Coexistence of predominantly nonculturable rhizobia with diverse, endophytic bacterial taxa within nodules of wild legumes

A previous analysis showed that Gammaproteobacteria could be the sole recoverable bacteria from surface-sterilized nodules of three wild species of Hedysarum. In this study we extended the analysis to eight Mediterranean native, uninoculated legumes never previously investigated regarding their root-nodule microsymbionts. The structural organization of the nodules was studied by light and electron microscopy, and their bacterial occupants were assessed by combined cultural and molecular approaches. On examination of 100 field-collected nodules, culturable isolates of rhizobia were hardly ever found, whereas over 24 other bacterial taxa were isolated from nodules. None of these nonrhizobial isolates could nodulate the original host when reinoculated in gnotobiotic culture. Despite the inability to culture rhizobial endosymbionts from within the nodules using standard culture media, a direct 16S rRNA gene PCR analysis revealed that most of these nodules contained rhizobia as the predominant population. The presence of nodular endophytes colocalized with rhizobia was verified by immunofluorescence microscopy of nodule sections using an Enterobacter-specific antibody. Hypotheses to explain the nonculturability of rhizobia are presented, and pertinent literature on legume endophytes is discussed.     

Resprouting of <Emphasis Type="Italic">Echinacea angustifolia</Emphasis> Augments Sustainability of Wild Medicinal Plant Populations

Resprouting of Echinacea angustifolia Augments Sustainability of Wild Medicinal Plant Populations. Overharvest of wild Echinacea species root has been a significant concern to the herbal industry. Harvesters of wild Echinacea angustifolia showed us that even after harvesting the top 15 to 20 cm of root, some plants resprout. We marked locations of harvested plants at sites in Kansas and Montana and reexamined them two years later to see if they resprouted from remaining root reserves. Approximately 50% of the roots resprouted at both Kansas and Montana sampling sites, despite droughty weather conditions in Montana. The length of root harvested significantly affected the ability of the plant to resprout. Those plants that were more shallowly harvested and had less root length removed were more likely to resprout. These data indicate that echinacea stands can recover over time from intensive harvest if periods of nonharvest occur. Our echinacea harvest study emphasizes that the entire biology of medicinal plants must be considered when evaluating their conservation status.     

Mexican Arnica Anti–Inflammatory Action: Plant Age is Correlated with the Concentration of Anti–Inflammatory Sesquiterpenes in the Medicinal Plant <Emphasis Type="Italic">Heterotheca inuloides</Emphasis> Cass. (Asteraceae)<Superscript>1</Superscript>

Mexican Arnica Anti–Inflammatory Action: Plant Age Is Correlated with the Concentration of Anti–inflammatory Sesquiterpenes in the Medicinal Plant Heterotheca inuloides Cass. (Asteraceae). Mexican árnica (Heterotheca inuloides Cass.) is a widely used anti–inflammatory medicinal plant in Mexican folk medicine. Although it has been suggested that plant age, fertilization, and harvesting regime influence the concentration of secondary compounds affecting the therapeutic activity of the plant, the effect of these variables on the concentration of the Mexican árnica anti–inflammatory compounds was not known. We quantified anti–inflammatory sesquiterpenes (caryolan–1, 9β–diol, cadalen–15–oic acid, 7–hydroxycadalene, 4–hydroxy–2–isopropyl–4, 7–dimethyl–1[4H] naftalinone, 7–hydroxy–4αH–3, 4–dihydrocadalene, β–caryophyllene, and β–caryophyllene epoxide) in Mexican árnica plants subjected to fertilization and successive harvests of flowering stems, conditions that mimic the cultivation and harvesting for árnica in México. Fertilization and successive harvesting and their interaction had no significant effect on the concentration of anti–inflammatory compounds. However, the concentrations of these compounds were 60% higher in flowering stems from 15–month–old plants than in those from 4– or 8–month–old plants and was independent of the number of harvests and fertilization regime applied.     

