水牛RNA聚合酶Ⅲ启动子的克隆与鉴定

[目的]获得有效的水牛RNA聚合酶Ⅲ启动子序列,为开展水牛源细胞的基因特异沉默研究奠定基础.[方法]通过启动子上、下游保守序列对水牛源启动子7SK、U6进行克隆和启动子关键顺式作用元件识别,利用一段针对EGFP的shRNA片段（shEGFP）对水牛7SK、U6启动子进行功能性分析,然后分别在水牛源及鼠源细胞中与pEGFP-N1共转染,转染48h后用荧光显微镜检测EGFP的表达情况,并用流式细胞仪和荧光实时定量PCR检测EGFP沉默表达情况.[结果]克隆获得水牛7SK、U6启动子序列分别为430和357 bp,其OCT-1（或CACCC盒）及TATA盒高度保守.连接shEGFP后与pEGFP-N1共转染细胞,通过荧光显微镜可观察到细胞发生了明显的荧光表达沉默现象;通过流式细胞分析,发现水牛7SK和U6启动子引导的shEGFP在水牛源细胞中沉默效率高达93.82％和87.45％;荧光实时定量PCR检测结果显示,在转染bu7SK-shEGFP和buU6-shEGFP的水牛源BFF细胞中,EGFP表达水平均显著低于其他物种启动子的细胞转染组（P＜0.05）,而在鼠源PT67细胞中,水牛启动子的启动效率与其他物种启动子差异不显著（P＞0.05）.[结论]水牛7SK和U6启动子可高效启动shRNA表达,且具有一定的物种特异性.     

鸡败血支原体pMGA多基因家族及其免疫逸机制的研究进展

pMGA多基因家族主要编码一类黏附素／血凝素蛋白，存在于鸡败血支原体的细胞表面，其主要功能是促进支原体黏附到宿主细胞上。近年来的相关研究发现，编码pMGA的基因数量从32－70不等，主要转录水平上通过（GAA）n 基序的数量改变引发pMGA基因的选择性转录，造成pMGA的抗原发生变异，从而干扰宿主正常免疫功能的发挥，使支原体对宿主产生严重的免疫逃逸。其中，（GAA）n基序已被证实在pMGA基因表达调控中起重要作用，这一三联体重复基序数目的多少直接影响到pMGA基因的ON／OFF不，本文通过对鸡败血支原体pMGA多基因家分子生物学研究进展浅要综述，以提出pMGA基因可能的转录调控机制，这对研究鸡败血支原体的分子致病机理及其免疫机制大有裨益。     

新城疫病毒ClassⅠ强毒株磷蛋白在细胞中的表达与定位

根据Class Ⅰ新城疫病毒分离株9a5b编码序列设计特异性引物扩增P蛋白基因,原核表达后利用纯化的蛋白免疫BALB/c小鼠制备单克隆抗体,经二次亚克隆后筛选到4株针对P蛋白的单克隆抗体,利用所得单抗进行P蛋白在细胞内定位分析,NDV 9a5b株按5 M.O.I感染量接种DF1单层细胞,当PI=4 h时,P蛋白呈点状弥散分布于细胞浆内;PI=6~8 h时,P蛋白聚集于细胞核周围;PI=12 h时,P蛋白呈单极化聚集于细胞核一侧;而PI=16 h后开始形成合胞体,P蛋白在单个细胞中仍呈现单极分布;当PI=48 h时,细胞核已发生碎裂,荧光强度有所减弱。本研究为深入开展P蛋白在病毒增殖以及细胞周期中的作用机制研究奠定了基础。     

鸡霍乱口服弱毒疫苗的研究

用经过选择的禽多杀巴氏杆菌自然弱毒菌株Ｒ１－２３菌株制成的鸡霍乱固体培养口服弱毒疫苗，其安全剂量接近于２００个免疫剂量。该口服苗两次饮水免疫的最适总剂量为每羽５０亿个活菌，最佳间隔时间为４８小时。免疫鸡第二次饮苗后第３天即可产生较好的免疫保护率（７／８）。免疫鸡对异型强毒株Ｐ１０５９（８∶Ａ）、Ｐ２７２３（９∶Ａ）和同型强毒株Ｃ４８－１（５∶Ａ）滴鼻攻击的近期保护率平均为５／１０、６／１０和９／１０。６个月免疫期的平均保护率为７６．７０％（４６／６０），一万多羽鸡田间试验结果表明，免疫后２个月的平均保护率为９３．１０％（２７／２９），６个月的平均保护率为７３．５０％（２５／３４），除对产蛋率有轻度影响（下降１．２７％）外，无其它不良反应。野外大面积使用结果表明，该口服苗不但性能稳定，免疫原性优良，而且安全可靠，适用于鸡口服免疫。     

适宜水稻花粉萌发的液体培养基的优化

花粉萌发率是检测花粉活力的一个重要指标,但实践中还缺少便捷、可靠的水稻花粉萌发率测定方法。本研究在前人研究基础上,采用液体培养基对相关培养条件进行了优化。结果表明,采用155g/L蔗糖+20mg/L H3BO3+40mg/L CaCl2+5 mg/L VB1相配合的培养基较适合两系杂交中稻两优818花粉的萌发,萌发率最高可达63.78%。此外,该培养方法具有培养条件要求不高、发芽结果稳定性好、培养基分布均匀等优点。     

Effects of land ownership and landscape-level factors on rare-species richness in natural areas of southern Ontario,Canada

