Leaf surface wetness in sorghum and resistance to shoot fly, Atherigona soccata: role of soil and plant water potentials

In experiments with potted plants, the relationships between soil matric potential, plant water potential and production of water droplets (leaf surface wetness) on the folded central whorl leaf of seedlings of sorghum genotypes that are either resistant or susceptible to shoot fly (Atherigona soccata) damage were investigated. Differences in soil matric potentials in the pots affected the plant water status, which in turn had profound effects on the production of water droplets on the central whorl leaf of the sorghum genotype susceptible to shoot fly. There was no consistent variation in the relationship between plant water potential and soil matric potential of resistant and susceptible sorghum genotypes. However, there was very little or practically no water droplets on the central whorl leaf of the resistant genotypes, indicating that the production of water droplets is not solely the result of internal water status of the plant. It is suggested that leaf surface wetness is genetically controlled and that an understanding of the mechanism by which water is transferred to the leaf surface will enhance breeding for resistance to shoot fly.     

Morphological factors of the central whorl leaf associated with leaf surface wetness and resistance in sorghum to shoot fly,Atherigona soccata*

Earlier studies showed that leaf surface water on the central whorl leaf of sorghum seedlings is associated with resistance to shoot fly. In this study, the results of an experiment to determine if leaf surface wetness (LSW) originates from atmospheric condensation or from the plant are described. Morphological structures: trichomes, stomata, leaf cuticle and quantity of surface wax of the central whorl leaf were also examined for their role in LSW production. The results suggest that LSW of the central whorl leaf originates from the plant and is not due to condensation of atmospheric moisture. The presence of trichomes was indirectly associated with LSW and resistance to shoot fly but stomatal density was not associated with LSW production. The amount of wax extracted per 100 mg of fresh weight varied significantly between genotypes and seedling age. It was more in susceptible than in resistant genotypes; however, cuticular thickness was not associated with resistance. It is suggested that LSW could be the result of some form of cuticular movement of water to the leaf surface.     

Reduction of shoot fly damage in irrigated post-rainy season sorghum by manipulating irrigation

Soil moisture was manipulated in an attempt to control shoot fly (Atherigona soccata Rondani) incidence in irrigated post-rainy season sorghum grown under a rainout shelter (ROS) and in field conditions. After uniform irrigation at sowing, the plants were subjected to water stress at young seedling stage (7–28 days after emergence, DAE) for different lengths of time. Soil water had profound effects on the production of water droplets on the surface of the central whorl leaf of seedlings (leaf surface wetness, LSW) of sorghum genotypes. LSW, which facilitates movement of the larvae, was more drastically affected in susceptible (CSH 5) than in moderately resistant (IS 1054) sorghum genotypes. Shoot fly oviposition (infestation) and deadhearts (crop damage) were much higher in treatments with full irrigation (control) than in treatments to which less water was applied during the first 3 wk after seedling emergence. This resulted in higher plant biomass and overall grain yield in the latter treatments than in the control. Using insecticides to control shoot fly infestation, it was shown that a simple cultural practice of inducing plant stress by reduced soil moisture content during early plant growth gave the same or better control of shoot fly damage and the same or higher grain yield than insecticide-protected plots with full irrigation. Thus the costs associated with irrigation requirement and insecticide can be greatly reduced in the former management option compared with the latter. It is suggested that manipulation of soil water content during the vulnerable early stages of crop growth can reduce shoot fly damage in irrigated post-rainy season sorghum.     

