Bioconversion of sodium dodecyl sulphate to rhamnolipids by transformed Escherichia coli DH5α cells-a novel strategy for rhamnolipid synthesis.

AIMS: Biological synthesis of rhamnolipids from SDS by Pseudomonas aeruginosa S15 is found to be a cost effective mode of rhamnolipid synthesis. This study aimed to attempt rhamnolipid synthesis by transformant Escherichia coli DH5α cells. METHODS AND RESULTS: Molecular analysis by curing experiments revealed that the properties of SDS based rhamnolipid synthesis were plasmid borne. Transformation of 10 kb plasmid to E. coli DH5α cells conferred rhamnolipid synthetic ability to transformant. Various genetic elements involved in SDS based rhamnolipid synthesis were analyzed using PCR based and restriction digestion based approaches. PCR amplification using primers specific for sdsA gene encoding alkylsulfatases yielded two significant amplicons viz, 1·2 kb fragment and 422 bp fragment, coding for putative dehydratase and ABC transporter respectively. Amplicon of sdsB gene lacked ability of SDS degradation and rhamnolipid synthesis. Rhamnolipid biosynthesis by transformant E. coli DH5α containing the whole of the 10 kb plasmid, was optimized to yield of 3·38 g l(-1) in 5 days of incubation. CONCLUSIONS: Plasmid encoded rhamnolipid synthesis from recombinant E. coli cells is novel and could serve as yet another promising approach among various steps adopted for safe and effective rhamnolipid synthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: SDS based rhamnolipid synthesis by S15 attained a high substrate (SDS) to product (Rhamnolipid) conversion ratio. However, the use of Pseudomonas strains is always discouraged as they are opportunistic pathogens and could sometimes turn infectious. Thus, transformation of genetic elements coding SDS based rhamnolipid synthesis to nonpathogenic strains could be promising.

Reflective mulch and acibenzolar-S-methyl treatments relative to thrips (Thysanoptera: Thripidae) and tomato spotted wilt virus incidence in tomato.

Management of thrips-transmitted tomato spotted wilt (TSW) virus typically relies on tactics that either reduce the thrips vector numbers or change the plant's response to the virus to reduce economic loss. We attempted to quantify the interaction between two such tactics, reflective mulch and the plant activator acibenzolar-S-methyl (Actigard), respectively, on a TSW-susceptible tomato hybrid. A split plot experiment was conducted in 2009 and 2010 where main-plots were three types of plastic mulch (two metalized reflective vs. black) and subplots consisted of a range of plant defense activator applications. TSW pressure varied over year with 80% of untreated plants having TSW in 2009 where as <7% of plants was infected in 2010. No significant interaction between mulch and subplots was found relative to thrips and marketable yield in either year. In 2009, the seasonal average of Frankliniella fusca (Hinds) populations and incidence of TSW were significantly lower and yield significantly higher on both reflective mulches than on black mulch. Seasonal averages of thrips and fruit yield differed significantly among treatments of acibenzolar-S-methyl. However, there was a significant acibenzolar-S-methyl by mulch interaction relative to TSW incidence. In 2009, a minimum of acibenzolar-S-methyl at transplant plus foliar treatments at 10 and 20 d after transplant was required to significantly reduce TSW incidence compared with untreated plants before harvest. Under lower TSW pressure in 2010, average TSW incidence was significantly less in all plots treated with acibenzolar-S-methyl treated plots compared with the check. Acibenzolar-S-methyl treatments functioned better with the thrips reducing tactic, ultraviolet-reflective mulch. We propose that acibenzolar-S-methyl is less effective than metalized reflective mulch in reducing the incidence of TSW in tomato.

Effects of fertilizer and low rates of imidacloprid on Adelges tsugae (Hemiptera: Adelgidae).

