Host recognition by a polyphagous lepidopteran (Helicoverpa armigera): Primary host plants, host produced volatiles and neurosensory stimulation

An important question in the host-finding behaviour of a polyphagous insect is whether the insect recognizes a suite or template of chemicals that are common to many plants? To answer this question, headspace volatiles of a subset of commonly used host plants (pigeon pea, tobacco, cotton and bean) and nonhost plants (lantana and oleander) of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) are screened by gas chromatography (GC) linked to a mated female H. armigera electroantennograph (EAG). In the present study, pigeon pea is postulated to be a primary host plant of the insect, for comparison of the EAG responses across the test plants. EAG responses for pigeon pea volatiles are also compared between females of different physiological status (virgin and mated females) and the sexes. Eight electrophysiologically active compounds in pigeon pea headspace are identified in relatively high concentrations using GC linked to mass spectrometry (GC-MS). These comprised three green leaf volatiles [(2E)-hexenal, (3Z)-hexenylacetate and (3Z)-hexenyl-2-methylbutyrate] and five monoterpenes (α-pinene, β-myrcene, limonene, E-β-ocimene and linalool). Other tested host plants have a smaller subset of these electrophysiologically active compounds and even the nonhost plants contain some of these compounds, all at relatively lower concentrations than pigeon pea. The physiological status or sex of the moths has no effect on the responses for these identified compounds. The present study demonstrates how some host plants can be primary targets for moths that are searching for hosts whereas the other host plants are incidental or secondary targets.

Development of a synthetic plant volatile-based attracticide for female noctuid moths. I. Potential sources of volatiles attractive to Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae).

This paper is the first of a series which will describe the development of a synthetic plant volatile-based attracticide for noctuid moths. It discusses potential sources of volatiles attractive to the cotton bollworm, Helicoverpa armigera (Hubner), and an approach to the combination of these volatiles in synthetic blends. We screened a number of known host and non-host (for larval development) plants for attractiveness to unmated male and female moths of this species, using a two-choice olfactometer system. Out of 38 plants tested, 33 were significantly attractive to both sexes. There was a strong correlation between attractiveness of plants to males and females. The Australian natives, Angophora floribunda and several Eucalyptus species were the most attractive plants. These plants have not been recorded either as larval or oviposition hosts of Helicoverpa spp., suggesting that attraction in the olfactometer might have been as nectar foraging rather than as oviposition sources. To identify potential compounds that might be useful in developing moth attractants, especially for females, collections of volatiles were made from plants that were attractive to moths in the olfactometer. Green leaf volatiles, floral volatiles, aromatic compounds, monoterpenes and sesquiterpenes were found. We propose an approach to developing synthetic attractants, here termed 'super-blending', in which compounds from all these classes, which are in common between attractive plants, might be combined in blends which do not mimic any particular attractive plant.

Increased attractiveness of honeybee hive product volatiles to adult small hive beetle, Aethina tumida, resulting from small hive beetle larval infestation

The small hive beetle, Aethina tumida Murray (Coleoptera: Nitidulidae), is a recent but significant pest of honeybee Apis mellifera L. (Hymenoptera: Apidae) hives in various regions throughout the world, including Eastern Australia. The larval stage of this beetle damages hives when they feed on brood, pollen, and honeycomb, leaving behind fermented wastes. In cases of extreme damage, hives collapse and are turned to an odorous mass of larvae in fermenting hive products. The yeast Kodamaea ohmeri (Etchells & Bell) Yamada et al. (Ascomycota) has been consistently isolated from the fermenting material as well as each life stage of this beetle. Various studies have noted that the small hive beetle is attracted to volatiles from hive products and those of the yeast K. ohmeri, although earlier studies have not used naturally occurring hive products as their source of fermentation. This study investigated changes through time in the attractiveness of natural honeybee hive products to the small hive beetle as the hive products were altered by the action of beetle larvae and fermentation by K. ohmeri. We used gas chromatography-mass spectrometry and choice-test behavioural assays to investigate these changes using products sampled from three apiaries. Attractiveness of the fermenting hive products (‘slime’) increased as fermentation progressed, and volatile profiles became more complex. Fermenting hive products remained extremely attractive for more than 30 days, significantly longer than previous reports. These results have strong implications for the development of an external attractant trap to assist in the management of this invasive pest.

