鲍分子遗传学研究进展

阐述了近年来鲍分子遗传学方面的研究进展,总结了核型分析、等位酶、微卫星和小卫星、随机扩增多态性DNA、限制性片断长度多态性、线粒体DNA、表达序列标签研究和基因序列等技术在鲍种群遗传多样性、遗传分化、遗传结构及种质鉴定等方面的应用。并指出今后应加强鲍蛋白质组学、功能基因组学、遗传连锁图谱、数量性状基因座和标记辅助选择等方面研究。     

沙田柚黄龙病病原16S rDNA片段的克隆与序列分析

采集田间表现斑驳症状的沙田柚叶脉,用CTAB法提取总DNA。根据柑橘黄龙病病原16S rDNA的核苷酸序列设计引物P1/P2,进行PCR扩增,获得1条大小为1 167 bp的片段。酶切分析显示,该片段可被切成大小分别约为640 bp和520 bp的2个片段。扩增产物经纯化,与pM D 18-T V ector连接,转化大肠杆菌(E scherich ia coli)DH 5α,筛选克隆重组子。对PCR产物进行测序及序列分析,结果表明,与柑橘黄龙病病原亚洲种16S rDNA的同源性为99%,与非洲种的同源性为97%,与美洲种的同源性为96%。认为,沙田柚的斑驳症状是由黄龙病病原引致的,称之为沙田柚黄龙病。该沙田柚黄龙病病原属于柑橘黄龙病病原亚洲种(L iberobacter as iaticus)中的一个成员。系统进化树分析显示,沙田柚黄龙病病原与中国柑橘黄龙病病原亲缘关系最近,推测是直接来自中国柑橘黄龙病病原。     

小菜蛾类钙粘蛋白cDNA片段的克隆和序列分析

鳞翅目昆虫中肠刷状缘膜囊泡（brush border membrane vesicles，BBMV）有两类能与Cryl毒素结合的受体蛋白，一类是氨基肽酶N（APN），另一类是类钙粘蛋白（cadherin-like protein）。Gahan等（2001）报道了烟芽夜蛾中肠类钙粘蛋白基因失活导致了烟芽夜蛾（Heliothis viresce）对Bt毒素CrylA产生了高抗性，引起了各国科研人员的关注。近年来，先后有多种昆虫类钙粘蛋白基因被克隆和测序（Gahan et al．，2001：Morin et al．，2003）。     

绿脓杆菌ATCC27853菌株外毒素A全长编码基因的克隆及其在 HEK 293T细胞中的表达

采用PCR方法从绿脓杆菌ATCC27853菌株基因组扩增其外毒素A（PEA）全长编码基因,并将PEA基因插入到pcDNA3.1A真核表达载体中,构建成pcDNA3.1/ PEA质粒表达载体。经磷酸钙介导将重组质粒转染 HEK293T 细胞进行表达。结果表明本文克隆的PEA基因与标准菌株PA103碱基序列基本相似,同源性达99%。表达产物经蛋白免疫印迹法（Western-blot）检测,表明本文构建的含有原基因信号肽的PEA基因能够在真核细胞进行分泌性表达,这为基于PEA的免疫毒素、疫苗佐剂和疫苗载体的研究奠定了基础。     

Worldwide genotyping of castor bean germplasm (<Emphasis Type="Italic">Ricinus communis</Emphasis> L.) using AFLPs and SSRs

Worldwide genetic diversity in 200 individuals comprising 41 castor bean accessions was assessed using amplified fragment polymorphisms (AFLPs) and simple sequence repeats (SSRs). We found that, despite surveying five continents and 35 countries, genetic diversity in castor bean germplasm is relatively low (overall H _e = 0.126 for AFLPs and 0.188 for SSRs) compared to estimates of genetic diversity in other plant species. Our data also show no geographic structuring of genotypes across continents or countries within continents. An assessment of the congruence between AFLP and SSRs indicates a low correlation (R ² = 0.19) between the two data sets, but each marker class nonetheless shows similar patterns of low-genetic diversity and a lack of geographic structure. Our data do suggest that SSRs yield a higher percentage of polymorphic loci, higher heterozyosity and a greater range of genetic distances, and are therefore more informative than are AFLPs on a locus-by-locus basis. Based on comparisons with numerous other plant species, we suggest that the lower genetic variation in this worldwide collection may be due to one or more factors including: sampling strategies that have not captured the full extent of genetic variation in the species; artifactual variation due to long-term germplasm storage and seed regeneration; or intense selection followed by domestic cultivation of a limited number of castor bean genotypes, which are widely propagated for their horticultural and agro-economic value.     

