Aspects of potato leafhopper (Homoptera: Cicadellidae) biology on Solanum berthaultii and other potato genotypes

Survival of nymphs and adult males and females of potato leafhopper, Empoasca fabae (Harris), on six potato (Solanum spp.) genotypes with varied levels of resistance was evaluated and related to type, density, and droplet/head size of foliar glandular trichomes. Survival was greater on Solanum tuberosum cultivars ('Allegany' and 'Elba') compared with accessions of the wild S. berthaultii (PI 473331 and PI 473334). The barrier provided by glandular trichomes may affect survival of E.fabae by restricting nymph and adult feeding. PI 473331 was the most pubescent host, bearing type A and type B trichomes and the most unsuitable host for E. fabae. Allegany possesses leaflets with low trichome densities and small gland diameter and was the most suitable host for adult E. fabae survival. Females generally had higher survival rates than males. Nymphal and adult leafhoppers differed greatly in their survival rates, particularly on S. tuberosum x S. berthaultii hybrids (Q174-2 and NY123). Interestingly, nymphs had longer (190 d) mean lethal time (LT50) than adults (31 d) when held on Q174-2, and a lower LT50 (19 d) when held on NY123 compared with adults (104 d). Starvation, dehydration, or both were likely associated with mortality of nymphs confined on PI 473331 and PI 473334. Nymphs on these accessions died at a comparable rate to those confined on a starvation treatment and more rapidly than those supplied with water only, sucrose solution or other potato genotypes. Darkened trichome exudates were observed on insects held on PI 473331 and PI 473334, consistent with previous reports that glandular trichomes of S. berthaultii contain phenolic oxidation chemistry. This study aids in the identification of the probable mechanisms of resistance to E. fabae used by S. berthaulti and its hybrids with S. tuberosum.     

Host acceptance behaviour of Colorado potato beetle larvae influenced by potato glandular trichomes

ABSTRACT. In no-choice tests, larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), avoided contact with, and were less frequently observed feeding on, excised foliage of the resistant wild potato Solanum berthaultii Hawkes than on leaves of the cultivated potato, S.tuberosum L. For older larvae, reduced feeding was a consequence of less frequent contact with S.berthaultii leaves. However, first instars appeared unable to penetrate the adhesive barrier presented by glandular trichomes of S. berthaultii , and those that remained on the leaflets fed less often than did first instars on S. tuberosum. Removal of the trichome barrier by wiping leaflets with tissue paper did not attenuate the apparent repellent effect of S. berthaulti foliage, but led to increased incidence of feeding by first instars. Inhibition of larval feeding may therefore rely on a mechanical barrier provided by the glandular trichomes of S.berthaultii.     

Purification and Characterization of Polyphenol Oxidase from Glandular Trichomes of Solanum berthaultii

Type A glandular trichomes of the wild potato (Solanum berthaultii Hawkes) entrap insects by rapidly polymerizing the trichome contents after breakage by insect contact. Polymerization of trichome exudate appears to be driven by a soluble polyphenol oxidase (PPO). PPO constitutes up to 70% of the protein in individually collected trichomes and reaches a concentration approaching 200 μm in these organs. Trichome PPO has been purified and shown to be a monomeric copper metalloprotein with an isoelectric point of 5.5, possessing only o-diphenol oxygen oxido-reductase activity, and is larger than most other reported PPOs, with relative molecular weight of 59,000. Chlorogenic and caffeic acid were the most readily oxidized of 14 phenolic substrates tested. Polyclonal antibodies raised against the relative molecular weight 59,000 S. berthaultii trichome PPO were used to show that S. tuberosum L. trichomes express low levels of a cross-reactive protein that lacks detectable PPO activity.     

Glandular trichomes of Solanum berthaultii and its hybrids with potato deter oviposition and impair growth of potato tuber moth

Glandular trichomes on foliage of the wild potato species, Solanum berthaultii Hawkes, deter oviposition by the potato tuber moth (PTM), Phthorimaea operculella Zeller and negatively affect other important performance parameters. Oviposition deterring factors are localized in the glandular trichomes of S. berthaultii. When mechanically transferred to foliage of a susceptible potato cultivar, trichome contents reduced egg laying by 97%. Removal of glandular trichomes from S. berthaultii foliage using a combination of chemical and mechanical procedures increased oviposition rates ca. 210-fold. Removal of trichomes also led to increased mobility of larvae on the leaf surface, more leaf feeding, shorter larval development and larger pupae. The resistance conferred by glandular trichomes of S. berthaultii provides an important genetic trait potentially useful for management of PTM.     