Relationships between food quality and fitness in the desert locust, Schistocerca gregaria, and its distribution over habitats on the Red Sea coastal plain of Sudan

The effect of millet, Pennisetum typhoideum Rich. (Poaceae), leaf nitrogen content on fitness parameters of the desert locust, Schistocerca gregaria Forsk. (Orthoptera: Acrididae), was studied under laboratory conditions. Locusts reared on high‐nitrogen leaves were larger, developed faster, had higher survival, reproduced more and earlier, and showed greater synchronization than those fed on low‐nitrogen leaves. Active and passive cannibalism contributed to mortality when locusts were reared on low‐nitrogen leaves, but not when reared on high‐nitrogen leaves. Elevated leaf nitrogen content of host plants increased net reproduction and intrinsic rate of increase, and lowered generation time. The findings show that nitrogen content of host plants affects the potential for population increase in the desert locust. Leaf samples of common plant species were collected in the Heliotropium arbainense (Fresen.) (Boraginaceae) and Panicum turgidum (Forssk.) (Poaceae) plant communities on the Red Sea coastal plain of Sudan during the winters of 1999 and 2000. The levels of leaf nitrogen in host plants were comparable to those in the laboratory studies and consistently higher in plant samples from the Heliotropium community than in samples from the Panicum community. Both in 1999 and 2000, locust densities were much higher in the Heliotropium than in the Panicum plant community. It should be assessed whether the desert locust would be attracted to sites where host plants have high leaf nitrogen content, as this would not only increase their fitness, but also the likelihood of gregarization and outbreaks.     

Uses and Conservation of Plant Species in a National Park—A Case Study of Ben En, Vietnam

Uses and Conservation of Plant Species in a National Park—A Case Study of Ben En, Vietnam. This paper surveys the use of wild and cultivated plants by local people in Ben En National Park, Vietnam, and analyzes its impact on the conservation status of some of the utilized species. A total of 208 species used for a range of nonmedicinal purposes are listed. See Hoang et al. (2008a) for 230 medicinal plants used in the park. Most species are used for food. The use of plants contributes very significantly to the livelihood of local people in the park, but the current use patterns are not sustainable and would lead to local extinction of rare and endangered species if no additional conservation measures are introduced. Men collect nonmedicinal plants more often than women. A total of 38 useful plant species are commercialized, and contribute 12% of the average income of individual households. Bamboo shoots of Schizostachyum funghomii (Poaceae) are the most important for income generation. The monetary equivalent of noncommercialized useful plants probably far exceeds the value of the traded plant products. Plant use is independent of the ethnicity of the different populations living in the park. Larger households make use of a greater variety of useful plant species than small families. Abundant species in the forest have a higher use index (UI) than less common species. Out of the 208 useful species, as many as 27 were found to be endangered locally, many more than the 11 or 8 endangered species included in national or global red lists. Currently, useful plants, especially important timber trees, are more abundant in the less disturbed parts of the park, far away from the villages, indicating the pressures of illegal logging and harvesting near villages on the ecosystems.     

The Houses That Matsutake Built<Superscript>1</Superscript>

The Houses That Matsutake Built. In the mountains of northwest Yunnan, China, a valuable mushroom, matsutake or song rong (Tricholoma matsutake) was commodified in the 1980s. Since that time, it has been exported in large quantities to Japan. The sale of matsutake now contributes more to the income of Shangri-la County (Diqing Autonomous Tibetan Prefecture) than any other crop, including timber and livestock. During the 1980s and 1990s, villagers in this remote region used their mushroom earnings to build spacious, beautiful new houses in the traditional local (Kham) style, and in some cases to buy motor vehicles or open businesses. In villages with access to productive matsutake habitat, virtually every household was able to build a new house; entire villages were transformed. During the 1990s, several villages developed locally based management regimes to enhance production and to address the problems and conflicts that arose from the harvest of such a valuable product. More recently, government agencies and NGOs have played a highly visible role in promoting “sustainable” harvest policies. The implications of their involvement are briefly examined and the future of matsutake harvest in Yunnan is discussed.     

