CN104160999A - Construction method and application of restorer line of hybrid scallops of purple scallops and Argopecten irradians concentricus - Google Patents
Construction method and application of restorer line of hybrid scallops of purple scallops and Argopecten irradians concentricus Download PDFInfo
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- CN104160999A CN104160999A CN201410429308.8A CN201410429308A CN104160999A CN 104160999 A CN104160999 A CN 104160999A CN 201410429308 A CN201410429308 A CN 201410429308A CN 104160999 A CN104160999 A CN 104160999A
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Abstract
The invention relates to a construction method and application of the restorer line of hybrid scallops of purple scallops and Argopecten irradians concentricus. The restorer line is Argopecten irradians concentricus strains capable of enabling the generations of the male sterility line to restore the characteristic of male sterility, and the hybrid scallops of the purple scallops and the Argopecten irradians concentricus are the hybrid generations of purple scallop ova and Argopecten irradians concentricus sperms. The construction method comprises the steps that excellent individuals in the male sterility line are selected, different varieties of Argopecten irradians concentricus are used for continuous backcross until all generations are hermaphrodite, and the restorer line of the male sterility line is obtained; finally, the restorer line and the corresponding male sterility line are used for hybridization to produce commodity offspring seeds of the hybrid scallops of the purple scallops and the Argopecten irradians concentricus. According to the construction method and application, the restorer line of the male sterility liner is firstly built in animals, the obstacles of autofertilization of hermaphrodite scallops in the hybridization process are effectively eliminated, and the yield of the produced hybrid scallops of the purple scallops and the Argopecten irradians concentricus is improved by more than 42 percent relative to that of the Argopecten irradians concentricus in the same year.
Description
Technical field
The invention belongs to shellfish breeding technique--scallop hybrid commodity offspring seed cultivation method, especially relates to a kind of construction method and application of purple black scallop hybrid (filial generation of purple scallop ovum and Gulf of Mexico scallop sperm) restorer.
Background technology
Gulf of Mexico scallop (Argopecten irradians concentricus) originates in U.S. Gulf of Mexico bank, introduces China in eighties of last century the nineties.Gulf of Mexico scallop fast growth, temperature wide accommodation, is especially relatively applicable to high temperature sea area, and strong stress resistance is applicable to the cultivation in south China marine site very much, is one of the main scallop culture kind in southern marine site at present.But due to long-term inbreeding after introducing, scallop germplasm in breeding process in the Gulf of Mexico is degenerated serious, show as commodity shellfish specification more and more less, in seedling cultivation process, lethality is higher, therefore scallop culture industry is in the urgent need to carrying out genetic improvement to Gulf of Mexico scallop, and wherein crossbreeding is one of the most effective germplasm improvement method.
For this reason, inventor has introduced purple scallop (Argopecten purpuratus) from Peru first from 2007.Purple scallop is the medium-sized scallop of a kind of fast-growing type that originates in the South Pacific Ocean, all belongs to Argopecten belong to Gulf of Mexico scallop, and chromosome number is identical, and is all hermaphroditism, but the two is in aspect proterties complementations such as Individual Size, growth and thermal adaptabilities.Therefore inventor, again by purple scallop and Gulf of Mexico scallop are carried out to interspecific cross, turns out orthogonal (purple scallop ovum and the fertilization of Gulf of Mexico scallop sperm) and two kinds of hybridization first filial generations of reciprocal cross (Gulf of Mexico scallop ovum and the fertilization of purple scallop sperm).The individual weight of hybridization first filial generation improves 72-85% than the Gulf of Mexico scallop of cultivating the same period, and (southern music is red etc., the Primary Study of 2012. purple scallops and Gulf of Mexico scallop interspecific to demonstrate huge industry potentiality.China's agronomy circular, 28:131-135).
Although above-mentioned hybridization first filial generation can produce larger hybrid vigour, but want spread still to exist larger difficulty, this is because Gulf of Mexico scallop and purple scallop are all hermaphroditic scallops, when breeding, discharge sperm and ovum simultaneously, the ovum of discharge is easily subject to the pollution of sperm of the same race, be difficult to obtain in a large number unfertilized egg for interspecific cross, thereby cause filial generation ratio real in obtained commodity seed greatly to reduce, thereby make the increase rate of its output greatly limited.
