Behavioral Responses of a Generalist Pest, Spilosoma Obliqua Walker, Towards the Leaf Surface Wax Chemicals of Three Types of Jute Crop and Implications in Pest Management

Behavioral responses of a generalist pest, Spilosoma obliqua Walker (Lepidoptera: Arctiidae), towards the leaf surface wax chemicals of three types of jute crops (white jute, Corchorus capsularis, tossa jute, C. olitorious,, and mesta jute or kenaf, Hibiscus cannabinus [Malvaceae]) and their implications in pest management was studied under laboratory conditions. The GC-MS and GC-FID analyses of the jute leaf epicuticular waxes indicated the presence of 27 n-alkanes, having chain lengths from n-C14 to n-C36 and 14 free fatty acids (FFAs) having chain lengths from C12:0 to C22:0. Among the identified n-alkanes and FFAs of selected jute cultivars n-C34 (144.397±6.971 µg leaf−1) and C16:1 (37.034±0.848 µg leaf−1) of tossa jute leaves were most abundant. The host preference (white jute> tossa jute > mesta jute) of S. obliqua was evaluated simultaneously by olfactory, visual, and tactile recognition, as valid for other lepidopteran species. For olfaction [females], oviposition [gravid females], and feeding [larvae]) in S. obliqua, the most stimulating combined-synthetic-mixture of epicuticular wax components was represented by 4 n-alkanes (n-C16, n-C18, n-C20, n-C22) and 5 FFAs (C16:1, C16:0, C18:0, C20:0, C22:0) in mixture at leaf equivalent (µg leaf−1) amounts (195.209±2.950 µg, 119.777±1.857 µg, and 50.567±3.508 µg, respectively) in white jute, tossa jute, and mesta jute, respectively. Thus, the present study suggests that the synthetic blends of 4 n-alkanes and 5 FFAs of respective jute crops can be used as lures to develop baited trap as a part of integrated pest management (IPM) of S. obliqua for sustainable jute cultivation.

Interaction of surfactants with barley leaf surfaces: time-dependent recovery of contact angles is due to foliar uptake of surfactants

Main conclusion Time-dependent contact angle measurements of pure water on barley leaf surfaces allow quantifying the kinetics of surfactant diffusion into the leaf. Abstract Barley leaf surfaces were sprayed with three different aqueous concentrations (0.1, 1.0 and 10%) of a monodisperse (tetraethylene glycol monododecyl ether) and a polydisperse alcohol ethoxylate (BrijL4). After 10 min, the surfactant solutions on the leaf surfaces were dry leading to surfactant coverages of 1, 10 and 63 µg cm−2, respectively. The highest surfactant coverage (63 µg cm−2) affected leaf physiology (photosynthesis and water loss) rapidly and irreversibly and leaves were dying within 2–6 h. These effects on leaf physiology did not occur with the lower surfactant coverages (1 and 10 µg cm−2). Directly after spraying of 0.1 and 1.0% surfactant solution and complete drying (10 min), leaf surfaces were fully wettable for pure water and contact angles were 0°. Within 60 min (0.1% surfactant) and 6 h (1.0% surfactant), leaf surfaces were non-wettable again and contact angles of pure water were identical to control leaves. Scanning electron microscopy investigations directly performed after surfactant spraying and drying indicated that leaf surface wax crystallites were partially or fully covered by surfactants. Wax platelets with unaltered microstructure were fully visible again within 2 to 6 h after treatment with 0.1% surfactant solutions. Gas chromatographic analysis showed that surfactant amounts on leaf surfaces continuously disappeared over time. Our results indicate that surfactants, applied at realistic coverages between 1 and 10 µg cm−2 to barley leaf surfaces, leading to total wetting (contact angles of 0°) of leaf surfaces, are rapidly taken up by the leaves. As a consequence, leaf surface non-wettability is fully reappearing. An irreversible damage of the leaf surface fine structure leading to enhanced wetting and increased foliar transpiration seems highly unlikely at low surfactant coverages of 1 µg cm−2.

