Influence of Canopy Height and Fertilization Levels on the Resistance of Lycopersicon Hirsutum to Aculops Lycopersici (Acari: Eriophyidae)

The objective of this work was to study the effect of NK fertilization levels and canopy height on the resistance of Lycopersicon hirsutum and Lycopersicon esculentum to Aculops lycopersici (Acari: Eriophydae). The effects of NK fertilization levels and canopy height in the leaf size and density of trichomes and their effects on tridecan-2-one (2-TD) and undecan-2-one (2-UD) limiting the attack of A. lycopersici on tomato plants were assessed. Different NK fertilization levels had no effect on the resistance of L. hirsutum to A. lycopersici. No significant differences were found in attack rates of this mite on leaves of the top and median parts of L. hirsutum canopy. The type and density of trichomes were the main determining factor of A. lycopersici attack on tomato plants. High trichome densities and type VI glandular trichomes which produce tridecan-2-one are important resistance factors on tomato plants. L. hirsutum showed a high resistance level to A. lycopersici due to high densities of type VI glandular trichomes and consequently higher levels of tridecan-2-one in its leaves.     

Role of canopy height in the resistance of Lycopersicon hirsutum f. glabratum to Tuta absoluta (Lep., Gelechiidae)

The objective of this work was to study the effect of plant canopy height on the resistance of the tomato species Lycopersicon hirsutum to the tomato leafminer Tuta absoluta . Determination of the levels of tridecan-2-one (2-TD) and undecan-2-one (2-UD) in L. hirsutum , as well as the leaf area, density and types of trichomes present in L. hirsutum and L. esculentum were made on apical, medium and basal parts of plant canopy. Correlation of these data were made with the following biological characteristics of T. absoluta : rates of oviposition and egg hatching; length of egg, larval and pupal stages; mortality of the larval and pupal stages; pupal weight; and sex ratio. Levels of 2-TD on leaves of L. hirsutum increased from bottom to top of canopy. Tuta absoluta had higher oviposition and egg hatching, as well as lower mortalities and smaller larval and pupal periods in L. esculentum than in L. hirsutum . Longer pupal period and greater number of small and large mines of T. absoluta were verified in the apical and medium parts of L. hirsutum plants. On the other hand, higher pupal mortality and longer larval periods were obtained in the apical and basal parts of the L. hirsutum plants. Larval mortality was higher in the basal part of those plants. In L. esculentum , leaves from the apical part of the plant promoted lower mortality, longer larval period and greater number of large mines of T. absoluta . However, a shorter pupal period was verified on leaves of the basal part. It appears that an increase in glandular trichome density in L. hirsutum leads to an increase in the levels of 2-TD, which, in turn slows down larval development.     

Trichomes of Lycopersicon spp. and their effect on Myzus persicae (Sulzer) (Hemiptera: Aphididae)

Abstract The green peach aphid ( Myzus persicae ) is becoming resistant to many of the pesticides used in the tomato ( Lycopersicon esculentum ) industry. A potential alternative method of control is host-plant resistance, which may be mediated by trichomes. Resistance to the green peach aphid was assessed for six accessions of L. hirsutum, three of L. pennellii and a L. esculentum control. Mortality, entrapment and number of aphids unaffected by trichomes were recorded 24, 48 and 72 h after placement. Using multiple regression against counts of glandular and non-glandular trichomes, the role of foliar pubescence in resistance to the aphid was established. Lycopersicon pennellii possessed the highest level of resistance after 72 h and type IV trichome density accounted for 26.5% of variance in mortality ( P  < 0.001). Low densities of type IV and high densities of type VII trichomes increased the number of aphids 'unaffected' by trichomes, a relationship accounting for 29% of variance ( P  < 0.001). Information on which types of trichomes most strongly affect resistance to insect pests will aid future breeding and L. pennellii shows potential for use in breeding programs to increase levels of host-plant resistance in L. esculentum.     

