Meena Haribal

Natural variation in maize aphid resistance is associated with 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside methyltransferase activity

Susceptibility to corn leaf aphid in maize is associated with changes in defense-inducing maize benzoxazinoid levels, with high HDMBOA-Glc levels and low DIMBOA-Glc levels leading to toxicity but a reduced plant aphid defense response of callose deposition. Variation in HDMBOA-Glc production is due to a transposon insertion that inactivates O-methyltransferases in lines with low HDMBOA-Glc levels. Plants differ greatly in their susceptibility to insect herbivory, suggesting both local adaptation and resistance tradeoffs. We used maize (Zea mays) recombinant inbred lines to map a quantitative trait locus (QTL) for the maize leaf aphid (Rhopalosiphum maidis) susceptibility to maize Chromosome 1. Phytochemical analysis revealed that the same locus was also associated with high levels of 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one glucoside (HDMBOA-Glc) and low levels of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside (DIMBOA-Glc). In vitro enzyme assays with candidate genes from the region of the QTL identified three O-methyltransferases (Bx10a-c) that convert DIMBOA-Glc to HDMBOA-Glc. Variation in HDMBOA-Glc production was attributed to a natural CACTA family transposon insertion that inactivates Bx10c in maize lines with low HDMBOA-Glc accumulation. When tested with a population of 26 diverse maize inbred lines, R. maidis produced more progeny on those with high HDMBOA-Glc and low DIMBOA-Glc. Although HDMBOA-Glc was more toxic to R. maidis than DIMBOA-Glc in vitro, BX10c activity and the resulting decline of DIMBOA-Glc upon methylation to HDMBOA-Glc were associated with reduced callose deposition as an aphid defense response in vivo. Thus, a natural transposon insertion appears to mediate an ecologically relevant trade-off between the direct toxicity and defense-inducing properties of maize benzoxazinoids.

Isothiocyanates Stimulating Oviposition by the Diamondback Moth, Plutella xylostella

Recognition of cabbage as a host plant for the diamondback moth (DBM) has previously been shown to depend on compounds that are extracted by soaking intact foliage in chloroform. Analysis of such chloroform extracts by open column chromatography has now resulted in the isolation of highly active fractions that elicit oviposition on treated filter papers. Further separation of these fractions by high-performance liquid chromatography revealed the presence of two distinct groups of active compounds that may be classified as volatile and non-volatile. The two prominent volatile components were separated and identified by mass spectrometry as the isothiocyanates, iberin (3-methylsulfinylpropyl isothiocyanate) and sulforaphane (4-methylsulfinyl-3-butenyl isothiocyanate). Subsequent bioassays of a range of isothiocyanates showed that iberin and sulforaphane were the most active of those tested. Other isothiocyanates with sulfur in the side chain were also active, whereas alkyl and phenyl isothiocyanates had only limited activity. In electrophysiological experiments, electroantennograms (EAGs) indicated positive responses of moth antennae to the isothiocyanates that were most active in behavioral assays. Since sulforaphane has been identified as a major inducer of anticarcinogenic activity in mouse tissue, a synthetic analog (exo-2-acetyl-5-isothiocyanatonorbornane) that shows similar inducer activity was tested on DBM. This bicyclic analog was highly active in both behavioral and EAG assays, suggesting similarity in receptor sites for the two types of biological activity.

DUAL CHEMICAL BARRIERS PROTECT A PLANT AGAINST DIFFERENT LARVAL STAGES OF AN INSECT

The host plants of the native American butterfly, Pieris napi oleracea, include most wild mustards. However, garlic mustard, Alliaria petiolata, a highly invasive weed that was introduced from Europe, appears to be protected from this insect. Although adults will oviposit on the plant, most larvae of P. n. oleracea do not survive on garlic mustard. We used feeding bioassays with different larval stages of the insect to monitor the isolation and identification of two bioactive constituents that could explain the natural resistance of this plant. A novel cyanopropenyl glycoside (1), alliarinoside, strongly inhibits feeding by first instars, while a flavone glycoside (2), isovitexin-6″-D-β-glucopyranoside, deters later instars from feeding. Interestingly, the first instars are insensitive to 2, and the late instars are little affected by 1. Furthermore, differential effects of dietary experience on insect responses suggest that 1 acts through a mechanism of post-ingestive inhibition, whereas 2 involves gustatory deterrence of feeding.

