Christian Nansen

The maize lipoxygenase, ZmLOX10, mediates green leaf volatile, jasmonate, and herbivore-induced plant volatile production for defense against insect attack

Fatty acid derivatives are of central importance for plant immunity against insect herbivores; however, major regulatory genes and the signals that modulate these defense metabolites are vastly understudied, especially in important agro-economic monocot species. Here we show that products and signals derived from a single Zea mays (maize) lipoxygenase (LOX), ZmLOX10, are critical for both direct and indirect defenses to herbivory. We provide genetic evidence that two 13-LOXs, ZmLOX10 and ZmLOX8, specialize in providing substrate for the green leaf volatile (GLV) and jasmonate (JA) biosynthesis pathways, respectively. Supporting the specialization of these LOX isoforms, LOX8 and LOX10 are localized to two distinct cellular compartments, indicating that the JA and GLV biosynthesis pathways are physically separated in maize. Reduced expression of JA biosynthesis genes and diminished levels of JA in lox10 mutants indicate that LOX10-derived signaling is required for LOX8-mediated JA. The possible role of GLVs in JA signaling is supported by their ability to partially restore wound-induced JA levels in lox10 mutants. The impaired ability of lox10 mutants to produce GLVs and JA led to dramatic reductions in herbivore-induced plant volatiles (HIPVs) and attractiveness to parasitoid wasps. Because LOX10 is under circadian rhythm regulation, this study provides a mechanistic link to the diurnal regulation of GLVs and HIPVs. GLV-, JA- and HIPV-deficient lox10 mutants display compromised resistance to insect feeding, both under laboratory and field conditions, which is strong evidence that LOX10-dependent metabolites confer immunity against insect attack. Hence, this comprehensive gene to agro-ecosystem study reveals the broad implications of a single LOX isoform in herbivore defense.

Considerations Regarding the Use of Hyperspectral Imaging Data in Classifications of Food Products, Exemplified by Analysis of Maize Kernels

Development of robust analytical procedures is critical when using hyperspectral imaging technology in food technology and agriculture. This study used near-isogenic inbred corn lines to address two basic questions: (1) To what extent is classification accuracy increased by grinding maize kernels? (2) Can the classification accuracy of two near-isogenic inbred lines be increased by using a spectral filter to classify only certain hyperspectral profiles from each image cube? Whole kernels and ground kernels in two particle intervals, 0.250-0.354 mm (size 1) and 0.354-0.841 mm (size 2), were examined. Spectral profiles acquired from ground kernels had higher spectral repeatability than data collected from whole kernels. The classification error of discriminant functions from whole kernels was >3 times lower than that of size 1 ground particles. Applying a spectral filter to input data had negligible effect on classifications of hyperspectral profiles from whole kernels and size 2 ground particles, but for size 1 ground particles a considerable increase in accuracy was observed. Independent validation confirmed that distinction between wild type and mutant inbred maize lines could be conducted with >80% accuracy after the proposed spectral filter had been applied to hyperspectral profiles of size 1 ground particles. A combination of discriminant analysis and regression analysis could be used to accurately predict mixture ratios of the two inbred lines. The use of spectral filtering to increase the level of spectral repeatability and the use of hyperspectral imaging technology in large-scale commercial operations are discussed.

Reflectance-based identification of parasitized host eggs and adult Trichogramma specimens

