用RAPD分析多年生黑麦草品种间遗传多样性

多年生黑麦草是世界上应用最广泛的冷季型草坪草兼牧草之一,具有建植快速,分蘖能力强,适口性良好等特点。为了评价其品种间的遗传多样性,选取了多年生黑麦草的14个草坪型品种和2个意大利黑麦草品种用RAPD进行遗传多样性分析,并且对每个品种的几项坪用特性进行了观察。61个引物共扩增出408条带,多态条带比率(PPB)为89．95％。聚类分析结果表明,多年生黑麦草品种之间的遗传多样性较低,相似系数分布于44．55％～95．69％之间。一年生黑麦草的2个品种TopOne和A,vance没有单独聚成一类,而是与14个多年生黑麦草品种混合聚在一起,而且Avance与其他多年生黑麦草之间的相似系数较高。聚类结果与形态特征没有明确的对应关系。     

牦牛白细胞介素2(IL2)基因cDNA 的分子克隆和表达研究

目的克隆牦牛免疫基因并研究其免疫特性。方法以伴刀豆球蛋白A（ConA）激活的体外培养的牦牛血液淋巴细胞,提取激活淋巴细胞的总RNA,用RT—PCR方法,从中克隆出白细胞介素2cDNA,连接到T载体上测序,并亚克隆到pC,EX4T-1表达质粒,在大肠杆菌进行了重组表达,纯化融合表达YIL2产物,MTT比色法测定其体外刺激牦牛血液淋巴母细胞增殖的免疫活性。结果YIL2cDNA序列分析显示：在其编码区442位点的一个碱基发生突变（由C突变为T）,从而导致终止密码子（TAA）出现,使YIL2蛋白表达提前终止,与其它牛IL2蛋白相比少了8个氨基酸。MTT比色法测定结果表明重组牦牛IL2蛋白体外能显著促进牦牛淋巴母细胞的增殖。结论本实验成功克隆了牦牛IL2基因,其原核表达产物具有显著的免疫活性,这为研制新型免疫增强剂来提高牦牛对各种疾病的抵抗力,增强牦牛疫苗的免疫保护率奠定了基础。     

不同危害状态下寄主萜烯挥发物含量的变化及松墨天牛对其组分的触角电位反应

用气相色谱-质谱（GC-MS）联用法对5种生理状态（健康、人为物理损伤、饵木、取食和产卵）的马尾松Pinus massoniana和取食危害状态的雪松Cedrus deodara的萜烯进行分析。研究结果显示,5种生理状态下,α-蒎烯在萜烯组分中均占有最高比例。健康状态下马尾松树干和树枝中的萜烯组分不同且含量有显著差异。随着饵木的衰弱进程,α-蒎烯的相对含量显著减少,β-蒎烯的含量显著增加,我们推测,α-蒎烯与β-蒎烯的相对比例越小则寄主树势衰弱的程度越深,其两者比例的变化是马尾松寄主树势衰弱程度变化的一个重要信号。人为物理损伤的马尾松树干与树枝的α-蒎烯、β-蒎烯和β-水芹烯含量差异显著。不同产卵危害程度的马尾松树干萜烯类组分不同,同一产卵危害程度的马尾松树干上部和下部的多数萜烯组分相对含量差异显著,但变化无明显规律。取食危害状态下马尾松和雪松萜烯种类不同且含量差异显著。重度产卵危害马尾松树干与取食危害树枝萜烯中,α-蒎烯、β-蒎烯、柠檬烯和β-水芹烯相对含量差异显著。第13天的饵木与人为物理损伤的马尾松树干的萜烯中,饵木的β-蒎烯和柠檬烯相对百分含量均比人为物理损伤状态的高,而α-蒎烯则相反。α-蒎烯和β-蒎烯在取食危害树枝中的含量显著高于健康树枝,而β-月桂烯、β水芹烯和反式石竹烯在健康状态树枝中的含量显著高于取食危害树枝。除了α-蒎烯和β蒎烯,健康状态与产卵危害状态的马尾松树干萜烯相对含量大都差异显著。α-蒎烯,β-蒎烯和β-水芹烯在人为物理损伤与虫害状态的马尾松树干及树枝中含量差异较大。总之,α-蒎烯、β-蒎烯和β-水芹烯很可能是影响和调节松墨天牛取食和产卵行为最为重要的信号物质。松墨天牛Monochamus alternatus Hope对寄主萜烯的触角电位（EAG）表明,该天牛对β-蒎烯的EAG活性高于其他萜烯,乙醇也能引起较高的EAG反应活性,但总体上对单一萜烯的EAG反应活性不强,且存在雌雄差异。     

