Agrobacterium‐produced and exogenous cytokinin‐modulated Agrobacterium‐mediated plant transformation

Agrobacterium tumefaciens is a plant pathogenic bacterium that causes neoplastic growths, called ‘crown gall’, via the transfer and integration of transferred DNA (T‐DNA) from the bacterium into the plant genome. We characterized an acetosyringone (AS)‐induced tumour‐inducing (Ti) plasmid gene, tzs (trans‐zeatin synthesizing), that is responsible for the synthesis of the plant hormone cytokinin in nopaline‐type A. tumefaciens strains. The loss of Tzs protein expression and trans‐zeatin secretions by the tzs frameshift (tzs‐fs) mutant is associated with reduced tumorigenesis efficiency on white radish stems and reduced transformation efficiencies on Arabidopsis roots. Complementation of the tzs‐fs mutant with a wild‐type tzs gene restored wild‐type levels of trans‐zeatin secretions and transformation efficiencies. Exogenous application of cytokinin during infection increased the transient transformation efficiency of Arabidopsis roots infected by strains lacking Tzs, which suggests that the lower transformation efficiency resulted from the lack of Agrobacterium‐produced cytokinin. Interestingly, although the tzs‐fs mutant displayed reduced tumorigenesis efficiency on several tested plants, the loss of Tzs enhanced tumorigenesis efficiencies on green pepper and cowpea. These data strongly suggest that Tzs, by synthesizing trans‐zeatin at early stage(s) of the infection process, modulates plant transformation efficiency by A. tumefaciens.     

Taxonomic notes on the genus Holepyris (Hymenoptera: Bethylidae) from Korea with descriptions of six new species

Six new species of the genus Holepyris (Hymenoptera: Bethylidae: Epyrinae) from Korea are described: H. brevicarinatus sp. nov., H. crinitus sp. nov., H. dimidiatus sp. nov., H. longicephalus sp. nov., H. mucro sp. nov. and H. multo sp. nov. They are described with illustrations of male genitalia and the biometric measurement data. A key to the Korean species of Holepyris based on males is provided.     

Systematics and phylogeography of the Taiwanese endemic minnow Candidia barbatus (Pisces: Cyprinidae) based on DNA sequence,allozymic, and morphological analyses

The phylogeographical patterns of the endemic minnow, Candidia barbatus, were examined using mitochondrial (mt) cytochrome b sequences, allozyme electrophoresis, and morphology. The 23 mtDNA haplotypes recovered were grouped into six lineages (A–F). According to a molecular clock of 1.7% per million years, we estimated that the colonization of C. barbatus to Taiwan took place three million years ago with several subsequent vicariance events. In the allozyme analysis, the Bayesian approach partitioned different populations into three clusters, with clusters I, II, and III mainly found in northern, central, and southern Taiwan, respectively. Morphologically, fish from north and south of the Tropic of Cancer had distinct numbers of lateral line scales and were separated into two morphs. The cohabitation of different mtDNA lineages with high allozymic and morphological similarities suggests that interbreeding amongst them is possible. Therefore, in spite of highly differentiated mtDNA, samples from different collecting sites should be considered a single species. Based on the current population distribution and levels of divergence, we propose that both orogenic activities and marine regression during glacial epoch were the two main forces shaping population differentiation of this species. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161 , 613–632.     

