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1.
探讨了不同浓度的胰岛素和葡萄糖对水牛体外受精早期胚胎体外发育的影响。结果表明,添加胰岛素浓度为5μg/mL时,水牛早期胚胎的囊胚发育率、囊胚细胞数及冷冻-解冻后胚胎的孵化率都显著高于对照组及其他浓度组(P<0.05);加入葡萄糖的浓度大于6 mm o l/L时显著影响水牛早期胚胎发育到囊胚及孵化囊胚的阶段(P<0.05);联合添加胰岛素和葡萄糖时能显著提高水牛早期胚胎的孵化囊胚率、总囊胚率和囊胚细胞数(P<0.05)。实验证实,浓度为5μg/mL胰岛素能促进水牛体外受精早期胚胎的发育;在TCM-199中单独添加葡萄糖是不必要的,联合添加葡萄糖和胰岛素能促进水牛体外受精早期胚胎的发育。  相似文献   

2.
以黄牛(Bos taurus)卵母细胞为受体,牦牛(Bos grummiens)耳成纤维细胞为核供体进行体细胞核移植,研究供体细胞性别、细胞周期同期化处理方式及冷冻对异种克隆牦牛胚胎发育的影响.结果显示,雄性供体细胞的异种克隆牦牛囊胚发育率显著高于雌性供体细胞(56.6% vs 39.5%,P<0.05),但卵裂率和囊胚细胞数差异不显著;血清饥饿和接触抑制提供的供体细胞对异种克隆牦牛胚胎发育率和胚胎质量的影响没有明显差别;供体细胞冷冻-解冻组的克隆胚胎卵裂率明显低于新鲜消化组(54.5% vs 78.2%,P<0.05),但两组之间的囊胚率和囊胚细胞数没有显著差异.说明供体细胞的性别对异种克隆牦牛囊胚发育有影响,而同期化处理和细胞冷冻对异种克隆牦牛囊胚发育影响很小.  相似文献   

3.
研究了生物频谱辐射对小鼠桑椹胚玻璃化冷冻后体内外发育的影响，并采用荧光染色法对经生物频谱辐射前后的胚胎进行囊胚细胞计数，以评估生物频谱对冷冻损伤的恢复程度。结果表明在EFS40 玻璃化溶液中处理1min一步法冷冻保存的胚胎，解冻后经生物频谱辐射，其发育率由对照组的54％提高到98％，差异极显著（P< 0.01）。冷冻胚胎辐射后移植于同期化处理的受体子宫中，妊娠率和产仔率与对照组相比差异均不显著（40％∶56％；50％∶42％）（P> 0.05）。将新鲜胚、经辐射的新鲜胚、冷冻胚、冷冻后经生物频谱照射共四组胚胎，用 H33342 染色，证实冷冻后胚胎的囊胚细胞数与对照组（新鲜胚）相比显著减少（24±3.4∶52±5.4）（ P< 0.01）。解冻后的胚胎经生物频谱辐射后，其囊胚细胞数显著高于不加辐射组（35±5.624±3.4）（P< 0.01）。另外，新鲜胚胎用生物频谱辐射后，其囊胚细胞数（49±3.1） 与对照组相比无显著差异（P> 0.05）。  相似文献   

