Cytogeographic Distribution and Genome Size Variation in Prairie Cordgrass (Spartina pectinata Bosc ex Link)

Prairie cordgrass plants (Spartina pectinata Bosc ex Link) were examined from 61 locations representing the geographic distribution of prairie cordgrass in the U.S. Using flow cytometry, the genome size of 183 individual plants of prairie cordgrass was determined, and the chromosome counts were obtained. Three distinct ploidy levels were observed: tetraploid ( $ \overline x = {1}.{56} {\text{pg}},{2}n = {4} \times = {4}0 $ ), hexaploid ( $ \overline x { = 2}.{33} {\text{pg}},{2}n = {6} \times = {6}0 $ ), and octoploid ( $ \overline {\text{x}} { = 3}.0{6} {\text{pg}},{2}n = {8} \times { = 8}0 $ ). In the sampled areas, the tetraploid populations extended from the East North Central to the New England regions of the U.S., while the octoploid cytotypes were mostly distributed in the West North Central regions. Populations of the tetraploids and octoploids were found in close proximity in the West North Central (Iowa and Kansas) and the West South Central (Oklahoma) regions. The hexaploid cytotype was found in one mixed population (4x?+?6x) occurring in Illinois. No statistically significant intraploidy genome size variation was found in the tetraploid populations, while significant intraploidy genome size variation was found in the octoploid populations. This study precisely defined the geographic distribution of cytotypes in prairie cordgrass throughout the different regions of the U.S. These results provide critical genome size and ploidy distribution information needed to design efficient breeding schemes for high yielding cultivars of prairie cordgrass with local adaption.     

Determination of laccase gene expression during degradation of 2,4,6-trinitrotoluene and its catabolic intermediates in Trametes versicolor

We have cloned a laccase gene fragment isolated from a Trametes versicolor strain in Korea. It showed high similarity in nucleotide sequences when compared with other fungal laccases. TNT (2,4,6-trinitrotoluene), a widely used explosive, was transformed rapidly by T. versicolor. When TNT and its catabolic intermediates were added to the fungal culture, they were transformed during the first few hours and the expression level of the laccase gene was increased during the early stage of cultivation.     

Early to mid-Pleistocene ostracod δO and δC in the central Tibetan Plateau: Implication for Indian monsoon change

As one of the pioneering projects of the Chinese Environmental Scientific Drilling (CESD) Program, a 206.5 m long sediment core (CE) was retrieved from Co Ngoin (Co = lake) Basin in the central Tibetan Plateau. Limnic records of the abundance of ostracod shells and their oxygen and carbon isotope measurements, together with the geochemical and the sedimentological investigations, were carried out on the interval between 143 and 52 m core depths, corresponding to ca. 2.01–0.84 Myr. Distinct oscillations of the abundance and the chemical compositions of ostracod shells have yielded the original information about variations in lakewater chemistry in association with the change in the Indian monsoon over the period of 2.01 to 0.84 Myr. The climate conditions during the Early Pleistocene were characterized by frequent oscillations and the ostracod records had expressed its synchronous response to these oscillations. A major climate change occurred in the Co Ngoin area at about 0.93 Myr. A pronounced fluctuation in ostracod δ¹⁸O and δ¹³C during the Mid-Pleistocene transition (MPT) is possibly associated with an increase in continental ice sheets. The compositional and the sedimentological characters of the immediately overlying sediments imply a shallow aquatic environment and also a harsh (salinity?) condition beyond the tolerance limits for ostracod survival, resulting in the disappearance of the Qinghaicypris crassa Huang 1979 around 0.84 Myr.     

Molecular cloning and expression pattern of 14-3-3ζ from the malaria vector, Anopheles sinensis

14-3-3 proteins are known to play a pivotal role in cell survival, apoptosis and signal transduction. The 14-3-3ζ isoform has been cloned and characterized from many eukaryotic organisms, including the fruit fly and silkworm. However, no study on mosquito 14-3-3 has been reported to date. In an attempt to investigate the function of 14-3-3 in midgut epithelial cells undergoing apoptosis, a cDNA library was generated from the malaria vector, Anopheles sinensis , which was treated with apoptosis-inducing Actinomycin-D. We were able to identify and obtain A. sinensis 14-3-3ζ cDNA ( Ansi14-3-3ζ ) from expressed sequence tags (EST) analysis after conducting massive sequencing of the A. sinensis cDNA library. Ansi14-3-3ζ has very high homology to 14-3-3 homologs of various insects, such as Anopheles gambiae (100%), Aedes aegypti (100%), Drosophila melanogaster (96%), Bombyx mori (93%), Apis mellifera (93%) and Mus musculus (81%), indicating that mosquito 14-3-3ζ is a highly conserved gene in diverse organisms. Analysis of temporal expression patterns showed that Ansi14-3-3ζ mRNA is highly expressed in egg, early pupae and adult stages and is also expressed, although at low levels, in fourth instar larvae and late pupae. In response to two immune elicitors (lipopolysaccharide and laminarin), no striking induction of 14-3-3ζ mRNA was observed in A. sinensis . Further studies of the precise biological function, inducibility and subcellular distribution of 14-3-3ζ are required in Plasmodium invasion-induced apoptotic midgut cells in A. sinensis in the context of the Time Bomb model.     