Research discovery, the hermaphroditism characteristic of above-mentioned two kinds of Argopecten scallops is similar to the monoecism characteristic of many crops.Also the problem that exists color pollination in hermaphrodite crop breeding, needs artificial emasculation, and this point is similar to the self-fertilization problem running in the scallop crossover process of the Gulf of Mexico to purple scallop.The key addressing this problem in agricultural depends on the discovery of male sterile plants, utilize male sterile plant and its male parent continuous backcross, just can select male sterile line, maintainer and restorer, build the three series mating breeding system in crop breeding with this, realize the large-scale production of crossbreed, thereby solve a hybrid seeding difficult problem.Up to now, in numerous crops set up three series mating breeding system both at home and abroad, and be widely used in, in the production of crossbreed, becoming the main path that increases substantially crop yield.
But in animal breeding, there is no up to now both at home and abroad and find male sterile and femalely educate individual report, thereby also cannot set up the male sterile line, maintainer and the restorer that are similar to crops, for crossbreeding.Obviously; if can build the three series mating breeding system that similarly comprises male sterile line, maintainer and restorer in animal breeding; also just can realize low cost, the scale interspecific cross of hermaphroditism type animal; improve hybridization efficiency; improve resistance and the output of breeding animal, thereby realize the important breakthrough of animal breeding technology.
Summary of the invention
The object of the invention is to realize in animal breeding, specifically in hermaphroditism type scallop, build first purple black scallop hybrid restorer, to overcome the deficiency of existing scallop hybrid breeding technique.
Another object of the present invention is to utilize above-mentioned restorer and corresponding male sterile line is extensive, the application method of the purple black scallop hybrid seed of low-cost production.
Above-mentioned three is to refer to scallop male sterile line and maintainer and restorer.It is undesired that male sterile line is wherein that male gonad is grown, and can not produce the sperm of function, but its egg development is normal, can accept eupyrene sperm and be fertilized, and can be by its male sterile characteristic hereditary the strain to offspring.Obviously male sterile line is the core strain in three series mating system, and its Main Function is the unfertilized ovum that provides a large amount of in hybrid seedlings is commercially produced, and it is bred and depends on the maintainer supporting with it simultaneously.Male sterile line is to utilize individuality and its male parent continuous backcross that male sterile serendipitous and ovum can educate to set up the family that backcrosses, when all individualities of the family that backcrosses are the scallop hybrid strain obtaining when male sterile and ovum can be educated.
Restorer claims again male sterile line restorer, and after the ovum fertilization with its sperm to male sterile line, the female and male fertile of the filial generation that produces all recovers normal.Restorer is the hermaphroditism strain that can normally breed, can breed restorer of future generation by selfing.In animal breeding, the effect of restorer is to provide the ovum fertilization of sperm and male sterile line, to realize the cultivation of best possible merchandise hybrid seedlings.
The present invention is based on following design and practice: 1) inventor studies the male sterile individuality of having found to exist in purple black scallop hybrid (filial generation of purple scallop ovum and Gulf of Mexico scallop sperm) first filial generation 5-8%, these individualities do not produce activated sperm, but its ovum can be fertilized by the sperm of Gulf of Mexico scallop, and the fertilized egg obtaining can normal development, has obtained the scallop that backcrosses, 2) form rear approximately 92% individuality and still show as hermaphroditism with the ovum of male sterile individuality in Gulf of Mexico scallop sperm and purple black scallop hybrid the cultivated scallop that backcrosses that backcrosses, therefore can be by setting up multiple methods that backcross family and carry out coordinate force test, selecting its offspring is all the male sterile line that hermaphroditism and proterties are good (female parent)-Gulf of Mexico scallop (male parent) combination, Gulf of Mexico scallop strain in this combination has the ability that makes male sterile line recover male fertile, obtain the restorer of purple black scallop hybrid male sterile line, 3) by the sperm of these restorers and corresponding male sterile kind shellfish are hybridized, can realize convenient, efficient, the purple black scallop hybrid commodity seed of large-scale production, and the scallop hybrid commodity seed obtaining has significant advantage aspect growth, Individual Size and temperature tolerance.