Individual and Interactive Temporal Implications of UV-B Radiation and Elevated CO2 on the Morphology of Basil (Ocimum basilicum L.)

Temporal and spatial variations in ozone levels and temporal changes in solar radiation greatly influence ultraviolet radiation incidence to crops throughout their growth, yet the interactive effects of CO2 and UV-B radiation on Basil production under sunlight environmental conditions has not been studied. Basil ‘Genovese’ plants grown under sunlit plant growth chambers were subjected to a combination of supplemental UV-B (0 and 10 kJ m−2d−1) and ambient (420 ppm) and elevated (720 ppm) CO2 treatments for 38 days after 14 days of germination. UV-B radiation treatments caused a decrease in basil stem branching, fresh mass, and stem dry mass under both CO2 treatments when harvested after 17 and 38 days of treatment. There was also an increase in basil leaf surface wax under UV-B (10 kJ m−2d−1) treatment compared to controls (0 kJ m−2d−1). Elevated CO2 treatments caused a decrease in morphological features, including specific leaf area and fresh mass. Interactive effects between UV-B and CO2 treatments existed for some morphological features, including plant height, root surface area, and average root diameter. Understanding the impacts that CO2 and UV-B radiation treatments have on basilcan improve existing varieties for increased tolerance while simultaneously improving yield, plant morphology, and physiology.

Short-range attraction and oviposition stimulant of a biocontrol agent, Galerucella placida Baly (Coleoptera: Chrysomelidae) toward weed leaf surface waxes

Two Polygonaceae weeds, Rumex dentatus L. and Polygonum glabrum Willd. are abundant in wheat- and rice-fields, respectively, in India. Galerucella placida Baly (Coleoptera: Chrysomelidae) is a biocontrol agent of these two weeds. The importance of long-chain alkanes and free fatty acids present in leaf surface waxes of these weeds was assessed as short-range attractant and ovipositional stimulant in G. placida females. Extraction, TLC, GC-MS and GC-FID analyses demonstrated 19 n-alkanes from n-C14 to n-C35 and 14 free fatty acids from C12:0 to C22:0 in leaf surface waxes. Hentriacontane was predominant among alkanes in both weeds, while oleic acid and docosanoic acid were predominant among free fatty acids in R. dentatus and P. glabrum, respectively. Females of G. placida were attracted toward one leaf equivalent surface wax of both weeds against the control solvent (petroleum ether) in a short Y-tube olfactometer bioassay. But, the insect could not differentiate between one leaf equivalent surface wax of R. dentatus and P. glabrum, indicating that both weed leaves were equally attractive in females. A synthetic blend of either 2.44, 35.57 and 23.58 μg ml−1 of octadecane, heptacosane and nonacosane, respectively, resembling the amounts present in one leaf equivalent surface wax of R. dentatus or 4.08, 19.54 and 23.58 μg ml−1 of octadecane, palmitoleic acid and docosanoic acid, respectively, resembling the amounts present in one leaf equivalent surface wax of P. glabrum acted as short-range attractant and ovipositional stimulant in G. placida. These results could be a basis for host plant specificity of the biocontrol agent.

Fatty Alcohols, a Minor Component of the Tree Tobacco Surface Wax, Reduce Insect Herbivory

Despite decades of research resulting in a comprehensive understanding of epicuticular wax biosynthesis and metabolism, the function of these almost ubiquitous metabolites in plant-herbivore interactions remains unresolved. To develop a better understanding of this role, we investigated plant-herbivore interactions in four Nicotiana glauca (tree tobacco) genome edited mutants. This included [eceriferum1 (cer1), eceriferum3 (cer3), beta-ketoacyl-coA synthase6 (kcs6), and fatty acyl-coA reductase (far)] displaying a wide range of alkane and fatty alcohol abundances. Three interaction classes were examined: chewing herbivory with seven caterpillar and one snail species, phloem feeding with Myzus persicae (green peach aphid), and egg laying with Bemisia tabaci (sweet potato whitefly). We found that high wax load and alkane abundance did not reduce caterpillar or snail herbivory. However, fatty alcohol content was negatively correlated with caterpillar growth, suggesting a role in reducing insect herbivory despite its lower levels. Aphid reproduction and feeding activity were not correlated with wax load and composition but are potentially affected by altered cutin composition of cer1 mutants. When examining non-feeding activities, wax crystal morphology could explain the preference of B. tabaci to lay eggs on wildtype plants relative to cer1 and far mutants. Accordingly, the fatty alcohol wax component reduces caterpillar herbivory on the chemical level, but oviposition is increased when wax crystals are dense. The results suggest that this varied response between herbivore classes and species, at times displaying increased and at times reduced fitness in response to altered wax composition is in part a consequence of co-evolution that shaped the specific effects of different N. glauca metabolites such as anabasine and fatty alcohols in plant-herbivore interactions.