A wide range of imaging and spectroscopy technologies is used in medical diagnostics, quality control in production systems, military applications, stress detection in agriculture, and ecological studies of both terrestrial and aquatic organisms. In this study, we hypothesized that reflectance profiling can be used to successfully classify animals that are otherwise very challenging to classify. We acquired hyperspectral images from adult specimens of the egg parasitoid genus Trichogramma (T. galloi, T. pretiosum and T. atopovirilia), which are ~1.0 mm in length. We also acquired hyperspectral images from host eggs containing developing Trichogramma instar and pupae. These obligate egg endoparasitoid species are commercially available as natural enemies of lepidopteran pests in food production systems. Because of their minute size and physical resemblance, classification is time consuming and requires a high level of technical experience. The classification of reflectance profiles was based on a combination of average reflectance and variogram parameters (describing the spatial structure of reflectance data) of reflectance values in individual spectral bands. Although variogram parameters (variogram analysis) are commonly used in large-scale spatial research (i.e. geoscience and landscape ecology), they have only recently been used in classification of high-resolution hyperspectral imaging data. The classification model of parasitized host eggs was equally successful for each of the three species and was successfully validated with independent data sets (>90% classification accuracy). The classification model of adult specimens accurately separated T. atopovirilia from the other two species, but specimens of T. galloi and T. pretiosum could not be accurately separated. Interestingly, molecular-based classification (using the DNA sequence of the internally transcribed spacer ITS2) of Trichogramma species published elsewhere corroborates the classification, as T. galloi and T. pretiosum are closely related and comparatively distant from T. atopovirilia. Our results emphasize the importance of using high-spectral and high-spatial resolution data in the classification of organism relatedness, and hyperspectral imaging may be of relevance to a wide range of commercial (i.e. producers of biocontrol agents), taxonomic and evolutionary research applications.

Estimating effect of augmentative biological control on grain yields from individual pearl millet heads

Pearl millet is the principal staple food crop in large portions of Western Sub‐Sahelian Africa and the millet head miner (Heliocheilus albipunctella) is one of its most devastating insect pests. Since 2006, augmentative mass releases of the larval ectoparasitoid, Habrobracon hebetor, have been conducted in Mali, Burkina Faso and Niger as part of minimizing pearl millet grain losses imposed by millet head miners. These ongoing mass releases are based on low‐cost mass‐rearing of both host larvae and parasitoids. A release of parasitoids consists of placing jute bags containing pearl millet grain and flour and parasitized host larvae near pearl millet fields. The total production costs of a single jute bag with parasitized rice moth larvae are US

Evolutionary refinement approaches for band selection of hyperspectral images with applications to automatic monitoring of animal feed quality

3–4. Based on a study of 6634 individual pearl millet heads collected at harvest in 12 farmers fields in southern Niger in 2010, we demonstrated (i) a strong negative correlation between pearl millet head damage (mining) and grain yield and (ii) that parasitism by H. hebetor reduced grain losses by, on average, 34% (comparison of infested millet heads with/without parasitism) within the given growing season. Additional benefits may include reduction in millet head miners in subsequent generations. Data from 900 pearl millet heads collected in nine farmers fields in 2011 were used to confirm data trends observed in the 2010 data and to characterize the dispersal of parasitoids in upwind and downwind directions from a release site. This study provided a quantitative description of the negative impact of millet head miner infestations on pearl millet grain yields and of benefits on grain yield of parasitism by H. hebetor. Our findings strongly support (i) intensification of mass‐rearing of H. hebetor, (ii) expansion of educational activities to increase local empowerment and understanding of the potential of augmentative biological control and (iii) optimization of H. hebetor mass release programmes among smallholders in Sub‐Sahelian Africa.

Geographical variation of Plutella xylostella (Lepidoptera: Plutellidae) populations revealed by mitochondrial COI gene in China

This paper presents methods for spectral band selection in hyperspectral image HSI cubes based on classification of reflectance data acquired from samples of livestock feed materials and ruminant-derived bonemeal. Automated detection of ruminant-derived bonemeal in animal feed is tested as part of an on-going research into development of automated, reliable fast and cost-effective quality control systems. HSI cubes contain spectral reflectance in both spatial dimensions and spectral bands. Support vector machines are used for classification of data in various domains. Selecting a subset of the spectral bands speeds processing and increases accuracy by reducing over-fitting. We developed two methods utilizing divergence values for selecting spectral band sets, 1 evolutionary search method and 2 divergence-based recursive feature elimination approach.