Morphophysiological and biochemical attributes influence intra-genotypic preference of shoot fly [Atherigona soccata (Rondani)] among sorghum genotypes

Shoot fly [Atherigona soccata (Rondani)] is a destructive pest of sorghum at the seedling stage and causes huge losses to grain yield and green fodder. The host-plant resistance mechanism is the best approach to reduce the attack of insects in plants. The damage parameters, morphophysiological traits, and biochemical metabolites had been investigated in the leaves and stem of contrasting sorghum genotypes, viz., resistant (IS18551, ICSV705, ICSV700), moderately resistant (PSC-4), and susceptible (SWARNA and SL-44) at 15 and 21 days after emergence (DAE) against shoot fly infestation. The resistant genotypes recorded lowest shoot fly oviposition and incidence (0.3–0.7 eggs plant⁻¹ and 10–15%) than the susceptible genotypes (2.4–3.0 eggs plant⁻¹ and 70–80%), respectively. The susceptible genotype SWARNA recorded 50% and 80% higher deadheart formation than the resistant genotype IS18551 at 15 and 21 DAE, respectively. Resistant genotypes exhibited higher trichome density at adaxial and abaxial part of leaf (118–145 and 106–131) with pink colored leaf sheath (scale 1.50–3.25), glossy leaves (scale1.00–1.25), and lower leaf surface wetness (scale1.25–2.00) compared with susceptible genotype with 49.3–73.3 and 25.3–64.0, scale 2.50–4.00, scale 2.75–3.50, and scale 3.25–4.25 for the respective parameters. Another defense response of sorghum toward the insect attack was modulation of plant metabolism. The infested genotypes responded to insect attack by upregulation of total soluble sugar, total phenol, prussic acid, and chlorophyll content by 1.2–2.1-fold, 1.5–2.0-fold, 1.2–1.3-fold, and 1.2–3.9-fold with more induction in susceptible genotypes at 21 DAE. On the whole, the present study indicates that morphophysiological and biochemical attributes contribute toward the resistance mechanism in sorghum against shoot fly infestation.

     

Effect of epicuticular wax crystals on the localization of artificially deposited sub-micron carbon-based aerosols on needles of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

Elucidation of the mechanism of adsorption of particles suspended in the gas-phase (aerosol) to the outer surfaces of leaves provides useful information for understanding the mechanisms of the effect of aerosol particles on the growth and physiological functions of trees. In the present study, we examined the localization of artificially deposited sub-micron-sized carbon-based particles on the surfaces of needles of Cryptomeria japonica, a typical Japanese coniferous tree species, by field-emission scanning electron microscopy. The clusters (aggregates) of carbon-based particles were deposited on the needle surface regions where epicuticular wax crystals were sparsely distributed. By contrast, no clusters of the particles were found on the needle surface regions with dense distribution of epicuticular wax crystals. Number of clusters of carbon-based particles per unit area showed statistically significant differences between regions with sparse epicuticular wax crystals and those with dense epicuticular wax crystals. These results suggest that epicuticular wax crystals affect distribution of carbon-based particles on needles. Therefore, densely distributed epicuticular wax crystals might prevent the deposition of sub-micron-sized carbon-based particles on the surfaces of needles of Cryptomeria japonica to retain the function of stomata.     

Resistance in apple to shoot infection by Venturia inaequalis

Eight apple genotypes, including cultivars and breeding selections resistant and susceptible to Venturia inaequalis on foliage, were screened for shoot infection and the development of wood pustules following inoculation of shoot tips of 1–year maiden trees in the greenhouse. Where genotypes were highly resistant in terms of foliar symptoms (cvs Prima and Gavin), no shoot infection was observed. Where genotypes were highly susceptible in terms of foliar symptoms, then shoot tissue was either resistant (cv. Shinko) or susceptible (cv. Starking). In one experiment, shoot tips were inoculated sequentially as shoots extended. No pustules developed where inoculations were made later than May. In a further experiment, two leaf internode positions were inoculated on one occasion. Pustules were only observed where inoculations were made above the youngest unrolled leaf. The results suggested that shoot tips were more susceptible during early extension growth of the shoot. Pustules were noted in abundance on petioles of susceptible cultivars, and these probably contributed to early leaf abscission.     