Volatile components associated with bacterial spoilage of tropical prawns

Analysis of headspace volatiles by gas chromatography/mass spectrometry from king (Penaeus plebejus), banana (P. merguiensis), tiger (P. esculentus/semisulcatus) and greasy (Metapenaeus bennettae) prawns stored in ice or ice slurry, which is effectively an environment of low oxygen tension, indicated the presence of amines at the early stages of storage (less than 8 days) irrespective of the nature of the storage media. Esters were more prevalent in prawns stored on ice (normal oxygen conditions) at the latter stages of storage (more than 8 days) and were only produced by Pseudomonas fragi, whereas sulphides and amines occurred whether the predominant spoilage organism was Ps.fragi or Shewanella putrefaciens. The free amino acid profiles of banana and king prawns were high in arginine (12–14%) and low in cysteine (0.1–0.17%) and methionine (0.1–0.2%). Filter sterilised raw banana prawn broth inoculated with a total of 15 cultures of Ps. fragi and S. putrefaciens and incubated for two weeks at 5°C, showed the presence of 17 major compounds in the headspace volatiles analysed using gas chromatography/mass spectrometry (GC/MS). These were mainly amines, sulphides, ketones and esters. Principal Component Analysis of the results for the comparative levels of the volatiles produced by pure cultures, inoculated into sterile prawn broth, indicated three subgroupings of the organisms; I, Ps. fragi from a particular geographic location; II, S. putrefaciens from another geographic location; and III, a mixture of Ps. fragi and S. putrefaciens from different geographic locations. The sensory impression created by the cultures was strongly related to the chemical profile as determined by GC/MS. Organisms, even within the same subgrouping classified as identical by the usual tests, produced a different range of volatiles in the same uniform substrate.

Chemical Characterization of Urinary Pheromones in Brown Antechinus, Antechinus stuartii

In the small dasyurid marsupial, Antechinus stuartii, males exhibit scent-marking in the form of cloacal marking of nesting areas during the breeding season. Females of this species show no such behavior. To characterize the potential male pheromonal scent signal, urine-derived volatiles from sexually active males were analyzed by GC-MS and compared to that of females and a castrated male. More than 10 urinary compounds were identified. A series of homologous methylketones was observed in both males and females, whereas aldehydes were present only in female urine. Urine from the castrate was virtually compound-free except for minute concentrations of a compound tentatively identified as 2,4-dithiapentane. This compound was also found in one of the sexually active males. The GC profiles of the sexually active males contained high concentrations of two pyrazine derivatives and four methylketones that were not detected in the profiles of either females or the castrate. These compounds may influence social communication in the brown antechinus, Antechinus stuartii.

Variability in odour reception in the peripheral sensory system of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Mixtures of single odours were used to explore the receptor response profile across individual antennae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Seven odours were tested including floral and green-leaf volatiles: phenyl acetaldehyde, benzaldehyde, b-caryophyllene, limonene, a-pinene, 1-hexanol, 3Z-hexenyl acetate. Electroantennograms of responses to paired mixtures of odours showed that there was considerable variation in receptor tuning across the receptor field between individuals. Data from some moth antennae showed no additivity, which indicated a restricted receptor profile. Results from other moth antennae to the same odour mixtures showed a range of partial additivity. This indicated that a wider array of receptor types was present in these moths, with a greater percentage of the receptors tuned exclusively to each odour. Peripheral receptor fields show variation in the spectrum of response within a population (of moths) when exposed to high doses of plant volatiles. This may be related to recorded variation in host choice within moth populations as reported by other authors.

Demographics, pollination syndrome and conservation status of Macrozamia platyrhachis (Zamiaceae), a geographically restricted Queensland cycad

To assess the International Union for Conservation of Nature (IUCN) status of Macrozamia platyrhachis F.M.Bailey, we surveyed this central Queensland cycad for its population abundance and health and its pollinator type and pollination syndrome (thermogenesis and volatile emissions). Plants are locally abundant within the 11 discrete populations surveyed, with an estimated population of 611 315 adult plants. Plants are highly restricted to a small area of occupancy, seed dispersal is nearly non-existent and extreme fires appear to have destroyed almost all seeds and seedlings and decimated the pollinators. Of known Macrozamia pollinators, only the thrips, Cycadothrips chadwicki Mound, were found on cones, and these were found in very low numbers. The pollination syndrome for this cycad appears to be unique, based on two cone traits. For one, thermogenesis peaks in early evening, a contrast with daytime peaks of other Cycadothrips-pollinated Macrozamia, but matches that of the Tranes weevil-pollinated Macrozamia machinii. In addition, cone volatiles include both previously unreported compounds as well as those reported exclusively on either Cycadothrips- or Tranes-pollinated species. Based on its small, fragmented area of occupancy, projected population declines and the unique pollination syndrome, we recommend that M. platyrhachis retain its current status as 'Endangered'. Habitat management plans should stipulate that controlled burns be avoided during cycad coning season and that wildfires be controlled to minimise damage to seedlings and pollinators.