Healthy hemlock trees, Tsuga canadensis (L.) Carrière, and hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), populations should favor retention and population growth of adelgid predators such as Laricobius nigrinus Fender (Coleoptera: Derodontidae) and Sasajiscymnus tsugae (Sasaji & McClure) (Coleoptera: Coccinellidae). Eastern hemlock trees between 15 and 38 cm diameter at breast height (dbh) were treated with 0, 10, or 25% of 1.5 g imidacloprid (Merit 75 WP) per 2.5 cm dbh and were either fertilized or not, in a 3 by 2 factorial design. After 2 yr, imidacloprid reduced the numbers of ovisacs and eggs found on trees in a dosage-dependent manner, while enhancing tree growth parameters such as new shoots or needles and the length of new shoots. Fertilized trees had greater adelgid fecundity, which was positively correlated with total foliar N in both winter generations. In February 2009 (27 mo after imidacloprid treatment), higher imidacloprid dosages to unfertilized trees resulted in reduced adelgid fecundity. Concentrations of N, P, and K were higher in the foliage of trees treated with insecticide, whereas foliar aluminum concentrations were consistently lower in trees with higher insecticide dosages. Trees treated with low rates of imidacloprid were healthier than untreated trees, but only trees treated with the 0.1 x dosage had sufficient adelgids to possibly sustain predators over extended periods.

Effects of fertilization of four hemlock species on Adelges tsugae (Hemiptera: Adelgidae) growth and feeding preference of predators.

Understanding how fertilization affects host resistance to hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is important because fertilizers are often used to grow resistant selections to a suitable size for testing. We evaluated four hemlock species (Tsuga) under three different fertilizer regimes to assess whether fertility affected resistance to the adelgid and to determine whether it affected feeding preferences of the adelgid predators Laricobius nigrinus Fender and Sasajiscymnus tsugae (Sasaji & McClure). Treatments were long-term fertilization (from June 2008 to June 2009), short-term fertilization (from March to June 2009), and no fertilizer. Fertilizer was applied biweekly with 240 ppm N by using water-soluble fertilizer (N-P-K, 20:20:20). Plants (>1 yr old) were artificially infested with adelgids on 31 March 2009. Among unfertilized hemlocks (n=10 per species), foliar N was highest in Tsuga mertensiana (Bong.) CarriBre and lowest in T. chinensis (Franch.) E. Pritz. Significantly more progredien ovisacs or sisten eggs were present on T. mertensiana than on the other hemlock species with none on unfertilized T. chinensis. A. tsugae adults on T. heterophylla (Raf.) Sarg. were unaffected by fertility, but densities of developing A. tsugae nymphs were higher on unfertilized T. heterophylla plants than on fertilized T. heterophylla plants regardless of fertilizer treatment. Both L. nigrinus and S. tsugae consumed more adelgid eggs that developed on fertilized T. canadensis than from unfertilized plants. The predators did not exhibit this preference for adelgid eggs from females that developed on T. heterophylla or T. mertensiana.

Distribution and abundance of Adelges tsugae (Hemiptera: Adelgidae) within hemlock trees.

We studied the distribution of hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), within hemlock trees for three summer (progrediens) and two winter (sistens) generations in northern Georgia. Eastern hemlock, Tsuga canadensis (L.) Carrière, trees were treated with 0, 10, or 25% of 1.5 g of imidacloprid per 2.5 cm of tree diameter at breast height and fertilized or not in a factorial design. Adelgid ovisacs per centimeter of branch were more abundant from June 2007 to June 2008 in the upper tree crown of insecticide untreated trees and when all trees were combined and that was the general trend for most comparisons. However, ovisacs were more abundant in the lower crown of insecticide treated trees in June 2008. More sistens nymphs settled on the upper crown branches than on the lower branches in summers 2007 and 2008. Higher eggs per ovisac were observed in the upper crown in February 2008 and in both the winter and summer 2009. In contrast, adelgids were more fecund in the lower crown in June 2008. On fertilized trees, eggs laid per adult were higher in the upper crown in February 2008. In summer 2008, eggs per ovisac were higher in the lower crown, but this reversed again to the upper crown by summer 2009. New growth of branches also varied among sample dates. These data demonstrate the variable distribution of adelgid and hemlock growth within trees over time and suggest that sampling only one crown area will not provide accurate estimates of adelgid densities.