Chemical composition of epicuticular wax crystals on the slippery zone in pitchers of five <Emphasis Type="Italic">Nepenthes</Emphasis> species and hybrids

Plants of the carnivorous genus Nepenthes efficiently trap insects in leaf pitchers, mostly employing epicuticular wax crystals on the pitcher walls to make them slippery for the prey. In the present study, the compositions and micromorphologies of the wax crystals of five Nepenthes species and hybrids were analysed in order to test whether the chemical principles underlying this ecological function are widespread within the genus. Three wax layers could be distinguished within the Nepenthes pitcher cuticles: (1) the outermost part of the crystals forming the platelets visible in standard scanning electron microscopy, (2) the bottom portion of the epicuticular wax crystals, and (3) an intracuticular wax layer. The composition of the intracuticular wax differed significantly from that of the neighbouring epicuticular layer. The compositions of corresponding wax mixtures from all five Nepenthes species and hybrids were very similar, with almost equal amounts of very long chain aldehydes and primary alcohols. While triacontanal (C₃₀ aldehyde) was prevailing in the epicuticular crystals of Nepenthes albomarginata and Nepenthes x intermedia, Nepenthes x superba and Nepenthes x henriana were found to have especially high percentages of dotriacontanal (C₃₂ aldehyde). Nepenthes “khasiana” had an intermediate aldehyde composition with almost equal amounts of both chain lengths.     

甘蓝型油菜叶表皮蜡质组分及结构与菌核病抗性关系

本试验选用6个抗病性不同的甘蓝型油莱品种,研究其叶表皮蜡质组成及结构与菌核病抗性的关系。结果表明,抗病品种在去除叶表皮蜡质后病情指数显著增加;感病品种无显著变化。不同抗性品种（系）间除酯类组分含量无显著差异外,其余蜡质组分含量差异显著。相关分析表明,蜡质组分中酯类含量与病情指数呈显著负相关关系,醇类、酮类含量与病情指数呈显著正相关,其余组分和蜡质总量与病情指数无显著相关关系。抗性品种叶表皮蜡质中烷类及酯类所占比重较高,而易感品种酮类比重较高。扫描电镜结果显示,抗病品种（系）的蜡质晶体主要为颗粒状、杆状、丝状;而感病品种（系）的蜡质晶体中不规则片状晶体所占比例较大。这些结果说明油菜叶表皮蜡质的组分及结构可能是抗病品种抵抗和延迟病原菌侵入的机制之一。     

The dynamics of leaf surface wetness of sorghum seedlings in relation to resistance to the shoot fly, Atherigona soccata

In quantitative measurements of leaf surface wetness (LSW) of the central whorl leaf of sorghum seedlings in August (rainy season) and November (post-rainy season), the highest amount (6.29 mg of water) was recorded in August in the shoot fly Atherigona soccata (Diptera: Muscidae), susceptible sorghum genotype CSH 1, while the lowest (0.07 mg) was recorded in November in the resistant genotype IS 18551. Studies on diurnal fluctuation revealed that LSW was lowest at sunset, was highest between 02.00 and 04.00 h (closely corresponding with hatching of shoot fly eggs) and dropped before sunrise. This fluctuation was associated with the evaporation of water from the plant during the night. More LSW accumulation occurred during the main crop season (June-October) than in the post-rainy season (November-April). Annual fluctuation of LSW followed trends similar to the population dynamics of shoot fly and crop infestation and were correlated with rainfall, temperature and relative humidity. Measurements of leaf temperature and the vapour pressure gradient between the leaf and the air indicated that leaf surface water originates from the plant. This was further supported by the different amounts of LSW on susceptible and resistant cultivars with similar microclimatic conditions.     

