Genetic variation in mitochondrial genes and intergenic spacer region in harmful algae <Emphasis Type="Italic">Chattonella</Emphasis> species

In this study, nuclear ribosomal RNA gene internal transcribed spacer regions and the cox2-cox1 fragment of the mitochondrial (mt) genome were sequenced in 24 strains of Chattonella spp. Variability in both regions showed that the mt genome sequences of Chattonella spp. have a higher evolutionary rate than the nuclear rRNA gene sequences. A maximum likelihood tree based on the mt sequence grouped the Japanese Chattonella strains into two groups (Groups A and B), although no correlation was observed amongst the phylogenetic groups, their morphologies, or the isolated areas. Groups A and B were clearly identified by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay using Fokl, without the need for a sequencing experiment. The PCR-RFLP assay revealed that Chattonella cells obtained from sea water in Oita, Japan, in 2004 and 2005 belonged to Group B. This is the first report showing the genetic variation in Chattonella spp. using a PCR-RFLP identification protocol.     

A Dominant Locus,qBSC-1, Controls β Subunit Content of Seed Storage Protein in Soybean（Glycine max（L.） Merri.）

Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F2 segregated population and residual heterozygous lines（RHL） derived from the cross between Yangyandou（low level of β subunit） and Zhonghuang 13（normal level of β subunit） were used for mapping of β subunit content. Our results showed that β subunit content was controlled by a single dominant locus, qBSC-1（β subunit content）, which was mapped to a region of 11.9 cM on chromosome 20 in F2 population of 85 individuals. This region was narrowed down to 2.5 cM between BARCSOYSSR_20_0997 and BARCSOYSSR_20_0910 in RHL with a larger population size of 246 individuals. There were 48 predicted genes within qBSC-1 region based on the reference genome（Glyma 1.0, Williams 82）, including the two copies of β subunit coding gene CG4. An InDel marker developed from a thymine（TT） insertion in one copy of CG4 promoter region in Yangyandou cosegregrated with BARCSOYSSR_20_0975 within qBSC-1 region, suggesting that this InDel marker maybe useful for marker-assisted selection（MAS）.     

禽大肠杆菌菌毛产生的特性及免疫原性研究

用电镜检查广西分离的鸡致病性大肠杆菌 O_(60),O_(7)和 O_(8)菌株,在37℃和43℃培养均可见到菌毛,低于28℃不产生菌毛。用营养肉汤,Minca 培养液或最低限量培养液培养,冰浴高速磁力搅拌和MgCl_2反复沉淀提取,可得到较纯的菌毛。经测定,O_(60),O_(7)和 O_(8)菌毛蛋白分子量分别是15500,17500和17800。1日龄雏鸡用 O_(60)菌毛亚单位油乳苗皮下注射和菌毛悬浮液气雾免疫,均可保护鸡免受大肠杆菌对气囊的攻击。O_(60),O_(7)和 O_(8)菌株菌毛之间缺乏显著的交互保护。雏鸡在1日龄和10日龄用 O_(60),O_(7)和 O_(8)多价菌毛油乳苗免疫,21日攻毒时,可抵抗3种菌株侵袭气囊造成的急性呼吸道感染。     

Taxonomic status and intraspecific phylogeography of two sibling species of Metaphire (Oligochaeta: Megascolecidae) in Taiwan

In this study the synonymy of Amynthas formosae (Michaelsen, 1922) and A. yuhsii (Tsai, 1964) are reevaluated and the intraspecific phylogeography between them is studied. The morphological comparison and phylogenetic analysis based on cytochrome c oxidase subunit I suggest that A. formosae and A. yuhsii are not synonyms and both of them belong to Metaphire. The phylogeographic analysis suggests that the formation of mountains and rivers during the uplift of the Western Foothills in Taiwan comprised the vicariance events responsible for the intraspecific genetic differentiation between these two species.     