Quantification of Flavonoid Compounds from Strawberry ( Fragaria ananassa) and Correlation Effects with Antioxidant Activities

The amounts of flavonoid (myricetin , quercetin and kaempferol), antioxidant contents ( total phenolic and total flavonoid) and antioxidant activities (DPPH and FRAP) were analyzed in strawberry. Quantitative analysis of conjugated flavonoid contents were obtained after different hydrolysis concentration and reaction period . The highest yields of myricetin and quercetin were found with 1.0M HCl and a reaction period of 0.5 hr, however , for kaempferol was 1.0M HCl and a reaction period of 1hr , respectively. In the study, we also estimated the correlation coefficient of TPC, TFC, DPPH and FRAP. In addition , a positive correlation was observed ( r = 0.980, P < 0.05) between TPC and TFC, TPC and TFC are correlated to the DPPH activity (r = 0.913, P < 0.05 and r = 0.899 , P < 0.05 , respectively) , and to the FRAP activity ( r = 0.958, P < 0.05 and r = 0.936, P < 0.05, respectively), respectively. These results pointed out that strawberry generally possesses a high level of antioxidant activities, which could be linked to the levels of phenolic compounds in the fruit.     

Liverwort genomes display extensive structural variations

PIKE, L. M., HU, A., RENZAGLIA, K. S. & MUSICH, P. R., 1992. Liverwort genomes display extensive structural variations. Analyses of the total genomic DNA of eight species of liverworts and two species of green algae by thermal denaturation and CsCl buoyant density gradient centrifugation reveal a high degree of structural complexity and interspecific heterogeneity. The hepatic taxa exhibit two or more DNA components of varying base composition. Average G4-C contents of total cellular DNA calculated from melting profiles are similarly variable, ranging from 38% to 53% G + C. The green alga Chara , a member of the ancestral line to land plants, shows similarities with liverworts in possessing multiple DNA components of comparable complexity, whereas Hydrodiciyon DNA displays a single component. Detailed hybridization analyses of individual density gradient fractions using α-tubulin, rRNA and ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene probes were performed to locate the low-copy number and moderately repetitive nuclear genes, and the chloroplast chromosome, respectively. The location of each gene within the density gradient is highly variable among the organisms examined; a-tubulin occurs in fractions ranging from 44–64% G + C, rDNA in 50–64% G + C fractions, and the RbcL gene is located in fractions from 30–59% G + C. For a given species, the two nuclear genes normally overlap in their distributions within the gradient. In most instances, neither gene occurs in the major DNA components, indicating that these components may contain repetitive DNAs. The observed variation in the density of the rbcL gene implies substantial reorganization of the chloroplast genome. The overall differences in the genomic components within and between taxa provide insight into the dynamics of DNA structure that have occurred during the extended evolutionary history of these organisms.     

A Role for Ecdysteroids in the Induction and Maintenance of the Pharate First Instar Diapause of the Gypsy Moth,Lymantria dispar

Several lines of evidence suggest a novel regulatory mechanism for diapause regulation in the gypsy moth. We propose that ecdysteroids play a role in the induction and maintenance of the pharate first instar larval diapause in this species. A 55 kDa gut protein that is indicative of diapause is expressed in intact and neck-ligated pharate larvae but is not expressed when a ligature is placed posterior to the prothorax, site of the prothoracic gland. Guts cultured in vitro for 12 h cease to synthesize the 55 kDa protein, but synthesis of the protein resumes if the culture medium is enriched with a prothorax extract from pharate larvae or a prothoracic gland extract from fifth instar larvae. Injection of 20-hydroxyecdysone or the ecdysteroid agonist, RH-5992, into isolated abdomens stimulates synthesis of the diapause-specific 55 kDa protein, suggesting that the essential factor from the prothorax is an ecdysteroid. KK-42, an imidazole derivative known to inhibit ecdysteroid biosynthesis, averts diapause when applied to prediapausing pharate first instar larvae, but this effect can be countered by application of 20-hydroxyecdysone or RH-5992, i.e. KK-42 treated pharate larvae that are exposed to an ecdysteroid or RH-5992 readily enter diapause. A chilling period (120 days at 5 degrees C) is normally adequate to prompt an immediate termination of diapause when pharate larvae are transferred to 25 degrees C, but if such larvae are held in hanging drop cultures with ecdysteroids they fail to terminate diapause. Together, these results suggest that ecdysteroids are essential for the induction and maintenance of diapause and imply that a drop in the ecdysteroid titer is essential for diapause termination. Copyright 1997 Elsevier Science Ltd. All rights reserved     