Morphological traits of wheat (Triticum aestivum L.) associated with resistance to shoot fly,Atherigona approximata Malloch

Shoot flies (Atherigona spp.) are the members of muscidae family which have got economic importance as pest of several crops of Gramineae family mostly cereals and millets. One of the most effective management strategies for controlling shoot fly is the use of resistant wheat cultivars carrying specific resistant traits. Among thirty wheat genotypes screened at the University of Agricultural Sciences, Dharwad-Karnataka (India), two advanced germplasms UAS BW-12417 and UAS BW-11110 recorded least oviposition and dead heart due to shoot fly and found to be less preferred by Atherigona approximata Malloch. The morphological traits of different wheat genotypes revealed a significant negative correlation of shoot fly oviposition and dead heart with leaf length, leaf length to breadth ratio, seedling height, average seedling growth rate, seedling vigour, leaf glossiness and trichome density of wheat leaves, which indicated the enhanced shoot fly infestation as decrease in the values of above-mentioned morphological traits. Meanwhile, increase in leaf breadth and leaf area of wheat genotype aggrandised the oviposition and dead heart damage by shoot fly. Under the changing climate where, the minor insect pests attaining major pest status, the present investigation would pave way for breeders to tailor future breeding programmes to evolve shoot fly resistant hybrids with high yielding traits.     

Some Physiological Characteristics in Resistant and Susceptible Cotton Cultivars Infected with Cotton Leaf Curl Virus

Eight cultivars/lines of Gossypium hirsutum (CIM-443, CIM-448, CIM-1100, FH-634, S-111, S-113, Cedix, and LRA 5166) resistant to cotton leaf curl virus (CLCuV), one moderately-resistant (cv. NIAB-Krishma), and one susceptible (cv. S-12) were used. All the resistant lines remained free of all disease symptoms, whereas in moderately-resistant and susceptible cvs. leaf curling and vein thickening occurred. Cultivars with varying degree of CLCuV-resistance had different pattern of accumulation of macronutrients. Leaf N content was lowest in S-12, but increased considerably due to disease. Leaf K and Ca contents of S-12 were lower in the diseased leaves than in healthy ones. Chlorophyll a and b contents were highest in lines S-111, S-113 and S-12. A marked reduction in chlorophyll b content was observed in the diseased leaves of S-12. Leaf water potential in S-12 and NIAB-Krishma was also decreased due to disease. The most distinctive characteristic to differentiate between lines was epicuticular wax content, since all the resistant lines had considerably higher wax content on their leaf surfaces than the moderately-resistant or susceptible cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Epidermal conductance in different parts of durum wheat grown under Mediterranean conditions: the role of epicuticular waxes and stomata

Abstract. Epidermal (non-stomatally-controlled) conductance from the fourth leaf, first node leaf, flag leaf and ear of durum wheat (Triticum turgidum var durum L.) grown under Mediterranean field conditions has been measured, along with leaf stomatal frequency and the amount and distribution of epicuticular waxes. Measurements were carried out on varieties and land-races from the Middle East, North Africa, ‘Institut National de la Recherche Agricole’ (INRA) and ‘Centra Internacional de Mejora de Maiz y Trigo’ (CIMMYT). Significant differences were observed among genotypes in the epidermal conductances (g_e) of the four organs. For each of the four organs tested, genotypes from the Middle East and CIMMYT showed higher g_e. values than those from North Africa and INRA. Ears showed epidermal conductances that were more than four times higher than those of leaves when g_e. values were expressed per unit dry weight. The amount of epicuticular waxes was higher in the fourth leaves, intermediate in the first node and flag leaves and lower in the ears. For each organ, g_e differences among genotypes were unrelated with the amount of epicuticular waxes. Removal of epicuticular waxes by dipping the organs into chloroform significantly increased the epidermal conductance for the fourth and first node leaves and the ear. However, this did not occur for the flag leaf. For the fourth leaf, g_e of intact leaves and g_e of leaves in which epicuticular waxes were removed were unrelated (r = -0.265). The regression coefficient of this relation for the first node and flag leaves showed values of 0.666 and 0.650 (P > 0.05), respectively, and values were even higher in the ear (r > m 0.892, P > 0.01). Scanning electron microscope analysis showed that wax bloom decreased from the fourth leaf to the flag leaf, whereas the extent of amorphous wax increased. Wax bloom in leaves consisted mainly of deposits of thin wax plates. In the ears and the adaxial surface of flag leaves, fibrillar waxes predominated. In the first node and flag leaves, the wax deposits on the adaxial side cover the surface of the leaf more densely and uniformly than those on the abaxial side. There was no significant correlation between g_e and total stomatal density, or between g_e and either adaxial or abaxial stomatal density for any sample of the three different leaves. The contribution of epicuticular waxes plus total stomatal frequency only explained 42.4, 11.8, 28.3 and 16% of g_e (per unit leaf area) variations for the fourth leaf, first node leaf, flag leaf and the combined variation of the three leaves together, respectively. From these results, it is concluded that complex interrelationship between different morphophysiological characteristics probably control g_e differences among genotypes and that these interrelationships differ for each different plant part.     

