus clarkii can solve the problem of asynchronous maturation time of the procambarus clarkii parents, provide key information for parent selection strategies of the procambarus clarkii, accurately select mating of mature female individuals and mature male individuals of the procambarus clarkii, achieve the goal of mating and spawning in a short period of parents and improving production efficiency, and promote sustainable development of procambarus clarkii aquaculture.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of a method for rapidly distinguishing mature parent of procambarus clarkia in an embodiment of the invention;
FIG. 2 is a thermal graph of Kendall correlation between morphological parameters and ovarian developmental stage of an embodiment of the invention;
FIG. 3 is a diagram showing the ranking of importance of morphological parameters of procambarus clarkia using a random forest algorithm (RF) in accordance with an embodiment of the present invention;
FIG. 4 is a graph showing the relationship between white plaque coverage (WC) and stage of ovarian development in accordance with an embodiment of the present invention;
FIG. 5 is a decision tree for predicting ovarian developmental stage using white plaque coverage (WC) in accordance with an embodiment of the invention;
FIG. 6 is a schematic diagram of a white plaque change feature map according to an embodiment of the present invention;
FIG. 7 is a schematic representation of a first pair and a second pair of abdominal limbs of a mature male procambarus clarkii of an embodiment of the present invention;
FIG. 8 is a schematic representation of the third and fourth pairs of feet of the mature male procambarus clarkia of an embodiment of the present invention;
wherein, the head is 1-head, the tail is 2-tail, and the barb is 3-.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In order that the above-recited objects, features and advantages of the present invention will become more readily apparent, a more particular description of the invention will be rendered by reference to the appended drawings and appended detailed description.
The embodiment provides a method for rapidly distinguishing mature parent of procambarus clarkii, as shown in figure 1, comprising the following steps:
judging the sex of procambarus clarkia;
if the procambarus clarkia is female procambarus clarkia, collecting the area of the tail limbs of the procambarus clarkia and the area of the white plaque on the surfaces of the tail limbs, calculating the coverage of the white plaque, and judging the ovarian development stage of the procambarus clarkia based on the coverage of the white plaque;
and if the procambarus clarkia is male, collecting morphological characteristics of a first pair of abdominal limbs and morphological characteristics of a third pair of foot seat sections and a fourth pair of foot seat sections of the procambarus clarkia, and judging the development stage of the testis of the procambarus clarkia based on the morphological characteristics of the first pair of abdominal limbs and the morphological characteristics of the third pair of foot seat sections and the fourth pair of foot seat sections.
Specifically, the discrimination of mature female procambarus clarkii comprises the following steps:
s1.1, cleaning tail sector areas of procambarus clarkia;
s1.2, checking whether tail limbs of procambarus clarkia are enriched with white plaques or not;
S1.3, measuring the area of the tail limb of the female procambarus clarkia;
s1.4, measuring the area of a white plaque on the tail limb surface of the female procambarus clarkia;
s1.5, calculating white plaque coverage, and calculating a formula: white plaque coverage (%) = white plaque area/caudal area x 100%;
S1.6, generating a white plaque change characteristic plate according to the coverage of the white plaque of the tail limbs of procambarus clarkia at different ovarian development stages;
S1.7, identifying the ovarian development stage of female procambarus clarkia according to a white plaque change characteristic plate;
S1.8, when white plaques of the tail limbs of the female procambarus clarkia are densely distributed in a cluster shape and the coverage of the white plaques is more than 45%, the ovaries of the female procambarus clarkia are in the mature stage (the 6-stage of ovaries development).
Wherein, the area of the tail limb of the female procambarus clarkia in S1.3 and the area of the white plaque in S1.4 are obtained by measuring the photographs of the tail limb and the photographs of the white plaque. These pictures were taken with a camera (Lightools 1000KPA, china) fixed on a tripod and a bubble level was placed in the horizontal direction so that a plane picture was taken vertically, avoiding any parallax effect. Photographs were measured using Lightools software (Lightools 3.7, china).