Evaluation of natural and engineered resistance mechanisms in potato against Colorado potato beetle in a no-choice field study

The Colorado potato beetle, Leptinotarsa decemlineata Say, is the major insect pest of potato, Solanum tuberosum L., in eastern North America and is renowned for resistance development, currently resistant to >40 insecticides worldwide. Host plant resistance may assist in delaying in resistance development to insecticides. We evaluated natural host plant resistance mechanisms (glandular trichomes and Solanum chacoense Bitter-derived resistance) and engineered resistance mechanisms (Bacillus thuringiensis [Bt] Berliner cry3A and cry1Ia1) in a no-choice cage study. Six different potato lines representing four host plant resistance mechanisms were evaluated over 2 yr. Egg masses were placed in each cage (one egg mass per plant). Almost no feeding was observed in the Bt-cry3A lines, and only minor feeding was observed in the Bt-cry1Ia1 lines in either year. On the S. chacoense-derived line, there was significantly less defoliation than on either the susceptible line or the glandular trichome line in 2003. In 2004, there was significantly higher defoliation on the S. chacoense-derived line than on the susceptible line or glandular trichome line. The defoliation of the Solanum chacoense-derived line was largely due to larvae clipping the petioles, rather than consumption of the leaves. Defoliation on the glandular trichome line did not differ significantly from the defoliation of the susceptible line, suggesting glandular trichomes may not be effective in controlling larvae and preventing defoliation. This study suggested that Bt can provide high levels of resistance, but the natural resistance mechanisms tested here are variable for control of Colorado potato beetle larvae in no-choice situations.     

Role of the glandular trichomes in resistance of perennial alfalfa to the potato leafhopper (Homoptera: Cicadellidae)

Experiments were performed to elucidate resistance of glandular-haired alfalfa, Medicago sativa L., to the potato leafhopper, Empoasca fabae (Harris). The primary objective was to examine the potential role of the glandular trichomes and stem lignification for imparting resistance to this pest. During free-choice foliar discoloration experiments, the resistant alfalfa clone FGplh13 expressed lower levels of injury than the susceptible P5373 after 13 d. No-choice tests performed using nymphal potato leafhoppers showed higher levels of mortality associated with the resistant glandular-haired clone FGplh13, and no nymphs survived after 48 h. Additional no-choice experiments using FGplh13 and P5373 alfalfa with the glandular and nonglandular trichomes intact or removed show that mortality of nymphs and adults decreased after removal of the glandular trichomes from FGplh13. Nymphal and adult mortality and nymphal development time did not differ on FGplh13 with the trichomes removed and P5373 with the trichomes intact or removed. Firstinstar, and possibly second-instar, potato leafhoppers were entrapped by an exudate produced by the glandular trichomes on the FGplh13 alfalfa. Stem lignification and the number of vascular bundles did not differ between FGplh13 and P5373 alfalfas. The glandular trichomes on FGplh13 alfalfa appeared to provide the major host resistance factor, with resistance to adults being chemically based and resistance to nymphs being chemically and mechanically based. To maintain levels of potato leafhopper resistance, breeders would appear to benefit by continuing to select for the expression of the glandular trichome phenotype.     

Molecular markers locate genes for resistance to the Colorado potato beetle,Leptinotarsa decemlineata,in hybrid Solanum tuberosum x S. berthaultii potato progenies

The wild Bolivian potato, Solanum berthaultii Hawkes, has been used as a source of resistance to the Colorado potato beetle (CPB), Leptinotarsa decemlineata Say, one of the most significant pests of potato. In this study, two reciprocal backcross S. tuberosum x S. berthaultii potato progenies, BCB and BCT, were mapped with RFLP markers and screened for resistance to CPB consumption, oviposition and defoliation. The genotypic and phenotypic data were combined and analysed to locate quantitative trait loci (QTLs) for resistance to CPB. Three QTLs on three chromosomes in BCB, and two QTLs on two chromosomes in BCT influenced resistance. The QTLs were generally additive but one instance of epistasis was noted. Each QTL accounted for 4–12% of the phenotypic variation observed in resistance. In the more resistant BCB population, a three QTL model explained ca. 20% of the variation in CPB oviposition. When alleles at the three QTLs were homozygous S. berthaultii, oviposition was reduced ca. 60% compared to the heterozygotes. The QTLs for resistance to CPB were compared to those previously identified for the type A and B glandular trichomes, which have been implicated in resistance in the same progenies. Generally, the QTLs for resistance to CPB coincided with loci associated with the glandular trichomes confirming the importance of the glandular trichomes in mediating resistance. However, a relatively strong and consistent QTL for insect resistance in both BCB and BCT on chromosome 1 was observed that was not associated with any trichome traits, suggesting the trichomes may not account for all of the resistance observed in these progenies.     

