Embryonic stem cells in fish: current status and perspectives   总被引：5，自引：0，他引：5  

Y. Hong  S. Chen  M. Schartl 《Fish physiology and biochemistry》2000,22(2):165-170

Totipotent embryonic stem (ES) cells represent a bridge that links in vitro and in vivo manipulations of animal genomes and have enormous potential for genetic engineering of livestock. We have recently established feeder cell-free conditions for culturing cells of midblastula embryos (MBE) of the medaka (Oryzias latipes) and obtained several stable cell lines that show all features of mouse ES cells in vitro. One of these lines, MES1, has been demonstrated to retain a diploid karyotype and can be induced to differentiate into various cell types in vitro. Upon microinjection into albino host blastulae, MES1 cells are able to form pigmented chimeras. Genotype-specific PCR analysis revealed that 90% of host blastulae transplanted with MES1 cells developed into chimeric fry. This high frequency was not compromised by cryostorage or DNA transfection of the donor cells. Transplantation of genetically labelled MES1 cells revealed a wide contribution to numerous organs derived from all three germ layers and differentiation into various types of functional cells. These ES properties of MES1 line was not abolished by stable gene transfer and long-term selection. Thus MES1 cells may represent a first promising cellular vehicle for the production of genetically modified fish. The genetic background has been found to have a profound effect on the efficacy of ES cell derivation and of chimera formation.  相似文献   

金冠苹果成熟胚离体诱导四倍体的研究   总被引：3，自引：0，他引：3  

李林光  何平  欧春青  李慧峰  张志宏 《园艺学报》2007,34(5):1129-1134

 以自然授粉的‘金冠’苹果种子为材料，研究了秋水仙素处理成熟胚离体诱导四倍体技术和嵌合体分离方法。结果表明：将完整胚在0.5%秋水仙素+1%二甲基亚砜的混合溶液中暗处理48 h，然后接种在MS+TDZ 1.0 mg·L^-1+IBA 0.5 mg·L^-1+蔗糖30 g·L^-1+琼脂6 g·L^-1的再生培养基中，变异率高达10.6%。与二倍体相比，四倍体试管苗生长缓慢，节间变短，叶片增大，叶形指数趋小化，气孔密度降低，保卫细胞增大。经压片法鉴定茎尖染色体数和流式细胞仪分析DNA含量，表明多数变异植株为同质四倍体。利用叶片离体再生器官技术，从诱导获得的嵌合体植株中分离出同质突变体。  相似文献   

花叶矮牵牛嵌合体再生体系的建立     

吕秀立  张冬梅  钱又宇 《上海农业学报》2007,23(1):34-38

以花叶矮牵牛为材料,研究了外植体、激素浓度、光照强度、叶片生理状态、暗培养时间等因素对诱导嵌合体不定芽的影响,建立了花叶矮牵牛嵌合体再生体系。结果表明:无菌苗在2000 lx光照条件下,置于MS 6-BA 0.2 mg/L培养基上,嵌合体诱导率最大达到90.2%;生理年龄为20 d的叶片,在MS 6-BA 0.4 mg/L IBA0.01 mg/L培养基中,嵌合体诱导率最大达到34.8%,暗培养会降低嵌合体的诱导率。  相似文献   

Induction of cytoplasmic male sterility in the seed progeny derived from artificially-synthesized interspecific chimera in Brassica     

Y. Hirata  T. Motegi  Y. Takeda  K. Morikawa 《Euphytica》2001,117(2):143-149

We produced artificial interspecific chimeras by in vitro grafting, and obtained cytoplasmic male sterile (CMS) variants in the seed progenies derived from backcrossing the chimera with one of the mother plants, B. campestris cv. Komatsuna. The induced CMS has been stably inherited by crossing it with `Komatsuna', not with `Ruby Ball' cabbage. The nuclear component of CMS is complete `Komatsuna' type in morphology, chromosome number (2n = 20) and Southern blot using ribosomal 17S RNA gene as a probe. PCR analysis by using mitochondrial atpA primer showed the complete `Ruby Ball' type, suggesting nuclear-cytoplasmic exchange. However, Southern blot patterns were different among those of the CMS and both parents by using atpA. Recombination or some unknown change is supposed in the mitochondrial genome via the processes of synthesis and propagation of the chimeras.  相似文献   

Vegetative and floral characteristics of interspecific Brassica chimeras produced by in vitro grafting     

Takashi Noguchi  Yutaka Hirata 《Euphytica》1993,73(3):273-280

Summary Interspecific Brassica chimeras (Brassica campestris+B. oleracea) were synthesized according to an in vitro graftculture method, and propagated by tissue culture of axillary buds or chimeric explants. A total of 127 regenerants obtained were investigated. The mericlinal chimera type was more easily produced than the other periclinal and revertant types. No sectorial chimera was produced. The flowering habit and inflorescence type of the chimeras were found to be controlled by the constitution of three tissue layers, but the petal shape and color were controlled only by the two outer tissue layers. Pollen viability of the chimeras were generally lower and more variable in parts than those of the parental types.  相似文献   