4.
通过小鼠(Mus musculus)胚胎体外培养,研究了培养基与不同氧分压环境之间是否存在偏好性;同时利用人体呼出的生理混合气体(肺气)对小鼠胚胎体外培养的可行性进行了探索;还比较了微孔(well of the well,WOW)培养法和成组培养法对胚胎发育的影响.结果表明:①在4%CO2+16%O2+78%N2+2%H2O(肺气)、5%CO2+5%O2+90%N2(5%O2,低氧)和5%CO2+20%O2+75%N2(20%O2,高氧)三种气相条件下,CZB和mKSOM两种培养基之间的胚胎卵裂率及囊胚率均差异不显著(P>0.05),但是,在肺气组中,CZB组的囊胚平均总细胞数明显高于mKSOM组(P<0.05);②低氧条件下,mKSOM组的囊胚率(22.6%)显著高于高氧和肺气条件下mKSOM组的囊胚率(P<0.05);③CZB培养基在低氧条件下获得的囊胚率与高氧或肺气条件下所获囊胚率差异不显著(P>0.05);④WOW培养法的囊胚发育率(74.6±5.1%)和囊胚平均总细胞数(76±2)显著高于成组培养法(分别为38.2±6.6%和58±4)(P<0.05).结果提示:培养基mKSOM和CZB在支持胚胎发育时对氧分压具有偏好性;肺气能有效支持胚胎的正常发育;WOW培养法有利于提高胚胎体外发育能力和囊胚质量.  相似文献   

5.
本文以黄牛卵母细胞胞质为受体,牦牛耳成纤维细胞为核供体进行体细胞核移植,研究供体细胞性别、细胞周期同期化处理方式及冷冻对牦牛异种克隆胚胎发育的影响。结果显示：雄性供体细胞的牦牛异种克隆囊胚发育率显著高于雌性供体细胞（56.6% vs 39.5％,P﹤0.05）,但卵裂率和囊胚细胞数差异不显著;血清饥饿和接触抑制提供的供体细胞对牦牛异种克隆胚胎发育率和胚胎质量的影响没有明显差别;供体细胞冷冻后解冻组的克隆胚胎卵裂率明显低于新鲜消化组（54.5％ vs 78.2％,P﹤0.05）,但两组之间的囊胚率和囊胚细胞数没有显著差异。说明供体细胞的性别对牦牛异种克隆囊胚发育有影响,而同期化处理和细胞冷冻对牦牛异种克隆囊胚发育影响很少。  相似文献   

6.
来源于体外成熟和体外受精的牛早期胚胎与不同细胞的单层细胞共同培养，以检测不同阶段胚胎的细胞数。本研究共检测了３１５３枚桑椹胚和囊胚。通过体外培养４～１０ｄ，不同阶段的胚胎，早期桑椹胚、紧凑型桑椹胚、早期囊胚、囊胚、待扩展囊胚、扩展囊胚、待孵化囊胚和孵化囊胚，经检测其细胞数分别为：４５，６９，７０，８８，１０３，１２７，１３９和１５１。第９ｄ的囊胚细胞数比第７ｄ或第８ｄ的囊胚细胞数无显著差异，然而，不同等级的囊胚的细胞数差异显著（Ｐ＜０．０５）；同时，细胞数少的胚胎发育也慢。通过囊胚免疫法检测，第７ｄ、第８ｄ和第９ｄ回收的扩展囊胚的细胞数分别为６６，６０和５２。此外，虽然在子宫内膜细胞或输卵管细胞－子宫内膜细胞的单层细胞上的囊胚发育慢，且发育率低，但其细胞数与在其它单层细胞上发育的囊胚相比无显著差异。试验结果表明，通过体外化培养出来的囊胚细胞数也能达到体内囊胚的水平。  相似文献   

7.
为研究无血清培养系统对山羊卵母细胞体外成熟及孤雌激活胚胎早期发育力的影响,分别以0.3%牛血清蛋白(BSA)(mV)、1%聚乙烯醇(PVA)(mV)和1%胰岛素转铁蛋白亚硒酸钠(ITS)(VV)代替成熟培养液中胎牛血清(FBS),以添加5?S的OM为对照组,对来源于屠宰场的卵巢卵母细胞复合体(COCs)进行体外培养。成熟率PVA添加组(50.00%)显著低于FBS组(83.23%)(P<0.01)、BSA组(71.19%)和ITS组(76.79%),与FBS组无显著差异(P>0.05);成熟卵母细胞用离子霉素联合6-DMAP激活,在SOFaa中进行孤雌胚培养,BSA、PVA、ITS及FBS各组的卵裂率依次为61.90%、46.97%、79.07%和83.58%,囊胚率分别为25.00%、19.70%、55.81%和60.45%。结果表明:在OM基础培养液中添加BSA或PVA,卵母细胞的成熟率明显低于添加FBS,且卵裂率和囊胚发育率也低于对照组(P<0.05或P<0.01);添加ITS,卵母细胞成熟率虽低于对照组,但孤雌胚卵裂率和囊胚发育率与对照组接近(P>0.05)。表明以ITS代替FBS,可用于山羊卵母细胞体外成熟培养。  相似文献   