Phenolic oxidase activities in glandular trichomes of Solanum berthaultii

Polyphenol oxidase and peroxidase activities were detected in foliar glandular trichomes of the wild, insect-resistant potato species, Solanum berthaultii. These enzyme activities may provide the basis for conversion of clear, viscous trichome exudate into a hard, brown substance which is formed rapidly after insect attack.     

龙葵腺毛发育的细胞学研究

利用光学显微镜、扫描电镜和透射电镜技术,观察了龙葵“四叶一心”期时叶片及茎表皮的腺毛的种类、分布,探究了不同类型腺毛的起源、生长、成熟、分泌、衰老等发育过程的细胞学特征;通过组织化学染色和荧光显微技术,观察了龙葵腺毛成分、分布,为龙葵的进一步开发利用提供参考。结果表明：（1）龙葵腺毛分为单细胞头腺毛和多细胞头腺毛两类,前者主要分布于茎表面和叶上下表皮,后者主要分布于茎表面的单细胞头腺毛之间、叶脉及叶边缘;（2）龙葵腺毛发育起始于表皮细胞突起,单细胞头腺毛行顶端生长,具1-4个柄细胞,四种类型;多细胞头腺毛可再分为一层、两层与三层多细胞头腺毛,另具三种特殊类型;（3）龙葵成熟腺毛具分泌能力,通过皮下空间的物质积累导致腺毛头细胞表面形成突起、包块、破口,最终释放分泌物;而头细胞与柄细胞随即皱缩、衰老。（4）超微结构显示,腺毛头细胞中内质网与高尔基体极为丰富,合成代谢及分泌活动活跃,产生大量包裹嗜锇物质的囊泡,囊泡与细胞壁融合,进而将嗜锇物质转移至细胞壁并积累,随后储存在角质层下的皮下空间直至分泌释放;（5）组织化学染色结果表明,腺毛含有萜类、生物碱、脂类、蛋白质、酚类和多糖。头细胞中主要含有萜类、生物碱、脂类、蛋白质、酚类和中性多糖;柄细胞中主要含有萜类、生物碱、脂类。     

Differences in distribution and performance of two sap-sucking herbivores on glandular and non-glandular Datura wrightii

1. Datura wrightii Regel (Solanaceae) is polymorphic with regard to trichome type. Some plants are densely covered with short, non-glandular trichomes, whereas other plants in the same populations possess glandular trichomes that excrete a sticky exudate. The hypothesis that glandular trichomes enhance resistance to all small insect herbivores is evaluated.
2. Field censuses in four southern Californian D. wrightii plant populations revealed that glandular plants are indeed resistant to whitefly spp. (Homoptera: Aleyrodidae). Whiteflies are almost exclusively found on non-glandular plants. In contrast, Tupiocoris notatus (Distant) (Heteroptera: Miridae), another sap-sucking herbivore of similar body size, is found predominantly on plants with glandular trichomes.
3. Laboratory experiments showed that whiteflies are unable to colonize glandular D. wrightii phenotypes. After the whitefly adults had landed on the leaves of these plants, they were trapped in the exudate and died.
4. Tupiocoris notatus adults, on the other hand, laid significantly more eggs on glandular plants. The presence of the exudate was shown to be the cue that determined their choice of glandular plants.
5. In no-choice experiments, T. notatus nymphs reared on glandular plants had significantly higher survival rates and had shorter developmental periods than those raised on non-glandular plants. This, combined with the higher oviposition rates, resulted in higher T. notatus population growth rates on glandular plants than on non-glandular plants.
6. Glandular trichomes are not therefore a universal protection against small herbivores. Differences in distribution over the two plant types within the natural herbivore guild on D. wrightii may, among other selection pressures, contribute to the maintenance of the observed trichome polymorphism.     