The tail limb area of female procambarus clarkia comprises the area of the inner limb and the area of the outer limb part above the transverse seam.
The ovarian development stage of female procambarus clarkia in S1.5 is divided into 7 stages: stage 1 (oogenic stage), stage 2 (immature stage), stage 3 (pre-yolk formation stage), stage 4 (yolk formation stage), stage 5 (mid-yolk formation stage), stage 6 (late yolk formation stage), and stage 7 (post-partum development stage).
The ovarian development stage of female procambarus clarkia identified in S1.5 according to the plate of white plaque change features includes:
(1) The tail limbs of female procambarus clarkia in the development stage of ovary of 1-3 are free of white plaque, and only muscles are arranged in the inner cavity of the tail limbs.
(2) The white plaque of the tail limb of the female procambarus clarkii in the development stage of the ovary is less than 30%, and the white plaque is only positioned around the muscle of the tail limb and distributed linearly.
(3) The coverage of white plaque of the tail limbs of female procambarus clarkia at stage 5 of ovarian development is more than 30%, and the white plaque is mostly located near the tail limb muscles and spreads downwards and distributed in a sheet shape.
(4) The coverage of white plaque of the tail limbs of female procambarus clarkii in the development stage of the ovary is more than 45%, and the white plaque is closely packed and wrapped and even covered on the tail limb muscles and distributed in a clustered manner.
(5) The white plaque of the tail of female procambarus clarkii at stage 7 of ovarian development disappeared.
Specifically, the discrimination of the mature male procambarus clarkii comprises the following steps:
S2.1, observing morphological changes of a first pair of abdominal limbs of the male procambarus clarkia;
S2.2, observing the morphological changes of the seat joints of the third pair and the fourth pair of feet of the male procambarus clarkia;
s2.3, identifying the development stage of the testis of the male procambarus clarkii according to the morphological characteristics of the first pair of abdominal limbs and the morphological characteristics of the seat joints of the third pair of feet and the fourth pair of feet;
S2.4, when the first pair of abdominal limbs of the male procambarus clarkii have differentiated obvious head processes and tail processes, and obvious barbs are arranged on seat joints of the third pair of feet and the fourth pair of feet, the male procambarus clarkii is in a mature period (a period of development 5 of spermary).
The development stage of the testis of the male procambarus clarkii in S2.3 is divided into 5 stages: stage1 (spermatogenic stage), stage2 (primary spermatocyte stage), stage3 (secondary spermatocyte stage), stage4 (sperm stage), stage 5 (sperm stage).
The specific implementation of the mature parent discrimination for female procambarus clarkii and male procambarus clarkii is as follows:
Firstly, judging mature female procambarus clarkii;
Discrimination of mature female procambarus clarkii is carried out from 7 months 30 days to 10 months 30 days, during which 30 female procambarus clarkii are sampled every 10 days.
First, morphological parameters of the female shrimp were measured, respectively, body weight, head and chest nail length, body width (body width refers to the width of the first body segment of the exoskeleton), abdominal girth length, and weight of the contents of the hindlimb (the contents of the hindlimb include white plaque and hindlimb muscles). After white plaque appeared on the tail limbs of the female shrimp, the tail limb area and the white plaque area of the female shrimp were measured. Then, the weight of the hindlimb contents (WW) and white plaque coverage (WC) of the relative body weight were calculated. The parameters were calculated as follows: relative body weight hindlimb content weight (%) = hindlimb content weight/body weight x 100%; white plaque coverage (%) = white plaque area/caudal area x 100%. Wherein the white plaque area and the hindlimb area are obtained by measuring the hindlimb photograph and the white plaque photograph. These pictures were taken with a camera (Lightools 1000KPA, china) fixed on a tripod and a bubble level was placed in the horizontal direction so that a plane picture was taken vertically, avoiding any parallax effect. Photographs were measured using Lightools software (Lightools 3.7, china).