Epicuticular wax crystals of Wollemia nobilis: morphology and chemical composition

BACKGROUND AND AIMS: The morphology of the epicuticular leaf waxes of Wollemia nobilis (Araucariaceae) was studied with special emphasis on the relationship between the microstructure of epicuticular wax crystals and their chemical composition. Wollemia nobilis is a unique coniferous tree of the family Araucariaceae and is of very high scientific value as it is the sole living representative of an ancient genus, which until 1994 was known only from fossils. METHODS: Scanning electron microscopy (SEM), gas chromatography (GC) combined with mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR) were used for characterizing the morphology and the chemical structure of the epicuticular wax layer of W. nobilis needles. KEY RESULTS: The main component of the leaf epicuticular wax of W. nobilis is nonacosan-10-ol. This secondary alcohol together with nonacosane diols is responsible for the tubular habit of the epicuticular wax crystals. Scanning electron micrographs revealed differences in the fine structure of adaxial and abaxial leaf surfaces that could be explained by gas chromatographic studies after selective mechanical removal of the waxes. CONCLUSIONS: SEM investigations established the tubular crystalline microstructure of the epicuticular wax of W. nobilis leaves. GC-MS and NMR experiments showed that nonacosan-10-ol is the major constituent of the epicuticular wax of W. nobilis leaves.     

A genetic basis for the manipulation of sink–source relationships by the galling aphid <Emphasis Type="Italic">Pemphigus batae</Emphasis>

We examined how the galling aphid Pemphigus batae manipulates resource translocation patterns of resistant and susceptible narrowleaf cottonwood Populus angustifolia. Using carbon-14 (¹⁴C)-labeling experiments in common garden trials, five patterns emerged. First, although aphid galls on resistant and susceptible genotypes did not differ in their capacity to intercept assimilates exported from the leaf they occupied, aphids sequestered 5.8-fold more assimilates from surrounding leaves on susceptible tree genotypes compared to resistant genotypes. Second, gall sinks on the same side of a shoot as a labeled leaf were 3.4-fold stronger than gall sinks on the opposite side of a shoot, which agrees with patterns of vascular connections among leaves of the same shoot (orthostichy). Third, plant genetic-based traits accounted for 26% of the variation in sink strength of gall sinks and 41% of the variation in sink strength of a plant’s own bud sinks. Fourth, tree susceptibility to aphid gall formation accounted for 63% of the variation in ¹⁴C import, suggesting strong genetic control of sink–source relationships. Fifth, competition between two galls was observed on a susceptible but not a resistant tree. On the susceptible tree distal aphids intercepted 1.5-fold more ¹⁴C from the occupied leaf than did basal aphids, but basal aphids compensated for the presence of a distal competitor by almost doubling import to the gall from surrounding leaves. These findings and others, aimed at identifying candidate genes for resistance, argue the importance of including plant genetics in future studies of the manipulation of translocation patterns by phytophageous insects.     