Sequestration of aristolochic acids by the pipevine swallowtail, Battus philenor (L.): evidence and ecological implications

Summary. It has long been assumed that the North American pipevine swallowtail, Battus philenor (L.) (Papilionidae, Troidini), is protected from natural enemies by aristolochic acids sequestered from its Aristolochia food plants. This study confirmed that populations of B. philenor from Virginia and east Texas sequester these compounds. A comparison of the aristolochic acid profiles of the Virginia butterflies and their A. macrophylla food plants revealed several differences. The aristolochic acid fraction of the foliage was dominated by aristolochic acids I and II, whereas the insects had a much lower proportion of aristolochic acid II and contained, in addition, substantial amounts of aristolochic acids Ia and IVa, which were not detected in the plants. The eggs, larval integument, osmeterial glands, pupal cuticle, and adults (wings and bodies) all contained aristolochic acids. These findings help explain the abundant ecological data indicating that both immature and adult B. philenor are unpalatable and protected from natural enemies.

Isovitexin 6″-O-β-d-glucopyranoside: A feeding deterrent to Pieris napi oleracea from Alliaria petiolata

Abstract Pieris napi oleracea, an indigenous butterfly to North America lays eggs on Alliaria petiolata, an invasive weed introduced from Europe, but the larvae generally do not survive. A new apigenin glycoside, isovitexin 6″-O-glucoside has been isolated from the leaves A. petiolata and identified as a feeding deterrent for P. napi fourth instar larvae. The structure was elucidated by UV, MS and NMR spectroscopy.

Seasonal and population variation in flavonoid and alliarinoside content of Alliaria petiolata.

Pieris napi oleracea, an indigenous butterfly in North America, lays eggs on Alliaria petiolata, an invasive weed that was introduced from Europe. However, larval development on plants from different sources varies considerably. A. petiolata is a compulsive biennial, and its foliage is rich in apigenin flavonoids. We compared the chemistry of different vegetative forms from different populations in the vicinity of Ithaca, NY throughout the year. Significant differences occurred in the number of apigenin derivatives in different populations and vegetative forms, and seasonal variations in the amounts of these compounds were found. We have previously isolated two major compounds, alliarinoside [(2Z)-4-(β-D-glucopyranosyloxy)-2-butenenitrile] (1) and isovitexin-6-O″β-D-glucoside (3), which negatively affect development of P. napi oleracea larvae. Comparative analyses of these compounds in two populations throughout the year showed that their concentrations reached maxima twice annually. Foliage is almost devoid of flavonoids in June-July. Thus, variation in the chemistry of the plant may account for observed variation in development rates and survival of the larvae. Several apigenin compounds were isolated and identified by spectral studies.

Chemistry of preen gland secretions of passerines: different pathways to same goal? why?

Summary.Passeriformes is the largest order of birds and includes one third of the bird species of the world, living in very diverse habitats. We investigated the chemistry of preen gland secretions of some groups of passerines from temperate regions found in diverse microhabitats. Some of the common components were mixtures of homologous monoesters made up of long chain acids and alcohols. Individual species had characteristic distribution of esters and was unique to a given species, although there were some individual variations. We compared the composition of acids and alcohols that formed same molecular weight esters in different species and we found that the combination of acids and alcohols to arrive at same molecular compositions varied distinctly between species. To compare compositions of over all acids and alcohols that formed the esters, the representative samples of secretions from the individual species were transesterfied the produce methyl esters and alcohols. We found that there were distinct differences in number of acids and alcohols that produced the combination of homologous mixtures of esters. Also they differed both qualitatively and quantitatively. There were also seasonal differences in the secretion components. Thus though the intact mixtures of esters in individual species had some similarities, they were very complex mixtures and differed characteristically for individual species. Here we discuss possible causes for evolution of these variations. We suggest that the evolution in variation of preen gland secretion is probably due to selective pressures caused by ectosymbionts such as feather-mites and feather-chewing lice that live on feathers and probably feed on the secretions and surrounding environments

