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.     

Trichome‐based host plant resistance of Lycopersicon species and the biocontrol agent Mallada signata: are they compatible?

Trichome‐based host plant resistance of Lycopersicon (Solanaceae) species offers the potential to reduce pesticide use in tomato production, but its compatibility with biocontrol agents is largely unknown. The effect of trichome‐based host plant resistance on the lacewing biocontrol agent, Mallada signata, was assessed for five accessions of L. cheesmanii, four accessions of Lycopersicon hirsutum, two accessions of Lycopersicon pennellii, and one Lycopersicon esculentum cultivar. An intact leaf was isolated from the whole plant using Tangletrap to coat the petiole and 20 green peach aphids [Myzus persicae (Sulzer) (Homoptera: Aphididae)] were placed on the leaf surface. After 24 h, 10 lacewings were placed on the leaf. The numbers of dead, trapped by exudates, untrapped and predated lacewings and aphids, and the numbers that had left the leaf were recorded a further 24 h later. Differences in insect designations between accessions were analysed using ANOVA. A General Linear Model (GLM), consisting of the densities of each trichome type and leaf area, was fitted to the data to determine the role of trichomes on the observed effects on lacewings and aphids. Lacewing mortality was greater on one accession of L. pennellii and one accession of L. hirsutum than on L. esculentum. The GLM indicated that type IV trichomes decreased the numbers of aphids predated, and increased cannibalism and, along with type III trichomes, increased entrapment‐related predator mortality. Although there were no differences in the numbers of predated aphids, with the majority predated for all accessions, the compatibility of trichome‐based host plant resistance of Lycopersicon species and the biocontrol of aphids by lacewings is questionable.     

Induced tolerance of neonate Heliothis zea to host plant allelochemicals and carbaryl following incubation of eggs on foliage of Lycopersicon hirsutum f. glabratum

Summary Incubation of Heliothis zea (Boddie) eggs on foliage of Lycopersicon hirsutum f. glabratum C.H. Mull (accession PI 134417) results in neonates with elevated levels of tolerance to the toxic effects of PI 134417 foliage attributable to 2-tridecanone found in the glandular trichomes which abound on that foliage. The neonates from such eggs are also shown to have elevated levels of tolerance to the carbamate insecticide carbaryl. Incubation of eggs in an atmosphere containing 2-tridecanone similarly produced elevated levels of tolerance to 2-tridecanone among resulting neonates, indicating that 2-tridecanone is the likely inducing agent and that exposure to 2-tridecanone vapor, which is known to emanate from PI 134417 foliage, is sufficient for induction. Analysis of the cytochrome P-450 content in gut microsomes of fifth instar larvae indicated that exposure of larvae to 2-tridecanone in artificial diet or to PI 134417 foliage resulted in significantly elevated levels of cytochrome P-450 relative to larvae fed diet without 2-tridecanone or foliage of L. esculentum which contains no 2-tridecanone. In addition, removal of the glandular trichomes from PI 134417 foliage eliminated the ability of that foliage to induce elevated levels of cytochrome P-450. These results provide circumstantial evidence that cytochrome P-450 may be involved in the induced tolerance to xenobiotics among neonates from eggs exposed to 2-tridecanone or PI 134417 foliage.Support for this research was provided by the USDA Competitive Research Grants Program in Biological Stress under Grant No. 83-CRCR-1-1241 and Grant No. 85-CRCR-1-1615, and the North Carolina Agricultural Research Service. Paper No. 10856 of Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC, USA 27650. Use of trade names does not imply endorsement of products named nor criticisms of similar ones not mentioned     

