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| 1 | Molecular basis and genetic improvement of economically important traits in aquaculture animals显示文摘Aquaculture has been believed to be a major Chinese contribution to the world. In recent 20 years, genome and other genetic technologies have promoted significant advances in basic studies on molecular basis and genetic improvement of aquaculture animals, and complete genomes of some main aquaculture animals have been sequenced or announced to be sequenced since the beginning of this century. Here, we review some significant breakthrough progress of aquaculture genetic improvement technologies including genome technologies, somatic cell nuclear transfer and stem cell technologies, outline the molecular basis of several economically important traits including reproduction, sex, growth, disease resistance, cold tolerance and hypoxia tolerance, and present a series of candidate trait-related genes. Finally, some application cases of genetic improvement are introduced in aquaculture animals, especially in China, and several development trends are highlighted in the near future. | GUI JianFang ZHU ZuoYan | 2012 | Chinese Science Bulletin2012，57，15： | 49 |
| 2 | Integration mechanisms of transgenes and population fitness of GH transgenic fish显示文摘It has been more than 20 years since the first batch of transgenic fish was produced. Five stable germ-line transmitted growth hormone (GH) transgenic fish lines have been generated. This paper reviews the mechanisms of integration and gene targeting of the transgene, as well as the viability, reproduction and transgenic approaches for the reproductive containment of GH-transgenic fish. Further, we propose that it should be necessary to do the following studies, in particularly, of the breeding of transgenic fish: to assess the fitness of transgenic fish in an aqueous environment with a large space and a complex structure; and to develop a controllable on-off strategy of reproduction in transgenic fish. | HU Wei & ZHU ZuoYan* State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China | 2010 | Science China(Life Sciences)2010，53，4： | 27 |
| 3 | Genetic analysis of 'all-fish' growth hormone gene transferred carp (Cyprinus carpio L.) and its F_1 generation显示文摘Recornbinant 'all-fish' growth hormone gene (GH) was microinjected into the fertilized eggs of carp. A comparison between the growth traits of transgenics and non-transgenics was carried out, and the transgenic individuals with significant 'fast-growing' effect were successfully gained. A comparison on the reproductivities was also given out between the transgenics and their non-transgenic siblings, and showed that the reproductive capacity of transgenics was substantially equivalent to those of the non-transgenics. On the other hand, the genetic separation and the characteristic distribution of the FI generation were genetically analyzed, which gave solid evidence for the hypothesis that 2-3 chromosomes are integrated with trans-gene. In addition, the distinct biological effects for multi-site-integrated transgenes were further discussed. The present study opens a door for the breeding of 'fast-growing' transgenic fish. | WANG Yaping, HU Wei, WU Gang,SUN Yonghua, CHEN Shangping, ZHANG Fuying,ZHU Zuoyan, FENG Jianxin & ZHANG Xirui1. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China 2. Henan Institute of Aquaculture, Zhengzhou 450044, China | 2001 | Chinese Science Bulletin2001，46，14： | 23 |
| 4 | Progress in the evaluation of transgenic fish for possible ecological risk and its containment strategies显示文摘Genetically improved transgenic fish possess many beneficial economic traits; however, the commer-cial aquaculture of transgenic fish has not been performed till date. One of the major reasons for this is the possible ecological risk associated with the escape or release of the transgenic fish. Using a growth hormone transgenic fish with rapid growth characteristics as a subject, this paper analyzes the following: the essence of the potential ecological risks posed by transgenic fish; ecological risk in the current situation due to transgenic fish via one-factor phenotypic and fitness analysis, and mathe-matical model deduction. Then, it expounds new ideas and the latest findings using an artificially simulated ecosystem for the evaluation of the ecological risks posed by transgenic fish. Further, the study comments on the strategies and principles of controlling these ecological risks by using a trip-loid approach. Based on these results, we propose that ecological risk evaluation and prevention strategies are indispensable important components and should be accompanied with breeding re-search in order to provide enlightments for transgenic fish breeding, evaluation of the ecological risks posed by transgenic fish, and development of containment strategies against the risks. | HU Wei WANG YaPing ZHU ZuoYan | 2007 | Science China(Life Sciences)2007，50，5： | 17 |