8.
绵羊(Ovis arise)卵母细胞经丁内酯-Ⅰ(BL-Ⅰ)抑制培养,然后转入成熟培养液培养不同时间,进行体外受精并观察胚胎发育情况.结果表明,解除8 h抑制再培养8 h组处于MⅡ期卵母细胞率6.38%,显著低于其它组(P<0.01).继续培养16和20 h的卵母细胞成熟率与对照组(常规培养24 h)差异不显著,培养24 h组卵母细胞成熟率(70.83%)稍高于对照组(66.18%),但差异不显著(P>0.05).培养16和20 h组囊胚率分别为2.99%和13.64%与对照组24.39%差异显著(P<0.05),培养24 h组的囊胚率20.33%与对照组差异不显著(P>0.05).表明BL-Ⅰ对绵羊卵母细胞的体外成熟有明显抑制作用,但其抑制作用是可逆的,而经BL-Ⅰ处理后未能提高绵羊卵母细胞体外受精和胚胎的整体发育能力.  相似文献   

9.
试验探索重离子束辐照对绵羊卵母细胞的体外成熟及体外受精卵早期胚胎发育的影响。用不同剂量的重离子束辐照绵羊卵母细胞和精液,观察卵母细胞体外成熟率及体外受精后早起胚胎的卵裂率、桑椹胚率及囊胚率。试验结果显示:与对照组相比,1.0、1.5Gy辐射可以明显提高绵羊卵母细胞的体外成熟率(p0.01);重离子辐照绵羊精液和卵母细胞并做体外受精后,早期胚胎发育有着显著变化。当用0.5、1.0Gy剂量辐照精液时,绵羊早期胚胎的卵裂率和桑葚胚率显著升高(p0.05),囊胚率无显著变化,1.5 Gy剂量,卵裂率显著提高(p0.05),2.0 Gy剂量,卵裂率、桑葚胚率、囊胚发育率均极显著降低(p0.01);当重离子束辐照卵母细胞时,0.5、1.0Gy剂量组的卵裂率、桑葚胚率和囊胚发育率显著升高(p0.05),而2.0Gy剂量组的卵裂率和桑葚胚发育率极显著降低(p0.01),而囊胚发育率为0。结果表明低剂量的重离子辐照绵羊精液、卵母细胞,对体外受精和早期胚胎发育具有明显的促进作用,而高剂量辐照则抑制细胞的生长发育。  相似文献   

10.
OPS法玻璃化冷冻牛卵母细胞和囊胚   总被引：13，自引：5，他引：13  
本文报道了利用拉细开口型细管 (Open Pulled Straw,OPS)方法进行玻璃化冷冻保存体外成熟(IVM)的牛卵母细胞及体外生产 (IVP)的牛囊胚的实验结果。实验 1的牛卵母细胞冷冻处理分 2组 ,组 1(G1 )是将卵母细胞在 IVM2 1 h时去除部分卵丘细胞后马上进行冷冻处理。组 2 (G2 )卵母细胞在 IVM6h时去除部分卵丘细胞 ,然后继续培养至 2 2 h冷冻 ;冷冻后的卵母细胞在液氮中保存 0 .5～ 2 h后解冻再继续 IVM1 h后与正常 IVM2 4h的对照组卵母细胞一起进行体外受精 (IVF)处理。实验 2则冷冻了来自IVP的 6、 7日龄的牛囊胚。结果显示 ,G1的卵母细胞 IVF后 8d的囊胚率 (2 2 .8% ,2 6/1 1 4)和孵化囊胚率 (1 4.9% ,1 7/1 1 4)都极显著地高于 G2 (分别为 2 .9% ,5 /1 71和 0 .5 % ,1 /1 71 ,P<0 .0 0 1 ) ;但两处理组的受精卵裂率差别不大 (分别为 48.2 %和 41 .5 % ,P>0 .3 )。冷冻第 6d和第 7d的体外囊胚 ,其冻后存活率和囊胚孵化率分别为 92 .7%、 89.2 %和 83 .6%、 66.7%。结果表明 ,利用 OPS法玻璃化冷冻经体外培养成熟的牛卵母细胞及体外生产的牛囊胚均能获得良好的效果  相似文献   

