Development and characterization of simple sequence repeat (SSR) markers in the water lotus (Nelumbo nucifera)

Simple sequence repeat (SSR) markers were developed in the water lotus (Nelumbo nucifera Gaertn.) from an SSR-enriched genomic library. Of the SSR markers tested, 11 primer pairs produced clearly distinguishable DNA banding patterns. Forty-three alleles were detected with the 11 markers. The allele number per locus ranged from 2 to 5 with an average of 3.9. Polymorphism values ranged from 0.11 to 0.66 with an average of 0.51. These primers were also applicable to another Nelumbo species, Nelumbo lutea (Willd.) Pers. (American lotus) and hybrids between N. nucifera and N. lutea. These results indicate that the SSR markers developed in this study are informative and will be useful for genetic analysis in Nelumbo species.     

Cannibalism and Predation as Paths for Horizontal Passage of Wolbachia between Terrestrial Isopods

The alpha-proteobacteria Wolbachia are the most widespread endosymbionts in arthropods and nematodes. Mainly maternally inherited, these so-called sex parasites have selected several strategies that increase their vertical dispersion in host populations. However, the lack of congruence between the Wolbachia and their host phylogenies suggests frequent horizontal transfers. One way that could be used for horizontal Wolbachia transfers between individuals is predation. The aim of this study was to test whether horizontal passage of Wolbachia is possible when an uninfected terrestrial isopod eats an infected one. After having eaten Armadillidium vulgare harbouring Wolbachia, the predator-recipients (the two woodlice A. vulgare and Porcellio dilatatus dilatatus) that were initially Wolbachia-free were tested positive for the presence of Wolbachia both by quantitative PCR and Fluorescence in situ Hybridization (FISH). Even if the titers were low compared to vertically infected individuals, this constitutes the first demonstration of Wolbachia occurrence in various organs of an initially uninfected host after eating an infected one.     

Cultivable bacteria isolated from apple trees cultivated under different crop systems: Diversity and antagonistic activity against Colletotrichum gloeosporioides

This study evaluated the diversity of cultivable plant growth-promoting (PGP) bacteria associated with apple trees cultivated under different crop management systems and their antagonistic ability against Colletotrichum gloeosporioides. Samples of roots and rhizospheric soil from apple trees cultivated in organic and conventional orchards in southern Brazil were collected, together with soil samples from an area never used for agriculture (native field). Bacteria were identified at the genus level by PCR-RFLP and partial sequencing of the 16S rRNA, and were evaluated for some PGP abilities. The most abundant bacterial genera identified were Enterobacter (27.7%), Pseudomonas (18.7%), Burkholderia (13.7%), and Rahnella (12.3%). Sixty-nine isolates presented some antagonist activity against C. gloeosporioides. In a greenhouse experiment, five days after exposure to C. gloeosporioides, an average of 30% of the leaf area of plants inoculated with isolate 89 (identified as Burkholderia sp.) were infected, whereas 60 to 73% of the leaf area of untreated plants was affected by fungal attack. Our results allowed us to infer how anthropogenic activity is affecting the bacterial communities in soil associated with apple tree crop systems, and to obtain an isolate that was able to delay the emergence of an important disease for this culture.     

Fulton's body condition factor K correlates with survival time in a thermal challenge experiment in juvenile Lahontan cutthroat trout (Oncorhynchus clarki henshawi)

Thermal tolerance was measured in an experimental setting using seven full-sibling family groups of hatchery reared Lahontan cutthroat trout (Oncorynchus clarki henshawi). Body weight and fork length were measured and Fulton's body condition factor (K) was estimated for each individual. Heritabilities were moderately high for body weight (0.50), upper temperature tolerance (0.44), and K factor (0.50). Estimates of genetic and phenotypic correlations between upper temperature tolerance and K factor were 0.53 and 0.41, respectively. K factor may be an indicator as to the ability of an individual to survive periods of increased water temperature during the summer months.     