| 5 | Rapid growth cost in “all-fish” growth hormone gene transgenic carp:Reduced critical swimming speed显示文摘Evidence has accumulated that there is a trade-off between benefits and costs associated with rapid growth. A trade-off between growth rates and critical swimming speed (Ucrit) had been also reported to be common in teleost fish. We hypothesize that growth acceleration in the F3 generation of 'all-fish' growth hormone gene (GH) transgenic common carp (Cyprinus carpio L.) would reduce the swimming abilities. Growth and swimming performance between transgenic fish and non-transgenic controls were compared. The results showed that transgenic fish had a mean body weight 1.4―1.9-fold heavier,and a mean specific growth rate (SGR) value 6%―10% higher than the controls. Transgenic fish,however,had a mean absolute Ucrit (cm/s) value 22% or mean relative Ucrit (BL/s) value 24% lower than the controls. It suggested that fast-growing 'all-fish' GH-transgenic carp were inferior swimmers. It is also supported that there was a trade-off between growth rates and swimming performance,i.e. faster-growing individuals had lower critical swimming speed. | LI DeLiang FU CuiZhang HU Wei ZHONG Shan WANG YaPing ZHU ZuoYan | 2007 | Chinese Science Bulletin2007，52，11： | 14 |
| 6 | Rapid growth and sterility of growth hormone gene transgenic triploid carp显示文摘Triploid carp(100%)with 150(3n=150)chromosomes were obtained by crossing the females of improved tetraploid hybrids(♀, 4n=200)of red crucian carp(♀)×common carp(♂)with the males of diploid yellow river carp(♂,2n=100).The crosses yielded transgenic triploid carp(positive triploid fish,44.2%of the progeny)and non-transgenic triploid carp(negative triploid fish). Histological examination of the gonads of 24-month-old positive triploid fish suggested they were sterile and the fish were not able to produce mature gametes during the breeding season.Morphologically,both the positive and negative triploid fish were similar.They had a spindle-shaped,laterally compressed,steel grey body with two pairs of barbells.Most of the quantifiable traits of the triploid carp were intermediate between those of the two parents.The positive and negative triploid fish were raised in the same pond for 2 years.The mean body weight of the positive triploid fish was 2.3 times higher than the negative triploid fish.The weight of the largest positive triploid fish was 2.91 times higher than that of the largest negative triploid fish.Thus,we produced fast-growing transgenic triploid carp that have a reduced ecological risk because of their inability to mate and produce progeny. | YU Fan XIAO Jun LIANG XiangYang LIU ShaoJun ZHOU GongJian LUO KaiKun LIU Yun HU Wei WANG YaPing ZHU ZuoYan | 2011 | Chinese Science Bulletin2011，56，16： | 11 |
| 7 | Progress in studies of fish reproductive development regulation显示文摘Mechanisms of the animal reproductive development are an important research field in life sciences.The study of the reproductive development and regulatory mechanisms in fishes is important for elucidating the mechanisms of animal reproduction.This paper summarizes recent advances in the mechanisms of fish sex determination and differentiation,of fish gonad development and maturation,and of fish germ cell development,as well as the according regulating strategies.Fishes comprise an evolutionary stage that links invertebrates and higher vertebrates.They include diversiform species,and almost all vertebrate types of reproduction have been found in fishes.All these will lead to important advances in the regulatory mechanisms of animal reproduction by using fishes as model organisms.It will also enable novel fish breeding techniques when new controllable on-off strategies of reproduction and/or sex in fishes have been developed. | CHEN Ji HU Wei ZHU ZuoYan | 2013 | Chinese Science Bulletin2013，58，1： | 10 |