The construction method of the black scallop hybrid restorer of purple of the present invention is as follows:
From the black scallop hybrid male sterile line of purple, select large individual scallop, after induced spawning, inseminate with the good different Gulf of Mexico scallop sperm of proterties respectively, set up a large amount of familys that backcrosses, to supply smart Gulf of Mexico scallop from numerous self-fertilization family (sff) of setting up simultaneously, measure growth and the survival condition of filial generation and also carry out coordinate force test; From the second year every year according to coordinate force test result and gonad development situation, select the family that backcrosses that proterties is wherein good and contain hermaphroditism scallop, relative male sterile family kind shellfish is backcrossed and sets up the family that backcrosses with the Gulf of Mexico scallop hybridization family sperm for smart continuously, until backcross, family is all that hermaphroditism and proterties are good,, by numerous Gulf of Mexico Process of Rearing Scallop Seeds method selfing expansion conventional Gulf of Mexico scallop self-fertilization family (sff) for smart, obtain the restorer of this male sterile line.The effect of restorer is to provide the ovum fertilization of sperm and male sterile line, to realize the cultivation of best possible merchandise hybrid seedlings.Recovery is the Gulf of Mexico scallop strain of hermaphroditism and normal development, therefore breeds restorer of future generation by conventional Gulf of Mexico scallop colony autocopulation method.
The application method of the black scallop hybrid restorer of purple of the present invention is as follows:
First select male sterile line and the restorer kind shellfish of purple black scallop hybrid according to the ratio of 5:1, and stimulate respectively after accelerating according to a conventional method male sterile line kind shellfish to lay eggs and the restorer kind shellfish ejaculation of laying eggs, again the smart ovum mixed liquor of this restorer is obtained to the seminal fluid of restorer with 500 order silk cover filterings, the seminal fluid of restorer is added in spawning pond and stirred, and add the amount of sperm to have 5-6 sperm to be advisable around with each ovum, finally after hatching, obtain purple black scallop hybrid commodity seed according to a conventional method.
Restorer construction method of the present invention is also applied to the breeding practice that the Argopecten such as violet black scallop hybrid (filial generation of Gulf of Mexico scallop ovum and purple scallop sperm), purple extra large scallop hybrid (filial generation of purple scallop ovum and bay scallop sperm) and extra large purple scallop hybrid (filial generation of bay scallop ovum and purple scallop sperm) belong to scallop hybrid, and is applied to the large-scale method for producing of setting up these scallop hybrid commodity seeds.
Method of the present invention is also applied to the interspecific cross breeding of other hermaphroditism type scallops, and for example Pecten belongs to the interspecific cross of scallop, and basic skills of the present invention is also applicable to the interspecific cross breeding of other hermaphroditism type invertebrates.
The purple China ink scallop hybrid restorer that the present invention builds is the restorer of the scallop set up in the animal kingdom first, should belong to the important breakthrough in animal breeding technology.The present invention has effectively overcome autogamous obstacle in hermaphroditism scallop crossover process; realize low cost and scale and obtained filial generation completely; therefore can make full use of hybrid vigour; reach the characteristic that makes cultivated purple China ink scallop hybrid commodity seed simultaneously there is Gulf of Mexico scallop and purple scallop; aspect growth, Individual Size and temperature tolerance, there is significant advantage than Gulf of Mexico scallop; therefore can be in southern sea area as the marine sites such as Guangdong, Guangxi and Fujian successfully cultivate, and improve more than 42% when the comparable Gulf of Mexico of annual production scallop.
Embodiment
The present invention has built the restorer of purple black scallop hybrid, and has set up the method for producing purple black scallop hybrid commodity seed with restorer and corresponding male sterile line hybridization, and concrete steps are as follows:
Embodiment 1
1) seed selection of purple black scallop hybrid restorer: select 50 of large individual scallops in purple black scallop hybrid male sterile line, sperm fertilization with different Gulf of Mexico scallop respectively after induced spawning, set up at least 20 familys that backcross, to supply smart Gulf of Mexico scallop from numerous self-fertilization family (sff) of setting up simultaneously, measure growth and the survival condition of backcross progeny and also carry out coordinate force test; Every year select according to coordinate force test result and gonad development situation the family that backcrosses that proterties is wherein good and contain hermaphroditism scallop from the second year; relative male sterile family kind shellfish is backcrossed and sets up the family that backcrosses with the Gulf of Mexico scallop hybridization family sperm for smart continuously; until backcross, family is all that hermaphroditism and proterties are good; the Gulf of Mexico scallop family selfing for smart is expanded numerous; obtain the restorer of this purple China ink scallop hybrid male sterile line, for the large-scale production of the black scallop hybrid commodity of purple seed.