棉铃虫鞣化激素基因克隆及分子特性分析

【目的】获得棉铃虫鞣化激素2个亚基(α亚基和β亚基)基因序列,分析其分子特性和表达模式,研究棉铃虫鞣化激素的作用机制奠定基础。【方法】通过生物信息学和分子生物学技术分别得到棉铃虫鞣化激素α和β亚基cDNA序列,将棉铃虫及NCBI中公布的已知其它昆虫鞣化激素α和β亚基氨基酸序列分别整理分析,利用MEGA7(7.0.14)软件的Jones-Taylor-Thornton(JTT)模型构建系统进化树并进行聚类分析,qRT-PCR分别检测两亚基在棉铃虫不同生长发育阶段的表达模式。【结果】分别获得了棉铃虫鞣化激素 α亚基与β亚基核苷酸序列。其中α亚基(GenBank登录号:AHM0247472.1)cDNA片段长694 bp,开放阅读框480 bp,编码159个氨基酸残基,预测该蛋白质分子量为17.7 kDa。该基因在卵期第3 d、1龄幼虫第1 d、3~5龄幼虫第1 d、2和3龄末蜕皮期(3M和4M)、蛹期第0 d、第3 d和第7 d表达量相对较高。β亚基(GenBank登录号:AHM0247473.1)cDNA片段长779 bp,开放阅读框420 bp,编码139个氨基酸残基,预测该蛋白质分子量为15.9 kDa。该基因在卵期至2龄末蜕皮期(3M)、3龄幼虫第2 d、4~5龄幼虫第1 d、蛹期第1 d至羽化前表达量相对较高。鞣化激素α亚基和β亚基基因均在棉铃虫胸神经节中相对表达量最高,咽下神经节次之,脑部和腹部相对表达量较弱。【结论】鞣化激素在鳞翅目昆虫中具有较高的保守性;α亚基和β亚基基因主要在棉铃虫卵期、初孵幼虫期、蛹期和新羽化成虫期发挥作用;鞣化激素在棉铃虫幼虫体内主要由胸部神经节转录合成。     

小麦种子贮藏蛋白研究进展

介绍了谷蛋白亚基的分类、结构和编码位点，谷蛋白聚集体的结构、体积和粒度分布及其之间的关系，概述了醇溶蛋白的分类和编码位点，探讨了贮藏蛋白研究存在的问题和趋势．     

人工合成六倍体小麦的高分子量谷蛋白亚基组成分析

利用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳技术(SDS-PAGE)对103份人工合成六倍体小麦(2n=6x=42,AABBDD)的高分子量谷蛋白亚基组成进行了分析。结果发现,所分析材料具有丰富的等位变异。其中,在Glu-A1住点上共有5种变异类型,在Glu-B1和Glu-D1位点上均出现了12种等住变异类型;有些材料在Glu-D1位．占、上只表达1Dy类型亚基,而在1Dx亚基位置上出现空位形式;同时还发现了一些新的亚基组合形式和一未知新亚基。这些人工合成六倍体小麦材料中丰富的高分子量谷蛋白亚基变异,可能对小麦品质改良具有重要的应用价值。     

丹毒丝菌SpaA基因免疫保护区的克隆及其在毕赤酵母中的表达

【目的】以毕赤酵母Pichia pastoris X-33为宿主表达猪丹毒丝菌Erysipelothrix rhusiopathiae Spa A基因氨基端的免疫保护区蛋白.【方法】以采集于广东某猪场的猪丹毒丝菌为模板,根据NCBI中已发表的Spa基因c DNA序列设计1对引物,通过PCR扩增得到Spa A-N,并将其连接到表达载体p PICZαC上,得到重组表达质粒p PICZαC-Spa A-N;用Sac I酶将重组表达质粒p PICZαC-Spa A-N线性化后电转化入毕赤酵母X-33,经含博来霉素ZencinTM抗性的YPDS平板筛选和PCR鉴定的阳性转化子,用含不同浓度博来霉素抗性的YPDS平板筛选出高拷贝子并进行甲醇诱导培养,分别于诱导48、72、96 h后离心收集上清液,立即做SDS-PAGE,并进行SDS-PAGE及Western-blot试验.【结果和结论】成功克隆并表达了Spa A-N基因,构建了重组表达质粒p PICZαC-Spa A-N,并以毕赤酵母X-33为宿主成功表达了猪丹毒丝菌Spa A基因氨基端的免疫保护区蛋白.丹毒丝菌Spa A-N作为免疫保护区在酵母宿主中得以成功表达.     

Subunit composition of proteins from seeds of Lupinus albus

All the main globulins in the seeds ofLupinus Albus are oligomeric glycoproteins. Legumins (33%) consist of two similar protein molecules which contain protomers linked by disulphide bridges. They result from a partial proteolytic breakdown of an original polypeptide chain. Vicilins (44%) consist of four similar protein molecules with several protomers linked together by non-covalent bonds. Globulin 1 (6%) has a native M.W. of 199 kd and is formed by four 45.0 kd subunits consisting of two smaller protomers (28.0 and 16.0 kd) linked by -S-S- bonds. Globulin 9b (12.5%) has the lowest M.W. (44.0 kd) and is made up of three protomers, two of which are linked by disulphide bonds.