11.
低氮和干旱胁迫对富士和秦冠生长及氮素利用的影响   总被引：2，自引：2，他引：0  
【目的】以富士（Fuji）、 秦冠（Qinguan）嫁接在平邑甜茶（Malus hupehensis Rehd.）上的当年生盆栽苗为试验材料,采用砂培方法,研究了缺氮胁迫和干旱对富士和秦冠生长情况、 光合参数、 植株各部位氮磷钾含量及氮素利用效率的影响,分析比较了低氮干旱条件下富士和秦冠生长及氮素利用的差异,以期为果树生产高效肥水利用提供理论指导。【方法】试验共设四个处理: 正常氮正常水（ZZ）、 低氮正常水（DZ）、 正常氮干旱（ZG）、 低氮干旱（DG）。氮素和水分均设置两个水平,分别为正常氮（6 mmol/L NO-3-N）、 低氮（0.3 mmol/LNO-3-N）、 正常供水（保持盆中砂子相对含水量为饱和含水量的80%~85%）、 干旱处理（保持盆中砂子相对含水量为饱和含水量的60%~65%）。【结果】富士和秦冠的生物量（茎和叶）、 株高茎粗等生长指标以及光合速率、 气孔导度、 蒸腾速率均为正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）,并且相对应处理下秦冠的以上指标均高于富士;正常供水下,缺氮处理使富士、 秦冠的根冠比比正常氮处理均有所增加,富士提高了2.05%,秦冠提高了22.40%。富士和秦冠的氮、 磷、 钾含量均表现出正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）; 氮、 钾元素含量在植株各部位的分布顺序依次是叶＞根＞茎,磷元素则是根＞叶＞茎;光合氮素利用效率（PNUE）和氮素利用效率表现为秦冠处理之间差异极显著,富士处理之间差异不显著;秦冠的PNUE和NUE明显高于富士,在低氮正常水（DZ）处理下,秦冠氮肥利用率比富士高42.07%,在低氮干旱（DG）处理下高64.14%;低氮胁迫下富士和秦冠的NUE显著提高,并且秦冠提高的幅度高于富士。【结论】施用氮肥能够显著提高富士与秦冠的干物质量,同等水肥条件下,秦冠生长优于富士;水分亏缺会减少叶片对氮的吸收,干旱条件下适度增施氮肥,可提高果树的抗旱能力;低氮干旱胁迫下秦冠的生长指标、 光合指标及氮素利用效率指标均优于富士,表现出较强的抗低氮干旱胁迫的能力。  相似文献   