| 8 | High efficient gene targeting in rice field eel Monopterus albus by transcription activator-like effector nucleases显示文摘Genome editing plays an important role in the research of gene functions,gene correction and cell replacement therapy.Traditionally,genome editing is mainly dependent on the technology of homologous recombination(HR)and embryonic stem(ES)cell lines in mammals[1].However,genome editing is severely limited by the low efficiency of HR and lack of ES cell lines in most organisms.Recently,the development of zinc finger nucleases(ZFNs), | Ke Feng Hongrui Luo Yongming Li Ji Chen Yaping Wang Yonghua Sun Zuoyan Zhu Wei Hu | 2017 | Science Bulletin2017，62，3： | 9 |
| 9 | Fish genome manipulation and directional breeding显示文摘Aquaculture is one of the fastest developing agricultural industries worldwide.One of the most important factors for sustainable aquaculture is the development of high performing culture strains.Genome manipulation offers a powerful method to achieve rapid and directional breeding in fish.We review the history of fish breeding methods based on classical genome manipulation,including polyploidy breeding and nuclear transfer.Then,we discuss the advances and applications of fish directional breeding based on transgenic technology and recently developed genome editing technologies.These methods offer increased efficiency,precision and predictability in genetic improvement over traditional methods. | YE Ding ZHU ZuoYan SUN YongHua | 2015 | Science China(Life Sciences)2015，58，2： | 9 |
| 10 | Cloning and expression analysis in mature individuals of two chicken type-Ⅱ GnRH (cGnRH-Ⅱ) genes in common carp (Cyprinus carpio)显示文摘Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differing cGnRH-II cDNAs of common carp, namely cGnRH-II cDNA1 and cDNA2, were firstly cloned from the brain by rapid amplification of cDNA end (RACE) and reverse transcription- polymerase chain reaction (RT-PCR). The length of cGnRH-II cDNA1 and cDNA2 was 622 and 578 base pairs (bp), respectively. The cGnRH-II pre-cursors encoded by two cDNAs consisted of 86 amino acids, including a signal peptide, cGnRH-II decapeptide and a GnRH-associated peptide (GAP) linked by a Gly-Lys-Arg proteolytic site. The results of intron trapping and Southern blot showed that two differing cGnRH-II genes in common carp genome were further identified, and that two genes might exist as a single copy. The multi-gene coding of common carp cGnRH-II gene offered novel evidence for gene duplica-tion hypothesis. Using semi-quantitative RT-PCR, expression and relative expression levels of cGnRH-II genes were detected in five dissected brain regions, pituitary and gonad of common carp. With the exception of no mRNA2 in ovary, two cGnRH-II genes could be expressed in all the detected tissues. However, expression levels showed an apparent difference in different brain regions, pituitary and gonad. According to the expression characterization of cGnRH-II genes in brain areas, it was presumed that cGnRH-II might mainly work as the neurotransmitter and neuromodulator and also operate in the regulation for the GnRH releasing. Then, the ex-pression of cGnRH-II genes in pituitary and gonad suggested that cGnRH-II might act as the autocrine or paracrine regulator. | LI Shuangfei HU Wei WANG Yaping ZHU Zuoyan | 2004 | Science China(Life Sciences)2004，47，4： | 8 |
| 11 | Heritable/conditional genome editing in C. elegans using a CRISPR-Cas9 feeding system显示文摘 | Pengpeng Liu Lijiang Long Kai Xiong Bo Yu Nannan Chang Jing-Wei Xiong Zuoyan Zhu Dong Liu | 2014 | Cell Research2014，24，7： | 8 |
| 12 | Regulation of transcriptionally active genes via the catalytically inactive Cas9 in C. elegans and D. rerio显示文摘 | Lijiang Long Hong Guo Di Yao Kai Xiong Yongjun Li Pengpeng Liu Zuoyan Zhu Dong Liu | 2015 | Cell Research2015，25，5： | 7 |