Pollen morphology of the tribe Cynoglosseae of Boraginoideae (Boraginaceae) in China

Pollen morphology of 16 Chinese species representing 7 genera in the tribe Cynoglosseae of Boraginoideae(Boraginaceae) was examined under LM and SEM, and 5 species under TEM. Pollen grains are cocoon-shaped, rarely subprolate, prolate or ovoid, very small, 7～ 15.7μm× 3.5 ~ 13.9 μm in size, P/E = 1.6～2.02; 3-colporate apertures alternate with 3-pseudocolpi, with equatorial endocingulus except those in Bothriospermum; exine surface is usually smooth, with or without perforations in two poles, rarely with tuberculate ornamentation; exine is rather thin and includes ectexine and endexine, while ectexine consists of imperforate tectum, columellae and foot-layer. The tribe Cynoglosseae has many common characters of pollen grains, but there are some differences among genera. A key to the genera is given based on pollen morphology. Bothriospermum, without endocingulus, may be a primitive genus in this tribe, and its pollen morphology is more similar to that of Eritrichieae than to Cynoglosseae, thus it seems more reasonable to put it into Eritrichieae. Solenanthus may be the most advanced genus in the tribe Cynoglosseae for its pollen grains of sub-isopolar and ovoid shape.     

兰州八里镇蔬菜基地春季土壤纤毛虫群落特征

2005年4月至2007年6月,用活体观察和固定染色方法对兰州八里镇蔬菜基地春季土壤纤毛虫的群落特征进行了研究.共鉴定出土壤纤毛虫67种,其中包括2个未定名种和14个中国土壤纤毛虫新纪录种.在八里镇蔬菜基地的土壤中鉴定到纤毛虫34种,隶属于3纲、7目、17科、18属;下毛目、肾形目、前口目为其优势类群;吻吴氏虫(Woodruffia rostrata)、近亲游仆虫(Euplotes afflnls)、苔藓圆纤虫(Strongylidium muscorum)、鱼形瘦尾虫(Uroleptus piscis)、似膜袋虫(Cyclidium simulans)等为其优势种.在对照样点(兰山)中鉴定到纤毛虫49种,隶属于3纲、9目、23科、30属;下毛目、盾纤目、肾形目和篮口目为其优势类群;苔藓游仆虫(E.muscicola)、近缘殖口虫(Gonostomum affine)、皮速体虫(Tachysoma pellionella)、食藻斜管虫(Cyclidium simulans)等为其优势种.八里镇蔬菜基地和对照样点的Gleason-Margalef物种多样性指数分别为3.672、5.393;与对照样点土壤纤毛虫的群落Jaccard相似系数为0.239.结果表明,八里镇蔬荣基地春季土壤纤毛虫的物种多样性与对照样点相比有所降低,其群落结构简单化.     