Surprisingly few studies have considered the extent to which the nature of the ownership of land is associated with differences in biodiversity. We analysed ownership and other landscape-level effects on rare-species richness for both globally- and regionally-rare biota (including birds, herpetofauna, butterflies, mammals, and plants) in 289 designated natural areas (NAs) in southern Ontario, Canada. Information about each NA −including area, number of plant communities, ownership status and details of species diversity were collected from published sources. Length of perimeter of NA, relative isolation, and an estimate of fragmentation were measured using image analysis and GIS techniques. NAs were in general relatively small, with mean area of 158 ha (median 85 ha, range from 0.9 to 1278 ha) for private NAs; public NAs had mean area of 132 ha (median 16 ha, range from 0.1 to 1481 ha). Mean number of plant communities was 4.6 (median 4, range 1- 13) at private NAs and 3.8 (median 3, range 1-16) at public NAs. Our results show that, of several landscape-level factors, area had the greatest effects on rare-species richness and other biotic indices. Effects of area were followed by effects of plant community diversity, however this was itself significantly affected by area and the extent of perimeter of the NA. Both these factors were followed by effects of ownership of the NA and by effects of isolation of the NA (represented by minimum distance to nearest NA and by number of NAs in 10 km radius). Other landscape- level factors did not appear to have overall significant effects. Variation in area accounted for 0.1% to 29% of variation in number of rare species, with lower values for globally-rare, than for regionally-rare taxa. For all biotic groups, public ownership of NAs was associated with significantly greater rare-species richness compared to private ownership, even after other factors such as area were controlled. For all globally-rare biota except butterflies, area of NA had greater effects on rare-species richness than did ownership. Richness of regionally- rare birds was more affected by plant community diversity than by area of NA. Number of recorded plant communities accounted from 2.1% of variation in number of globally-rare plant species to as high as 31% of variation in regionally-rare butterflies. The diversity of plant communities was itself influenced by total site area (accounting for 45% of variation), extent of elongation of the NA, and both external- and interior- edge perimeters. Public NAs had greatest numbers of rare biota and so should be a significant focus for conservation programs. Smaller, privately-owned patches of natural area dominate (by number and area) in this densely populated region and their significance should not be overlooked. This revised version was published online in July 2006 with corrections to the Cover Date.     

森林经营方案编制及实施效果评价的微机辅助系统研究

本文以微型计算机为工具，选用ｄＢＡＳＥⅢ关系型数据库和ＢＡＳＩＣＡ语言，研制了国营林场森林经营方案编制及实施效果评价的辅助系统，并以福建林学院莘口教学林场的森林资源数据试运行．经过运行表明，该系统结构合理、功能较全、性能可靠、科学实用．     

Emergence of anoxia in the California current large marine ecosystem

Eastern boundary current systems are among the world's most productive large marine ecosystems. Because upwelling currents transport nutrient-rich but oxygen-depleted water onto shallow seas, large expanses of productive continental shelves can be vulnerable to the risk of extreme low-oxygen events. Here, we report the novel rise of water-column shelf anoxia in the northern California Current system, a large marine ecosystem with no previous record of such extreme oxygen deficits. The expansion of anoxia highlights the potential for rapid and discontinuous ecosystem change in productive coastal systems that sustain a major portion of the world's fisheries.     

山东省青岛市畜禽屠宰管理现状

为了解山东省青岛市畜禽屠宰管理发展现状,优化管理政策措施,开展了畜禽屠宰管理现状调查。调查发现:青岛市肉食消费以猪肉为主,牛、羊、禽、兔肉为副;全市现有生猪定点屠宰企业20家,年屠宰量约220万头;家禽屠宰企业4家,年屠宰量2亿只;牛羊屠宰企业6家,年屠宰量2万头。通过对青岛市畜禽屠宰企业监管体系和追溯体系建设,以及无害化处理等内容进行调研分析,指出当前存在代宰模式隐患多、监管责任难厘清、规范管理待完善等问题,并由此提出严格代宰管理,探索"两检合一"措施,改善产品追溯体系,完善行业管理规定,引导健康消费及加大资金投入等发展建议。     

葡萄锰中毒症及其相关营养元素分析

以玉溪市葡萄生产上出现的锰中毒症株及正常株为研究对象,采用田间调查与室内检测分析、锰中毒株与正常株比较相结合的方法,研究了葡萄锰中毒株及正常株叶片的N、P、K、Ca、Mg、Mn、Fe等指标,以及根际土壤的pH、有效态Mn、Fe、Ca、Mg等指标,以期为玉溪市葡萄生产上锰中毒症的防控提供技术依据。结果表明:与正常株比较,葡萄发生锰中毒株的叶片锰含量平均高达1 501.3 mg·kg^-1,比正常株的552.6 mg·kg^-1提高了1.7倍之多;P、K含量也极显著提高1倍多,而Ca、Mg含量则极显著降低近40%;锰毒株的Ca/Mn、Mg/Mn很低,分别为17.7、2.1,均显著低于正常株的98.3、10.5。葡萄发生锰中毒株的根际土壤的pH为4.29~4.35(为强酸性),有效Mn含量高达159.8~178.2 mg·kg^-1;土壤的Ca/Mn、Mg/Mn也很低,分别为7.1、0.6,均显著低于正常株的180.2、13.3。在玉溪市葡萄生产上发生的以"叶片主脉、支脉及叶脉附近的叶肉组织呈褐色状"为典型症状的锰毒株,其叶片锰吸收量过高,分析其原因是由于根际土壤pH较低为强酸性,土壤有效Mn含量过高,有效态Ca、Mg含量过低,Ca/Mn、Mg/Mn也很低所致。为葡萄生产上选择适宜土壤及其pH调节、合理平衡施肥提供参考依据。