Hardwood Plantation P&D Semiochemistry

Insects can cause considerable damage in hardwood plantations and because pesticide use is controversial, future pest management may rely on manipulating insect behaviour. Insects use infochemical cues to identify and locate mates and host plants and this can be used to manipulate their behaviour and reduce pest impacts in plantations. Infochemicals include chemical signals produced by insects, such as pheromones and kairomones, or those produced by host plants as odours or volatiles that are attractive to insects. This research is learning how insects perceive and interact with chemical cues or infochemicals in their environment and how these interactions can be manipulated for monitoring and control. Pest species being investigated include the giant wood moth (Endoxyla cinerea), Culama wood moths, the eucalypt leaf beetle (Paropsis atomaria), red cedar tip moth (Hypsipyla robusta) and several longicorn wood borers. The project will contribute to new strategies for minimising damage and controlling pest densities in Queensland's hardwood plantations.

Improved understanding of the damage, ecology, and management of mirids and stinkbugs in Bollgard II

In recent years mirids and stinkbugs have emerged as important sucking pests in cotton. While stinkbugs are causing damage to bolls, mirids are causing damage to seedlings, squares and bolls. With the increasing adoption of Bollgard II and IPM approaches the use of broad-spectrum chemicals to kill Helicoverpa has been reduced and as a result mirids and stinkbugs are building to levels causing damage to bolls later in crop growth stages. Studies on stinkbugs by Dr Moazzem Khan revealed that green vegetable bug (GVB) caused significant boll damage and yield loss. A preliminary study by Dr Khan on mirids revealed that high mirid numbers at later growth stages also caused significant boll damage and that damage caused by mirids and GVB were similar. Mirids and stinkbugs therefore demand greater attention in order to minimise losses caused by these pests and to develop IPM strategies against these pests to enhance gains in IPM that have been made with Bt-transgenic cotton. Progress in this area of research will maintain sustainability and profitability of the Australian cotton industry. Mirid damage at early growth stages of cotton (up to squaring stage) has been studied in detail by Dr Khan. He found that all ages of mirids cause damage to young plants and damage by mirid nymphs is cumulative. Maximum damage occurs when the insect reaches the 4th and 5th nymphal stages. He also found that mirid feeding causes shedding of small and medium squares, and damaged large squares develop as ‘parrot beak’ bolls. Detailed studies at the boll stage, such as which stage of mirids is most damaging or which age boll is most vulnerable to feeding, is lacking. This information is a prerequisite to developing an IPM strategy for the pest in later crop growth stages. Understanding population change of the pest over time in relation to crop development is an important aspect for developing management strategies for the pest which is lacking for mirids in BollgardII. Predators and parasitoids are integral components of any IPM system and play an important part in regulating pest populations. Some generalist predators such as ants, spiders, damsel bugs and assassin bugs are known to predate on mirids. Nothing is known about parasitoids of mirids. Since green mirid (GM), Creontiades dilutus, is indigenous to Australia it is likely that we have one or more parasitoids of this mirid in Australia, but that possibility has not been investigated yet. The impact of the GVB adult parasitoid, Trichopoda giacomelli, has been studied by Dr Khan who found that the fly is established in the released areas and continues to spread. However, to get wider and greater impact, the fly should be released in new locations across the valleys. The insecticides registered for mirids and stinkbugs are mostly non-selective and are extremely disruptive to a wide range of beneficial insects. Use of these insecticides at stage I and II will minimise the impact of existing IPM programs. Therefore less disruptive control tactics including soft chemicals for mirids and stinkbugs are necessary. As with soft chemicals, salt mixtures, biopesticides based on fungal pathogens and attractants based on plant volatiles may be useful tools in managing mirids and stinkbugs with less or no disruption. Dr Khan has investigated salt mixture against mirids and GVB. While salt mixtures are quite effective and less disruptive, they are quite chemical specific. Not all chemicals mixed with salt will give the desired benefit. Therefore further investigation is needed to identify those chemicals that are effective with salt mixture against mirids and 3 of 37 GVB. Dr Caroline Hauxwell of DPI&F is working on fungal pathogen-based biopesticides against mirids and GVB and Drs Peter Gregg and Alice Del Socorro of Australian Cotton CRC are working on plant volatile-based attractants against mirids. Depending on their findings, inclusion of fungal-based biopestcides and plant volatile-based attractants in developing a management system against mirids and stinkbugs in cotton could be an important component of an IPM approach.