Studies on stomatal function, epicuticular wax and stem-root transition region of polyethylene glycol-treated and nontreatedin vitro grape plantlets

Summary Scanning electron microscopy, light microscopy, and gravimetric analysis was used to evaluate stomatal function, epicuticular wax, and the stem-root transition region of grape (Vitis sp. ‘Valiant’) plantlets grownin vitro, polyethylene glycoltreatedin vitro, and greenhouse-grown plants. Scanning electron microscopic studies of leaf surfaces ofin vitro-grown plants showed widely open stomata as compared to leaf stomata of polyethylene glycol-treatedin vitro-cultured and greenhouse-grown plants. Ultrastructurally, leaf epicuticular wax ofin vitro plants was less dense than in their polyethylene-treated and greenhouse counterparts. Quantitatively,in vitro-grown plants had reduced epicuticular was as compared to polyethylene glycol-treated and greenhouse-grown plants. Light microscopic studies showed no obvious differences in the vascular connections in the stem-root transition region ofin vitro-cultured, polyethylene glycol-treatedin vitro-cultured, and greenhouse-grown plants. It is therefore likely that the rapid wilting and desiccation observed after transplantingin vitro grape plantlets is due to their defective stomatal function and reduced epicuticular wax and may not be due to poor water transport associated with vascular connection.     

What do microbes encounter at the plant surface? Chemical composition of pea leaf cuticular waxes

In the cuticular wax mixtures from leaves of pea (Pisum sativum) cv Avanta, cv Lincoln, and cv Maiperle, more than 70 individual compounds were identified. The adaxial wax was characterized by very high amounts of primary alcohols (71%), while the abaxial wax consisted mainly of alkanes (73%). An aqueous adhesive of gum arabic was employed to selectively sample the epicuticular wax layer on pea leaves and hence to analyze the composition of epicuticular crystals exposed at the outermost surface of leaves. The epicuticular layer was found to contain 74% and 83% of the total wax on adaxial and abaxial surfaces, respectively. The platelet-shaped crystals on the adaxial leaf surface consisted of a mixture dominated by hexacosanol, accompanied by substantial amounts of octacosanol and hentriacontane. In contrast, the ribbon-shaped wax crystals on the abaxial surface consisted mainly of hentriacontane (63%), with approximately 5% each of hexacosanol and octacosanol being present. Based on this detailed chemical analysis of the wax exposed at the leaf surface, their importance for early events in the interaction with host-specific pathogenic fungi can now be evaluated. On adaxial surfaces, approximately 80% of Erysiphe pisi spores germinated and 70% differentiated appressoria. In contrast, significantly lower germination efficiencies (57%) and appressoria formation rates (49%) were found for abaxial surfaces. In conclusion, the influence of the physical structure and the chemical composition of the host surface, and especially of epicuticular leaf waxes, on the prepenetration processes of biotrophic fungi is discussed.     

Infestation of cassava genotypes by Neosilba perezi (Romero & Ruppell) (Diptera: Lonchaeidae)

The objectives of this work were to assess the infestation of ten genotypes of cassava by the shoot fly Neosilba perezi (Romero & Ruppell) and to investigate effects of plant age, temperature or precipitation on cassava plants infestation by the shoot fly. Thirty-two individuals of each cassava genotype were planted and analyzed every two weeks in order to calculate the percentage of plants infested by shoot fly larvae at each sampling event and per genotype. Infestation by the fly was different across the genotypes. Genotype IAC Caapora 105-66 and genotype IAC Cascuda were resistant to shoot fly infestation, whereas the genotype IAC 15 was the most susceptible to this insect. Plant age may have an influence on infestation by shoot flies. Advanced plant age apparently favors lower or even inexistent infestation rates. However, infestation rate does not seem to be affected by temperature or precipitation.     