After the above parameters are determined and calculated, female procambarus clarkii is dissected. Ovarian tissue of female shrimp was washed with physiological saline and fixed in Bouin's solution (Wohan service biosciences, wohan, china) at 10 times the tissue volume for 24 hours at 5 ℃. Ovarian samples were dehydrated in a series of graded ethanol solutions, embedded in paraffin mixtures, sectioned on a microtome (section thickness 4 μm, leica RM2016, USA), and finally stained with hematoxylin and eosin counterstained. Slice samples were examined under an optical microscope (Olympus BX53, japan) to obtain ovarian developmental stages.
Ovarian development was divided into 7 stages: stage 1 (oogenic stage), stage 2 (immature stage), stage 3 (pre-yolk formation stage), stage 4 (yolk formation stage), stage 5 (mid-yolk formation stage), stage 6 (late yolk formation stage), and stage 7 (post-partum development stage).
Then, analyzing the change rule of each morphological parameter in the ovarian development process through Kendall correlation analysis and random forest algorithm, and searching morphological parameters capable of representing the mature ovarian development, so as to judge the mature female procambarus clarkia.
As shown in fig. 2, the ovarian development stage (OS) and the abdominal annular groove length (AL), the Body Width (BW), the body weight (W), the brachiocephalus length (CL), the weight of the caudal content (WW) relative to the body weight, and the white plaque coverage (WC) were analyzed by Kendall correlation analysis, wherein "×" indicates P >0.05, P <0.01, and P <0.001. It can be seen that the positive correlation between body weight (W), relative body weight of the tail content (WW), white plaque coverage (WC) and the ovarian developmental stage is strong (all R > 0.2), with the highest correlation between white plaque coverage (WC) and the ovarian developmental stage (r=0.49, p=0.000) and the second correlation between relative body weight of the tail content (WW) and the ovarian developmental stage (r=0.35, p=0.000).
As shown in fig. 3, parameters of morphological characteristics such as the abdominal annular groove length (AL), the Body Width (BW), the body weight (W), the brachiocephalus length (CL), the weight of the content of the hindlimb relative to the body weight (WW), and the white plaque coverage (WC) were scored based on a random forest algorithm using MEAN DECREASE GINI (MDG). The importance of morphological features is ordered in descending order of score, with the first three being white plaque coverage (WC), weight of the content of the hindlimb relative to body weight (WW), body weight (W). Clearly, white plaque coverage (WC) best reflects ovarian developmental changes in female procambarus clarkia.
Considering that the tail limbs of the female shrimps in stages 1-3 and 7 of the ovarian development are free of white plaques, the present example classifies the female shrimps free of white plaques into a group, and further evaluates morphological parameters capable of judging the mature state of the ovaries. In this case, as shown in fig. 4, where N represents the ovarian developmental stage (stage 1, stage 2, stage 3, and stage 7) of the crayfish without white plaque, Y represents the ovarian developmental stage (stage 4, stage 5, stage 6) of the crayfish with white plaque, and Kendall correlation analysis showed that there was a very strong positive correlation (r=0.95, p=0.000) between the white plaque coverage (WC) and the ovarian maturation status (ovarian developmental stages other than stages 1-3 and 7). Further decision tree analysis was performed, as shown in FIG. 5, and it was found that female procambarus clarkia having a white patch and a white patch coverage (WC) of less than 30% was in stage 4 of ovarian development, female procambarus clarkia having a white patch coverage (WC) of greater than 30% and less than 43% was in stage 5 of ovarian development, and female procambarus clarkia having a white patch coverage (WC) of greater than 43% was in stage 6 of ovarian development. Considering that the discrimination parameters need to be compatible with reliability and intuitiveness, the key node information of the white plaque coverage (WC) is determined to be 30% and 45%.