Ecotope mapping for landscape ecological assessment of habitat and ecosystem

An ecotope (spatial eco-space) map that considers topography and bio-organism-relevant variables emerges as an important basic framework when landscape-scale characteristics for ecosystem management and wildlife conservation are needed. A spatio-geoecological framework based on geographic information systems (GIS) and a vegetation survey were developed for wildlife habitat evaluation of national parks and applied to a representative rugged valley area of Mt. Sorak National Park in Korea. An ecotope map was classified into hundreds of types and dozens of groups by combining biological and geophysical variables. Variables included: forest vegetation type, topographic solar radiation, normalized difference vegetation index (NDVI), elevation, and anthropogenic factors, such as, streams and roads. Layers of GIS variables were produced by field surveys, modeling, satellite images, or digitalization. Vegetation surveys were carried out to identify finer-scale distribution of vegetation types in the rugged valley area. Digital forest vegetation maps from the Forestry Administrator were then modified using the field-surveyed vegetation maps. Topographic solar radiation was predicted with a daily topographic radiation model. The NDVI was calculated from the satellite imagery of a Landsat Thematic Mapper. A digital elevation model (DEM) was used and the other layers were digitized using topographical maps with a scale of 1:25000. The aim of this study is to determine the geoecological factors relating to the spatial pattern of plant community. It was cleared by the spatial pattern of environmental variables and vegetation characteristics by detrended correspondence analysis using plant species and the environmental variables of each plot. The ordination component value of the first axis shows significant regression to some environmental variables. A case study of habitat evaluation was carried out using the resultant ecotope map. The spatial distribution of potential goral habitat and vegetation characteristics were predicted and the impact of human trails on the neighboring vegetation was also examined for restoration planning. The GIS-based framework developed for wildlife habitat evaluation is useful for natural resource management and human activity control in national parks in Korea.     

Enhanced litter input rather than changes in litter chemistry drive soil carbon and nitrogen cycles under elevated CO2: a microcosm study

Elevated CO₂ has been shown to stimulate plant productivity and change litter chemistry. These changes in substrate availability may then alter soil microbial processes and possibly lead to feedback effects on N availability. However, the strength of this feedback, and even its direction, remains unknown. Further, uncertainty remains whether sustained increases in net primary productivity will lead to increased long‐term C storage in soil. To examine how changes in litter chemistry and productivity under elevated CO₂ influence microbial activity and soil C formation, we conducted a 230‐day microcosm incubation with five levels of litter addition rate that represented 0, 0.5, 1.0, 1.4 and 1.8 × litterfall rates observed in the field for aspen stand growing under control treatments at the Aspen FACE experiment in Rhinelander, WI, USA. Litter and soil samples were collected from the corresponding field control and elevated CO₂ treatment after trees were exposed to elevated CO₂ (560 ppm) for 7 years. We found that small decreases in litter [N] under elevated CO₂ had minor effects on microbial biomass carbon, microbial biomass nitrogen and dissolved inorganic nitrogen. Increasing litter addition rates resulted in linear increase in total C and new C (C from added litter) that accumulated in whole soil as well as in the high density soil fraction (HDF), despite higher cumulative C loss by respiration. Total N retained in whole soil and in HDF also increased with litter addition rate as did accumulation of new C per unit of accumulated N. Based on our microcosm comparisons and regression models, we expected that enhanced C inputs rather than changes in litter chemistry would be the dominant factor controlling soil C levels and turnover at the current level of litter production rate (230 g C m⁻² yr⁻¹ under ambient CO₂). However, our analysis also suggests that the effects of changes in biochemistry caused by elevated CO₂ could become significant at a higher level of litter production rate, with a trend of decreasing total C in HDF, new C in whole soil, as well as total N in whole soil and HDF.