Insecticide’s Disappearance after Field Treatment and during Processing into Byproducts

Insecticide\'s disappearance after field treatments could be ascribed to different factors such as sunlight photodegradation, dilution effect due to fruit growth, co-distillation during fruit respiration and evaporation. Moreover, the epicuticular waxes could speed or slow down the degradation rate, and the cultivation in an open field or greenhouses could affect the residues dramatically. After harvest, the processing techniques to produce byproducts deeply influence insecticide residues. For example, fruit drying, winemaking, the industrial processing of tomatoes to produce purée, triple-concentrated paste, fine pulp, diced, olive processing to obtain table olive and olive oil, and other industrial applications on fruits affect residues and their half-life time. The scope of this chapter is to highlight the major factors responsible for the disappearance of insecticides after treatment. Moreover, the chapter intends to review the influence of the industrial processes on insecticide behaviour when the raw material is transformed into its byproducts.

Leaf Cuticular Transpiration Barrier Organization in Tea Tree Under Normal Growth Conditions

The cuticle plays a major role in restricting nonstomatal water transpiration in plants. There is therefore a long-standing interest to understand the structure and function of the plant cuticle. Although many efforts have been devoted, it remains controversial to what degree the various cuticular parameters contribute to the water transpiration barrier. In this study, eight tea germplasms were grown under normal conditions; cuticle thickness, wax coverage, and compositions were analyzed from the epicuticular waxes and the intracuticular waxes of both leaf surfaces. The cuticular transpiration rates were measured from the individual leaf surface as well as the intracuticular wax layer. Epicuticular wax resistances were also calculated from both leaf surfaces. The correlation analysis between the cuticular transpiration rates (or resistances) and various cuticle parameters was conducted. We found that the abaxial cuticular transpiration rates accounted for 64–78% of total cuticular transpiration and were the dominant factor in the variations for the total cuticular transpiration. On the adaxial surface, the major cuticular transpiration barrier was located on the intracuticular waxes; however, on the abaxial surface, the major cuticular transpiration barrier was located on the epicuticular waxes. Cuticle thickness was not a factor affecting cuticular transpiration. However, the abaxial epicuticular wax coverage was found to be significantly and positively correlated with the abaxial epicuticular resistance. Correlation analysis suggested that the very-long-chain aliphatic compounds and glycol esters play major roles in the cuticular transpiration barrier in tea trees grown under normal conditions. Our results provided novel insights about the complex structure–functional relationships in the tea cuticle.

Comparative floral surface micromorphology helps to discriminate between species of Paphiopedilum (Orchidaceae: Cypripedioideae) from Peninsular Malaysia

The floral micromorphology of critically endangered Paphiopedilum Pfitzer [P. barbatum (Lindl.) Pfitzer, P. callosum var. sublaeve (Rchb.f.) P.J.Cribb and P. niveum (Rchb.f.) Stein] were analyzed concerning either infrageneric taxonomy or physioecological demands. The first two species are phylogenetically close and superficially identical but occur with distinct phytogeographical distributions in the region. The third species is a phylogenetically distant congener that inhabits limestone areas in the northern part of Peninsular Malaysia. Using scanning electron microscopy (SEM), we investigated the surface of the dorsal sepal, synsepal, lateral petals, pouch or labellum, and staminode. Amongst the investigated features were epicuticular waxes, epicuticular ornamentation, trichome distribution and type, pustular glands, and papillae. Our study supports the distinction of P. barbatum from P. callosum var. sublaeve, which belong to subgenus Paphiopedilum, and from P. niveum, a species belonging to subgenus Brachypetalum, a separated monophyletic clade. Comparatively, P. barbatum has Type III non-glandular trichomes on the margin of its lateral petals, which are absent in P. callosum var. sublaeve. Paphiopedilum callosum var. sublaeve and P. niveum are distinguishable from P. barbatum by a confined distribution of papillae. The epicuticular ornamentation and distribution of trichomes on staminode discriminate between P. barbatum and P. callosum var. sublaeve and differentiates them from P. niveum. Compared to P. barbatum and P. niveum, stomata in P. callosum var. sublaeve were superficial with prominently raised guard cells. From the physioecological view, the absence of glandular trichomes, and the low occurrence of papillae and stomata on the floral parts explain the unscented flowers of P. barbatum and P. callosum var. sublaeve. A combination of the features examined is taxonomically valuable for delimitation of the species at the infrageneric level, although the diagnostic characters are far inadequate for a generic taxonomic revision. A study with a more extensive sampling from the three subgenera of Paphiopedilum, including subgenus Parvisepalum, is anticipated to elucidate the level of variation of the analyzed microcharacters. Key Words: physioecological importance, Peninsular Malaysia, subgenus Brachypetalum, subgenus Paphiopedilum, scanning electron microscopy, taxonomic delimitation