Cuticular Wax Composition is Essential for Plant Recovery Following Drought with Little Effect under Optimal Conditions

Despite decades of extensive study, the role of cuticular lipids in sustaining plant fitness is far from being understood. To answer this fundamental question, we employed genome editing in tree tobacco (Nicotiana glauca) plants and generated mutations in 16 different cuticular lipids-related genes. We chose tree tobacco due to the abundant, yet simply composed epicuticular waxes deposited on its surface. Five out of 9 different mutants that displayed a cuticular lipids-related phenotype were selected for in depth analysis. They had either reduced total wax load or complete deficiency in certain wax components. This led to substantial modification in surface wax crystal structure and to elevated cuticular water loss. Remarkably, under non-stressed conditions, mutant plants with altered wax composition did not display elevated transpiration or reduced growth. However, once exposed to drought, plants lacking alkanes were not able to strongly reduce their transpiration, leading to leaf death and impaired recovery upon resuscitation, and even to stem cracking, a phenomenon typically found in trees experiencing drought stress. In contrast, plants deficient in fatty alcohols exhibited an opposite response, having reduced cuticular water loss and rapid recovery following drought. This deferential response was part of a larger trend, of no common phenotype connecting plants with a glossy appearance. We conclude that alkanes are essential under drought response and much less under normal non-stressed conditions, enabling plants to seal their cuticle upon stomatal closure, reducing leaf death and facilitating a speedy recovery.

Interrelation between Surface Wax Alkanes from Red Kidney Bean (Phaseolus vulgaris L.) Seeds and Adzuki Bean Weevil [Callosobruchus chinensis (F.)] (Coleoptera: Bruchidae)

Background: Callosobruchus chinensis (Fabricius) (Coleoptera: Bruchidae) is one of the major insect pests of Phaseolus vulgaris L. grains, commonly known as rajma seeds, in Europe and Asia including India. Infestations of these insects destroy majority of legume seeds including rajma which causes a great economic loss. Hence, a proper sustainable pest management measures are necessary for storage of rajma seeds. For this, the study aims to identify and quantify the n-alkane profile from the surface waxes of rajma seeds and their role as olfactory cue in C. chinensis. Individual synthetic alkane followed by the synthetic blends mimicking rajma seed surface wax n-alkanes as olfactory cue was also evaluated.Methods: Collected rajma seeds were solvent extracted to isolate surface waxes. The extract then fractioned by thin-layer chromatography and followed by gas chromatography-mass spectrometry analyses to purify, quantify and identify n-alkanes.Result: Rajma seeds’ surface waxes analysis revealed 18 n-alkanes between n-C15 and n-C33. The predominant alkanes were n-octacosane and n-hexadecane. n-Octadecane was the least abundant alkane in seeds. Total alkane content was 3502.67±12.82µg from 100 g (number 200 ± 5.13) seeds. Adult female C. chinensis elicited attraction towards the surface wax alkanes at concentrations of 0.5, 1, 2, 4 and 6 seed(s) equivalent of rajma seed(s) in the Y-tube olfactometer bioassay, but the highest attraction was observed at 6 seeds equivalent. Hence, a synthetic alkane blend resembling of 6 seeds equivalent, present in seeds’ surface wax alkane or a combination of nine (which elicited positive response) synthetic alkane blend resembling 6 seeds equivalent could be used as lures in developing baited trap in insect pest management programme.