Sex Determination of Bovine Embryo Biopsies Using in situ Hybridization*

A male specific bovine DNA fragment was used as probe to determine the sex of bovine interphase cells by in situ hybridization. This method also proved useful for determining the sex of bovine embryos.     

Detection of papillomavirus-DNA in mesenchymal tumour cells and not in the hyperplastic epithelium of feline sarcoids

Abstract We examined 12 formalin-fixed paraffin-embedded feline skin tumours which had the histopathological features of fibropapillomas for the presence of papillomavirus (PV) DNA using touchdown polymerase chain recation (PCR), DNA sequencing and nonradioactive in situ hybridization. Nine of the tumours contained a 102-bp PCR product demonstrated using consensus PV primers that amplify a portion of the L1 gene. The nucleotide sequences are closely related, but not identical to that of ovine PV type 2, rabbit oral PV and reindeer PV. The deduced amino acid sequences had strong homologies with the major capsid protein L1 of deer PV, bovine papillomavirus (BPV) 1 and BPV 2, and European elk PV. Although PV antigens were not detected in any of the tumours by immunohistochemistry, PV DNA was demonstrated in individual mesenchymal cells or cell nests of 4/12 tumours by in situ hybridization. A nonproductive infection of mesenchymal fibroblast-like tumour cells with a papillomavirus would explain the lack of PV antigen expression and the absence of PV DNA in the hyperplastic epithelium. Because these tumours and their pathogenesis are similar to equine sarcoids, we suggest that they should be reclassified as 'feline sarcoids' instead of fibropapillomas.     

山羊卵巢无腔卵泡的体外发育

在三维琼脂凝胶培养体系中 ,山羊无腔卵泡的生长模式类似于体内。在一定培养时间内能善为保持其完整的立体结构和形态。原始卵泡能在体外启动 ,并得到一定程度的生长。初级卵泡能发育为次级卵泡 ,次级卵泡发育为有腔卵泡。卵泡在体外发育过程中 ,表现出了明显的优势化。本试验首次初步揭示了山羊无腔卵泡的体外发育基本过程和规律。     

Voluntary intake of silages in dairy cows depending on chemical composition and in vitro gas production characteristics

A study was conducted to investigate the possibilities to develop models for predicting the relative silage dry matter intake (SDMI) in dairy cows utilising information on chemical composition and in vitro gas production (GP) kinetics of silages. In five experiments, each with an average of 38 lactating dairy cows, SDMI was recorded for 15 grass silages made from primary growth and regrowth swards of timothy (Phleum pratense L.). The silages were characterised by chemical analysis and by utilising an automated in vitro GP recording technique with end point measurements of substrate residues. The silage samples were analysed both as dried and wet samples to evaluate the effects of sample preparation techniques on GP kinetics and their relations to SDMI. Relationships between feed variables and SDMI were investigated utilising simple linear and multiple regression. The wet silage samples had higher cumulative GP and different GP curves compared to the dried samples. The linear relationships between, GP variables, harvest number (first or second cut) of the grass, chemical characteristics of the silages and the relative SDMI show that the GP technique is a powerful tool to detect silage quality. By using the parameters from the dried samples the multiple regression analysis resulted in a relationship, relative SDMI (kg per 100 kg live weight (LW)) per day = 0.071 + 0.0029 × NDFD − 0.266 × C (R² = 0.82, S.D. = 0.06). NDFD is the degradability of the neutral detergent fibre (NDF) (g/kg NDF) and C is the variable that regulates the switching characteristics of the GP profiles. By using the wet silage samples the multiple relationship did not include any GP variable; relative SDMI (kg per 100 kg LW per day) = 1.86 − 0.008 × acetic acid (g/kg DM) + 0.024 × ethanol (g/kg DM) (R² = 0.62, S.D. = 0.08). The results from the regression analysis and the experience of the laborious sample preparation technique for wet samples, give the conclusion that dried silage samples are recommended for determining feed characteristics using the GP technique in intake studies.     