Synthesis and biological evaluation of 2-(5-methyl-4-phenyl-2-oxopyrrolidin-1-yl)-acetamide stereoisomers as novel positive allosteric modulators of sigma-1 receptor

Novel positive allosteric modulators of sigma-1 receptor represented by 2-(5-methyl-4-phenyl-2-oxopyrrolidin-1-yl)-acetamide enantiomers were synthesised using an asymmetric Michael addition of 2-nitroprop-1-enylbenzene to diethyl malonate. Following the chromatographic separation of the methyl erythro- and threo-4-nitro-3R- and 3S-phenylpentanoate diastereoisomers, target compounds were obtained by their reductive cyclisation into 5-methyl-4-phenylpyrrolidin-2-one enantiomers and the attachment of the acetamide group to the heterocyclic nitrogen. Experiments with electrically stimulated rat vas deference contractions induced by the PRE-084, an agonist of sigma-1 receptor, showed that (4R,5S)- and (4R,5R)-2-(5-methyl-4-phenyl-2-oxopyrrolidin-1-yl)-acetamides with an R-configuration at the C-4 chiral centre in the 2-pyrrolidone ring were more effective positive allosteric modulators of sigma-1 receptor than were their optical antipodes.     

Flavonoids of Plagiochila asplenioides

Besides an apigenin- and a luteolin-di-C-glycoside, 5 previously unknown di-C-glycosides of tricetin were identified in the gametophytic and sporophytic tissues of Plagiochila asplenioides. Two of them were new 6,8-di-C-hexopyranosyltricetins, and two were new 6-C- hexopyranosyl-8-C-pentopyranosyltricetins. 6-C-Hexopyranosyl-8-C-pentopyranosyltricetin-5′-methyl ether was also found.     

Distribution of Nothanguina phyllobia and Its Potential as a Biological Control Agent for Silver-leaf Nightshade

The nematode Nothanguina phyllobia Thorne was found within large foliar galls on the perennial weed Solanum elaeagnifolium Cav. in west Texas. A two-year survey of a 6400 sq-km area in west Texas showed extensive distribution of the nematode. No hosts other than S. elaeagnifolium were observed. Densities of juvenile nematodes in the soil were high. N. phyllobia spread rapidly after small numbers of infective juveniles were applied in a foliar spray to an S. elaeagnifolium population. The host plant declined in vigor and frequently died. Artificial inoculation of an S. elaeagnifolium population with large numbers of the nematodes by broadcasting infected plant tissue resulted in high infection incidence.     

Jointly silencing BoDWARF,BoGA20ox and BoSP (SELF-PRUNING) produces a novel miniature ornamental Brassica oleracea var. acephala f. tricolor variety

The ornamental Brassica oleracea var. acephala f. tricolor is a good winter and spring foliage plant. Plant architecture is an important agronomic trait of plants, especially for ornamental plants with high ornamental and economic value. In this study, three miniature-related genes, BoDWARF, BoGA20ox and BoSP (SELF-PRUNING), were cloned and their tissue-specific expression patterns were analyzed. The results showed that the three genes were all highly expressed in young leaves and flowers, followed by the lateral roots, seeds and stems. To further achieve the purpose of miniaturization of plants, an RNAi expression vector, jointly targeting BoDWARF, BoGA20ox and BoSP, was constructed and transformed into kale plants. Smaller plant size and slower growth and development speed of flowers and roots were observed in jointly silenced kales. Brassinosteroids and gibberellin contents in leaves and flower buds of transgenic plants were significantly decreased. Furthermore, the expressions of brassinolide-, gibberellin- and flowering-related genes were down-regulated by varying degrees in silenced plants. These results suggest that BoDWARF, BoGA20ox and BoSP play important roles in plant architecture, and that brassinolide and gibberellin are important hormones controlling plant growth and architecture. This miniaturization strategy of kale provides an efficient approach for cultivation of new varieties of ornamental plants and crops.     

Chelators in cupressaceae as iron transport agents for Salmonella typhimurium

Strains of Salmonella typhimurium which are unable to synthesize their own iron transport agents and require an erogenous chelator were used to examine extracts of the wood of species of Cupressaceae for the presence of iron chelators. Wood from 19 species of five genera were examined and all were found to contain substances that would function as iron transport agents for S. typhimurium. The biological activity of most of these species could be explained by the known presence and activity of the thujaplicins. Juniperus virginiana and J. occidentalis were found to contain a non-tropolone substance that functioned as chelators in S. typhimurium. The tropolone nootkatin from Chamaecyparis nootkatensis was ineffective as an iron transport agent.     