Finally, a white plaque change profile was generated, as shown in fig. 6, where a and B are stages 1-3 of ovarian development, C and D are stage 4 of ovarian development, E and F are stage 5 of ovarian development, G and H are stage 6 of ovarian development, and I and J are stage 7 of ovarian development. The white plaque change characteristic plate specifically comprises: the tail limbs of the female procambarus clarkia in the development stage of the ovary is 1-3 stages without white plaque, and the inner cavities of the tail limbs are only provided with muscles; white plaque exists in the tail limbs of female procambarus clarkii in the development stage of the ovary, the coverage of the white plaque of the tail limbs of the female procambarus clarkii in the stage is less than 30%, and the white plaque is only positioned around the muscles of the tail limbs and distributed linearly; the coverage of white plaque of the tail limb of the female procambarus clarkii in the development stage of the ovary is more than 30% and less than 45%, and the white plaque is positioned near the tail limb muscles and spreads downwards and is distributed in a sheet shape; the coverage of white plaque of the tail limbs of the female procambarus clarkii in the development stage of the ovary is more than 45%, and the white plaque is closely packed and wrapped and even covered on the tail limb muscles and distributed in a clustered manner; the white plaque of the tail of female procambarus clarkii at stage 7 of ovarian development disappeared.
And (3) referring to the generated white plaque change characteristic plate, and judging the accuracy of the mature female procambarus clarkia (ovarian development stage 6) according to the white plaque change characteristic is up to 86.4%.
(II) judging mature male procambarus clarkii;
Discrimination of mature male procambarus clarkii is carried out from 7 months 30 days to 10 months 30 days, and 30 male procambarus clarkii are sampled every 10 days.
The weight of the male shrimp was first measured, and then the morphology of the first pair of abdominal limbs and the morphology of the seat joints of the third and fourth pairs of feet of the male shrimp were observed. As shown in FIGS. 7 and 8, when the first pair of abdominal limbs of the male procambarus clarkia has differentiated into distinct head processes 1 and tail processes 2, and the third and fourth pairs of feet have distinct barbs 3 on the base nodes, the male procambarus clarkia is dissected. The testis tissue of male shrimps was washed with physiological saline and fixed in Bouin's solution (Wohan service biosciences, wohan, china) at a volume 10 times of the tissue at 5℃for 24 hours. The testis samples were dehydrated in a series of graded ethanol solutions, embedded in paraffin mixtures, sectioned on a microtome (section thickness 4 μm, leica RM2016, USA), and finally stained with hematoxylin and eosin counterstained. Slice samples were examined under an optical microscope (Olympus BX53, japan) to obtain the development stage of the testis.
The development of the testis is divided into 5 stages: stage 1 (spermatogenic stage), stage 2 (primary spermatocyte stage), stage 3 (secondary spermatocyte stage), stage 4 (sperm stage), stage 5 (sperm stage).
The accuracy of judging the mature male procambarus clarkii (stage 5 of the development of the testis) according to the characteristics of the head process and the tail process of the first pair of abdominal limbs and the barbs on the third pair of feet and the fourth pair of feet is up to 100 percent.
The above embodiments are merely illustrative of the preferred embodiments of the present invention, and the scope of the present invention is not limited thereto, but various modifications and improvements made by those skilled in the art to which the present invention pertains are made without departing from the spirit of the present invention, and all modifications and improvements fall within the scope of the present invention as defined in the appended claims.
Claims (8)
1. A method for rapidly distinguishing a mature parent of procambarus clarkia is characterized by comprising the following steps:
judging the sex of procambarus clarkia;
If the procambarus clarkia is female procambarus clarkia, collecting the area of the tail limbs of the procambarus clarkia and the area of the white plaque on the surfaces of the tail limbs, calculating the coverage of the white plaque, and judging the ovarian development stage of the procambarus clarkia based on the coverage of the white plaque;
And if the procambarus clarkia is male procambarus clarkia, collecting morphological characteristics of a first pair of abdominal limbs and morphological characteristics of a third pair and a fourth pair of foot seat sections of the procambarus clarkia, and judging the development stage of the testis of the procambarus clarkia based on the morphological characteristics of the first pair of abdominal limbs and the morphological characteristics of the third pair and the fourth pair of foot seat sections.
