Production and Identification of Clone Sheep Using Bone Marrow Mesenchymal Stem Cells as Donor Cell

In order to obtain the cloned sheep using bone marrow mesenchymal stem cells（BMSCs）as the donor cells,the BMSCs of sheep were chosen and reconstructed embryos were built to transfer.10 published microsatellite markers were chosen,and the DNA samples from clone sheep,donor cells and surrogate ewes were amplified,and the relationship of father-son（RCP）was analyzed using the Quantity One for genotyping.The results showed that the reconstructed embryos were successfully built for electric fusion using sheep BMSCs as nuclear donor,and making nuclear transplantation into enucleated mature oocytes of which the fusion rate was 80.62%.20 surrogate ewe were chosen to be implanted with the reconstructed embryos at morula stage by implant surgery,and 5 lambs were born and only 3 were survived.The genotype of cloned sheep was in line with the dornor cell and the RCP were more than 99.999%.In conclusion,the first clone sheep were obtained successfully by using BMSCs as a nuclear donor in this experiment.     

Relationships among condition indices, feeding and growth of walleye in Lake Erie

Abstract  Condition indices are often used as surrogates of fish health, growth, and feeding and to compare ecological well-being among fish populations. In an effort to identify easily measured indices, growth and food consumption were compared with gonadal-somatic index, liver-somatic index (LSI), fat-somatic index and relative weight ( W _r) for ages 1–3 walleye, Sander vitreus (Mitchill), in Lake Erie from 1986 to 1988. The LSI and W _r were significantly correlated with growth rate or food consumption, but correlations were too small to be considered biologically meaningful. Furthermore, no consistent relationships between condition indices and growth or consumption were found among combinations of fish age and season. None of the indices are considered reliable surrogates for more laborious estimates of growth and food consumption for Lake Erie walleye. Significant relationships between W _r and relative abundance of key prey species warrant further investigation.     

Grain Protein Content of Interspecific Progenies Derivedfrom the Cross of African Rice (Oryza glaberrima Steud.) and Asian Rice (Oryza sativa L.)

Abstract

The protein contents of the grain of 50 interspecific progenies developed from the cross between WAB56-104, an Oryza sativa variety, and CG 14, an Oryza glaberrima line, were investigated. In contrast to the higher protein content of O. glaberrima than O. sativa on the average, the protein content of CG 14 was always lower than that of WAB56-104. However, judging from the average of three seasons, 72% of the interspecific progenies had a higher protein content than the mid-parent and 50% of them had a higher protein content than WAB56-104. Although the actual values of protein content of the interspecific progenies were significantly different among the seasons, a highly significant correlation was always observed in protein content between any two of the three seasons. Protein content therefore was considered character of each interspecific progeny though it was also affected by environment. A significant correlation was not observed between paddy yield and protein content in any season; several interspecific progenies showed higher protein content and paddy yield than the mid-parents. A low paddy yield is likely to be associated with high protein content through physiological regulation without a genetic linkage between the two traits. However, the results suggest that the transgressive segregation of protein content observed in the interspecific progenies is attributed not to this physiological regulation but to a certain mechanism to concentrate protein in grains with a genetic background.     

Development and application of fluorescence in situ hybridization (FISH) method for simple and rapid identification of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella in cultured and natural seawater

ABSTRACT:   The toxic dinoflagellates Alexandrium tamarense (Lebour) Balech and A. catenella (Whedon and Kofoid) Balech produce potent neurotoxins, such as saxitoxin and gonyautoxin and have been mainly responsible for paralytic shellfish poisoning (PSP) in Japan. To prevent a negative effect on the fishery industry, it is necessary to identify these toxic species precisely and rapidly before and during the bloom. In this paper, a rapid and simple protocol of a fluorescence in situ hybridization (FISH) method using ribosomal RNA (rRNA)-targeted probes has been established for identifying the cultured strains and natural cells of A. tamarense and A. catenella . Using the FISH method established in this study, it was possible to identify these toxic species species-specifically and rapidly, within 30 min. The procedure of detection constituted three steps: (i) fixation/dehydration; (ii) hybridization; and (iii) washing; this made the identification simple. Moreover, this method did not require either special techniques or equipment, and the cost for detection was low. The specificity, rapidity, and simplicity of the developed method suggest that it might be useful for routine monitoring of these toxic microalgae.     