Novel Class 1 Integrons in Multi-drug Resistant Isolates from Eastern China

Integrons are mobile genetic elements able to capture, express and excise resistance genes, playing an important role in the spread of bacterial resistance. The present study was to investigate the occurrence and diversity of integrons in 120 clinical multi-drug resistant Gram-negative isolates from eastern China. Screening of integrons was performed by PCR and gene cassettes were further characterized by PCR–RFLP and sequencing. Class 1 integrons were detected in 70.8 % of isolates and no class 2 and class 3 integrons were detected in any isolates. A total of 19 resistant gene cassettes were identified, four representative of novel gene cassettes: an aacA3 variant (aacA3c), an aacA4 variant (aacA4′-17), a bla _OXA variant (bla _OXA-251 ), and a catB8 gene cassette interrupted by an insertion sequence IS10 (catB8::IS10). In addition, 14 cassette arrays were detected, including three novel integrons: gcuD1-aacA4′-17-gcu38B-catB8::IS10 (In712), aacA3c-aadA13-bla _OXA-251 (In713) and dfrA1-gcu37-aadA5 (In714). The presence of novel integron structures in clinical isolates suggests hospital environments may favor the formation of novel combination of gene cassettes. Moreover, the high prevalence of integrons in multi-drug resistant isolates highlights the urgent need to employ effective means to avoid dissemination of drug-resistant bacteria.     

Biologically active diterpene esters from euphorbia peplus

By means of partition and preparative TLC, two pro-inflammatory diterpene esters were isolated from Euphorbia peplus. These compounds were identified as 20-deoxyingenol 3-O-angelate which exhibited an ID₅₀ of O.18 μg on mice and the new ester ingenol 20-O-octanoate which exhibited an ID50 of 1.0 μg also on mice skin.     

Expression of xyloglucan endotransglucosylase/hydrolase (XTH) genes and XET activity in ethylene treated apple and tomato fruits

Xyloglucan endotransglucosylase/hydrolase (XTHs: EC 2.4.1.207 and/or EC 3.2.1.151), a xyloglucan modifying enzyme, has been proposed to have a role during tomato and apple fruit ripening by loosening the cell wall. Since the ripening of climacteric fruits is controlled by endogenous ethylene biosynthesis, we wanted to study whether XET activity was ethylene-regulated, and if so, which specific genes encoding ripening-regulated XTH genes were indeed ethylene-regulated. XET specific activity in tomato and apple fruits was significantly increased by the ethylene treatment, as compared with the control fruits, suggesting an increase in the XTH gene expression induced by ethylene. The 25 SlXTH protein sequences of tomato and the 11 sequences MdXTH of apple were phylogenetically analyzed and grouped into three major clades. The SlXTHs genes with highest expression during ripening were SlXTH5 and SlXTH8 from Group III-B, and in apple MdXTH2, from Group II, and MdXTH10, and MdXTH11 from Group III-B. Ethylene was involved in the regulation of the expression of different SlXTH and MdXTH genes during ripening. In tomato fruit fifteen different SlXTH genes showed an increase in expression after ethylene treatment, and the SlXTHs that were ripening associated were also ethylene dependent, and belong to Group III-B (SlXTH5 and SlXTH8). In apple fruit, three MdXTH showed an increase in expression after the ethylene treatment and the only MdXTH that was ripening associated and ethylene dependent was MdXTH10 from Group III-B. The results indicate that XTH may play an important role in fruit ripening and a possible relationship between XTHs from Group III-B and fruit ripening, and ethylene regulation is suggested.     