Kinase-dependent adhesion to fibronectin: regulation by calreticulin

We studied the phosphorylation (activation status) of c-Src and CaMKII in MEFs either wild type for calreticulin, calreticulin-null, or rescued with full-length calreticulin. We found that calreticulin-null cells were poorly spread on the substratum and formed few, if any, focal contacts. Fibronectin expression and deposition were lower in calreticulin-null MEFs compared to calreticulin-expressing cells, which also exhibited increased c-Src and CaMKII phosphorylation (activity). Plating MEFs on preformed fibronectin rescued the poor adhesive phenotype of calreticulin-null cells, and caused a decrease in c-Src Y418 phosphorylation (activity). c-Src inhibition caused the calreticulin-null MEFs to become well spread on the substratum and to make many prominent focal contacts. Calmodulin and CaMKII inhibition caused similar results, along with a notable increase in paxillin phosphorylation (activation). To test if the calcium storage function of calreticulin was responsible for these effects, we manipulated intracellular [Ca(2+)]. Lowering [Ca(2+)](ER) caused an increase in c-Src phosphorylation and a decrease in fibronectin abundance. Conversely, increasing [Ca(2+)] caused opposite effects. These results suggest that calreticulin regulates both the c-Src and calmodulin/CaMKII pathways, enabling cells to be better spread on the substratum by allowing greater fibronectin deposition and increased focal contact formation.     

Unscrambling thermal stability and temperature adaptation in evolved variants of a cold-active lipase

Directed evolution by error-prone PCR was applied to stabilize the cold-active lipase from Pseudomonas fragi (PFL). PFL displays high activity at 10 degrees C, but it is highly unstable even at moderate temperatures. After two rounds of evolution, a variant was generated with a 5-fold increase in half-life at 42 degrees C and a shift of 10 degrees C in the temperature optimum, nevertheless retaining cold-activity. The evolved lipase displayed specific activity higher than the wild type enzyme in the temperature range 29-42 degrees C. Biophysical measurements did not indicate any obvious difference between the improved variant and the wild type enzyme in terms of loss of secondary structure upon heat treatment, nor a shift in the apparent melting temperature.     

Dimerization of N-methyltransferases involved in caffeine biosynthesis

Caffeine is synthesized from the precursor xanthosine through three methylation and one nucleoside removal steps. Methylation is catalyzed by N-methyltransferases, designated as CaXMT1, CaMXMT1 and CaDXMT1, which, respectively, convert xanthosine into 7-methylxanthosine, 7-methylxanthine into 3,7-dimethylxanthine, and 3,7-dimethylxanthine into 1,3,7-trimethylxanthine (caffeine). In the present study, we examined their cytological and biochemical properties using fusion proteins with fluorescent proteins. All three enzymes were found to localize in cytosol as visualized by green fluorescence protein fusions. The possibility of dimer formation among these enzyme proteins was examined in vivo by transient expression of bimolecular fluorescence complementation of yellow fluorescent protein (YFP) using onion epidermal cell layers. Results showed that each enzyme protein formed a homo-dimer in cytosol as seen by a clear reconstituted YFP fluorescence. In addition, each enzyme also formed a hetero-dimer with each of the other two enzymes in cytosol. The biological significance of dimerization among structurally resembling methyltransferases involved in caffeine biosynthesis is discussed.     

Evaluation of antimicrobial activity of certain Chinese plants used in folkloric medicine

Six selected plants, belonging to 3 families from Nanjing of China, were extracted with the solvent 95% (v/v) ethanol to yield 11 extracts. The extracts were evaluated for their effects on the growth of eight clinical bacteria, two fungi and one yeast using a modified agar diffusion method. The results showed that the majority of the extracts investigated showed greater activities against the Gram-positive bacteria than against the Gram-negative bacteria, the fungi and the yeast. The strongest antimicrobial activity was exhibited by the stem extracts of Mahonia fortunei against multiresistant Staphylococcus aureus strains, followed by the stem extracts of Mahonia bealei, while Bacillus thuringiensis was the most sensitive to all extracts.