Comparison of polyphenol oxidase expression in glandular trichomes of solanum and lycopersicon species

Tetralobulate glandular trichomes are present on the foliage of many solanaceous species. Resistance of many of these species to insects is conditioned by the ability of trichomes to rupture upon contact and to rapidly polymerize their contents, resulting in entrapment of insects in hardened trichome exudate. In the wild potato, Solanum berthaultii, polymerization of trichome exudate is initiated by a soluble M_r 59,000 polyphenol oxidase (PPO), which is a dominant protein constituent of the organ. PPOs, although ubiquitous in angiosperms, typically display great heterogeneity in molecular weight and are found at low levels in plant cells. Because of the unusually high accumulation and tissue-specific expression of the M_r 59,000 PPO in S. berthaultii glandular trichomes, we analyzed trichome proteins of a number of Lycopersicon and Solanum species to assess the extent to which possession of the M_r 59,000 PPO is conserved. Trichomes were collected manually and examined for PPO activity, immuno-cross-reactivity with S. berthaultiiM_r 59,000 PPO, and protein content. In addition, N-terminal amino acid sequences were obtained for five trichome PPOs. All species analyzed possessed trichome PPOs similar in structure and level of expression to that of S. berthaultii. The relationship between sequences and structures of these conserved PPOs and the variable PPOs of leaf is discussed.     

The Ultrastructure of the Glandular Trichomes of Solanum tuberosum

The potato plant has two types of glandular trichomes whichwere investigated by electron microscopy. One type has a eight celled globular head on a neck cell anda stalk cell Each glandular cell has many rather large vacuoles,a large nucleus, many ribosomes and mitochondria, a few Golgibodies, and darkly coloured, often irregular plastids (chloroplasts).The plastids are mostly located near the axial cell wall borderinga large central intercellular space filled with secretion materialThe plastids are assumed to participate in the formation ofthe secretion material, which reacts positively to esterasetests. The outer wall is covered by a thin cuticle. The other type has a club-shaped multicellular head on a singlestalk cell. The cytoplasmic features in the cells are similarto those of the globular-headed trichome, except that they possesslarge central vacuoles and randomly distributed plastids. Centricendoplasmic reticulum has been observed in young cells. Intercellularspaces develop between the cells and into the outer wall, whichis thus split into two. Whereas the older glandular cells reactpositively to tests for esterase, the secretion material itselfis pectinaceous and reacts negatively. The outer wall is cutinizedand covered by a cuticle. Solanum tuberosum L., potato, glandular trichomes, ultrastructure     

Seasonal occurrence and relative abundance of aphids on potato plants with classical and transgenic characters of resistance to Colorado potato beetle Leptinotarsa decemlineata (Say)

The seasonal abundance and dynamics of potato-infesting aphids were studied in two seasons, on potato Solanum tuberosum L. with various types of transgenic and classical resistance to primary pests, especially the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say). The lines tested were the NewLeaf potato, a transgenic expressing the Bacillus thuringiensis CryIIIA toxin (BTT); NYL 235-4 a parental line derived from S. berthaultii (NYL) with a high density of glandular trichomes; and a transgenic line expressing oryzacystatln I (OCI), a proteinase inhibitor gene from rice; potatoes from two commercial cultivars, Superior and Kennebec, served as controls. Over the two seasons, Macrosiphum euphorbiae (Thomas) and, to a lesser extent, Myzus persicae (Sulzer) were by far the most abundant aphids observed in the experimental plots. M. persicae was observed relatively late in the season, and was most abundant on the Kennebec-OCI potato. Macrosiphum euphorbiae was more regular over seasons, and its highest densities were observed on the NYL 235-4 line. The population of M. euphorbiae showed markedly different patterns on the five lines studied, which was clearly related to potato plant phenology. Density of M. euphorbiae was observed on the NYL 235-4 line, in late July and early August.     

Aphid repellent sesquiterpenes in glandular trichomes of Solanum berthaultii and S. tuberosum

Exudate from glandular trichomes of the wild potato species, Solanum berthaultii Hawkes, and the cultivated potato, S. tuberosum L., contained volatile substances including sesquiterpenes when examined with gas chromatography/mass spectrometry procedures. Sesquiterpenes were the major volatile constituents of Type A trichome exudate but were a minor constituent of Type B exudate. Sesquiterpene mixtures of the two potato species differed qualitatively and quantitatively. -Caryophyllene and E--Farnesene were major components identified in Type A trichome exudate of the cultivated potato. Sesquiterpene mixtures of both potato species were repellent to the green peach aphid, Myzus persicae (Sulzer). The role of trichome sesquiterpenes as a determinant of aphid behavior on the two potato species and in the expression of host resistance is discussed.