也门铁及其嵌合体叶绿素合成特性研究     

刘和平  何业华  林剑波  林顺权  胡桂兵  杨转英 《北方园艺》2011,(14):120-122

以2 a生的也门铁及其嵌合体变异品种‘金心也门铁’和‘金边也门铁’为试材,研究了也门铁及其嵌合体的主要叶绿素合成前体物质。结果表明:‘金边也门铁’叶片变异(黄色)部分叶绿素合成在PBG与UrogenⅢ之间受阻,‘金心也门铁’叶片变异(混合)部分的叶绿素合成在UrogenⅢ与原卟啉之间受阻,导致颜色发生变化;‘金边也门铁’叶片绿部、‘金心也门铁’叶片绿部及也门铁三者之间的叶绿素a、b及叶绿素总含量差异不大,所以三者都呈绿色。  相似文献   

Development of avian embryo manipulation techniques and their application to germ cell manipulation     

Mitsuru NAITO 《Animal Science Journal》2003,74(3):157-168

The development of chicken embryo culture techniques, from single‐cell stage to hatching, makes it possible to manipulate developing embryos at any developmental stage. Production of germline chimeric chickens by the transfer of stage X blastodermal cells or primordial germ cells enables the manipulation of germline cells in vitro. Production of transgenic chickens has been attempted by the retroviral vector method, microinjection of DNA into a fertilized ovum at the single‐cell stage, use of chimeric intermediates produced by the transfer of stage X blastodermal cells or primordial germ cells, manipulation of spermatozoa, and in vivo manipulation of gonads. So far, the only non‐viral method that has successfully produced transgenic chickens is microinjection of DNA into a fertilized ovum. Manipulation of primordial germ cells could become an efficient system for producing transgenic chickens by combining it with the highly efficient transfection method or the in vitro culture system for primordial germ cells. Preservation of avian genetic resources has now become possible by cryopreservation of stage X blastodermal cells or primordial germ cells as well as spermatozoa. The development of nuclear transfer techniques for avian species is necessary.  相似文献   

供体PGCs在鸡-鸭嵌合体胚胎性腺外的发育   总被引：1，自引：0，他引：1  

马玉忠  李赞东  沙金  刘春海  王宁 《中国兽医学报》2002,22(4):397-398

通过微注射法将鸡原始生殖细胞 (PGCs)注入到鸭的胚盘下腔中 ,用鸡 W染色体 DNA探针通过原位杂交的方法对供体 PGCs在嵌合体胚胎性腺外的发育作了研究。结果表明 ,所取的 4 8枚胚胎中 ,有 18.8%的脑内和 2 3.9%的神经管周围出现了阳性信号  相似文献   

苹果试管染色体组工程育种   总被引：7，自引：0，他引：7  

石荫坪  王强生 《落叶果树》1993,25(4):1-6

用秋水仙０．４％和０．８％的浓度，诱变苹果７个二倍品种自然授粉的胚，通过试管培养，抑制胚芽成梢，促使染色体组突变的四倍体细胞分化不定芽，形成植株。经染色计数梢端切片，鉴定出一批同质的四倍体株系，部分植已栽植于果园。此项研究了简易可靠的诱变，有效地避免或减轻了浣诱变中嵌合体的干扰，为人工创造四倍种质，有计划地进行多倍体育种开辟了新途径。  相似文献   

鸡胚原始生殖细胞的分离和鸡鸭嵌合体的制备研究   总被引：1，自引：0，他引：1  

刘春海  李赞东  黄劲松  沙金  危华  赵晨  孙明军 《河北农业大学学报》2001,24(3):62-66

探索了 12～ 17期鸡胚血液中的原始生殖细胞 (PGCs)迁移数量变化规律 ,将其在液氮中冷冻保存。并以Fi coll密度梯度离心、MiniMACS磁气分离、滤膜 3种方法对PGCs进行分离 ,结果发现 ,12～ 17期血液中均有PGCs存在 ,13期达到高峰 (4 7 1± 10 5 )个·μL-1,在血液中比例为 0 0 12 6 %。冷冻保存 3个月后解冻成活率达 80 %以上。3种分离方法所得的分离效果分别为 95 7%、39 2 %、6 3 0 % ;纯度为 2 7 5 %、8 4%、3 1%。将分离的原始生殖细胞以微注射法转移至 14～ 15期麻鸭胚胎中制备了鸡鸭种间嵌合体 ,获得 8只雏鸭 (8/ 110 )。以鸡W特异性DNA探针原位杂交法在早期鸭胚性腺中检测到鸡原始生殖细胞 ,嵌合率达 84 2 % (16 / 19)。表明鸡原始生殖细胞能够迁移定居到鸭胚性腺中 ,并有可能增殖分化成有功能的配子  相似文献