山东临朐人群HLA等位基因多态性 与幽门螺杆菌感染关系的研究

为了研究山东临朐地区健康人白细胞抗原（HLA）Ⅰ类等位基因多态性与幽门螺杆菌（Hp）感染的关系,运用序列特异性引物聚合酶链反应（PCR SSP）的方法,检测Hp阳性人群（90例）和Hp阴性人群（49例）的HLA Ⅰ类和Ⅱ类等位基因。结果在HLA Ⅰ类（A、B、CW）的共68个等位基因多态中,发现在感染及非感染人群中存在4个显著性差异的位点;在HLA Ⅱ类（DRB1、DQB1和DRB3、DRB4、DRB5）的共22个等位基因多态中,没有发现显著性差异的位点。A*02等位基因频率,Hp阳性低于阴性人群（OR,0.56;95%CI,0.33～0.94;P,0.029）;B*48等位基因频率,Hp阳性低于阴性人群（OR,0.15;95%CI,0.03～0.72;P,0.007）;CW*08〖STBZ〗等位基因频率,Hp阳性低于Hp阴性人群（OR,0.32;95% CI,0.15～0.69;P,0.003）;CW*15等位基因频率,Hp阳性高于Hp阴性人群（OR,5.11;95%CI,0.63～40.90;P,0.024）。结果表明HLA Ⅰ类等位基因的多态性可能与山东临朐地区Hp的易感性有关;HLA Ⅱ类等位基因的多态性可能与其无关。HLA Ⅰ类等位基因中,CW*15可能是Hp感染的易感基因;A*02、B*48和CW*08可能是保护性基因。Abstract: In order to analyze the relationship of HLA polymorphisms and the infection of H.pylori in the population of Linqu County in Shandong Province,polymerase chain reaction with sequence specific primers(PCR SSP) was used to determine the alleles of HLA typeⅠand Ⅱ in 90 Hp positive persons and 49 Hp negative controls.The results showed that among the 68 alleles of HLA typeⅠ,4 alleles were found significantly different between Hp positive and Hp negative population,while no significant difference was found among the 22 alleles of HLA typeⅡ.Hp positive persons had a lower allele frequency of A*02（OR=0.56,95% CI=0.33～0.94;P=0.029）, B*48（OR=0.15,95% CI=0.03～0.72;P=0.007）,CW*08（OR=0.32,95% CI=0.15～0.69;P=0.003）and a higher allele frequency of CW*15（OR=5.11,95% CI=0.63～40.90;P=0.024）compared with Hp negative controls.Our results indicated that the polymorphisms of HLA typeⅠis involved in the genetic susceptibility of Hp infection in Linqu County,while the polymorphisms of HLA typeⅡ may have no relationship with the genetic susceptibility of Hp infection.It was shown that among the alleles of HLA typeⅠ,CW*15might be a susceptible gene of Hp infection while A*02,B*48 and CW*08might be protective genes.     

褶合光谱分析法在农业科学分析中的应用

褶合光谱分析法是一种新的光谱信号分析法,刚刚开始应用于农业科学分析。本文详尽地介绍了它的产生,测定原理和在农业上适测领域,旨在推动此分析技术在农业上的应用。     

中国紫草科破布木属花粉形态和外壁超微结构

为了深入探讨紫草科（Boraginaceae）的分类问题,用光学显微镜和扫描电子显微镜观察了该科破布木属（CordiaL.) 10种植物的花粉形态和外壁超微结构。发现该属花粉具三孔、三孔沟、三拟孔沟和三合沟4种萌发孔类型。外壁表面具微刺状纹饰、刺状纹饰、网状纹饰和不规则的条纹网状纹状。破布木属的花粉特征表明,该属花粉在紫草科中既是独特的分类群,又是比较原始的属种。     

Development of a loop-mediated isothermal amplification assay for porcine circovirus type 2

In this study,the loop-mediated isothermal amplification(LAMP)method was used to develop a rapid and simple detection system for porcine circovirus type 2(PCV2).According to the PCV2 sequences published in GenBank,multiple LAMP primers were designed targeting conserved sequences of PCV2.Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template,LAMP reactions in a PCV2 LAMP system was performed,the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye.The results showed highly-efficient and specific amplification in 30 min at 63℃ with a LAMP real-time turbidimeter.Furthermore,PCV2 DNA templates,with a detection limit of 5.5×10-5ng of nucleic acid,indicated that this assay was highly sensitive.The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter,showing the potential simplicity of interpretation of the assay results.The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples.In addition it offers higher specificity and sensitivity,shorter reaction times and simpler procedures than the currently available methods of PCV2 detection.It is therefore a promising tool for the effective and efficient detection of PCV2.