小鼠H-Y单克隆抗体ELISA检测方法的建立   总被引：2，自引：0，他引：2  

杭苏琴  刘为敏  牛树理  冯紫云 《中国兽医学报》2004,24(6):547-549

以纯系 BAL B/ c雄性小鼠脾细胞腹腔注射免疫同系雌性小鼠 9次 ,获得的抗血清经雌、雄鼠脾细胞吸收后用于精子细胞毒性试验 ,测得 H- Y抗血清效价为 1/ 16 0。选取免疫应答最好的雌鼠脾细胞与 SP2 / 0骨髓瘤细胞融合 ,用精子细胞毒性方法筛选效价较高的细胞株制备腹水 ,建立优化的 EL ISA反应条件。优化后的 EL ISA反应条件为 :抗原4℃过夜 ,加 H- Y抗血清 37℃反应 12 0 m in,加 HRP- Ig G 37℃反应 30 min,加 TMB 2 5℃ 30 min。优化后的 EL ISA与常规 EL ISA比较 ,可以明显降低阴性吸光值 ,检测灵敏度从 1/ 32 0上升为 1/ 12 80  相似文献   

Evaluation of a combined penetrometer for simultaneous measurement of penetration resistance and soil water content     

Yurui Sun  Peter Schulze Lammers  Daokun Ma 《植物养料与土壤学杂志》2004,167(6):745-751

A combined penetrometer is an appropriate tool to measure the soil cone resistance and the water‐content profile. As a relatively new technique, a combined capacitance‐penetrometer for the simultaneous measurement of cone index and soil water content was developed at the Department of Agricultural Engineering of Bonn University in 2002. The objective of this study was the evaluation of the effectiveness and applicability of the innovated penetrometer with a focus on three aspects: (1) A capacitance sensor with two electrode configurations was calibrated for silt loam, sandy loam, and sand. The calibration results show that both electrode configurations have sufficient water‐content sensitivity, but soil‐specific calibrations seem necessary. (2) Under laboratory conditions, the dynamic resolution and response of the capacitance‐penetrometer were validated, and its radius of influence was determined. (3) The field measurement results demonstrate that this measurement technique can be used to improve the interpretation quality of soil cone index data.  相似文献   

貉受精的细胞学和X射线微区分析研究     

冯怀亮  陈大元 《中国兽医学报》1994,14(2):108-113

应用电子显微镜和Ｘ射线微分析技术，对貉受精过程和受精卵膜元素研究的结果表明，貉卵子的卵丘细胞具有吞噬和过滤功能；卵丘细胞间的精子顶体尚未发生囊泡化，而附着于透明带的精子发生了顶体反应，并以８０°角穿入透明带，在其穿入的前方打开一个通道，最后穿过。穿过透明带的精子以赤道段或顶体后区同卵膜融合，并激发皮质颗粒释放，接着穿入卵内，最终发育成雌、雄原核。原核期受精卵质膜上具有高含量的钙，并呈集团分布，它在皮质颗粒胞吐释放中起着重要作用。  相似文献   

Differential effects of -mannose and 2-deoxy- -glucose on attempted powdery mildew fungal infection of inappropriate and appropriate ramineae     

R. J. Zeyen  W. M. Kruger  M. F. Lyngkjr  T. L. W. Carver 《Physiological and Molecular Plant Pathology》2002,61(6)

Barley, oat and wheat were used as both inappropriate hosts (IH) and appropriate hosts (AH) for three formae speciales of the fungus Blumeria graminis, the causal agent of powdery mildew disease. Treatment with either the glucose analog 2-deoxy- -glucose (DDG) or with -mannose dramatically suppressed penetration resistance in IH and to a much lesser extent in AH combinations. Other effects of DDG and -mannose were strikingly dissimilar. DDG greatly reduced localized autofluorescence at fungal attack sites on epidermal cells, and prevented hypersensitive epidermal cell death (HR). -mannose had little effect on autofluorescence or HR. DDG arrested the development of fungal haustoria and apparently prohibited biotrophy leading to secondary hyphae. -mannose allowed haustorial development and functional biotrophy leading to the production of elongating secondary hyphae. This suggests that B. graminis is in some way capable of utilizing -mannose as a carbon substrate. Results with IH combinations paralleled those of known mlo -barley responses to DDG and -mannose. Results are discussed in relation to specific physiological processes known to be influenced by either DDG or by -mannose, or by both compounds.  相似文献   

Effects of temperature on ascospore germination and penetration of oilseed rape (Brassica napus) leaves by A- or B-group Leptosphaeria maculans (phoma stem canker)     

