表达HAV结构基因的重组腺病毒鉴定及细胞病变分析

利用RT-PCR方法,从HAV-L8株的RNA中扩增出结构基因(vp3 vp1),克隆到穿梭质粒pXCX2NotI上。采用磷酸钙-DNA共沉淀技术,将腺病毒载体pJM17与pXCX2-CMV-HAV共转染293细胞。在光学显微镜下观察到明显的细胞病变(CPE);经RT-PCR、免疫荧光染色和蛋白印迹鉴定证明HAV的结构基因已插入到腺病毒基因组中;在电子显微镜下观察发现有二十面体的病毒颗粒。以上结果表明获得了重组腺病毒rAdHAV,纯化后的rAdHAV滴度为1×109.0TCID50/mL。     

优氯净对苏云金杆菌伴孢晶体消毒效果及机理的研究

苏云金杆菌(Bacillus thuringiensis,简称BT菌)的伴孢晶体被优氯净液处理后,在相差显微镜下观察,失去暗蓝色光泽,呈暗黑色,难溶于碱性缓冲液。虽能溶于昆虫的胃液,却失去致病性。用扫描电镜和SDS—聚丙烯酰胺凝胶电泳进一步研究,结果表明:优氯净能破坏伴孢晶体蛋白的一级和立体结构,使晶体蛋白失活,达到消毒的目的。消毒能力测定结果表明:1毫升0.03％有效氯优氯净液在3分钟内能消毒50微升BT菌液(3μg伴孢晶体/μl菌液)。消毒BT菌污染桑叶,用0.01％有效氯优氯净液浸泡3分钟即可。考虑到生产上的安全性,我们建议用0.05％有效氯优氯净消毒3—5分钟。     

小兴安岭蕨类植物生态特性的研究

采用样线与样方相结合的调查方法，根据点相关系数及Ｘ＾２值连续性较正公式，测定了小兴安岭凉水自然保护区具有环境梯度的７个森林类型内主要蕨类植物的种间联结。结果表明：各林型均有其适生的蕨类类群，与该类群呈正联结的蕨类种其数量分布也多，反之刚少；具有环境梯度的不同林型序列，反映出蕨类植物适生生境的不同，据此，将本区主要蕨类植物分为４种生态类型。     

结球莴苣“烧边”成因及其调控措施的研究进展

结球莴苣烧边病是生理性病害，严重影响其品质，制约生产。国内外多数研究者认为钙缺乏导致结球莴苣烧边病的发生，但是现在许多研究表明其他因素也能导致烧边病的出现，例如环境因素、生长速率、内源激素等，所以研究烧边病发生的机制及其与环境因素的关系是防御结球莴苣烧边病发生的关键。     

不同生态型大白菜抽薹时内源激素含量比较

大白菜〔Brassicacampestrissubspeciespekinesis(Lour. )Olsson〕先期抽薹一直困扰着春大白菜和高纬度、高海拔地区的大白菜生产。为育成冬性较强不易抽薹的品种,大白菜晚抽薹特性及利用的研究逐渐成为新的热点。不同生态型的大白菜品种,在相同的环境条件下冬性越强越不易抽薹。国外对控制或减轻未熟抽薹的环境条件有较多报道〔1~3〕,本研究主要探讨了低温处理后不同生态型的大白菜品种抽薹时内源生长素类 (IAA、IPA)、赤霉素类(GA3、GA4 )、细胞分裂素类 (ZR、DHZR)和脱落酸(ABA)含量差异,以期为选育具有较强冬性的大白菜品…     

国宝--金梅枣

金梅枣为河南省南时大枣研究所培育的特晚熟鲜食极品。金梅枣果实特别大,近圆形,成熟期为金红色,形似李梅。平均单果重30克左右,最大过重65克,色泽光洁美丽,皮薄肉脆,细嫩多汁,甜似蜂蜜,枣味香浓郁,品质顶级。可食率93.8%,含水率65.4%,含糖量32.2%,维生素C含量352mg,蛋白质1.65%,含人体所需的氨基酸19种。金梅枣药用价值也特别高,每百克果肉中含环磷酸腺苷50mg,这种物质对人体癌细胞具有极强的破坏和抑制作     

Comparison of morphology, viability, and function between blood and milk neutrophils from peak lactating goats

The morphological features of blood and milk neutrophils from peak lactating goats were compared using light microscopy, scanning electron microscopy and flow cytometry in order to investigate the cytological changes of neutrophils after migration into the mammary gland. The kinetics of reactive oxygen intermediates (ROI) generation and gelatinase release of blood and milk neutrophils, with or without stimulation of phorbol 12-myristate, 13-acetate ester (PMA), were used to characterize their responses to inflammatory stimuli. Neutrophils isolated from goat milk were highly segmented and contained multi-lobed nuclei. Ultrastructurally, milk neutrophils were more ruffled on the surface compared to blood neutrophils. Approximately 30% of milk neutrophils were undergoing cell death, either necrosis or apoptosis, in contrast to 8% of blood neutrophils. The ROI production of activated milk neutrophils peaked earlier than blood neutrophils, but the duration and the intensity were much less. Neutrophils from both sources augmented the release of gelatinase in response to PMA (1 ng/mL). However, the amount of gelatinase released from milk neutrophils was lower (P < 0.05) than that of blood neutrophils. In summary, more neutrophils become apoptotic and necrotic in the mammary gland, presumably due to spontaneous aging, the process of diapedesis, and the interaction with milk components. Milk neutrophils have impaired functionalities in comparison with blood neutrophils. The information is relevant when studying mammary gland immunity and related diseases, such as mastitis.     

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.