Resistance of wild Lycopersicon species to the potato moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae)

Abstract  Trichome-based host plant resistance of Lycopersicon species to potato moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), was examined in a laboratory bioassay. Neonate P. operculella were placed on the foliage of accessions of L. hirsutum , L. hirsutum f. glabratum , L. pennellii , L. cheesmanii f. minor and L. esculentum for 48 h . At the end of this period, larval mortality, the numbers of larvae emigrating from, and mining into, the leaf surface were recorded as were leaf area and the densities of trichomes. Analysis by categorical logistic regression found accessions differed in levels of resistance to P. operculella ( P  < 0.001). Further analysis using generalised linear models showed mortality was associated with high densities of type VI and low densities of type V trichomes; emigration from the leaf was associated with high densities of type I and type VII trichomes; and the numbers of mines associated with low densities of type I and type VI trichomes and increased leaf area. Although results indicate that certain accessions of L. hirsutum and L. hirsutum f. glabratum may be most appropriate for inclusion in a breeding program aimed at introducing trichome-based host plant resistance of wild Lycopersicon species into the tomato, further research is necessary to fully understand the mechanisms.     

Organ-Level Analysis of Idioblast Patterning in Egeria densa Planch. Leaves

Leaf tissues of plants usually contain several types of idioblasts, defined as specialized cells whose shape and contents differ from the surrounding homogeneous cells. The spatial patterning of idioblasts, particularly of trichomes and guard cells, across the leaf epidermis has received considerable attention as it offers a useful biological model for studying the intercellular regulation of cell fate and patterning. Excretory idioblasts in the leaves of the aquatic monocotyledonous plant Egeria densa produced light blue autofluorescence when irradiated with ultraviolet light. The use of epifluorescence microscopy to detect this autofluorescence provided a simple and convenient method for detecting excretory idioblasts and allowed tracking of those cells across the leaf surfaces, enabling quantitative measurement of the clustering and spacing patterns of idioblasts at the whole leaf level. Occurrence of idioblasts was coordinated along the proximal–distal, medial–lateral, and adaxial–abaxial axes, producing a recognizable consensus spatial pattern of idioblast formation among fully expanded leaves. Idioblast clusters, which comprised up to nine cells aligned along the proximal–distal axis, showed no positional bias or regularity in idioblast-forming areas when compared with singlet idioblasts. Up to 75% of idioblasts existed as clusters on every leaf side examined. The idioblast-forming areas varied between leaves, implying phenotypic plasticity. Furthermore, in young expanding leaves, autofluorescence was occasionally detected in a single giant vesicle or else in one or more small vesicles, which eventually grew to occupy a large portion of the idioblast volume as a central vacuole. Differentiation of vacuoles by accumulating the fluorescence substance might be an integral part of idioblast differentiation. Red autofluorescence from chloroplasts was not detected in idioblasts of young expanding leaves, suggesting idioblast differentiation involves an arrest in chloroplast development at a very early stage, rather than transdifferentiation of chloroplast-containing epidermal cells.     

Leaf anatomy ofDombeya andNesogordonia (Sterculiaceae), emphasizing epidermal and internal idioblasts

We studied leaf anatomy, using clearings, resin sections, and scanning electron microscopy, from mostly herbarium specimens of 123 species ofDombeya and 11 species ofNesogordonia (Sterculiaceae). Species were placed in seven idioblast categories, ranging from those without any to single and bicelled epidermal forms to multicelled nodules and single mesophyll idioblasts. Idioblast contents are possibly mucilaginous, but were not identified. In these two genera the range of foliar idioblast morphology surpasses that known previously for the entire family. Leaves are dorsiventral with mostly abaxial anomocytic stomata and typical palisade and spongy layers; paraveinal mesophyll is lacking. Miniature glandular (clavates, capitates) and nonglandular (mostly stellate) trichomes occur. Prismatic crystals predominate; epidermal prismatics and mesophyll druses are rare.     