Genetic Analysis of the Unique Epicuticular Wax Profile of ‘Odourless Greenleaf’ Onion

The amounts and types of epicuticular waxes on onion (Allium cepa) leaves affect the severity of feeding damage by onion thrips (Thrips tabaci), a serious insect pest of onion. Onion plants with light green leaves are referred to as “glossy” and accumulate less epicuticular wax relative to the blue–green (“waxy”) foliage of wild-type onion. The onion cultivar Odourless Greenleaf (OGL) has visually glossy foliage, shows resistance to thrips feeding damage, and has the unique profile of accumulating waxes with 28 or fewer carbons. Plants of glossy OGL were crossed with the glossy inbred B9885 and waxy inbred lines DH2107, DH066619, and B8667. Hybrid progenies from glossy OGL by waxy plants had waxy foliage, indicating recessiveness of the glossy OGL phenotype relative to the waxy phenotype. Hybrids from the cross of glossy OGL with glossy B9885 were also waxy, revealing different genetic bases for the glossy phenotype in these two onions. Hybrid plants were self-pollinated and segregations in F2 families from OGL × waxy crosses fit the expected 3:1 ratio for the single locus at which the homozygous recessive genotype conditions glossy foliage. Segregations in F2 families from crosses of glossy 9885 × glossy OGL fit the 9:7 ratio, supporting two independently segregating loci, where the recessive genotype at either locus conditions the glossy phenotype. Amounts and types of epicuticular waxes on leaves of F2 progenies from crosses of OGL × waxy B8667 and glossy B9885 × OGL were determined using gas chromatography-mass spectrometry. Single-nucleotide polymorphisms were genotyped and genetic maps were constructed. The visually glossy phenotype from OGL and its unique profile of epicuticular waxes were conditioned by one locus on chromosome 6, for which we propose the name glogl. Onion populations such as OGL with unique epicuticular wax profiles will be important germplasms for the development of onion cultivars that suffer less feeding damage from onion thrips compared with waxy onion.

Plasticity of the Cuticular Transpiration Barrier in Response to Water Shortage and Resupply in Camellia sinensis: A Role of Cuticular Waxes

The cuticle is regarded as a non-living tissue; it remains unknown whether the cuticle could be reversibly modified and what are the potential mechanisms. In this study, three tea germplasms (Wuniuzao, 0202-10, and 0306A) were subjected to water deprivation followed by rehydration. The epicuticular waxes and intracuticular waxes from both leaf surfaces were quantified from the mature 5th leaf. Cuticular transpiration rates were then measured from leaf drying curves, and the correlations between cuticular transpiration rates and cuticular wax coverage were analyzed. We found that the cuticular transpiration barriers were reinforced by drought and reversed by rehydration treatment; the initial weak cuticular transpiration barriers were preferentially reinforced by drought stress, while the original major cuticular transpiration barriers were either strengthened or unaltered. Correlation analysis suggests that cuticle modifications could be realized by selective deposition of specific wax compounds into individual cuticular compartments through multiple mechanisms, including in vivo wax synthesis or transport, dynamic phase separation between epicuticular waxes and the intracuticular waxes, in vitro polymerization, and retro transportation into epidermal cell wall or protoplast for further transformation. Our data suggest that modifications of a limited set of specific wax components from individual cuticular compartments are sufficient to alter cuticular transpiration barrier properties.