Influence of plant parameters on occurrence and abundance of arthropods in residential turfgrass.

The effect of taxa [common Bermuda grass, Cynodon dactylon (L.); centipedegrass, Eremochloa ophiuroides Munro Hack; St. Augustinegrass, Stenotaphrum secundatum [Walt.] Kuntze; and zoysiagrass, Zoysia spp.], density, height, and weed density on abundance of natural enemies, and their potential prey were evaluated in residential turf. Total predatory Heteroptera were most abundant in St. Augustinegrass and zoysiagrass and included Anthocoridae, Lasiochilidae, Geocoridae, and Miridae. Anthocoridae and Lasiochilidae were most common in St. Augustinegrass, and their abundance correlated positively with species of Blissidae and Delphacidae. Chinch bugs were present in all turf taxa, but were 23-47 times more abundant in St. Augustinegrass. Anthocorids/lasiochilids were more numerous on taller grasses, as were Blissidae, Delphacidae, Cicadellidae, and Cercopidae. Geocoridae and Miridae were most common in zoysiagrass and were collected in higher numbers with increasing weed density. However, no predatory Heteroptera were affected by grass density. Other beneficial insects such as staphylinids and parasitic Hymenoptera were captured most often in St. Augustinegrass and zoysiagrass. These differences in abundance could be in response to primary or alternate prey, or reflect the influence of turf microenvironmental characteristics. In this study, Simpson's diversity index for predatory Heteroptera showed the greatest diversity and evenness in centipedegrass, whereas the herbivores and detritivores were most diverse in St. Augustinegrass lawns. These results demonstrate the complex role of plant taxa in structuring arthropod communities in turf. An increased understanding of how turf species and cultivars help shape pest and beneficial arthropod communities will enhance predictive abilities and further pest management objectives.

Predatory potential of Geocoris spp. and Orius insidiosus on fall armyworm in resistant and susceptible turf.

Predatory potential and performance of the predaceous heteropterans, Geocoris punctipes (Say), G. uliginosus (Say) (Geocoridae), and Orius insidiosus (Say) (Anthocoridae), were evaluated using fall armyworm, Spodoptera frugiperda (J. E. Smith), as prey on different turfgrass taxa (resistant zoysiagrasses, 'Cavalier' and 'Palisades'; moderately resistant Bermuda grass, 'TifSport'; and susceptible seashore paspalum, 'Sea Isle 1') through laboratory and field studies. When background mortality was taken into account, in small arena trials in the laboratory, the greatest mortality by predators occurred on TifSport. The predator impact on TifSport by O. insidiosus was 92.6% above the mortality in the no-predator treatment on that grass. Predator induced mortality was rarely significant on the highly resistant zoysiagrass cultivar Cavalier because mortality, even in the absence of predators, was so high. Survival of larvae on TifSport Bermuda grass was significantly reduced by the addition of just two O. insidiosus per pot in laboratory pot trials. An increase in predator density to 4, 6, 8, or 10 further suppressed larval survival. O. insidiosus reduced larval survival on Sea Isle 1 at all densities. On Sea Isle 1, a density of two O. insidiosus resulted in > 50% reduction in live fall armyworms compared with the no predator treatment in laboratory trials. However, addition of O. insidiosus did not significantly reduce survival of fall armyworm larvae on this cultivar in the field in the presence of alternative prey and predators. O. insidiosus densities of six or higher per 181.4 cm2 did significantly reduce larval survival on TifSport Bermuda grass by as much as 80% during a 5-d trial period in the field. Predator-induced mortality among all trials was most consistent on a grass of intermediate resistance, TifSport Bermuda grass.