The Application of Near Infrared Spectroscopy for the Assessment of Avocado Quality Attributes, Infrared Spectroscopy. Chapter 14

Quality and safety evaluation of agricultural products has become an increasingly important consideration in market/commercial viability and systems for such evaluations are now demanded by customers, including distributors and retailers. Unfortunately, most horticultural products struggle with delivering adequate and consistent quality to the consumer. Removing inconsistencies and providing what the consumer expects is a key factor for retaining and expanding both domestic and international markets. Most commercial quality classification systems for fruit and vegetables are based on external features of the product, for example: shape, colour, size, weight and blemishes. However, the external appearance of most fruit is generally not an accurate guide to the internal or eating quality of the fruit. Internal quality of fruit is currently subjectively judged on attributes such as volatiles, firmness, and appearance. Destructive subjective measures such as internal flesh colour, or objective measures such as extraction of juice to measure sweetness (oBrix) or assessment of dry matter (DM) content are also used, although obviously not for every fruit – just a sample to represent the whole consignment. For avocado fruit, external colour is not a maturity characteristic, and its smell is too weak and appears later in its maturity stage (Gaete-Garreton et al., 2005). Since maturity is a major component of avocado quality and palatability, it is important to harvest mature fruit, so as to ensure that fruit will ripen properly and have acceptable eating quality. Currently, commercial avocado maturity estimation is based on destructive assessment of the %DM, and sometimes percent oil, both of which are highly correlated with maturity (Clark et al., 2003; Mizrach & Flitsanov, 1999). Avocados Australia Limited (AAL (2008)) recommend a minimum maturity standard for its growers of 23 %DM (greater than 10% oil content) for the ‘Hass’ cultivar, although consumer studies indicate a preference for at least 25 %DM (Harker et al., 2007).

Short-range resource location by Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) demonstrates a strong preference for fungi associated with cotton seed

Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is a common stored grain pest for which a wide range of suitable resources has been recorded. These beetles are facultatively fungivorous and their resource range may extend to fungi associated with non-grain resources (e.g. cotton seed) and other decaying plant matter. Little is known with respect to fungi in terms of resource location by these beetles in the field. We, therefore, conducted a series of experiments in laboratory arenas, glasshouse cages and the field to determine how beetles respond to grain resources in relation to cotton seed (together with its lint stubble and associated fungal flora). Results from the tests conducted in relatively small arenas and cages in the laboratory and glasshouse reveal that the responses of T. castaneum adults to food resources were twice as strong when walking as when flying (as measured by the proportion of the released beetles that were trapped). Also, a clear preference for linted cotton seeds was evident in walking T. castaneum, especially in small-scale arenas in the laboratory, where at least 60% of beetles released preferred linted cotton seeds over wheat and sorghum. Similarly, in cages (1 m3) they responded five times more strongly to linted cotton seed than to conventional grain resources. However, this pattern was not consistent with those obtained from field trapping over 20 m and the beetles did not show any particular preference to any of the resources tested above. Our results suggest a focus on walking beetles in trapping studies for population estimations and, for developing effective food-based trapping lures, the potential use of active volatiles from the fungi associated with linted cotton seed. © 2012 Elsevier Ltd.

Responses of tribolium castaneum to olfactory cues from cotton seeds, the fungi associated with cotton seeds, and cereals

We tested, in an olfactometer, whether or not Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) responds preferentially to the volatiles that emanate from the fungi associated with cotton [Gossypium hirsutum L. (Malvaceae)] seed over those that emanate from cereals, because cereals are usually portrayed as the primary resources of these beetles. Pairwise comparisons were conducted between cotton seed, wheat (Triticum aestivum L.), and sorghum [Sorghum bicolor (L.) Moench] (both Poaceae); volatiles were tested from intact seeds and from both water and ethanol extracts. The results demonstrate that T. castaneum is attracted more strongly to cotton seeds with its lint contaminated with fungi, than to the conventional resources of this species (i.e., wheat and sorghum). Further tests prove that it is the fungus on the lint that produces the active volatiles, because the beetles did not respond to sterilized cotton lint (i.e., without the fungi typically associated with it when cotton seed is stored). Tests with five fungal cultures (each representing an unidentified species that was isolated from the field-collected cotton lint) were variable across the cultures, with only one of them being significantly attractive to the beetles. The others were not attractive and one may even have repulsed the beetles. The results are consistent with the beetles having a strong ecological association with fungi and suggest it would be worth investigating the ecology of T. castaneum from this perspective. © 2012 The Netherlands Entomological Society.