The impact of epicuticular wax on gas-exchange and photoinhibition in Leucadendron lanigerum (Proteaceae)

This study investigated the seasonal modification of wax deposition, and the impact of epicuticular wax on gas-exchange as well as photoinhibition in Leucadendron lanigerum, a species from the Proteaceae family with wax-covered leaf surfaces and the stomata also partially occluded by wax. The results of this study demonstrated that the deposition of epicuticular wax in L. lanigerum is dependent on the age of the leaf as well as the season, and generation and regeneration of wax occur mostly in spring while transformation and also degeneration of wax crystals occur in winter. Epicuticular waxes decreased cuticular water loss, but had little impact on leaf reflectance. The temperature of leaves without wax was lower than that of wax-covered leaves, indicating that the rate of transpiration impacted more on leaf temperature than reflectance of light in the PAR range in L. lanigerum. The wax coverage at the entrance of stomata in L. lanigerum increased resistance to gas diffusion and as a consequence decreased stomatal conductance, transpiration and photosynthesis. Also, the results indicated that epicuticular waxes do help prevent photodamage in L. lanigerum, and so this property could benefit plants living in arid environments with high solar radiation.     

Genomic diversity among sorghum genotypes with resistance to sorghum shoot fly, Atherigona soccata

Host plant resistance is one of the important components for management of sorghum shoot fly, Atherigona soccata. The levels of resistance in cultivated germplasm are low to moderate, and therefore, it is important to identify sorghum genotypes with diverse mechanisms of resistance based on physico-chemical and or molecular markers. We assessed the genetic diversity of 15 sorghum genotypes with different levels of resistance/susceptibility to shoot fly, A. soccata using 93 sorghum simple sequence repeat (SSR) primer pairs and simultaneously characterized for 15 morpho-biochemical traits associated with shoot fly resistance. Of these 93 SSR primer pairs, amplification products from 79, thought to correspond to single-copy loci distributed across all ten sorghum chromosome pairs, showed good polymorphism across the 15 sorghum genotypes. The polymorphic information content (PIC) values of these 79 SSR markers ranged from 0.06 to 0.86. The Principal Coordinate Analyses (PCoA) and cluster analyses based on dissimilarity matrices derived from SSR based allelic variation (Neighbor-Joining distance) and variation in 15 morpho-biochemical traits (based on Gower??s distance), revealed grouping of most susceptible genotypes in single cluster. The improved breeding lines grouped with resistant or susceptible genotypes, based on shared pedigree. Based on these results, three resistant accessions viz., IS 1054, IS 1057 and IS 4664 were found diverse to IS 18551, which is widely used as shoot fly resistance donor. These diverse sources, after further characterization for resistance mechanisms, can be used in breeding programs for improving shoot fly resistance.     

Chemical composition of the epicuticular and intracuticular wax layers on the adaxial side of Ligustrum vulgare leaves

Previous research has shown that cuticular triterpenoids are exclusively found in the intracuticular wax layer of Prunus laurocerasus. To investigate whether this partitioning was species-specific, the intra- and epicuticular waxes were identified and quantified for the glossy leaves of Ligustrum vulgare, an unrelated shrub with similar wax morphology. Epicuticular wax was mechanically stripped from the adaxial leaf surface using the adhesive gum arabic. Subsequently, the organic solvent chloroform was used to extract the intracuticular wax from within the cutin matrix. The isolated waxes were quantified using gas chromatography with flame ionization detection and identified by mass spectrometry. The results were visually confirmed by scanning electron microscopy. The outer wax layer consisted entirely of homologous series of very-long-chain aliphatic compound classes. By contrast, the inner wax layer was dominated (80%) by two cyclic triterpenoids, ursolic and oleanolic acid. The accumulation of triterpenoids in the intracuticular leaf wax of a second, unrelated species suggests that this localization may be a more general phenomenon in smooth cuticles lacking epicuticular wax crystals. The mechanism and possible ecological or physiological reasons for this separation are currently being investigated.