Cloning and blood‐meal dependent induction pattern of apolipophorin‐III from Anopheles sinensis

Apolipophorin‐III is known to play a role in transporting lipids in insects, and much attention has been paid to lepidopteran insects' apolipophorin. Thus, we were interested in examining the effects of blood‐meal on the expression pattern of apolipophorin‐III in mosquitoes. This led us to clone and partially characterize the full‐length cDNA of apoLp‐III (AnsiApoLp‐III) from Anopheles sinensis. Analysis of AnsiApoLp‐III cDNA shows that the 728‐bp sequence has a 582‐bp protein‐coding region with 94 bp of putative 5′ untranslated region and 152 bp of 3′ untranslated region. The deduced amino acid sequence begins with a methionine codon at position 95 and extends to position 674, encompassing a polypeptide of 193 amino acids. AnsiApoLp‐III has the highest identity (63%) to Culex quinquefasciatus apoLp‐III. Temporal expression pattern analysis shows that although AnsiApoLp‐III was expressed at all developmental stages, it was highly detected at egg and adult stages in the female mosquitoes. In addition, we found out that AnsiApoLp‐III was induced in An. sinensis adult females after uptaking a blood‐meal. Spatial expression patterns of AnsiApoLp‐III shows that AnsiApoLp‐III mRNA was strongly induced at day 1 and gradually decreased from day 1 to day 4 in the ovaries. Most interestingly, AnsiApoLp‐III mRNA in the Malpighian tubule was strongly induced at day 1, decreased during days 1–3, and then became elevated again at day 4. These data suggest that blood‐meal influences AnsiApoLp‐III mRNA induction in ovaries and Malpighian tubules. It remains to further elucidate the biological roles of AnsiApoLp‐III in these organs.     

DNA标记的种类、特点及其研究进展

生命的遗传信息存储于DNA序列之中,基因组DNA序列的变异是物种遗传多样性的基础。目前已发展出的DNA标记技术不下十几种,它们各具特色,并被广泛地应用于作物遗传育种、基因定位、亲缘关系鉴别、基因克隆研究等方面。对DNA标记技术进行分类,并作了初步概述。     

Tomato ringspot virus proteins containing the nucleoside triphosphate binding domain are transmembrane proteins that associate with the endoplasmic reticulum and cofractionate with replication complexes

Replication of all known positive-strand RNA viruses occurs in replication complexes associated with intracellular membranes. The putative nucleoside triphosphate binding (NTB) protein of Tomato ringspot virus (ToRSV) contains a stretch of hydrophobic residues at its C terminus, suggesting that it may act as a membrane anchor for the replication complex. Anti-NTB antibodies detected two predominant proteins in membrane-enriched fractions (the 66-kDa NTB and 69-kDa NTB-VPg proteins) along with other, larger proteins. The proteins containing the NTB domain cofractionated with markers of the endoplasmic reticulum (ER) and with ToRSV-specific RNA-dependent RNA polymerase activity in sucrose gradients. ToRSV infection induced severe changes in the morphology of the ER in plants expressing an ER-targeted green fluorescent protein (ER-GFP), and proteins containing the NTB domain colocalized with ER-GFP in indirect immunofluorescence assays. The proteins containing the NTB domain have properties of integral membrane proteins. Proteinase K protection assays using purified membranes from infected plants revealed that although the central portion of the NTB domain is exposed to the cytoplasmic face of the membranes, an 8-kDa fragment, recognized by anti-VPg antibodies, is protected by the membranes. This fragment probably consists of the 3-kDa VPg and the 5-kDa stretch of hydrophobic residues at the C terminus of the NTB protein, suggesting a luminal location for the VPg in at least a portion of the molecules. These results provide evidence that proteins containing the NTB domain are transmembrane proteins associated with ER-derived membranes and support the hypothesis that one or several of the proteins containing the NTB domain anchor the replication complex to the ER.     

Evaluation of cellulose-binding domain fused to a lipase for the lipase immobilization

A cellulose-binding domain (CBD) fragment of a cellulase gene of Trichoderma hazianum was fused to a lipase gene of Bacillus stearothermophilus L1 to make a gene cluster for CBD-BSL lipase. The specific activity of CBD-BSL lipase for oil hydrolysis increased by 33% after being immobilized on Avicel (microcrystalline cellulose), whereas those of CBD-BSL lipase and BSL lipase decreased by 16% and 54%, respectively, after being immobilized on silica gel. Although the loss of activity of an enzyme immobilized by adsorption has been reported previously, the loss of activity of the CBD-BSL lipase immobilized on Avicel was less than 3% after 12 h due to the irreversible binding of CBD to Avicel.