Due to sexual dimorphism in the growth of certain cultured fish species,the production of monosex fishes is desirable for the aquaculture industry.Nowadays,the most widely practiced technique available for the mass pr...Due to sexual dimorphism in the growth of certain cultured fish species,the production of monosex fishes is desirable for the aquaculture industry.Nowadays,the most widely practiced technique available for the mass production of monosex fish populations is sex steroid-induced sex reversal.Here,a novel strategy for the successful production of all-female(AF)common carp(Cyprinus carpio L.),to take advantage of the sexual dimorphism in growth documented in this species,has been developed using genetic engineering via single gene-targeting manipulation without any exogenous hormone treatments.Male and female heterozygous cyp17a1-deficient common carp were first obtained using the clustered regularly interspaced short palindromic repeats/CRISPR-associated endonuclease 9(CRISPR/Cas9)technique.An all-male phenotype for homozygous cyp17a1-deficient carp,regardless of the individuals’sexdetermination genotypes(XY or XX),has been observed.A male-specific DNA marker newly identified in our laboratory was used to screen the neomale carp population with the XX genotype from the cyp17a1-deficient carp.These neomale carp develop a normal testis structure with normal spermatogenesis and sperm capacity.The neomale common carp were then mated with wild-type(WT)females(cyp17a1^(+/+)XX genotype)using artificial fertilization.All the AF offspring sample fish from the neomale-WT female mating were confirmed as having the cyp17a^(1+/-)XX genotype,and normal development of gonads to ovaries was observed in 100.00%of this group at eight months post-fertilization(mpf).A total of 1000 carp fingerlings,500 from the WT male and female and 500 from the neomale and WT female mating,were mixed and reared in the same pond.The average body weight of cyp17a1^(+/-)XX females was higher by 6.60%(8 mpf)and 32.66%(12 mpf)than that of the control common carp.Our study demonstrates the first successful production of a monosex teleost population with the advantages of sexual dimorphism in growth using genetic manipulation targeting a single locus.展开更多
基金the National Key Research and Development Program,China(2018YFD0900205)to Zhan Yinthe National Natural Science Foundation,China(31972779 and 31530077)to Gang Zhai and Zhan Yin+2 种基金the Pilot Program A Project from the Chinese Academy of Sciences(XDA24010206)to Zhan Yinthe Youth Innovation Promotion Association of CAS(20200336)to Gang Zhaithe State Key Laboratory of Freshwater Ecology and Biotechnology(2016FBZ05)to Zhan Yin.
文摘Due to sexual dimorphism in the growth of certain cultured fish species,the production of monosex fishes is desirable for the aquaculture industry.Nowadays,the most widely practiced technique available for the mass production of monosex fish populations is sex steroid-induced sex reversal.Here,a novel strategy for the successful production of all-female(AF)common carp(Cyprinus carpio L.),to take advantage of the sexual dimorphism in growth documented in this species,has been developed using genetic engineering via single gene-targeting manipulation without any exogenous hormone treatments.Male and female heterozygous cyp17a1-deficient common carp were first obtained using the clustered regularly interspaced short palindromic repeats/CRISPR-associated endonuclease 9(CRISPR/Cas9)technique.An all-male phenotype for homozygous cyp17a1-deficient carp,regardless of the individuals’sexdetermination genotypes(XY or XX),has been observed.A male-specific DNA marker newly identified in our laboratory was used to screen the neomale carp population with the XX genotype from the cyp17a1-deficient carp.These neomale carp develop a normal testis structure with normal spermatogenesis and sperm capacity.The neomale common carp were then mated with wild-type(WT)females(cyp17a1^(+/+)XX genotype)using artificial fertilization.All the AF offspring sample fish from the neomale-WT female mating were confirmed as having the cyp17a^(1+/-)XX genotype,and normal development of gonads to ovaries was observed in 100.00%of this group at eight months post-fertilization(mpf).A total of 1000 carp fingerlings,500 from the WT male and female and 500 from the neomale and WT female mating,were mixed and reared in the same pond.The average body weight of cyp17a1^(+/-)XX females was higher by 6.60%(8 mpf)and 32.66%(12 mpf)than that of the control common carp.Our study demonstrates the first successful production of a monosex teleost population with the advantages of sexual dimorphism in growth using genetic manipulation targeting a single locus.
文摘为比较全雌翘嘴鳜Siniperca chuatsi与自交翘嘴鳜Siniperca chuatsi胚胎发育及仔幼鱼生长速度,采用人工繁殖的方法获得全雌翘嘴鳜和自交翘嘴鳜后代的受精卵,在水温为26~28℃条件下孵化,对全雌翘嘴鳜和自交翘嘴鳜后代的胚胎发育进行观察与比较,并详细记录了受精卵受精、卵裂、囊胚、原肠胚、神经胚、器官形成和出膜7个连续发育阶段特征,结合胚胎发育特征进一步比较了二者在仔幼鱼阶段的生长状况。结果表明:在水温、孵化方式等外部条件相同的情况下,全雌翘嘴鳜、自交翘嘴鳜的受精卵从受精到孵化出膜所需的总时间分别为39 h 12 min和38 h 44 min,各时期的胚胎发育特征基本一致;在仔幼鱼生长阶段,21日龄后全雌翘嘴鳜与自交翘嘴鳜的生长呈显著性差异(P<0.05);仔幼鱼全长-体质量生长结果显示,全雌翘嘴鳜的生长方程为W_(全雌鳜)=0.016L^(2.971)(R^(2)=0.991),说明在仔幼鱼阶段全雌翘嘴鳜略呈匀速生长,自交翘嘴鳜的生长方程为W_(自交鳜)=0.020L^(3.008)(R^(2)=0.991),说明在该阶段自交翘嘴鳜体质量生长快于全长生长。研究表明,全雌翘嘴鳜与自交翘嘴鳜的胚胎发育时间及其特征基本一致,但全雌翘嘴鳜仔幼鱼的生长速度显著快于自交翘嘴鳜。