2) produce the method for purple black scallop hybrid commodity seed by restorer: when grow seedlings spring, select after purple black scallop hybrid male sterile line and restorer kind shellfish accelerating according to the ratio of 5:1, lay eggs spawning pond moderate stimulation male sterile line kind shellfish, in bucket moderate stimulation restorer kind shellfish ejaculation, and obtain the restorer Gulf of Mexico scallop seminal fluid that does not contain ovum with 500 order silk cover filterings; The seminal fluid of restorer is evenly splashed and stirred in spawning pond and immediately, and fertilization situation is examined under a microscope in sampling at any time, and seminal fluid addition has 5-6 sperm to be advisable around with each ovum; Fertilized egg is hatched according to conventional method, obtains purple black scallop hybrid business seed after growth.
Embodiment 2
Within 2011, utilize the purple scallop of introducing and the Gulf of Mexico scallop of taking from Zhanjiang, cultivate altogether each 1 of interspecific cross first filial generation positive crossing and negative crossing family.Through actual measurement, the average shell high of purple black scallop hybrid (purple scallop ovum and the hybridization of Gulf of Mexico scallop sperm) is 52.38 millimeters, body weight is 27.40 grams, post weighs 2.46 grams, and the scallop average shell in the Gulf of Mexico high of the cultivation same period is 42.02 millimeters, body weight is 14.80 grams, and post is heavily 1.13 grams.Purple China ink hybridization first filial generation improves 24.7% than the Gulf of Mexico scallop shell of the same period is high, and body weight improves approximately 85.1%, and post is brought up again high by 117.7%.Spring in 2012, by the scallop of therefrom selecting individual great Qie Jinci district to grow after the scallop accelerating of above-mentioned hybridization first filial generation, after drying in the shade 30 minutes, scallop being placed in to the container moderate stimulation that fills 23 DEG C of seawater lays eggs, with the sperm insemination of single Gulf of Mexico scallop, cultivate 2 of the familys of backcrossing of purple black scallop hybrid first filial generation and Gulf of Mexico scallop.Find after accelerating that a family all shows as hermaphroditism in spring in 2013, in another family, most of individuality shows as male sterile.
Continue above-mentioned male sterile individuality its male parent Gulf of Mexico scallop self progeny's sperm to backcross, just can obtain offspring for seed selection at the most through a generation is all male sterile line (female parent)-maintainer (male parent) combination that male sterile and ovum can be educated, thereby obtains respectively male sterile line and maintainer.After gonad maturity accelerating, can obtain a large amount of unfertilized ovums with the conventional Gulf of Mexico scallop abductive approach male sterile line scallop of hastening parturition of laying eggs; used the sperm fertilization of maintainer, can be obtained on a large scale the male sterile line kind shellfish producing for purple black scallop hybrid commodity seed.
Sperm fertilization by the ovum of the male sterile line scallop in combinations thereof with different irrelevant Gulf of Mexico scallops, set up at least 20 familys carry out coordinate force test of backcrossing, the family of selecting offspring to contain hermaphroditism scallop, its maternal male sterile line kind shellfish is continued to the sperm fertilization with the Gulf of Mexico scallop self-fertilization family (sff) for smart, and according to growth traits and the continuous seed selection of gonad development situation, until backcross progeny is all that hermaphroditism and proterties are good, just obtaining after fertilization makes male sterile line offspring be hermaphroditism and the good restorer Gulf of Mexico scallop family of proterties.
Finally by after the male sterile line kind shellfish gonad maturity accelerating of the purple China ink scallop hybrid of above-mentioned acquisition, with the conventional a large amount of unfertilized ovums of Gulf of Mexico scallop induced spawning method acquisition, used the sperm fertilization of restorer, purple black scallop hybrid commodity seed just can be mass-produced.