Frankia共生固氮基因的分离与克隆

利用ＤＮＡ－ＤＮＡ杂交方法，分离细枝木麻黄的Ｆｒａｎｋｉａ菌株Ｃｏ０１的ｎｉｆ克隆ｐＣｃ１ＧＸ，赤杨内生菌株Ａｔ４的ｎｉｆ克隆ｐＡｔ１ＧＸ及沙棘的ＦｒａｎｋｉａＨｒ１８的ｎｉｆ克隆ｐＨｒ１８ＧＸ、ｐＨｒ１１－③ＧＸ。用基因功能互补法，从Ｆｒａｎｋｉａ菌株Ａｔ４的基因文库中分离到二个可能可互补豌豆根瘤菌ｎｏｄ基因功能的克隆ｐＡｔ２ＧＸ、ｐＡｔ３ＧＸ，并制作了ｐＡｔ２ＧＸ、ｐＡｔ３ＧＸ的亚克隆，予进一步实验，获得充分的证据证明ｐＡｔ２ＧＸ、ｐＡｔ３ＧＸ是否带有Ｆｒａｎｋｉａ菌株Ａｔ４的ｎｏｄ基因．     

聚乙二醇细胞融合技术在牛手工体细胞克隆中的应用初探

本研究分3个实验。实验1对比了不同浓度的离子霉素对牛去透明带卵母细胞孤雌激活的效果;实验2比较了手工半卵切割法去核与显微半卵切割法去核的效果;实验3对聚乙二醇（PEG）细胞融合技术在牛手工体细胞克隆的应用进行了初步探讨。结果表明,对于去透明带牛卵母细胞孤雌激活,离子霉素浓度为2．0μmol／L组的激活效果最好,其囊胚发育率（29．4％）极显著地高于浓度为5．0μmol／L的对照组和0．5μmol／L组（囊胚率分别为8．3％和9．4％,P〈0．01）;采用半卵切割法去核,手工切割的半卵存活率（85．6％）与显微操作（90．3％）差异不大,切割速率（约100枚卵／h）相仿。对双半卵与体细胞进行同步融合处理时,应用50％浓度的PEG融合率（39．3％）极显著好于45％、55％和60％的PEG（融合率分别为0％、16．2％和5．3％,P〈0．01）。     

利用远缘杂交创造核果类果树新种质的研究 I.不同处理对核果类果树远缘杂交亲和性的效应研究

 以桃、杏、李、樱桃、杏梅等核果类果树的9个品种为亲本共69个组合进行了远缘杂交,研究了不同处理对核果类果树远缘杂交亲和性的效应。结果表明,铃铛花期授粉的坐果率显著高于初花期授粉的坐果率;同一杂交组合,正反交坐果率差异显著;母本对远缘杂交亲和性影响很大,父本对远缘杂交亲和性的影响较小;适宜场强的静电场、He-Ne激光处理及60Coγ射线与He-Ne激光联合处理花粉均能显著提高花粉离体萌芽率,用上述处理的花粉进行远缘杂交,坐果率明显高于对照;60Coγ射线处理则降低了花粉离体萌芽率,远缘杂交的坐果率也低于对照。     

用荧光原位杂交技术检测黑麦染色质

荧光原位杂交技术（fluorescence in situ hybridization ,FISH）是一种快速、准确、直接检测和定位外源染色质的新技术。本研究荧光原位杂交用毛地黄毒苷-11-dUTP标记黑麦总DNA作探针,以检测小麦中黑麦染色质。结果如下：八倍体小黑麦（2n=8x=56）有丝分裂中期有14条染色体显示黄色,42条染色体显示红色,红色间期核显示有14个黄色亮点,表明这八倍体小黑麦含14条黑麦染色体。八倍体小黑麦中黄色黑麦染色体显C-带和H-带的末端有绿色的带。这是由于黑麦染色体末端结构异染色质含量高所致。黑麦附加系2R的间期细胞核中有1 ～ 2个黄色亮点。这表明2R含有1至2条黑麦染色体。     

PCR技术在住肉孢子虫虫种鉴定上的应用

采用 RAPD 方法对水牛梭形住内孢子虫、待定种及黄牛枯氏住内孢子虫缓殖子基因组 DNA 进行了分析,待定种 PCR 产物的电泳带型与枯氏住肉孢子虫的差异较梭形住内孢子虫大,这与二者表型性状一致的研究结果不吻合.对3种包囊种特异性的 PCR 指纹进行了筛选,回收和纯化了3种包囊种特异性的 DNA 片段,待定种0.7kb 片段经α-~(32)PdATP 标记后用作探针,只与同源 DNA 杂交,不与其它包囊、宿主细胞和艾美耳球虫杂交,用 pGEM-T Easy Vector 对该片段克隆成功,并测出其全长序列,为进一步设计特异引物建立标准 PCR 诊断方法和用作核酸探针奠定了基础。     

中介亲本在黑白杨派间杂交中的应用

该文利用黑杨与青杨的 4个杂种作中介亲本 ,做了 12个组合的黑白杨杂交试验并与 10个由各派内的杂种或原种组成的黑白杨杂交组合进行杂交亲和性与杂种子代高生长分化程度的对比试验 .结果表明 :①中介亲本的杂交亲和性优于派内树种亲本 ;②不同中介亲本或同一中介亲本在不同组合中亲和性差异较大 ;③部分派内杂种亲本的亲和性高于中介亲本 ;④杂交子代的高生长分化程度最大为中介亲本组合 ,最高 /最低达 7倍 ,其次为派内杂种亲本组合 ,最小为派内原种亲本组合 ;⑤中介亲本与派内杂种亲本的花粉管比原种亲本更易进入柱头