Interaction of fertilizer regime with host-plant resistance in tomato

The effect of fertilizer regime on trichome- and lamellar-based resistance in the wild tomato species, Lycopersicon hirsutum f. glabratum C.H. Mull accession PI 134417, to three insect pests of tomato, the tobacco hornworm, Manduca sexta (L.), the colorado potato beetle, Leptinotarsa decemlineata (Say), and the tomato fruitworm, Heliocoverpa zea (Boddie), was examined. Increasing the rate at which NPK fertilizer was applied, from 1.8 to 19.6 g/plant/week, reduced the trichome-based resistance of PI 134417 to M. sexta and L. decemlineata by lowering both the density of type VI (sensu Luckwill, 1943) glandular trichomes and the amount of 2-tridecanone contained in the tips of these trichomes. 2-Tridecanone is a toxic methyl-ketone responsible for glandular trichome-mediated resistance in PI 134417 to M. sexta and L. decemlineata. A similar increase in the application rate of NPK fertilizer reduced the lamellarbased resistance of PI 134417 to L. decemlineata and H. zea. The meachanisms for this reduction of resistance are unknown, but may be related to improved nutritional quality of hosts at higher fertilizer regimes.     

Interaction ofManduca sexta resistance in tomato with insect predators ofHelicoverpa zea

Resistance in accession PI 134417 ofLycopersicon hirsutum f.glabratum toManduca sexta L. results from the presence of the methyl ketones 2-tridecanone and 2-undecanone in the type VI trichome glands on foliage of these plants. Short (24 h) and long term (neonate to adult) laboratory experiments usingM. sexta-resistant and susceptible foliage and methyl ketone treated filter paper disks were conducted to evaluate the direct (plant-mediated), and indirect (prey-mediated) effects of this resistance on two predators ofHelicoverpa zea eggs,Coleomegilla maculata (DeGeer) andGeocoris punctipes (Say). Direct effects of resistance were manifest as reduced egg consumption and increased mortality for both predators. Indirect effects were manifest as reduced egg consumption byG. punctipes, but were not observed forC. maculata. Results of experiments using methyl ketone-treated filter paper disks instead of foliage were similar.     

Effect of plant variety, plant age and photoperiod on glandular pubescence and host-plant resistance to potato moth (Phthorimaea operculella) in Lycopersicon spp.

The effect of plant age and daylength on glandular pubescence was determined for two lines of tomato derived from Lycopersicon hirsutum (BTN 979 and LA 1777A) and a variety of L. esculentum (N 91‐1‐1‐1‐1). Densities of type I, IV, VI and VII glandular trichomes were lowest in N 91‐1‐1‐1‐1 and, over all varieties, were more dense on plants aged greater than 6 wk. Daylength interacted with variety to significantly affect densities of type VII trichomes only. Host‐plant resistance to Phthorimaea operculella was determined in preliminary tests using insects cultured from founders from a potato crop and in confirmatory tests using (less readily available) insects recovered from foliage of a tomato crop. Mortality of ex‐potato neonates on LA 1777 A and BTN 979 foliage was higher 18 h after placement than for N 91‐1‐1‐1‐1, with no effect of day length or plant age. Mortality for ex‐tomato neonates followed a similar trend. Ten days later, two‐thirds of ex‐tomato larvae had established mines on N 91‐1‐1‐1‐1 but fewer (16.7%) were live on other varieties. Stepwise multiple regression using variety as the sole factor was significant (P<0.001) in accounting for 61.4% of the variation in ex‐tomato larval survival but addition of other factors to the regression model was not significant. BTN 979 supported fewer, smaller adults to develop than did N 91‐1‐1‐1‐1, whilst no adults developed on LA 1777 A. In a non‐choice test using ex‐potato adults, significantly more eggs were laid on N 91‐1‐1‐1‐1 than on L. hirsutum varieties and 9‐wk‐old plants were preferred over plants three weeks older or younger. The same variety and plant age trends were evident in a free‐choice test using ex‐tomato adults.     

S-related protein can be recombined with self-compatibility in interspecific derivatives ofLycopersicon

Stylar proteins involved in the self-incompatible (SI) response ofLycopersicon hirsutum have been identified and mapped to the locus that controls SI (S locus).L. esculentum, a self-compatible (SC) species of cultivated tomato, does not display these proteins. Hybrids between SCL. esculentum and SIL. hirsutum are self-sterile despite these individuals bearing pollen containing theS allele ofL. esculentum. In progeny derived from backcrossing the hybrids toL. esculentum, there was a strong correlation between the presence of theS allele fromL. hirsutum and self-infertility. However, this relationship was uncoupled in a number of backcross (BC) progeny. The SI response appeared to be nonexistent in two self-fertile BC individuals that were heterozygous for theS allele ofL. hirsutum, based on Mendelian segregation of a tightly linked DNA marker,CD15, in selfed progeny. Among these progeny self-fertile individuals that were homozygous for theL. hirsutum allele of the linked marker were also determined to be homozygous for anS-related protein ofL. hirsutum through test crosses withL. esculentum. Therefore, plants were produced that were homozygous for a functionalS allele but were self-fertile. This result and other evidence suggest that theS-related proteins are not sufficient to elicit a self-incompatible response inL. esculentum and that there is a mutation(s) inL. esculentum somewhere other than theS locus that leads to self-compatibility.     