| 13 | Surrogate production of genome-edited sperm from a different subfamily by spermatogonial stem cell transplantation显示文摘The surrogate reproduction technique,such as inter-specific spermatogonial stem cells(SSCs)transplantation(SSCT),provides a powerful tool for production of gametes derived from endangered species or those with desirable traits.However,generation of genome-edited gametes from a different species or production of gametes from a phylogenetically distant species such as from a different subfamily,by SSCT,has not succeeded.Here,using two small cyprinid fishes from different subfamilies,Chinese rare minnow(gobiocypris rarus,for brief:Gr)and zebrafish(danio rerio),we successfully obtained Gr-derived genome-edited sperm in zebrafish by an optimized SSCT procedure.The transplanted Gr SSCs supported the host gonadal development and underwent normal spermatogenesis,resulting in a reconstructed fertile testis containing Gr spermatids and zebrafish testicular somatic cells.Interestingly,the surrogate spermatozoa resembled those of host zebrafish but not donor Gr in morphology and swimming behavior.When pou5f3 and chd knockout Gr SSCs were transplanted,Gr-derived genome-edited sperm was successfully produced in zebrafish.This is the first report demonstrating surrogate production of gametes from a different subfamily by SSCT,and surrogate production of genome-edited gametes from another species as well.This method is feasible to be applied to future breeding of commercial fish and livestock. | Fenghua Zhang Yongkang Hao Xianmei Li Yi Li Ding Ye Ru Zhang Xiaosi Wang Mudan He Houpeng Wang Zuoyan Zhu Yonghua Sun | 2022 | Science China(Life Sciences)2022，65，5： | 7 |
| 14 | Designing future farmed fishes using genome editing显示文摘Aquaculture,the most rapidly growing food production sector,has exhibited tremendous expansion over the past several decades and it currently accounts for more than half of the global fish production for human consumption.To ensure sustainable aquaculture,it is necessary to breed farmed fish species(strains)with valuable economic traits,such as high production,high quality,disease resistance,and stress tolerance.Modem fish genetic breeding techniques largely rely on key processes including understanding the genetic basis of economically important traits to the generation and selection of favorable genotypes. | Yonghua Sun Zuoyan Zhu | 2019 | Science China(Life Sciences)2019，62，3： | 6 |
| 15 | Reverse Genetic Approaches in Zebrafish显示文摘Zebrafish (Danio rerio ) 是一个生长得很好的脊椎动物动物模型。反向的遗传工具的一个全面集合为在这个有用有机体学习基因功能被开发了。Morpholino 是最广泛地使用的试剂 post-transcriptionally 将目标基因表示击倒。很长时间,指向了染色体修正重重地在大规模传统的向前基因的屏幕上被依靠了,例如 ENU (N-ethyl-N-nitrosourea ) mutagenesis 导出的 TILLING (指向在染色体导致了本地损害) 策略和伪 typed retrovirus 调停了 insertional mutagenesis。最近,设计了 endonucleases,包括 ZFN (锌手指核酸酶) 和 TALEN (抄写像使活跃之物的受动器核酸酶) ,提供新、有效的策略直接由在目标基因导致两倍海滨裂缝产生地点特定的 indel 变化。这里,我们为使用的 loss-of-function 研究总结主要反向的基因途径,包括基于为地点特定的基因指向的染色体宽的 mutagenesis 和方法的策略并且在 zebrafish 新兴。未来方向和期望将也被讨论。 | Peng Huang Zuoyan Zhu Shuo Lin Bo Zhang | 2012 | Journal of Genetics and Genomics2012，39，9： | 6 |
| 16 | Time course of foreign gene integration and expression in transgenic fish embryos显示文摘Using a nuclear transplantation approach, the integration and expression of the green fluorescent protein (GFP) gene in the embryogenesis of transgenic loach (Misgurnus anguillicaudatus Cantor) have been studied. The GFP gene expression is first observed at the gastrula stage, which is consistent with the initiation of cell differentiation of fish embryos. The time course of the foreign gene expression is correlated with the regulatory sequences. The expression efficiency also depends on the gene configuration: the expression of pre-integrating circular plasmid at early embryos is higher than that of the linear plasmid. The integration of the GFP gene is first detected at the blastula stage and lasts for quite a long period. When two types of different plasmids are co-injected into fertilized eggs, the behavior of their integration and expression is not identical. | Haobin Zhao Shangping Chen Yonghua Sun Yaping Wang Zuoyan Zhu | 2000 | Chinese Science Bulletin2000，45，8： | 5 |
| 17 | Effect of Shot Peening and Plasma Electrolytic Oxidation on the Intergranular Corrosion Behavior of 7A85 Aluminum Alloy显示文摘The effects of shot peening(SP) and plasma electrolytic oxidation(PEO) on the intergranular corrosion behavior of the novel high strength aluminum alloy 7A85(AA 7A85) were investigated by electrochemical polarization and electrochemical impedance tests.The intergranular corrosion mechanism of SP,PEO and PEO combined with sealingtreated AA 7A85 was studied by the metallographic analysis,residual stress testing,X-ray diffractometer analysis and scanning electron microscopy.The results show that AA 7A85-T7452 is very sensitive to intergranular corrosion.SP would significantly improve its intergranular corrosion resistance.This is attributed to the combination action of residual compressive stress and grain refinement.PEO would reduce the largest corrosion depth by 41.6%.Moreover,PEO without sealing did not eliminate the intergranular corrosion due to the existence of the micropores and microcracks in the oxide coating.However,PEO combined with the SiO2sol–gel sealing treatment could effectively protect the AA 7A85-T7452 from intergranular corrosion because of the good corrosion resistance and barrier function of the sealed coating. | Zuoyan Ye Daoxin Liu Chongyang Li Xiaoming Zhang Zhi Yang Mingxia Lei | 2014 | Acta Metallurgica Sinica(English Letters)2014，27，4： | 4 |