Embodiment 3
2014, utilize the purple scallop introduced and Gulf of Mexico scallop to build together and found 14 of the hybridization first filial generation familys of purple scallop ovum and Gulf of Mexico scallop sperm, and set up the self-fertilization family (sff) for smart Gulf of Mexico scallop, according to the method for embodiment 1, male sterile line family and corresponding maintainer and the restorer that can in 1-3, select purple black scallop hybrid, realize commercially producing of purple black scallop hybrid seed.
Embodiment 4
The present invention is for the construction and application of violet black scallop hybrid restorer
The construction method of the restorer that the present invention sets up is also applied to the breeding of violet black scallop hybrid (filial generation of Gulf of Mexico scallop ovum and purple scallop sperm), and is applied to the large-scale method for producing of setting up violet black scallop hybrid commodity seed.
First select the some large individual scallop in violet black scallop hybrid male sterile line, stimulate the sperm fertilization with multiple irrelevant purple scallops after laying eggs, if backcross, family is all hermaphroditic scallop, only needs to select a generation just to obtain the combination of male sterile line-restorer; If backcross, family only has part for hermaphroditism, continues to backcross and set up the family that backcrosses with the sperm of corresponding purple scallop self-fertilization family (sff) with male sterile scallop, continues to set up the self-fertilization family (sff) of the purple scallop of male parent simultaneously, carries out seed selection of future generation; Through a generation or after how for seed selection, in the time that backcross progeny is all hermaphroditism, just obtain the restorer of violet black scallop hybrid male sterile line.
With the male sterile line ovum of above-mentioned violet black scallop hybrid and the hybridization of the sperm of maintainer, can realize efficient, convenient, large-scale production violet black scallop hybrid male sterile kind shellfish; With the male sterile line ovum of above-mentioned violet black scallop hybrid and the hybridization of the sperm of restorer, can realize efficient, convenient, large-scale production violet black scallop hybrid seed.
Embodiment 5
The present invention is for the construction and application of purple extra large scallop hybrid restorer
The construction method of the restorer that the present invention sets up is also applied to purple extra large scallop hybrid (filial generation of purple scallop ovum and bay scallop sperm) breeding, and is applied to the large-scale method for producing of setting up purple extra large scallop hybrid commodity seed.
First select the some large individual scallop in purple extra large scallop hybrid male sterile line, stimulate the sperm fertilization with multiple irrelevant bay scallops after laying eggs, if backcross, family is all hermaphroditic scallop, only needs to select a generation just to obtain the combination of male sterile line-restorer; If backcross, family only has part for hermaphroditism, continues to backcross and set up the family that backcrosses with the sperm of corresponding bay scallop self-fertilization family (sff) with male sterile scallop, continues to set up the self-fertilization family (sff) of male parent bay scallop simultaneously, carries out seed selection of future generation; Through a generation or after how for seed selection, in the time that backcross progeny is all hermaphroditism, just obtain the restorer of purple extra large scallop hybrid male sterile line.
With the male sterile line ovum of the extra large scallop hybrid of above-mentioned purple and the hybridization of the sperm of maintainer, can realize efficient, convenient, the purple extra large scallop hybrid male sterile kind shellfish of large-scale production; With the sperm hybridization of the extra large scallop hybrid male sterile of purple kind shellfish and restorer, can realize efficient, convenient, the purple extra large scallop hybrid seed of large-scale production.
Embodiment 6
The present invention is for the construction and application of extra large purple scallop hybrid restorer
The construction method of the restorer that the present invention sets up is also applied to extra large purple scallop hybrid (filial generation of bay scallop ovum and purple scallop sperm) breeding practice, and is applied to the large-scale method for producing of setting up extra large purple scallop hybrid commodity seed.
First select the some large individual scallop in extra large purple scallop hybrid male sterile line, stimulate the sperm fertilization with multiple irrelevant purple scallops after laying eggs, if backcross, family is all hermaphroditic scallop, only needs to select a generation just to obtain the combination of male sterile line-restorer; If backcross, family only has part for hermaphroditism, continues to backcross and set up the family that backcrosses with the sperm of corresponding purple scallop self-fertilization family (sff) with male sterile scallop, continues to set up the self-fertilization family (sff) of the purple scallop of male parent simultaneously, carries out seed selection of future generation; Through a generation or after how for seed selection, in the time that backcross progeny is all hermaphroditism, just obtain the restorer of extra large purple scallop hybrid male sterile line.