2-Undecanone, a pupal mortality factor in Heliothis zea: sensitive larval stage and in planta activity in Lycopersicon hirsutum f. glabratum

2-Undecanone, a compound found in the tips of the Type VI glandular trichomes of the wild tomato species Lycopersicon hirsutum f. glabratum C. H. Mull (accession PI 134417), when incorporated in artificial diet, was previously shown to cause pupal deformity and mortality among Heliothis zea (Boddie). In the present study, the developmental stage of H. zea sensitive to the effects of dietary 2-undecanone was determined by rearing larvae through the fourth instar on diet with or without 2-undecanone, then transferring them to new diet either with or without 2-undecanone. Dietary levels of 2-undecanone (0.055% wet wt) were representative of concentrations found in PI 134417 foliage. Ingestion of 2-undecanone during the fifth instar was both necessary and sufficient to cause pupal mortality. Rearing fifth instar H. zea on PI 134417 foliage with glandular trichomes intact (2-undecanone present at 0.084±0.026% wt wt) or with the trichomes, and hence 2-undecanone, removed indicated that these effects occur in planta. Potential implications of these findings for using 2-undecanone-mediated resistance for insect pest management are discussed.

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Résumé Il a été démontré précédemment que l'undécanone-2, présente dans les extrémités des trichomes glandulaires de type VI de la tomate sauvage Lycopersicon hirsutum glabratum C. H. Mull (accession PI 134417), provoque déformations et mortalité chez les chrysalides de Heliothis zea (Boddie) lorsqu'elle est incorporée dans une nourriture artificielle. Dans cette étude, le stade de développement de H. zea sensible aux effets de l'undécanone-2 a été déterminé en élevant les larves jusqu'au 4e stade sur une diète contenant ou non de l'undécanone-2, puis en les transférant sur une nouvelle diète avec ou sans undécanone-2. Les concentrations en undécanone-2 utilisées (0.055% du poids frais) étaient comparables aux concentrations présentes dans les feuilles du PI 134417. L'ingestion de l'undécanone-2 durant le 5e stade s'est avérée nécessaire et suffisante pour provoquer une mortalité au stade chrysalide. L'élevage du 5e stade de H. zea sur des feuilles de PI 134417 dont les trichomes sont intacts (l'undécanone-2 est présente à la concentration de 0.084±0.025% du poids frais) ou sur des feuilles dont les trichomes, et par conséquent l'undécanone-2, ont été éliminés, a démontré que les effets toxiques se produisent in planta.L'undécanone-2 pourrait être un facteur de résistance efficace des tomates, mais probablement uniquement dans les régions où H. zea est polyvoltime sur la tomate et où les premiéres générations se développent sur le feuillage pendant toute la période larvaire avant la maturation des fruits. Normalement, les premiers stades larvaires de H. zea se nourrissent de feuilles et les derniers stades des fruits. Les fruits ne contenant pas d'undécanone-2, les chenilles qui se développeraient sur les plants en fructification pourraient échapper aux effets toxiques.

     

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.     