Pseudomonas cichorii as the causal agent of midrib rot,an emerging disease of greenhouse-grown butterhead lettuce in Flanders

Bacterial midrib rot of greenhouse-grown butterhead lettuce (Lactuca sativa L. var. capitata) is an emerging disease in Flanders (Belgium) and fluorescent pseudomonads are suspected to play an important role in the disease. Isolations from infected lettuces, collected from 14 commercial greenhouses in Flanders, yielded 149 isolates that were characterized polyphasically, which included morphological characteristics, pigmentation, pathogenicity tests by both injection and spraying of lettuce, LOPAT characteristics, FAME analysis, BOX-PCR fingerprinting, 16S rRNA and rpoB gene sequencing, as well as DNA–DNA hybridization. Ninety-eight isolates (66%) exhibited a fluorescent pigmentation and were associated with the genus Pseudomonas. Fifty-five of them induced an HR⁺ (hypersensitive reaction in tobacco leaves) response. The other 43 fluorescent isolates were most probably saprophytic bacteria and about half of them were able to cause rot on potato tuber slices. BOX-PCR genomic fingerprinting was used to assess the genetic diversity of the Pseudomonas midrib rot isolates. The delineated BOX-PCR patterns matched quite well with Pseudomonas morphotypes defined on the basis of colony appearance and variation in fluorescent pigmentation. 16S rRNA and rpoB gene sequence analyses allowed most of the fluorescent isolates to be allocated to Pseudomonas, and they belonged to either the Pseudomonas fluorescens group, Pseudomonas putida group, or the Pseudomonas cichorii/syringae group. In particular, the isolates allocated to this latter group constituted the vast majority of HR⁺ isolates and were identified as P. cichorii by DNA–DNA hybridization. They were demonstrated by spray-inoculation tests on greenhouse-grown lettuce to induce the midrib rot disease and could be re-isolated from lesions of inoculated plants. Four HR⁺ non-fluorescent isolates associated with one sample that showed an atypical midrib rot were identified as Dickeya sp.     

Empty loricas and the dynamics of Kellicottia longispina in a subalpine,oligotrophic lake

The seasonal dynamics of Kellicottia longispina in Lake Tahoe and an isolated embayment of Lake Tahoe, Emerald Bay, were investigated for an 18-month period in 1977–79. Population birth and death rates were similar in the two systems, although productivity and Mysis relicta densities were higher in Emerald Bay. The timing of population changes was also similar. A major population increase in late winter 1978 was preceded by an increase in egg ratio; the subsequent spring decline of K. longispina was concurrent with decreased birth rates and increased death rates. Empty loricas of K. longispina were occasionally abundant in the plankton samples and seemed to result from K. longispina deaths when densities were high and when egg ratios were declining; it is possible that population senescence was responsible for the high densities of empty loricas observed. A potentially important predator, M. relicta, defecates K. longispina remains in compact fecal pellets; however, it is unlikely that the observed empty loricas resulted from Mysis-related deaths.     

Effects of Crop Rotation and Nonfumigant Nematicides on Peanut and Corn Yields in Fields Infested with Criconemella Species

The effects of nematicide treatments and corn-peanut cropping sequences on the population development of Criconemella ornata, and C. sphaerocephala and the related impact on crop yields were investigated at two North Carolina locations. Criconemella ornata and C. sphaerocephala were present at the Norman Perry farm, Bertie County (BERTIE); however, only C. ornata was found at the Central Crops Research Station, Johnston County (CCRS). An untreated control was compared to aldicarb 15G, carbofuran 15G, ethoprop 10G, and terbufos 15G granular formulations applied at a rate of 2.2 kg a.i./ha. The cropping sequences were monocuhured corn (C-C-C); monocultured peanut (P-P-P); and two corn-peanut (C-P-C; P-C-P) rotations. Nematicides were inconsistent in controlling C. sphaerocephala and C. ornata. Nematicide treatments enhanced corn yields in the monoculture-cropping cycle in the final year of the experiment at CCRS. Peanut yields were greater in the rotated cropping sequence than under monoculture at BERTIE, but rotation had less effect on peanut yields at CCRS. Declining yields were correlated with an increase in numbers of nematodes. Corn was an intermediate host for C. sphaerocephala and a moderate to poor host for C. ornata. Peanut was an excellent host for C. ornata and a poor host for C. sphaerocephala.