Sesquiterpene carboxylic acids from a wild tomato species affect larval feeding behavior and survival of Helicoverpa zea and Spodoptera exigua (Lepidoptera: Noctuidae).

The sesquiterpene carboxylic acids (SCA), (+)-(E)-alpha-santalen-12-oic, (-)-(E)-endo-alpha-bergamoten-12-oic, and (+)-(E)-endo-beta-bergamoten-12-oic acid, are produced in glandular trichomes of Lycopersicon hirsutum f. typicum Humb. & Bonpl. accession (LA) 1777, which is highly resistant to a range of pests of cultivated tomatoes. L. esculentum Mill. Exposure of the larvae of two key tomato pests, tomato fruitworm [Helicoverpa zea (Boddie)] and beet armyworm [Spodoptera exigua (Hübner)], to these compounds in diets and on leaf surfaces resulted in reduced development rates and survival and deterred feeding. These effects were observed when levels of SCA, in artificial insect diet, applied to leaflets of susceptible cultivars, or synthesized in trichomes of leaves of plants, exceeded 2 mg SCA/g of diet or fresh leaf weight. This study suggests that cultivated tomatoes capable of synthesizing SCA, at 2 mg SCA/g of leaf tissue or greater, on their leaves and fruit would display enhanced host plant resistance to H. zea and S. exigua and other insect pests.     

Isolation and Analysis of Differential Expressed ESTs from Stem Trichomes of Lycopersicon esculentum (Solanaceae)

Glandular trichomes are special organs involved in plant defense response and synthesis of volatile secondary metabolites, analyzing trichome specific expressed sequence tags will help us further understand the specific function of plant trichomes. In this paper, suppression subtractive hybridization（SSH） based on magnetic beads technology was used to isolate differential expressed genes of the glandular trichomes in Lycopersicon esculentum. The differential expressing cDNA library was constructed using the glandular trichomes cDNA as tester and the cDNA from the stem without glandular trichomes as driver. After randomly sequencing 108 differential ESTs, Blast2go program was used to do blastx, functional annotation and metabolism analysis. The results show that most ESTs are related to substance metabolism, response to stress, biotic or abiotic stimulus, and have binding and catalytic function. These differential genes lay the foundation for further research on defense mechanism of the tomato trichomes.     

Calcium Deposition in Idioblasts of Mulberry Leaves

Large, rounded idioblasts were observed in adaxial leaves ofmulberry plants; they were clearly distinguishable from epidermal,trichome and parenchyma cells. The size and density of idioblastsvaried according to leaf age. Cytological features of idioblastswere as follows: the outermost region (‘cap’) ofidioblasts was situated on the adaxial surface as a dome-likeprotrusion; a cylindrical protuberance extended from the capregion to the inner part of the idioblast; in idioblasts frommature leaves a crystal mass was suspended from the lower tipof the cylindrical protuberance. Elemental analysis of idioblastsdemonstrated that silicon (Si) was localized in both the capregion and the cylindrical protuberance but calcium (Ca) waspresent in the large crystal, indicating site-specific cellularlocalization of Ca and Si within an idioblast. Histochemicalassays showed that a distinct Ca crystal filled the vacuolesof idioblasts in mature leaves, while immature leaves had manyidioblasts without Ca deposition. The increase in the Ca contentof leaves was directly proportional to the increase in leafage and appeared to be closely related to the Ca sink capacityof the developing idioblast vacuoles. The maximum sink capacitywas quantified to be approximately 40 ng per idioblast whenmulberry plants were grown hydroponically with excess Ca.Copyright1999 Annals of Botany Company Morus alba, idioblast, Ca deposition, Ca sink capacity, silicon, X-ray microanalysis, histochemistry, scanning electron microscopy.     