Accumulation and turnover of 2-tridecanone in Tetranychus urticae and its consequences for resistance of wild and cultivated tomatoes

In this study we assessed the dynamic changes of 2-tridecanone in a herbivorous mite (Tetranychus urticae) on tomato (Lycopersicon esculentum, cv. Moneymaker), a plant with methyl ketones in the tetracellular tips of the glandular trichomes (Type VI). We showed that spider mites accumulate 2-tridecanone when foraging on cultivated tomato. Thus, the rate of mite–trichome contact multiplied by the amount of toxin per trichome tip exceeded the relative rate of toxin turnover multiplied by the amount of toxin per mite. The relative rate of toxin turnover was estimated to be 1.1 per day on cucumber, a plant without this toxin. The amount per trichome tip varied from 0.33 ng for middle-leaf trichomes to 1.26 ng for main-stem trichomes. Hence, to achieve a static level of 2-tridecanone equal to 8–17 ng per mite – representing the level we found in mites on middle leaves – the rate of mite–trichome contact should be 26–57 per day. Because methyl ketone apparently accumulates in the spider mites on tomato, the rate of mite–trichome contact is probably higher than that. We expect the accumulation of ketones to occur especially on the stems of cultivated tomato, since this is the area most densely occupied with glandular hairs and because here the hairs have higher levels of the methyl ketones.Using dose–response relationships assessed earlier (Chatzivasileiadis and Sabelis, 1997, 1998), we estimated that the number of mite–trichome contacts causing 50% mortality per day is equal to 88 on a tomato stem, whereas it equals 70 for another strain of spider mites collected from cucumber. On wild tomato, L. hirsutum f. glabratum (PI 134417), just one to two contacts would suffice to cause 50% mortality per day. We suggest that methyl ketones from glandular hairs on tomato are an important mortality factor for spider mites on wild tomato and probably also on cultivated tomato.     

Growth, food consumption and mortality of Heliothis zea larvae on foliage of the wild tomato Lycopersicon hirsutum f. glabratum and the cultivated tomato, L. esculentum

Larvae of Heliothis zea (Boddie) were fed foliage of the resistant wild tomato Lycopersicon hirsutum f. glabratum C. H. Mull (PI 134417) and susceptible L. esculentum (cultivar Walter), either with the glandular trichomes removed or with the trichomes intact. Response variables measured included larval mortality, larval weight, consumption rate (CR), and efficiency of conversion of ingested material (ECI). Resistant foliage was found to contain factors in both the trichomes and leaf lamellae which increased larval mortality, reduced larval weight, reduced CR, and, except in second instars, reduced ECI. Susceptible foliage also contained trichome-based factors which increased larval mortality and decreased weight, as well as CR of second instars and ECI of fifth instars. Implications of these findings for breeding resistant tomato cultivars are discussed.

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Résumé La tomate sauvage, Lycopersicon hirsutum f. glabratum C. H. Mull (PI 134417) est fortement résistant à Heliothis zea. Les études préalables ont indiqués que la résistance est associée avec la présence de 2-tridécanone dans les extrémités des poils glandulaires de la tomate sauvage, et que les facteurs dans la lamelle des feuilles sont peut-être impliqués. À déterminer la présence des facteurs lamellaires et à caractériser de plus les facteurs de résistance dans les extrémités des poils glandulaires aussi, nous nourrissions les larves du feuillage résistant de PI 134417 et du feuillage susceptible L. esculentum (lignée Walter) les deux avec les extrémités des poils glandulaires enlèvés ou avec les extrémités des poils glandulaires intacts. Nous mesurions la mortalité des larves, les poids des larves, la taux de consommation relative et l'efficacité de conversion de matière mangé.On se trouvait le feuillage résistant à contenir les facteurs, tous deux dans le feuillage lamellaire et dans les extrémités des poils glandulaires, qui causaient l'augmentation de mortalité, les poids plus basses, la taux de consommation relative plus basse, et, hormis dans le cas des larves en phase deuxième, l'efficacité de conversion réduite. Le feuillage susceptible contenait aussi les facteurs dans les extrémités des poils glandulaires qui augmentaient la mortalité, diminuaient les poids et la taux de consommation relative des larves en phase deuxième, et diminuaient l'efficacité de conversion des larves en phase cinquième. Cettes conclusions indiquent la présence des facteurs de résistance en outre de 2-tridécanone et peuvent fournir les indices à l'identité de ces facteurs.