Foraging on Individual Leaves by an Intracellular Feeding Insect Is Not Associated with Leaf Biomechanical Properties or Leaf Orientation

The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is one of the most serious pests for cruciferous vegetable growers worldwide. To determine the relations of local and regional populations, we performed a mitochondrial COI gene analysis of eight P. xylostella populations from different locations in or around the Qinling Mountains and two other populations, one from Beijing and the other from Guangdong. The mtDNA divergences among the 10 populations were high, and 32 haplotypes were detected in 149 adults. The mean haplotype divergence was 1.7% (range 0.04–4.1%). Haplotype diversity in the 10 populations varied from 0.571 (AK) to 0.885 (HZ), and the nucleotide diversity varied from 0.00286 (AK) to 0.0117 (HZ). The results also did not show significant correlation between genetic and geographical distance. Also, the effective number of migrants between populations (Nm) ranged from 1.43 to infinite, suggesting that population exchange and gene flow among the P. xylostella populations occurred. However, principal component analysis (PCA) showed that the TB and TC populations were differentiated from other populations, indicating possible across‐mountain barrier to migration and gene flow.

The Influence of Maturity and Variety of Potato Plants on Oviposition and Probing of Bactericera cockerelli (Hemiptera: Triozidae)

Nearly all herbivorous arthropods make foraging-decisions on individual leaves, yet systematic investigations of the adaptive significance and ecological factors structuring these decisions are rare with most attention given to chewing herbivores. This study investigated why an intracellular feeding herbivore, Western flower thrips (WFT) Frankliniella occidentalis Pergande, generally avoids feeding on the adaxial leaf surface of cotton cotyledons. WFT showed a significant aversion to adaxial-feeding even when excised-cotyledons were turned up-side (abaxial-side ‘up’), suggesting that negative-phototaxis was not a primary cause of thrips foraging patterns. No-choice bioassays in which individual WFT females were confined to either the abaxial or adaxial leaf surface showed that 35% fewer offspring were produced when only adaxial feeding was allowed, which coincided with 32% less plant feeding on that surface. To test the hypothesis that leaf biomechanical properties inhibited thrips feeding on the adaxial surface, we used a penetrometer to measure two variables related to the ‘toughness’ of each leaf surface. Neither variable negatively co-varied with feeding. Thus, while avoiding the upper leaf surface was an adaptive foraging strategy, the proximate cause remains to be elucidated, but is likely due, in part, to certain leaf properties that inhibit feeding.

Using proximal remote sensing in non-invasive phenotyping of invertebrates

ABSTRACT n The ecological theory on host plant choice by herbivores suggests that mothers should choose plants that will maximize their offsprings success. In annual host plants, physiology (and therefore host suitability) is sometimes influenced by maturity and growth stage, which may influence female choice. Potato plants were grown under greenhouse conditions and used in choice and no-choice bioassays to determine the effect of plant maturity and variety on oviposition and number of stylet sheaths (which approximate stylet insertions) by tomato/potato psyllids. No-choice bioassays suggested that maturity (time since planting) did not influence oviposition behavior, but oviposition varied significantly among potato plant varieties. There was a significant effect of both maturity and variety on the number of stylet sheaths, which peak toward the middle of the growing season. We also examined tomato/potato psyllid responses to plants grown in a commercial field and again found no effect on oviposition but differences in stylet sheaths. The results suggest that differential susceptibility to zebra chip disease may be associated with unequal feeding rates. Future studies should examine whether the maturity of plants influences larval fitness. Finally, potato variety has an influence on both oviposition and “probing,” and has implications for management strategies and the development of resistant potato varieties.

Journal impact factors and the influence of age and number of citations

Proximal imaging remote sensing technologies are used to phenotype and to characterize organisms based on specific external body reflectance features. These imaging technologies are gaining interest and becoming more widely used and applied in ecological, systematic, evolutionary, and physiological studies of plants and also of animals. However, important factors may impact the quality and consistency of body reflectance features and therefore the ability to use these technologies as part of non-invasive phenotyping and characterization of organisms. We acquired hyperspectral body reflectance profiles from three insect species, and we examined how preparation procedures and preservation time affected the ability to detect reflectance responses to gender, origin, and age. Different portions of the radiometric spectrum varied markedly in their sensitivity to preparation procedures and preservation time. Based on studies of three insect species, we successfully identified specific radiometric regions, in which phenotypic traits become significantly more pronounced based on either: 1) gentle cleaning of museum specimens with distilled water, or 2) killing and preserving insect specimens in 70% ethanol. Standardization of killing and preservation procedures will greatly increase the ability to use proximal imaging remote sensing technologies as part of phenotyping and also when used in ecological and evolutionary studies of invertebrates.