12.
Staff members of the Department of Botany of Palacký University in Olomouc and Gene Bank Department – Workplace Olomouc, Research Institute of Crop Production in Prague, Czech Republic, conducted an expedition in seven European countries (Austria, Czech Republic, France, Germany, Italy, the Netherlands, Switzerland) in August/September 1999 to collect wild Lactuca spp. germplasm and study its geographic distribution, ecology and biodiversity. During the mission, more than 600 locations were visited resulting in the collection of 602 seed samples (accessions) of wild Lactuca species and 13 seed samples of related genera (Chondrilla and Mycelis). Lactuca serriola f. serriola, L. serriola f. integrifolia, L. saligna and L. viminea subsp. chondrilliflora were prevalent in southern Europe (Italy, France), however, only L. serriola was common in central and western Europe (Austria, Czech Republic, Germany, Netherlands, Switzerland). The greatest diversity of Lactuca species was found in France, where also the most seed samples (165) were collected. The most characteristic habitats with a high density of Lactuca spp. populations were observed along roads and highways, grassy ditches, ruderal communities, and dust-heaps. Natural infections by powdery mildew (Erysiphe cichoracearum) and downy mildew (Bremia lactucae) on some wild Lactuca spp. were observed. Recent observations concerning the geographic distribution, population structure, habitats, and natural occurrence of diseases of Lactuca spp. are discussed. This assemblage of genetic resources of Lactuca spp. can serve as the basis of future studies of species diversification, spatial population structure, plant microevolution, domestication processes, and genetic variability of host-parasite interactions.  相似文献   

13.
Laser-induced breakdown spectroscopy (LIBS) is a new technique for the analysis of plant material. This study investigates the application of LIBS to pasture-based plant samples. The LIBS measurements were obtained from pelletized pasture samples (100 samples) that had been also analyzed by inductively coupled plasma–optical emission spectroscopy (ICP-OES) following microwave digestion for calibration and comparison purposes. Comparisons for elements sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), boron (B), phosphorus (P), and sulfur (S) showed that LIBS could be used for almost all the standard profile total elements with concentrations down to low mg/kg levels (observed error of Na: 0.024 percent, K: 0.18 percent, Mg: 0.016 percent, Ca: 0.073 percent, P: 0.017 percent, Mn: 31 mg/kg, Fe: 150 mg/kg, Zn: 6.6 mg/kg, and B: 1.1 mg/kg). Elemental analysis at less than mg/kg levels was not possible using LIBS. The elements S and Cu were particularly difficult to analyze with reliability using LIBS at the concentration levels found in the plant samples. Replacing microwave digestion and subsequent ICP analysis with a direct analysis of dried plant samples using LIBS has the potential to improve the productivity and reduce the cost of testing.  相似文献   

14.
We have studied the effect of anaerobically digested (ADMSW) and composted municipal solid waste (CMSW) on mineralization and foodweb dynamics to verify the hypothesis that ADMSW would immobilize N right after addition to soil in contrast to addition with CMSW. Another hypothesis was that the mesofauna (enchytraeids and microarthropods) would stimulate N release from the decomposer community. We measured excretion of -N and urea-N from the mesofauna and hypothesized that enchytraeids would release urea. ADMSW and CMSW were amended to pots with sandy loam and barley. The pots were divided into treatments with or without mesofauna. Mesofauna, plant N and biomass, soil N and ergosterol (fungal biomass) were measured over a 113-day period of four equidistant samplings. Soil respiration, N mineralization and N release by the mesofauna were modelled from concurrent studies. ADMSW- and CMSW-treated soils initially (<20 days) immobilized N. The amendments did not increase plant growth substantially, and this was probably due to N-limitation in the early stages of plant growth. Enchytraeid abundance was about three times higher in ADMSW- than CMSW-treated soils, indicating that ADMSW contained more labile compounds, bacteria, and microfauna. The mesofauna did not affect N-content, but the cumulated -N excreted by the mesofauna was estimated to be substantial and about one-fifth of total plant N in ADMSW. An explanation for this discrepancy might be that in the absence of the mesofauna, other members of the detrivore and microbivore community performed the mesofauna’s function. Neither enchytraeids nor microarthropods excreted urea.  相似文献   