Identities of and Phenotypic Variation for Epicuticular Waxes among Leaves and Plants from Inbred Onion Populations

The amounts and types of epicuticular waxes on onion (Allium cepa) leaves affect feeding damage by onion thrips (Thrips tabaci). This study used gas chromatography mass spectrometry (GCMS) to establish the identities of waxes and measure over time wax amounts on leaves of inbred onion plants with glossy, semiglossy, and waxy foliage. Nine waxes were detected on leaves of all inbreds, and higher coefficients of variation (cv) were observed for less abundant waxes on foliage of doubled haploid onions. Older leaves had higher amounts of waxes compared with younger leaves on the same plant. Except for one minor wax, amounts of individual waxes on leaves were not significantly different for plants of different ages. There was a significant inbred by sampling date interaction due to lower amounts of waxes on the leaves of older plants from the semiglossy inbred. These results indicate that there is little advantage to multiple samplings of leaves over time from the same plant and resources may be better used to evaluate more plants. The relatively large cvs for amounts of specific waxes may reduce response to selection for unique epicuticular wax profiles to develop onion populations that suffer less feeding damage by onion thrips.

Cuticular wax composition of mature leaves of species and hybrids of the genus Prunus differing in resistance to clasterosporium disease

The interaction of a host plant with pathogen implies an extremely complex process involving the outer waxy layer of the cuticle, cutin, cell membrane, and intracellular structures. However, the initial contact between plants and pathogens takes place in cuticular waxes covering the surface of leaves, stems and fruits. Despite many findings on the role of plant epicuticular waxes, there is a gap in the understanding of the relationship between individual compounds and their functions. The pathogenic fungus Clasterosporium carpohilum (Lev.) Aderh. parasitizes the tissues of many stone fruit trees, damaging leaf and fruit surface. The aim of this work was to find out if the quantity and composition of leaf epicuticular wax could be responsible for the resistance to clasterosporium disease. The study of differences of plants in fungal resistance was carried out on species and hybrids of the genus Prunus from the collection of the Botanical Garden of Oles Honchar Dnipro National University (Dnipro city, Ukraine). The chloroform extracts of epicuticular waxes from the surface of mature leaves were analyzed by gas chromatography connected to mass-spectrometry. GC/MS assay was performed using Shimadzu GCMS-QP 2020 El equipped with capillary column (5% diphenyl/95% dimethyl polysiloxane), and helium as a carrier gas. Mass Spectrum Library 2014 for GSMS was used to identify the separated compounds of the wax extracts. The maximum total wax amount on the adaxial and abaxial leaf surface of hybrid 2 was twice the minimum wax accumulation for hybrid 4. Overall, 20 individual compounds belonging to six hydrocarbon classes were identified. Leaf epicuticular wax composition both in Prunus persica (L.) Batsch and P. dulcis (Mill.) D. A. Webb, and hybrids was dominated by long-chain n-alkanes with even carbon number (77.6–90.9% of total sum). The alkenes’ class was represented only by 17-pentatriacontene detected in the wax of both Prunus species. Octadecanaldehyde was found in epicuticular wax of P. dulcis while absent in wax of the more resistant species P. persica. Prime alcohols 1-tetradecanol and 1-hexacosanol were detected in leaf waxes of hybrid 4 and P. dulcis respectively. The ester class contained seven compounds found in leaf epicuticular waxes of both plant species and all hybrid forms. The identification of phthalic acid esters in leaf wax extracts was unexpected, and the phthalates’ origin is discussed. Strong positive correlation between leaf damage and tetrapentacontane content in epicuticular waxes could presumably be the result of infection-induced metabolism reprogramming in epidermal cells of infected leaves.