     

Variability of Lycopersicon hirsutum f. typicum and possible compounds involved in its resistance to Tuta absoluta

1 The resistance of Lycopersicon hirsutum f. typicum and L. esculentum to the leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) was investigated in two experiments. The first experiment assessed the resistance of L. hirsutum f. typicum (accession LA 1777) to T. absoluta, the second assessed the variability of the resistance among plants of this accession. 2 It was found that L. hirsutum f. typicum was resistant to the leafminer, which showed longer larval phase, higher larval mortality, smaller numbers of large mines and greater numbers of small mines/leaf than on L. esculentum. Seven plants of the accession LA 1777 showed lower number of small mines/leaf, but their results of larval mortality and length of larval and pupal phase were similar. Twenty-two peaks were observed in total ion chromatograms of L. hirsutum f. typicum hexane extracts of leaves subjected to a gas chromatographic-mass spectrometric analysis (CG-MS). However, the matches obtained between the peak spectra and the MS database were too low to justify even preliminary identification of the compounds.     

Variable responses of tuber moth to the leaf trichomes of wild potatoes

This study examines the response of tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), during the initial stages of attack, to variability in trichome density and composition on foliage of Solanum berthaultii (Hawkes) and Solanum tarijense (Hawkes) (Solanaceae). Solanum berthaultii bears two types of glandular trichome (type A and type B) that together reduced oviposition by the moth. Females were often completely deterred from ovipositing on foliage with >300 trichomes per cm². In contrast, neonate establishment on S. berthaultii was generally positively related to trichome densities, indicating that trichomes may be a poor defense against P. operculella when the moth oviposits in soil and neonate larvae select the host plant. Solanum tarijense has only one type of glandular trichome (type A) and eglandular hairs. Most eggs were deposited on the adaxial leaf surfaces that had lower trichome densities. Although the density of type A trichomes was negatively related to oviposition, high densities of hairs on the abaxial and adaxial leaf surfaces appeared to stimulate oviposition, leading to stronger positive relations between hair densities and oviposition. Larvae generally established on the abaxial surface where hair densities were greatest. Relationships between the abaxial densities of leaf hairs and neonate establishment on S. tarijense were positive. The results indicate that the responses by P. operculella to the types and density of trichomes are complex. Whereas type A and type B trichomes may act synergistically to reduce oviposition by the moth, leaf hairs do not defend against oviposition and neither leaf hairs nor type A and B trichomes reduce neonate establishment by this herbivore species.     

Feeding behavior of potato aphid affected by glandular trichomes of wild tomato

Mortality of the potato aphid, Macrosiphum euphorbiae (Thomas), on Lycopersicon pennellii (Corr.) D'Arcy and its F₁ hybrid with Lycopersicon esculentum Mill. was significantly greater than that on L. esculentum. Physical entrapment was not the sole mechanism of resistance in L. pennellii since few late instar aphids were found trapped in the sticky glandular exudate of the type IV trichomes; entrapment could, however, affect survival of early instars. Aphid settling on L. pennellii was dramatically less than that on L. esculentum, suggesting that starvation may have contributed to high mortality. Compared to L. esculentum, aphid feeding behavior on L. pennellii and the F₁ was characterized by a delay in the time to first probe, a reduction in the number of probes, and a decrease in the total proportion of time spent feeding. Removal of the glandular exudate of the type IV trichomes from L. pennellii resulted in a decrease in preprobe time and an increase in both the number of probes and the percent of time spent probing. Transfer of glandular trichome exudate of L. pennellii to leaflets of L. esculentum resulted in an increase in resistance as measured by these three parameters.