| 18 | Nuclear transplantation in different strains of zebrafish显示文摘Single later blastula nuclei from AB strain of zebrafish (Danio rerio) were transplanted into enucleated unfertilized eggs of Long fin strain. Of 1119 cloning embryos, 14 reconstructed embryos developed into fry. DNA fingerprinting systems of the cloned fish were similar to those of the nuclear donor fish, but were distinctly different from those of the nuclear recipient fish. It confirmed that the genetic material originated from nuclear donor cell other than from nuclear recipient egg. The research suggested that the basic technique for nuclear transplantation performed with different strains of zebrafish has made a breakthrough. It should be helpful for the study of some important developmental problems such as gene function, the regulation of gene expression during animal development, the developmental potential of a nucleus and the interactions between the donor nucleus and the recipient cytoplasm, etc. | Wei Hu Yaping Wang Shangping Chen Zuoyan Zhu | 2002 | Chinese Science Bulletin2002，47，15： | 4 |
| 19 | Acute and chronic un-ionized ammonia toxicity to ‘all-fish’ growth hormone transgenic common carp(Cyprinus carpio L.)显示文摘Ammonia is toxic to fish in natural and artificial waters.We evaluated the acute(96 h) and chronic(21 d) toxicity of un-ionized ammonia to GH transgenic common carp(Cyprinus carpio L.) and non-transgenic common carp using a static-renewal bioassay.The 24,48,72 and 96 h median lethal concentrations(LC50) of un-ionized ammonia were slightly lower in transgenic carp(2.64,2.44,2.28 and 2.16 mg N/L,respectively) than in non-transgenic carp(2.70,2.64,2.52 and 2.33 mg N/L,respectively).Similarly,the median lethal time(LT50) was significantly shorter for transgenic carp(1.41,7.91 and 117.42 h) than for non-transgenic common carp(2.53,14.06 and 150.44 h) following exposure to 3.86,3.29,or 2.09 mg N/L,respectively.Moreover,the mortality of transgenic carp was significantly higher than that of non-transgenic carp at all un-ionized ammonia concentrations((0.91 ± 0.12),(0.48 ± 0.06) and(0.12 ± 0.01) mg N/L) during the 21 d chronic toxicity test.Our results suggest that GH transgenic carp are less tolerant of unionized ammonia than non-transgenic carp.Our data are useful for evaluating potential environmental risk,optimizing stocking density in intensive aquaculture and establishing water quality criteria for ammonia in aquaculture. | GUAN Bo HU Wei ZHANG TangLin DUAN Ming LI DeLiang WANG YaPing ZHU ZuoYan | 2010 | Chinese Science Bulletin2010，55，35： | 4 |
| 20 | The integration characteristics of the exogenous growth hormone gene in a transgenic common carp(Cyprinus carpio L.) with fast-growth performance显示文摘The genetic stability and expression efficiency of exogenous genes in transgenic animals are closely related to integration site and copy number. In our laboratory, by transgenic manipulation and subsequent test crosses, we established an ‘‘all-fish'' growth hormone(GH)transgenic common carp family that exhibits fast growth.In this present study, genome walking, real-time quantitative polymerase chain reaction, and fluorescence in situ hybridization techniques were applied to identify the integration characteristics of the exogenous grass carp GH gene in the transgenic common carp. The exogenous GH genes, in the form of two complete and one incomplete tandem repeats, were found to have integrated into an ATrich region near the end of a chromosome pair. We hypothesize that the high efficiency of exogenous GH gene expression might be due to the low copy number in the genome and the AT-rich integration site. | Ji Chen Qing Luo Hairong Bao Lanjie Liao Yongming Li Zuoyan Zhu Yaping Wang Wei Hu | 2015 | Science Bulletin2015，60，19： | 3 |
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