Andosol to Luvisol evolution in Central Mexico: timing, mechanisms and environmental setting

Surface and buried Andosols and buried Luvisols of the Nevado de Toluca Late Quaternary tephra-paleosol sequence (Central Mexico) were studied to show whether these soils present an evolutionary sequence and to determine the pedogenic mechanisms and environmental factors involved in the evolutionary process. Micromorphological observations and mineralogical composition of fine sand and clay fractions were used to detect type and succession of soil-forming process. Some of the buried Andosols, defined as “intergrade” Andosols, have a predominantly blocky structure, humus-depleted areas, redoximorphic features and thin clay coatings in Ah horizons. Clay fractions of buried Andosols contain halloysite besides amorphous components, whereas in modern Andosols, allophane is dominant. Luvisols have micro-areas with granular structure and abundant phytoliths in the groundmass of Bt horizons assumed to be the relict Andosol features. Luvisol clay fractions are dominated by halloysite and kaolinite. Primary minerals show micromorphological weathering features in all studied soils being stronger in Luvisols; however, even in Luvisols, sand fractions consist mostly of unstable volcanic silicates. We hypothesise that the studied profiles form an evolutionary sequence: Andosols–“intergrade” Andosols–Luvisols; the soil transformation is supposed to be linked to progressive crystallisation of 1:1 clay minerals. Comparing the Nevado de Toluca paleosol properties with the existing data on volcanic soil climo- and chronesequences and assessing the regional paleopedological and lacustrine records of Quaternary paleoclimates, we concluded that wet/dry climatic oscillations took place during the formation of the studied paleosols. Rapid crystallisation of 1:1 minerals occurred during dry phases, which speeded up the Andosol to Luvisol transformation and made it independent from the primary mineral weathering status. The Andosol to Luvisol transformation accelerated by climatic fluctuations is thought to be a common soil evolutionary pathway in the subtropical and tropical regions of recent volcanism, which suffered contrasting precipitation oscillations in the Quaternary.     

Stoichiometry of base cations and silicon during weathering of a deep soil profile derived from granite

Evaluation of the stoichiometry of base cations (BCs, including K⁺, Na⁺, Ca²⁺, and Mg²⁺) and silicon (Si) (BCs:Si) during soil mineral weathering is essential to accurately quantify soil acidification rates. The aim of this study was to explore the differences and influencing factors of BCs:Si values of different soil genetic horizons in a deep soil profile derived from granite with different extents of mineral weathering. Soil type was typic acidi-udic Argosol. Soil samples were collected from Guangzhou, China, which is located in a subtropical region. To ensure that the BCs and Si originated from the mineral weathering process, soil exchangeable BCs were washed with an elution treatment. The BCs:Si values during weathering were obtained through a simulated acid rain leaching experiment using the batch method. Results showed that soil physical, chemical, and mineralogical properties varied from the surface horizon to saprolite in the soil profile. The BCs:Si values of soil genetic horizons during weathering were 0.3-3.7. The BCs:Si value was 1.7 in the surface horizon (A), 1.1-3.7 in the argillic horizon (Bt), and 0.3-0.4 in the cambic (Bw) and transition (BC) horizons, as well as in horizon C (saprolite). The general pattern of BCs:Si values in the different horizons was as follows: Bt > A > Bw, BC, and C. Although BCs:Si values were influenced by weathering intensity, they did not correlate with the chemical index of alteration (CIA). The release amounts of Si and BCs are the joined impact of soil mineral composition and physical and chemical properties. A comprehensive analysis showed that the BCs:Si values of the soil derived from granite in this study were a combined result of the following factors: soil clay, feldspar, kaolinite, organic matter, pH, and CIA. The main controlling factors of BCs:Si in soils of different parent material types require extensive research. The wide variance of BCs:Si values in the deep soil profile indicated that H⁺ consumed by soil mineral weathering was very dissimilar in the soils with different weathering intensities derived from the same parent material. Therefore, the estimation of the soil acidification rate based on H⁺ biogeochemistry should consider the specific BCs:Si value.     

水稻南方黑条矮缩病毒湖南鼎城株系S10片段的基因组序列分析

运用RT-PCR技术克隆了水稻南方黑条矮缩病毒（southern rice black-streaked dwarf virus,SRBSDV）湖南鼎城株系的基因组S10片段（SRBSDV-HuNDCS10）,并对其全序列进行了测定和生物信息学分析。结果显示,SRBSDV-HuNDC S10片段全长为1797bp（登录号：JQ337964）,含有1个ORF,编码557个氨基酸残基的衣壳蛋白,推测分子量约62.6kD,推测等电点为7.62,与已报道的广东、海南和云南分离物病毒的S10作比较,它们的核苷酸相似性分别为99.7%、99.0%和98.4%,氨基酸相似性分别为100.0%、99.5%和99.3%。对SRBSDV-HuNDCS10及部分Fijiviruses病毒对应片段在5＇URT与3＇URT存在的保守序列和互补序列进行了归纳,对其ORF编码的氨基酸序列进行了motif查找,得到该属（Fijiviruses）氨基酸序列的10个保守区段。此外,进行了糖基化位点、磷酸化位点及B细胞抗原表位预测,发现了3个可能的N端豆蔻酰基化位点,可能与病毒的侵染机制有关。     

植物EST-SSR标记开发及其应用

随着生物科技的进步,大量的植物表达序列标签（expressed sequence tags,ESTs）已经成为开发SSR标记的重要资源。EST-SSR作为一种新型分子标记,其多态性可能与基因功能直接相关,而且在相近植物间具有良好通用性,使得EST-SSR标记在实际应用中更具价值。采用计算机方法大规模发掘EST-SSR多态性位点极大地提高了SSR标记开发效率。尤其随着新一代测序技术的成熟以及测序成本的急剧下降,利用新一代测序技术产生大量的转录组数据进行EST-SSR多态性标记的计算机大规模发掘将对SSR标记的开发带来深远影响。本文简要介绍了植物EST-SSR分布特点,并对EST-SSR标记开发现状以及相关应用作了综合评述,此外,还对EST-SSR标记的计算机开发新策略进行了展望。