番茄茎腺毛差异表达序列分离与分析

腺毛是参与植物防御性反应与次生代谢挥发物合成的特异化器官,了解腺毛专一性表达序列标签将有助于我们进一步认识植物腺毛的特异性功能.本文应用磁珠介导的抑制消减杂交方法(SSH),以番茄茎腺毛为检测子(tester),去除腺毛的番茄茎为驱动子(driver),构建番茄茎腺毛差异表达cDNA文库,分离腺毛差异表达基因.随机挑选了108个差异ESTs进行测序,测序结果使用Blast2go程序进行blastx比对、功能注释和KEGG代谢路径分析.结果表明,绝大部分ESTs功能与胁迫响应、物质代谢、生物及非生物刺激反应相关,具有结合、催化功能.这些分离的差异ESTs为进一步研究番茄腺毛的植物防御性机制奠定了基础.     

Leucine uptake by tomato leaf tissue under low temperature: Preincubation dependence of uptake reduction

The uptake of ³H-leucine by leaf fragments of Lycopersicon esculentum Mill. cv. Rutgers and L. hirsutum Humb. & Bonpl., a wild tomato, was studied. Two altitudinal races of L. hirsutum were used which differed in chilling tolerance. The temperature dependence of uptake was initially similar for all plant varieties. However, at temperatures below about 11°C, uptake progressively decreased in the more chilling-sensitive varieties ( L. esculentum , Low-altitude L. hirsutum ), but not in the more chilling-tolerant (high-altitude L. hirsutum ) with increasing preincubation time. More than 60 min preincubation was required for this effect, and it was greatest at the lower temperatures. When leaf fragments, chilled for short periods of time (>22 h), were returned to 22°C, initial rates of uptake were recovered within 2 h. The relationship between membrane lipid changes and membrane protein activity under chill stress is discussed.     

Toxicity of methyl ketones from tomato trichomes to Tetranychus urticae Koch

The two methyl ketones, 2-tridecanone and 2-undecanone, are constituents of type VI glandular trichomes of the wild tomato, Lycopersicon hirsutum f. glabratum (PI 134417). They are known to cause mortality in several herbivorous insect species. In this study we investigated the effects of these chemicals on two strains of the two-spotted spider mite, Tetranychus urticae Koch, collected from tomato and cucumber crops in Dutch greenhouses. The two ketones were tested separately, in combination in the ratio found in L. hirsutum f. glabratum and in several other ratios to detect any synergistic interaction between them. We measured both the direct contact and residual toxicity, as well as the viability of the eggs produced by ketone-treated females. The methyl ketones can be considered as quite toxic to spider mites because their LC50 values were comparable to that found for the formulated acaricide amitraz. 2-Tridecanone was slightly more toxic than 2-undecanone but only in the tomato strain. Synergistic effects were not detected. In the bio assays for the residual effects of the two methyl ketones no significant mortality occurred but the mites avoided feeding on the treated surface and the eggs were laid almost exclusively on the untreated area. Furthermore, there was no significant egg viability for most of the treatments and when some egg viability occurred, it was not correlated to the dose of the chemical. Based on published data on the levels of methyl ketones in the trichome tips of wild and cultivated tomato (L. hirsutum f. glabratum and Lycopersicon esculentum Mill, respectively) it was calculated how many contacts are required to reach the LD50 (ng a.i. per mite). It appeared that on wild tomato just a few contacts suffice, but many are needed on cultivated tomato. Nevertheless, it seems justified to conclude that even in cultivated tomato, 2-tridecanone represents a naturally occurring acaricide. The cucumber and tomato strains of the two-spotted spider mite did not differ in their response to the methyl ketones. Because this is surprising, given the selection pressures on their respective host plants, we formulated two hypotheses, one explaining the absence of a difference due to cross-resistance and another one proposing that bioaccumulation obscures the underlying differences between the strains. This revised version was published online in November 2006 with corrections to the Cover Date.     