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Zusammenfassung Die Absterberate der Kartoffellaus, Macrosiphum euphorbiae Thomas, auf Lycopersicon pennellii (Corr.) D'Arcy, sowie auf der Kreuzung L. esculentum Mill. und L. pennellii, war deutlich grösser als auf L. esculentum. Das mechanische Verfangen der Läuse war nicht der Hauptgrund der Resistenz von L. pennellii. Wenige tote Läuse wurden in dem klebrigen Sekret der Typus IV Trichome gefunden. Auf L. pennellii siedelten sich die Läuse in viel geringerer Zahl an als auf L. esculentum. Dies führte zum Schluss, dass Verhungern eine der Ursachen der hohen Mortalität der Läuse war. Im Vergleich zum Saugverhalten auf L. esculentum war das Saugverhalten auf L. pennellii, wie auch auf F₁, durch Folgendes gekennzeichnet 1) Verspätung des ersten Stichversuchs, 2) Verminderung der Stichversuche pro Zeiteinheit und 3) Verminderung des Zeitanteils, der zum Saugen verwendet wurde. Die Entfernung des Sekrets der Typus IV Trichomen auf L. pennellii verursachte 1) eine kürzere Zeitspanne vor dem ersten Stichversuch, 2) eine Vergrösserung der Anzahl Stichversuche pro Zeiteinheit, 3) eine Verlängerung der Saugzeit. Die Uebertragung des Sekretes von L. pennellii auf Blätter von L. esculentum verbesserte deren Resistenz gegen Blattläuse gemessen mit den genannten drei Kriterien.

     

The development of bridge lines for interspecific gene transfer between Lycopersicon esculentum and L. peruvianum

Summary Using a modified embryo callus culture technique, hybrids between Lycopersicon esculentum and L. peruvianum were developed and their usefulness as bridge lines for facilitating interspecific gene transfer was evaluated. Four of these lines showed a high level of sexual compatibility with several other L. peruvianum var. typicum accessions, as well as with accessions of L. peruvianum var. humifusum and L. peruvianum var. glandulosum and L. esculentum. These bridge line x L. peruvianum hybrids could be crossed with L. esculentum to introgress genes from L. peruvianum into L. esculentum.     

Genome composition of asymmetric hybrids in relation to the phylogenetic distance between the parents. Nucleus-chloroplast interaction

Summary A series of fusion experiments were performed between protoplasts of a cytoplasmic albino mutant of tomato, Lycopersicon esculentum (ALRC), and gamma-irradiated protoplasts of L. hirsutum and the Solanum species S. commersonii, S. etuberosum and S. nigrum. These species were chosen for their different phylogenetic relationships to tomato. In all fusion combinations except from those between ALRC and S. nigrum, green calli were selected as putative fusion products and shoots regenerated from them. They were subsequently analyzed for their morphology, nuclear DNA composition and chloroplast DNA origin. The hybrids obtained between ALRC and L. hirsutum contained the chloroplasts of L. hirsutum and had the flower and leaf morphology of L. esculentum. After Southern blot analysis, using 13 restriction fragment length polymorphisms (RFLPs) randomly distributed over all chromosomes, all hybrids showed L. esculentum hybridization patterns. No chromosomes of L. hirsutum were found. These results indicate that these hybrids were true cybrids.The putative asymmetric hybrids, obtained with S. commersonii and S. etuberosum, showed phenotypic traits of both parents. After hybridization with species-specific repetitive nuclear DNA probes it was found that nuclear material of both parents was present in all plants. In the case of S. nigrum, which combination has the greatest phylogenetic distance between the fusion parents, no hybrid plants could be obtained. The chloroplast DNA of all hybrid plants was of the donor type suggesting that chloroplast transfer by asymmetric protoplast fusion can overcome problems associated with large phylogenetic distances between parental plants.     

Resistance to powdery mildew (Oidium lycopersicum) in Lycopersicon hirsutum is controlled by an incompletely-dominant gene Ol-1 on chromosome 6

The inheritance of resistance to powdery mildew (Oidium lycopersicum) in Lycopersicon hirsutum was investigated by disease tests in segregating populations obtained by hybridising tomato (L. esculentum) cv Moneymaker with the wild relative L. hirsutum G1.1560. One incompletely dominant gene Ol-1 was found to largely control resistance to the disease. To map Ol-1, DNA pools from seven resistant and ten susceptible F₂ plants were analyzed for random amplified polymorphic DNA (RAPD). With 32 primers tested, one RAPD, primed with the sequence 5-GACGTGGTGA-3, was observed between the susceptible and the resistant bulks, which cosegregated with resistance in the F₂ population of L. esculentum × L. hirsutum G1.1560. This RAPD was mapped on chromosome 6 by using an F₂ (L. esculentum × L. pennellii) already mapped for 49 RFLPs. RFLP analysis of the F₂ from L. esculentum cv Moneymaker × L. hirsutum G1.1560 demonstrated that Ol-1 maps near the Aps-1 region on chromosome 6, in the vicinity of the resistance genes to Meloidogyne spp. (Mi) and to Cladosporium fulvum (Cf-2/Cf-5).     