Identification and distribution of oviposition stimulants for monarch butterflies in hosts and nonhosts

Flavonol glycosides that act as oviposition stimulants for monarch butterflies were surveyed from a range of asclepiad hosts and some nonhosts. Major stimulants also were identified as quercetin-3-O-(2″-O-β-xylosyl)-β-D-galactoside and quercetin-3-O-β-D-galactoside from Asclepias syriaca and A. incarnata, respectively. The flavonol glycosides in A. curassavica, A. tuberosa, A. incarnata, A. syriaca, A. humistrata, A. albicans, A. eriocarpa, Calotropis procera, Cynanchum acutum, Vincetoxicum (Cynanchum) nigrum and in nonhosts Hoya australis and Nerium oleander were compared and characterized by HPLC and spectral studies. There was great variation in quercetin glycoside content. On the basis of the sugar moieties attached to quercetin, the asclepiad glycosides were classified broadly as those containing: (1) galactose, glucose, and rhamnose; (2) galactose, glucose, and xylose; and (3) galactose, glucose, xylose, and rhamnose. In most cases, galactose was attached to the 3-O-position (1″) of quercetin and other sugars were attached either to the 2″ or 6″ position of galactose. The sugars of triglycosides were attached at both 2″ and 6″ positions. A geographical pattern of flavonol distribution that may have affected the evolution of host recognition by the butterflies is suggested.

A Caffeoylcyclohexane-1-Carboxylic Acid Derivative From Asimina Triloba

Abstract Three caffeoyl derivatives of 1,3,4,5-tetrahydroxycyclohexane 1-carboxylic acid, one of them an oviposition stimulant for the zebra swallowtail butterfly, Eurytides marcellus , were isolated from Asimina triloba . The structure of the active isomer, 3-caffeoyl- muco -quinic acid, was determined by extensive NMR studies.

Combined roles of contact stimulant and deterrents in assessment of host-plant quality by ovipositing zebra swallowtail butterflies.

Zebra swallowtail (Eurytides marcellus) butterflies are stimulated to oviposit by a single compound, 3-caffeoyl-muco-quinic acid (1). Analysis of the aqueous extracts of the leaves of the host, Asimina triloba, showed that they contained stimulant 1, its isomer (2), and the flavonoids rutin (3) and nicotiflorine (4) as major components. We compared the concentrations of compounds 1–4 in terminal leaves (TL) and expanded leaves (EL) of the host plants at four different times throughout the growing season. In spring, the concentration of 1 was highest in TLs, and flavonoids were not detectable or present at low levels. As the season progressed, however, the concentrations of flavonoids increased, reached maxima by late summer, and then decreased as the plants started senescing. There were also significant differences in the concentrations of these compounds between TLs and ELs. In a choice assay with model leaves, we tested equivalent amounts of post-dichloromethane aqueous extracts made in spring (May) and in fall (September). September extracts received significantly fewer approaches and eggs. In greenhouse experiments with potted A. triloba plants, the butterflies chose some leaves to lay eggs, while others were rejected or ignored. Analyses showed that the concentrations of compound 1 were not significantly different in the three kinds of leaves. The flavonoids (3 and 4), however, were significantly higher in the leaves that were ignored. Multiple-choice tests using model plants suggested that concentrations of both flavonoids and stimulant were important in assessing host suitability. There was a gradual decrease in approaches as the concentration of 1 decreased. Higher amounts of flavonoids deterred egg laying even in the presence of high concentrations of stimulant 1. At lower concentrations of 1, the addition of low doses of flavonoids deterred egg laying. Thus, the results suggest that the butterflies use both qualitative and quantitative information about these compounds to assess host quality. This behavior may have evolved to take advantage of seasonal variation in the chemistry of their host, A. triloba.