Inhibition of Telenomus sphingis an egg parasitoid of Manduca spp. by trichome/2-tridecanone-based host plant resistance in tomato

The resistance of accession PI 134417 of the wild tomato Lycopersicon hirsutum f. glabratum C. H. Mull to Manduca sexta (L.) (Lepidoptera: Spingidae) and Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) is conditioned by the high densities of 2-tridecanone-containing, glandular trichomes associated with the foliage. In laboratory experiments, rates of parasitism of M. sexta eggs by Telenomus sphingis (Ashmead) (Hymenoptera: Scelionidae) were lower among eggs on PI 134417 foliage than among eggs on foliage of the cultivated tomato L. esculentum Mill. (cv. Better Boy). The latter is characterized by a significantly lower density of type VI glandular trichomes than PI 134417 and an absence of 2-tridecanone. Parasitism by T. sphingis was also reduced among eggs on foliage of the F₁ hybrid between PI 134417 and L. esculentum. The hybrid foliage lacks 2-tridecanone but has a density of type VI glandular trichomes that is intermediate between those of PI 134417 and L. esculentum, indicating that elevated densities of type VI glandular trichomes adversely affect T. sphingis. This conclusion was further substantiated by the finding that there were no differences among plant lines in the levels of parasitism of M. sexta eggs when the eggs were on foliage that had been divested of glandular trichomes.In bioassays in which T. sphingis adults or immatures in host eggs were exposed to filter paper treated with 2-tridecanone at rates comparable to those associated with PI 134417 foliage, 2-tridecanone was acutely toxic and caused high levels of mortality. In addition, at high concentrations, 2-tridecanone vapors were repellent to T. sphingis adults. However, when exposed to PI 134417 foliage, few T. sphingis adults were killed.Parasitism of M. sexta eggs was unaffected when the eggs were deposited by moths reared as fifth instar larvae on diet containing 2-tridecanone and/or 2-undecanone at levels comparable to those associated with PI 134417 foliage.     

Release of hormones from conjugates: chloroplast expression of β-glucosidase results in elevated phytohormone levels associated with significant increase in biomass and protection from aphids or whiteflies conferred by sucrose esters

Transplastomic tobacco (Nicotiana tabacum) plants expressing β-glucosidase (Bgl-1) show modified development. They flower 1 month earlier with an increase in biomass (1.9-fold), height (1.5-fold), and leaf area (1.6-fold) than untransformed plants. Trichome density on the upper and lower leaf surfaces of BGL-1 plants increase by 10- and 7-fold, respectively, harboring 5-fold more glandular trichomes (as determined by rhodamine B staining), suggesting that BGL-1 lines produce more sugar esters than control plants. Gibberellin (GA) levels were investigated because it is a known regulator of flowering time, plant height, and trichome development. Both GA(1) and GA(4) levels are 2-fold higher in BGL-1 leaves than in untransformed plants but do not increase in other organs. In addition, elevated levels of other plant hormones, including zeatin and indole-3-acetic acid, are observed in BGL-1 lines. Protoplasts from BGL-1 lines divide and form calli without exogenous hormones. Cell division in protoplasts is enhanced 7-fold in the presence of exogenously applied zeatin-O-glucoside conjugate, indicating the release of active hormones from their conjugates. Whitefly (Bemisia tabaci) and aphid (Myzus persicae) populations in control plants are 18 and 15 times higher than in transplastomic lines, respectively. Lethal dose to kill 50% of the test population values of 26.3 and 39.2 μg per whitefly and 23.1 and 35.2 μg per aphid for BGL-1 and untransformed control exudates, respectively, confirm the enhanced toxicity of transplastomic exudates. These data indicate that increase in sugar ester levels in BGL-1 lines might function as an effective biopesticide. This study provides a novel strategy for designing plants for enhanced biomass production and insect control by releasing plant hormones or sugar esters from their conjugates stored within their chloroplasts.