Leaflet blade epidermis and its taxonomic significance in 13 species of Bignonieae (Bignoniaceae) from Pico do Jabre, Paraíba, Northeast of Brazil

Bignonieae is the largest tribe of Bignoniaceae, with 21 genera and 393 species of lianas and shrubs, 2–3-foliolate with the terminal leaflet modified as tendrils. We examined the micromorphologies of the leaflet blade epidermises of 13 species of Bignonieae belonging to Amphilophium, Anemopaegma, Bignonia, Cuspidaria, Dolichandra, Fridericia, Pyrostegia, Tanaecium, and Xylophragma, from Pico do Jabre, Paraíba, Brazil. These are lianas except for Tanaecium parviflorum (shrubby). We sought to identify epidermal leaflet parameters to support their taxonomy subject to great similarities between their vegetative characters, mainly in species of the same genus and related genera. Analyses were performed using light and scanning electron microscopy, and showed five types of epicuticular waxes, four cuticle types, three epidermal cell anticlinal wall types, and non-glandular and glandular trichomes. Hypostomatic leaves showed ten different types of stomata, with stomatal indices from 6.21% (Bignonia ramentacea) to 23.52% (Tanaecium parviflorum) and stomatal densities from 76 stomata/mm² (Pyrostegia venusta) to 752.9 (T. parviflorum). The presence of raphides in Amphilophium crucigerum and styloids in Fridericia pubescens constitute the first records for these genera. Epidermal micromorphology provided a set of distinctive characters to separate these species, representing an additional tool to support their taxonomies, as well as that of tribe Bignonieae.

Reflectance Spectroscopy for Non-Destructive Measurement and Genetic Analysis of Amounts and Types of Epicuticular Waxes on Onion Leaves

Epicuticular waxes on the surface of plant leaves are important for the tolerance to abiotic stresses and plant–parasite interactions. In the onion (Allium cepa L.), the variation for the amounts and types of epicuticular waxes is significantly associated with less feeding damage by the insect Thrips tabaci (thrips). Epicuticular wax profiles are measured using used gas chromatography mass spectrometry (GCMS), which is a labor intensive and relatively expensive approach. Biochemical spectroscopy is a non-destructive tool for measurement and analysis of physiological and chemical features of plants. This study used GCMS and full-range biochemical spectroscopy to characterize epicuticular waxes on seven onion accessions with visually glossy (low wax), semi-glossy (intermediate wax), or waxy (copious wax) foliage, as well as a segregating family from the cross of glossy and waxy onions. In agreement with previous studies, GCMS revealed that the three main waxes on the leaves of a wild type waxy onion were the ketone hentriacontanone-16 (H16) and fatty alcohols octacosanol-1 (Oct) and triacontanol-1 (Tri). The glossy cultivar “Odourless Greenleaf” had a unique phenotype with essentially no H16 and Tri and higher amounts of Oct and the fatty alcohol hexacosanol-1 (Hex). Hyperspectral reflectance profiles were measured on leaves of the onion accessions and segregating family, and partial least-squares regression (PLSR) was utilized to generate a spectral coefficient for every wavelength and prediction models for the amounts of the three major wax components. PLSR predictions were robust with independent validation coefficients of determination at 0.72, 0.70, and 0.42 for H16, Oct, and Tri, respectively. The predicted amounts of H16, Oct, and Tri are the result of an additive effect of multiple spectral features of different intensities. The variation of reflectance for H16, Oct, and Tri revealed unique spectral features at 2259 nm, 645 nm, and 730 nm, respectively. Reflectance spectroscopy successfully revealed a major quantitative trait locus (QTL) for amounts of H16, Oct, and Tri in the segregating family, agreeing with previous genetic studies. This study demonstrates that hyperspectral signatures can be used for non-destructive measurement of major waxes on onion leaves as a basis for rapid plant assessment in support of developing thrips-resistant onions.