Y. J. Huang  C. Toscano-Underwood  B. D. L. Fitt †  X. J. Hu  A. M. Hall 《Plant pathology》2003,52(2):245-255

Ascospores of both A-group and B-group Leptosphaeria maculans germinated at temperatures from 5 to 20°C on leaves of oilseed rape. Germination of ascospores of both groups started 2 h after inoculation and percentage germination reached its maximum about 14 h after inoculation at all temperatures. Both the percentage of A-/B-group ascospores that had germinated after 24 h incubation and germ tube length increased with increasing temperature from 5 to 20°C. Germ tubes from B-group ascospores were longer than those from A-group ascospores at all temperatures, with the greatest difference at 20°C. Hyphae from ascospores of both groups penetrated the leaves predominantly through stomata, at temperatures from 5 to 20°C. A-group ascospores produced highly branched hyphae that grew tortuously, whereas B-group ascospores produced long, straight hyphae. The percentage of germinated ascospores that penetrated stomata increased with increasing temperature from 5 to 20°C and was greater for A-group than for B-group L. maculans after 40 h incubation.  相似文献   

水牛精子在胞质内注射后的早期形态变化   总被引：1，自引：0，他引：1  

孟凡丽  韦精卫  石德顺  卢晟盛 《中国兽医学报》2004,24(6):609-612

应用胞质内精子注射 (intracytoplasmic sperm injection,ICSI)技术探讨了水牛卵母细胞胞质与注射精子间的相互作用以及 ICSI精子的形态变化。ICSI后 13h,5 9.4 %的精子头部已膨大 ,且有 18.8%的精子进入解聚状态。雌雄原核发育不同步 ,雄原核在 ICSI后 16 h和 19h的形成率分别为 3.2 %和 4 0 .0 % ;雌原核在 ICSI后 13h已达到 71.9% ,16 h时提高到 90 .3%。经离子霉素与 6 - DMAP联合激活 ,ICSI后 19h产生的胚胎核型以 2 PN PB1 为主 ,其雌原核形成 (激活 )率为 91.3% ,雄原核形成率为 4 0 .0 % ,5 1.3%为孤雌胚。用 5 mm ol/ L 的 DTT预处理精子 1h,可提高精子的解聚率 (30 .9%比 12 .7% ,P<0 .0 5 ) ,但对雄原核的形成率无显著影响 (33.3%比 32 .7% ,P>0 .0 5 )。结果表明 ,水牛精子 ICSI后的雄原核形成时间晚于卵子雌原核形成时间 ,且其比率低于卵子 ,DTT处理精子能提高其解聚率 ,但对雄原核形成无影响  相似文献   

Resistance to Botrytis cinerea in Solanum lycopersicoides is Dominant in Hybrids with Tomato, and Involves induced Hyphal Death   总被引：2，自引：0，他引：2  

Rejane L. Guimarães  Roger T. Chetelat  Henrik U. Stotz 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(1):13-23

Botrytis cinerea causes gray mold disease and affects hundreds of plant species, including tomato (Lycopersicon esculentum). The wild nightshade, Solanum lycopersicoides, is cross compatible with tomato and is more resistant to B. cinerea, thus representing a potential source for crop improvement. Tests involving droplet inoculation of detached leaves and spray inoculation of entire seedlings demonstrated that resistance to B. cinerea varies among S. lycopersicoides accessions, with S. lycopersicoides LA2951 being the most resistant accession tested. Expression of resistance in the intergeneric hybrid (L. esculentum cv. 'VF36' × S. lycopersicoides LA2951) suggested that resistance is at least partially dominant in tomato. A green fluorescent protein-tagged B. cinerea strain was used for confocal microscopic comparison of infection in leaves of S. lycopersicoides and tomato. Even though S. lycopersicoides supported spore germination, there was evidence for hyphal lysis and death 3 days after inoculation, at a time when lesions were expanding on susceptible tomato plants. The reduced frequency of B. cinerea lesion spread on S. lycopersicoides explains why this fungus produced fewer spores in this wild nightshade than in tomato.  相似文献   

In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network          下载免费PDF全文

AH Alm‐Kristiansen  ER Gaustad  G Bai  FB Standerholen  G Klinkenberg  E Kommisrud  KE Waterhouse 《Reproduction in domestic animals》2018,53(2):365-370

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time‐period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital^® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post‐thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital^® (SV) technology, the first commercialized production line of SpermVital^® (C) and by conventional procedure applying Biladyl^® extender (B). Post‐thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital^® technology is improved and is more efficient in conserving post‐thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.  相似文献   

Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation          下载免费PDF全文

P Li  H Du  XM Qiao  ZG Liu  QW Wei 《Reproduction in domestic animals》2018,53(2):472-483

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.  相似文献   

侵彻引信半实物仿真系统设计     

闫鹏杰  张亚 《湖南农机》2016,(2):35-36

鉴于侵彻引信实际作战环境的复杂多变性，设计一种半实物仿真系统对于引信综合性能的评估具有积极意义。首先重点分析了侵彻过载数据库的建立和侵彻引信的起爆控制模块的设计。为检验半实物仿真系统的实际工作能力，在现有的试验条件以及实现硬目标侵彻引信半实物仿真系统设计的基础之上，对引信的计时起爆控制模式进行了仿真试验研究。  相似文献