Low temperature storage and in vitro germination of cherimoya (Annona cherimola Mill.) pollen

Due to the protogynous dichogamy of cherimoya and to the absence of proper pollinating vectors, hand-pollination with fresh pollen is a common practice for cherimoya commercial production. In order to optimize the process of hand-pollination, in this work we have studied the conservation of cherimoya pollen at −20, −80 and −196 °C for up to 3 months. In vitro pollen germination of fresh pollen was 57.1% and it was progressively reduced with conservation time at the three temperatures studied reaching a minimum after 3 months of storage of 10.4%, 14.2% and 13.6% at −20, −80 and −196 °C, respectively. Differences in germination among temperatures were only significant during the first 2 weeks of storage. Field pollinations with pollen stored for up to 3 months at the three temperatures show no yield differences compared to pollinations performed with fresh pollen. The results indicate that pollen collected and stored at sub-zero temperatures at the beginning of the cherimoya blooming season can be used along the whole blooming season avoiding the need of collecting fresh pollen daily.     

苹果叶片离体培养中秋水仙素加倍效应的研究

用不同浓度的秋水仙素溶液处理苹果的栽培品种嘎拉试管苗的离体叶片,发现以０．５％的秋水仙素溶液处理４ｄ效果最佳,诱变频率达５６．１％。诱变后获得的四倍体植株在形态学和细胞学上均发生了明显变异。与对照株比较,变异株茎矮、粗壮、节间短、叶片厚、叶色浓绿;叶背面的气孔体积增大,单位面积气孔数目减少、茸毛数量增加;染色体数目加倍,２ｎ＝４ｘ＝６８（对照２ｎ＝２ｘ＝３４）,细胞核核仁数目增加。梢端组织学切片证明,变异株中同质突变体的频率达７３％。     

多效唑对芋种质离体保存影响的初步研究

以荔浦芋无菌试管苗为材料,研究了试管苗保存过程中不同浓度多效唑（PP粥）对试管苗保存效果的影响。研究结果表明,多效唑对芋种质资源的离体保存有显著影响,在光照时间14～16h／d,光照强度1500lx,室温（26＋2）℃的培养条件下,在MS＋6-BA2．5mg／L＋NAA0．02mg／L＋30g/L蔗糖的培养基中添加0．6mg／LPP粥,试管苗可保存240d,存活率为89．7％,且恢复生长后试管苗形态正常、长势良好,与对照株无明显差异。     

普通西瓜3个变种45S rDNA和5S rDNA的染色体定位

【目的】为了探明普通西瓜种(Citrullus lanatus)3个变种间的亲缘关系和进化与驯化过程,【方法】利用双色荧光原位杂交技术(fluorescence in situ hybridization,FISH),将核糖体45S rDNA和5S rDNA在11份栽培变种(Citrulluslanatus subsp.vulgaris)、2份Egusi变种(C.lanatus subsp.mucosospermus)和3份Citroides变种(C.lanatus subsp.lana-tus)有丝分裂中期染色体上进行了定位研究。11份栽培变种西瓜和2份Egusi类型西瓜都含有2对45S rDNA位点和1对5S rDNA位点,3份Citroides变种都含有1对45S rDNA位点和2对5S rDNA位点。【结果】结果表明,普通西瓜种3个变种内部各基因型间具有相同的45S rDNA和5S rDNA分布模式,栽培变种和Egusi变种具有相同的分布模式,但它们均与Citroides变种存在明显差异,说明栽培变种与Egusi变种之间的亲缘关系更近一些。【结论】rDNA序列分布分析为在分子细胞学水平开展栽培西瓜的进化与驯化分析提供了证据。     

耕层以下包气带中氮素蓄积的原位修复治理

耕层以下包气带土层中的氮素蓄积再释放是引起地下水污染的主要原因之一。将已优化制备的生物脱氮菌剂与白糖碳源共同施用，并于常规的田间施肥后在一定灌水量下以一定时段(3个月左右)观测，进行了现场原位修复治理试验研究。结果显示，实施该方法可有效去除耕层以下2．0m以浅主要发生氮蓄积的土壤层段中的高含量氮素，为防范地下水污染，消除潜在隐患，提供了科学依据。