15.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

16.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

17.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

18.
“Wicked” problems are those that are complex and that change when solutions are applied. Many conflicts in conservation fall into this category. The study approached the problem of how to constrain the apparent wickedness of a problem in the conservation management of a species by using simple empirical indicators to carry out iterative assessment of the risk to a population and to document how this risk evolves in relation to the addition of new data and the implementation of management actions. Effects of high levels of uncertainty within data and also concerning population structure were examined through stochastic simulation and by exploration of scenarios. Historical trends in the example used, the Steller sea lion, showed rapid declines in abundance in some regions during the 1980s. The current total population is 130,000-150,000 Steller sea lions through Alaska and British Columbia and this number has been stable since about 1990 in spite of regional differences in population dynamics. Regional differences in the sequence of changes in the number of pups and non-pups, suggested that an internal re-distribution of juveniles could have happened between 1980 and 1990. Current productivity also appears close to the long-term mean. Stochastic population projection using various scenarios showed that, based upon this history, the risk of extinction for the population has declined and is below reasonable thresholds for considering the population to be endangered. The trends in risk suggest that management actions taken since 1990 have probably been effective. Consequently, the conservation management objectives for the Steller sea lion are probably being met. The approach provides a mechanism, based upon experience and scenario analysis, for exploring future policy options and may help to constrain the debate amongst stakeholders about the cost-benefit trade-offs associated with different options.  相似文献   

19.
Nutrient distributions under no tillage (NT) compared with conventional disk-and-bed tillage (CT) management in the warm, humid region of the southeastern USA need to be assessed so that future placement, quantity, and type of fertilizers can be altered, if necessary, to efficiently match crop demands. We determined soil-profile distributions of pH, N, P, S, K, Ca, Mg, Na, Zn, Fe, Mn, and Cu to a depth of 0.9 m at the end of 8.5 years of continuous CT and NT management on a Weswood silty clay loam (fine, mixed, thermic Fluventic Ustochrept) in southcentral Texas. Most dramatic changes occurred within the 0–0.05 m depth, where soil under NT had lower pH, Fe, and Cu than under CT, but greater P, K, Zn, and Mn. Greater P and K under NT than under CT also occurred below the till-zone (0.15–0.3 m). At a depth of 0–0.3 m, soil under NT contained greater amounts of extractable P, K, Zn, Fe, Mn, and Cu than under CT. Nitrogen fertilization had little effect on nutrient distributions, except resulting in greater extractable K at 0–0.05 m and greater nitrate at 0–0.15 m. Few changes in soil-profile distributions were observed for extractable S, Ca, Mg, and Na. Long-term continuous use of NT on this fine-textured, high-fertility (except for N) soil had no apparent adverse effects on nutrient distributions relative to CT, but enhanced conservation and availability of P, K, Zn, Fe, Mn, and Cu near the soil surface where crop roots proliferate.  相似文献   

20.
The responses of tallgrass prairie plant communities and ecosystem processes to fire and grazing are well characterized. However, responses of invertebrate consumer groups, and particularly soil-dwelling organisms, to these disturbances are not well known. At Konza Prairie Biological Station, we sampled soil macroinvertebrates in 1994 and 1999 as part of a long-term experiment designed to examine the effects and interactions of annual fire, mowing, and fertilization (N and P) on prairie soil communities and processes. For nearly all taxa, in both years, responses were characterized by significant treatment interactions, but some general patterns were evident. Introduced European earthworms (Aporrectodea spp. and Octolasion spp.) were most abundant in plots where fire was excluded, and the proportion of the total earthworm community consisting of introduced earthworms was greater in unburned, unmowed, and fertilized plots. Nymphs of two Cicada genera were collected (Cicadetta spp. and Tibicen spp.). Cicadetta nymphs were more abundant in burned plots, but mowing reduced their abundance. Tibicen nymphs were collected almost exclusively from unburned plots. Treatment effects on herbivorous beetle larvae (Scarabaeidae, Elateridae, and Curculionidae) were variable, but nutrient additions (N or P) usually resulted in greater densities, whereas mowing usually resulted in lower densities. Our results suggest that departures from historical disturbance regimes (i.e. frequent fire and grazing) may render soils more susceptible to increased numbers of European earthworms, and that interactions between fire, aboveground biomass removal, and vegetation responses affect the structure and composition of invertebrate communities in tallgrass prairie soils.  相似文献   

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