With the male sterile line ovum of the purple scallop hybrid in above-mentioned sea and the hybridization of the sperm of maintainer, can realize efficient, convenient, the extra large purple scallop hybrid male sterile kind shellfish of large-scale production; With the sperm hybridization of the purple scallop hybrid male sterile in sea kind shellfish and restorer, can realize efficient, convenient, the extra large purple scallop hybrid seed of large-scale production.
Claims (9)
1. the black scallop hybrid restorer of purple, is characterized in that this restorer is to make male sterile line offspring recover the Gulf of Mexico scallop strain of male-fertile characteristic, and wherein purple black scallop hybrid is the filial generation of purple scallop ovum and Gulf of Mexico scallop sperm.
2. the construction method of the purple of claim 1 China ink scallop hybrid restorer, is characterized in that:
Select the defect individual in purple black scallop hybrid male sterile line, after induced spawning, use respectively the Gulf of Mexico scallop sperm insemination from Different Individual, set up 20 familys that backcross above, to supply smart Gulf of Mexico scallop from numerous self-fertilization family (sff) of setting up simultaneously, measure growth and the survival condition of filial generation and also carry out coordinate force test; From the second year every year according to coordinate force test result and gonad development situation, select the family that backcrosses that proterties is wherein good and contain hermaphroditism scallop, the male sterile family kind shellfish relevant to the family that backcrosses backcrossed with the Gulf of Mexico scallop hybridization family sperm for smart continuously, the foundation family that backcrosses, until backcross, family is all that hermaphroditism and proterties are good,, by numerous Process of Rearing Scallop Seeds method selfing the expansion conventional Gulf of Mexico scallop self-fertilization family (sff) for smart, obtain the restorer of this male sterile line.
3. the purple of the claims 1 and claim 2 China ink scallop hybrid male sterile line restorer is applied to and produces purple black scallop hybrid commodity seed, hybridize with the sperm of restorer and the ovum of corresponding purple black scallop hybrid male sterile line, obtain purple black scallop hybrid commodity seed.
4. the construction method of the purple China ink scallop hybrid restorer that claim 2 is set up is also applied to the breeding of following scallop hybrid:
(1) the violet black scallop hybrid of cultivating with Gulf of Mexico scallop ovum and the fertilization of purple scallop sperm;
(2) the purple sea scallop hybrid of cultivating with purple scallop ovum and the fertilization of bay scallop sperm;
(3) the purple scallop hybrid in sea of cultivating with bay scallop ovum and the fertilization of purple scallop sperm.
5. the application method of the purple China ink scallop hybrid male sterile line restorer that claim 3 is set up is also applied to the breeding of following scallop hybrid:
(1) the violet black scallop hybrid of cultivating with Gulf of Mexico scallop ovum and the fertilization of purple scallop sperm;
(2) the purple sea scallop hybrid of cultivating with purple scallop ovum and the fertilization of bay scallop sperm;
(3) the purple scallop hybrid in sea of cultivating with bay scallop ovum and the fertilization of purple scallop sperm.
6. the construction method of the purple China ink scallop hybrid male sterile line restorer that claim 2 is set up is also applied to other hermaphroditism type scallops, and for example Argopecten genus and Pecten belong to the interspecific cross breeding of scallop.
7. the purple China ink scallop hybrid male sterile line restorer application method that claim 3 is set up is also applied to other hermaphroditism type scallops, and for example Argopecten genus and Pecten belong to the interspecific cross breeding of scallop.
8. the basic construction method of the purple China ink scallop hybrid male sterile line restorer that claim 2 is set up is also applied to the interspecific cross breeding of other hermaphroditism type invertebrates.
9. the application method of the purple China ink scallop hybrid male sterile line restorer that claim 3 is set up is also applied to the interspecific cross breeding of other hermaphroditism type invertebrates.
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| CN116391654A (en) * | 2023-03-30 | 2023-07-07 | 中国海洋大学 | Large-scale hybridization breeding method for hermaphrodite scallops |
| CN116391654B (en) * | 2023-03-30 | 2023-11-03 | 中国海洋大学 | Large-scale hybridization breeding method for hermaphrodite scallops |
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