Micromorphological and chemical characterisation of Stachys recta L. subsp. serpentini (Fiori) Arrigoni in comparison to Stachys recta L. subsp. recta (Lamiaceae)

Stachys recta L. is a very polymorphous species in which numerous subspecies were recognised. S. recta L. subsp. serpentini (Fiori) Arrigoni is a typical endemism growing on serpentine soils in northern Apennines and particularly in Tuscany (Italy). In order to contribute to a better knowledge of this plant and to its differentiation with respect to S. recta L. subsp. recta, the micromorphological characters (non-glandular and glandular trichomes) and the essential oil composition of the two subspecies were investigated. Micromorphological characters were studied using scanning and transmission electron microscopy, while light microscopy was used for histochemical observations. Essential oil analysis was carried out by gas chromatography and mass spectrometry.In the two examined taxa, the morphology and distribution of glandular and non-glandular trichomes, and the different essential oil composition, may be considered distinctive characters at subspecies level. This is consistent with the taxonomic classification considering S. recta subsp. serpentini a subordinate taxon of S. recta.     

Pollen selection for low temperature adaptation in tomato

Summary Pollen selection experiments were conducted in tomato to determine the effects of low temperature conditions during pollination on the rate of root elongation of the progeny. Pollen was harvested from an F₁ interspecific hybrid between a high altitude Lycopersicon hirsutum accession and the cultivated tomato L. esculentum. The pollen was applied to stigmas of malesterile L. esculentum plants maintained in growth chambers set at either 12°C/7°C or 24°C/18°C. BC₁ seeds from the low and normal temperature crosses were germinated and root elongation rate was measured at either 9°C or 24°C. At 9°C, the rate of root elongation for progeny of the low temperature crosses was higher than for progeny of crosses at normal temperatures; at 24°C the rate of root elongation was similar for the two crossing treatments. To compare the temperature responses of the two backcross populations we also calculated the relative inhibitory effect of low temperature on the rate of root elongation: the ratio between the rate of root elongation at 9°C to that at 24°C. Root elongation of seedlings from the low temperature crosses was less inhibited by the cold than root elongation for progeny of the normal temperature crosses. These results suggest a relationship between pollen selection at low temperatures and the expression of a sporophytic trait under the same environmental stress.     

Trichoderma species form endophytic associations within Theobroma cacao trichomes

Trichoderma species are usually considered soil organisms that colonize plant roots, sometimes forming a symbiotic relationship. Recent studies demonstrate that Trichoderma species are also capable of colonizing the above ground tissues of Theobroma cacao (cacao) in what has been characterized as an endophytic relationship. Trichoderma species can be re-isolated from surface sterilized cacao stem tissue, including the bark and xylem, the apical meristem, and to a lesser degree from leaves. SEM analysis of cacao stems colonized by strains of four Trichoderma species (Trichoderma ovalisporum-DIS 70a, Trichoderma hamatum-DIS 219b, Trichoderma koningiopsis-DIS 172ai, or Trichoderma harzianum-DIS 219f) showed a preference for surface colonization of glandular trichomes versus non-glandular trichomes. The Trichoderma strains colonized the glandular trichome tips and formed swellings resembling appresoria. Hyphae were observed emerging from the glandular trichomes on surface sterilized stems from cacao seedlings that had been inoculated with each of the four Trichoderma strains. Fungal hyphae were observed under the microscope emerging from the trichomes as soon as 6 h after their isolation from surface sterilized cacao seedling stems. Hyphae were also observed, in some cases, emerging from stalk cells opposite the trichome head. Repeated single trichome/hyphae isolations verified that the emerging hyphae were the Trichoderma strains with which the cacao seedlings had been inoculated. Strains of four Trichoderma species were able to enter glandular trichomes during the colonization of cacao stems where they survived surface sterilization and could be re-isolated. The penetration of cacao trichomes may provide the entry point for Trichoderma species into the cacao stem allowing systemic colonization of this tissue.