Effect of Packaging Materials and Storage on Major Volatile Compounds in Three Australian Native Herbs

Lemon myrtle, anise myrtle, and Tasmanian pepper leaf are commercial Australian native herbs with a high volatile or essential oil content. Packaging of the herbs in high- or low-density polyethylene (HDPE and LDPE) has proven to be ineffective in preventing a significant loss of volatile components on storage. This study investigates and compares the effectiveness of alternate high-barrier property packaging materials, namely, polyvinylidene chloride coated polyethylene terephthalate/casted polypropylene (PVDC coated PET/CPP) and polyethylene terephthalate/polyethylene terephthalate/aluminum foil/linear low-density polyethylene (PET/PET/Foil/LLDPE), in prevention of volatile compound loss from the three native herbs stored at ambient temperature for 6 months. Concentrations of major volatiles were monitored using gas chromatography?mass spectrometry (GC-MS) techniques. After 6 months of storage, the greatest loss of volatiles from lemon myrtle was observed in traditional LDPE packaging (87% loss) followed by storage in PVDC coated PET/CPP (58% loss) and PET/PET/Foil/LLDPE (loss of 23%). The volatile loss from anise myrtle and Tasmanian pepper leaf stored in PVDC coated PET/CPP and PET/PET/Foil/LLDPE packaging was <30%. This study clearly indicates the importance of selecting the correct packaging material to retain the quality of herbs with high volatile content.

Postharvest physiology and volatile production by flowers of Ptilotus nobilis

Ptilotus nobilis (Lindl.) F. Muell. has potential in the floriculture industries as a cut flower crop. Ethylene production and respiration rates, fresh weight changes and volatile scent production from cut inflorescences of P. nobilis cultivars Passion (dark pink flowers) and Purity (white-green flowers) were measured during vase life. Inflorescence weight loss was significant (P<0.001) during vase life with wilting and colour loss being the primary reasons for loss of vase life. Inflorescences ready for the cut market stored and at 22 degrees C had vase lives of >12 d. Ethylene production by inflorescences was low to negligible. Treatment with silverthiosulphate (STS) and ethylene had no effects on vase life. Evidently, ethylene did not play a role in determining the postharvest longevity of cut P. nobilis flowers. Respiration rates of inflorescences were high at harvest (>700 mg CO2 kg(-1) FW h(-1)) and declined gradually there-after during vase life. Total volatile emissions followed a similar pattern. For Passion, respiration rates of immature florets were significantly greater (P=0.02) than florets from other developmental stages while the calyx produced the most CO2. For Purity, respiration rates of florets of different maturities did not differ and the reproductive tissue produced the most CO2. Only fully opened mature florets with their stigma and anthers revealed, emitted significant quantities of volatiles (P<0.001) and primarily from the calyx tissue for both cultivars. The individual volatiles differed somewhat for the two cultivars. However, both produced significant quantities of benzaldehyde, 3,5-dimethoxytoluene and benzyl alcohol. These. compounds have previously been associated with desirable floral scent. (C) 2013 Elsevier B.V. All rights reserved.

Increase in β-Ionone, a Carotenoid-Derived Volatile in Zeaxanthin-Biofortified Sweet Corn

Carotenoids are responsible for the yellow color of sweet corn (Zea mays var. saccharata), but are also potentially the source of flavor compounds from the cleavage of carotenoid molecules. The carotenoid-derived volatile, -ionone, was identified in both standard yellow sweet corn (Hybrix5) and a zeaxanthin-enhanced experimental variety (HZ) designed for sufferers of macular degeneration. As -ionone is highly perceivable at extremely low concentration by humans, it was important to confirm if alterations in carotenoid profile may also affect flavor volatiles. The concentration of -ionone was most strongly correlated (R2 > 0.94) with the -arm carotenoids, -carotene, -cryptoxanthin, and zeaxanthin, and to a lesser degree (R2 = 0.90) with the α-arm carotenoid, zeinoxanthin. No correlation existed with either lutein (R2 = 0.06) or antheraxanthin (R2 = 0.10). Delaying harvest of cobs resulted in a significant increase of both carotenoid and -ionone concentrations, producing a 6-fold increase of ?-ionone in HZ and a 2-fold increase in Hybrix5, reaching a maximum of 62g/kg FW and 24g/kg FW, respectively.