2. The rapid discrimination method of the mature parent of procambarus clarkii of claim 1, wherein collecting the area of the tail limb and the area of the white plaque on the surface of the tail limb of the female procambarus clarkii comprises:
Checking whether the cleaned female procambarus clarkia is enriched with white plaques, and if the female procambarus clarkia is not enriched with white plaques, directly discarding the judgment; and if the white plaque is enriched, taking a tail limb photo and a white plaque photo of the female procambarus clarkii, and determining the tail limb area and the white plaque area of the tail limb surface of the female procambarus clarkii based on the tail limb photo and the white plaque photo.
3. The rapid discrimination method of procambarus clarkia mature parents of claim 1, wherein the method for calculating the white plaque coverage is as follows: white plaque coverage (%) = white plaque area/caudal area x 100%.
4. The rapid discrimination method of a mature parent of procambarus clarkii of claim 1, wherein the ovarian development stage of female procambarus clarkii comprises: stage 1-egg primordial stage, stage 2-immature stage, stage 3-pre-vitelline formation stage, stage 4-vitelline formation stage, stage 5-mid vitelline formation stage, stage 6-post vitelline formation stage, and stage 7-post partum development stage.
5. The method for rapid determination of a mature parent of procambarus clarkia of claim 4, wherein determining the ovarian developmental stage of said female procambarus clarkia based on said white plaque coverage comprises:
Judging the ovarian development stage of the female procambarus clarkii according to the white plaque coverage by referring to a preset white plaque change characteristic plate, and when the ovarian development stage is in the later stage of the 6-phase yolk formation stage, the ovary of the female procambarus clarkii is mature; the white plaque change characteristic plate is constructed according to white plaque coverage of tail limbs of female procambarus clarkia at different ovarian development stages.
6. The rapid discrimination method of procambarus clarkia mature parents of claim 5, wherein the white plaque coverage of the female procambarus clarkia tails at different ovarian development stages comprises:
the tail limbs of the female procambarus clarkia with the ovarian development stage of 1-3 stages have no white plaque;
white plaque exists on the tail limbs of female procambarus clarkii with the ovarian development stage of 4, and the coverage of the white plaque is less than 30%;
The tail limbs of the female procambarus clarkia with the ovarian development stage of 5 stages have white plaques, and the coverage of the white plaques is more than 30% and less than 45%;
white plaque exists on the tail limbs of female procambarus clarkii with the ovarian development stage of 6 phases, and the coverage of the white plaque is more than 45%;
the white plaque of the tail limbs of female procambarus clarkii with the ovarian development stage of 7 stages disappears.
7. The rapid discriminating method of mature parent of procambarus clarkia of claim 1, wherein the testis development stage of male procambarus clarkia comprises: stage 1-spermatogenic stage, stage 2-primary spermatocyte stage, stage 3-secondary spermatocyte stage, stage 4-spermatocyte stage, and stage 5-spermatocyte stage.
8. The rapid determination method of the mature parent of procambarus clarkii of claim 7, wherein determining the testis development stage of the male procambarus clarkii based on the morphological features of the first pair of abdominal limbs and the morphological features of the third pair of and the fourth pair of foot-seat nodes comprises:
Observing morphological characteristics of the first pair of abdominal limbs and morphological characteristics of the third pair and the fourth pair of foot seat joints of the male procambarus clarkii, wherein when the first pair of abdominal limbs of the male procambarus clarkii have head processes and tail processes with preset sizes and the third pair and the fourth pair of foot seat joints have barbs with preset sizes, the testis development stage of the male procambarus clarkii is in the 5-stage sperm stage, and the testis of the male procambarus clarkii is mature.
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