地高辛标记cDNA探针检测苹果茎痘病毒

 Partial sequence(314 bp) of ASPV was cloned and used as a probe labelled with digoxigenin-11dUTP. The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA were detected by dot blot hybridization. The results showed that the probe was sensitive and specific. The probe couldn't hybridize with total RNA of Apple stem grooving virus, Apple mosaic virus and Apple chlorotic leaf spot virus samples as well as negative control, only hybridized with that extracted from dormant shoot infected with ASPV. The sensitivity for detection of plasmid contained ASPV-cDNA was 1.64 μg.     

茄子种子萌发过程中表皮粘膜对种子萌发的影响

通过对六叶茄和中长1号种子吸水率的测定,不同水分梯度种子萌发试验,不同发芽床种子萌发试验,垂直板种子萌发试验以及种子萌发过程中产生的表皮粘膜提取液处理,结果表明,粘膜不阻碍水分进入种子内部;随着种子表皮多余水分的增加,粘膜增厚,种子发芽率降低;以砂床作发芽床,种子不易产生粘膜,发芽率较高。     

猪附红细胞体PCR-微孔板杂交-酶免疫检测方法的研究

以猪附红细胞体和多种血营养菌16S rRNA基因的保守区设计合成引物HM-f/HM-r,在反向引物的5′端用生物素标记,以猪附红细胞体广东株16S rRNA基因的可变区序列设计种特异性寡核苷酸探针,探针的5′端用地高辛标记,成功地建立了猪附红细胞体PCR-微孔板杂交-酶免疫检测方法。通过测定不同浓度PCR产物对应的微孔板杂交-酶反应显色OD值,用SPSS统计软件进行回归分析,得到回归方程为y=2.1012-0.4492×logx,其相关系数R为0.977,显示对未知样品可进行定量检测。该方法与猪的多种细菌性病原、猫血巴尔通氏体CA株和E.wenyonii无交叉反应,其敏感性比常规PCR琼脂糖电泳检测提高了近100倍。用建立的方法对38份临床样品进行检测,结果有12份检测结果为阳性。     

猪蛔虫感染期幼虫抑制消减cDNA文库的构建

为筛选与线虫感染性相关的基因,本研究以猪蛔虫为对象,构建猪蛔虫感染期幼虫差异表达消减cDNA文库,为研究线虫期特异性发育的分子机制奠定基础。分别提取感染期幼虫和其它各期幼虫及成虫的总RNA,纯化mRNA后,采用Clontech公司PCR-selectTM试剂盒进行反转录合成cDNA并进行抑制消减杂交(SSH),构建猪蛔虫感染期幼虫差异表达的消减cDNA文库,并采用Southern斑点杂交进行消减效率的检测。随机从文库中抽取45个克隆进行测序及在线BLAST分析。试验结果表明,感染期幼虫差异表达的消减cDNA文库具有较强的特异性;在得到的41个表达序列标签(ESTs)中,有40个ESTs与已报道的基因有较高的相似性,主要代表猪蛔虫第三期幼虫基因和成虫头部基因,有1个cDNA片段可能代表新基因。猪蛔虫感染期幼虫差异表达消减cDNA文库的成功构建,为进一步研究幼虫发育差异表达基因的功能奠定了基础。     

Induction of interspecific allo-tetraploids of Megalobrama amblycephala♀×Megalobrama terminalis♂ by heat shock

Interspecific allo-tetraploidy was induced by a combination of hybridization of Megalobrama amblycephala ♀× Megalobrama terminalis ♂ and thermal shock. A thermal shock at 40 °C for 2 min applied to eggs 33 or 26 min (τ₀: 1.40) after fertilization resulted in a significantly ( P <0.01) higher yield of allo-tetraploids than hybrid crosses without heat shock. The yield of allo-tetraploids in embryos from mid-gastrula to hatching stages was 15.7–24.4% in different thermal shock treatment groups. The allo-tetraploidy rate in 330-day-old juveniles was 9.3% and 6.7% in different year treatment groups. After 2 years of rearing, allo-tetraploidy males attained sexual maturity with milk-like milt at the age of 2+, while allo-tetraploidy females at the age of 2+, and even at 3+, did not show any signs of sexual maturation exhibiting obviously delayed maturity. Only a few allo-tetraploidy females (three individuals) showed maturation characters at the age of 4+ and these females were induced to spawn.     

中华鳖脾脏雄激素受体mRNA 的原位杂交检测

采用原位杂交技术（In situ hybridization，ISH）研究雄激素受体（Androgen receptor，AR）mRNA在中华鳖（Pelodiseus Sinensis）脾脏的存在及细胞定位。取5只健康成年中华鳖的脾脏，进行石蜡切片。用地高辛标记的寡核苷酸探针对组织切片进行原位杂交（In situ hybridization，ISH），检测雄激素受体mRNA在脾脏免疫细胞内的表达定位。结果显示，大部分雄激素受体mRNA阳性细胞呈圆形或椭圆形，数量少胞体大且呈不规则形。阳性细胞在脾脏动脉周围淋巴鞘（Pefiatefial lymphatic sheath，PALS）和红髓（Red pulp，RP）内分布密集。椭球周围淋巴鞘（Periellipsoidal lymphatic sheath，PELS）内AR mRNA阳性细胞分布极少。杂交阳性信号物质在细胞质和细胞核均有分布。在标记为阳性的中华鳖脾脏中，推测圆形的阳性细胞可能为B淋巴细胞，而另一部分胞体大且形状不规则的阳性细胞则可能为巨噬细胞。脾脏内AR mRNA的存在进一步证实雄激素可能是调节脾脏等免疫器官功能的因素之一。[中国水产科学，2008，15（2）：337—341]     

Identification of differentially expressed genes associated with bud dormancy release in tree peony （Paeonia suffruticosa） by suppression subtractive hybridization

A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies.In order to identify genes that are highly expressed in buds released from dormancy,588 clones were examined by differential screening.Of these,185 clones were selected to be sequenced.A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database.Semi-quantitative RT-PCR was used to confirm further the expression...     

牡丹抑制栽培技术要领

通过对抑制栽培中苗木品质、储藏条件、温控设施、温度、湿度、光照等条件的论述，总结出牡丹抑制栽培过程中需注意的各种技术要领。     

新型换流变压器谐波模型及其配套滤波装置设计

阐述了新型换流变压器及其滤波系统的接线方案和谐波抑制机理.建立了新型换流变压器谐波模型,分析计算了新型换流变压器各绕组的谐波电流分布,实验验证了该模型的正确性.对直流输电开发平台进行了滤波装置实例设计,仿真和实验结果基本一致.证明了新型换流变压器及其滤波系统原理正确,滤波效果好,平台参数选择合理,新系统具有良好的应用前景.