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Résumé L'examen en chromatographie gazeuse et en spectométrie de masse a révélé la présence de substances volatiles contenant des sesquiterpènes dans l'exsudat des trichômes glandulaires des pommes de terre sauvage (Solanum berthaultii) et cultivée (S. tuberosum). Les sesquiterpènes étaient le principal composé volatile de l'exsudat de type A, tandis qu'ils n'étaient que faiblement représentés dans l'exsudat de type B. Les mélanges de sesquiterpènes des deux espèces différaient qualitativement et quantitativement. Le -Caryophyllène et le E--Farnesène constituaient les principaux composés identifiés dans l'exsudat de trichôme de type A de S. tuberosum. Les mélanges sesquiterpéniques des deux espèces ont repousée Myzus persicae Sulzer. La discussion a porté sur l'influence des sesquiterpènes des trichômes dans la définition du comportement des pucerons sur ces deux plantes et sur la manifestation de la résistance de l'hôte.

     

Glandular trichomes ofSolanum berthaultii alter host preference of the Colorado potato Beetle,Leptinotarsa decemlineata

Choice and no-choice studies were conducted to determine how the glandular trichomes of the wild potato,Solanum berthaultii Hawkes, affect host preference of the Colorado potato beetle,Leptinotarsa decemlineata (Say). Given a feeding choice betweenS. tuberosum andS. berthaultii, larvae and adults preferred the foliage ofS. tuberosum, but adults were more discriminating. When foliage ofS. berthaultii was appressed toS. tuberosum leaflets, fewer adults fed on the appressed leaflets. When given a choice between ‘trichome-intact’ and ‘trichome-removed’S. berthaultii foliage, adults preferred to feed on the latter. The preference for ‘trichome-removed’ foliage and the percent of adults initiating feeding, increased with the degree of trichome removal. These studies provide evidence that the resistance ofS. berthaultii is associated with feeding deterrents localized in the glandular trichomes, thatS. berthaultii possesses more than one mechanism of resistance to the Colorado potato beetle, and that the expression of resistance is dependent on the developmental stage of the insect.     

Location of resistance factors in the leaves of potato and wild tuber-bearing Solanum species to the aphid Myzus persicae

Analysis of electrically recorded feeding behaviour of aphids was combined with colony‐development tests to search for sources of resistance to Myzus persicae (Sulzer) (Homoptera: Aphididae) in tuber‐bearing Solanum species (Solanaceae), aiming at a reduction of potato leaf roll virus (PLRV) transmission. Twenty genotypes, originating from 14 gene bank accessions, representing 13 wild tuber‐bearing Solanum spp., three Solanum tuberosum L. (potato) cultivars, and one S. tuberosum breeding line, were selected. Colony‐development tests were carried out in no‐choice experiments by placing adult aphids on plants of each genotype and counting numbers of nymphs and adults on young plants after 8 and 15 days, and on flowering plants after 14 and 30 days. Large differences were observed among genotypes: some developed small colonies and others developed large ones. Also, in a few genotypes, resistance in mature plants was different for leaves of different ages; young leaves were resistant to aphids whereas old senescent leaves were susceptible. The electrical penetration graph (DC‐EPG system) technique was used to study aphid feeding behaviour on each Solanum genotype for 6 h. Electrical penetration graph (EPG) results also showed large differences among the genotypes, indicating resistance at the leaf surface and at three different levels of plant tissue (epidermis, mesophyll, and phloem). Therefore, it was concluded that different mechanisms of resistance to M. persicae exist among the genotypes analysed. EPGs recorded from aphids on Solanum berthaultii Hawkes and Solanum tarijense Hawkes with and without glandular trichomes showed that strong surface resistance can bias EPG parameters associated with resistance located in deeper tissues. Experimental evidence is presented that the resistance to aphids in the genotypes with glandular trichomes strongly depends on these morphological structures.     

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.     

RFLP Analysis of Chromosomal Segregation in Progeny from an Interspecific Hexaploid Somatic Hybrid between Solanum Brevidens and Solanum Tuberosum

Segregation of restriction fragment length polymorphism (RFLP) loci was monitored to determine the degree of homeologous pairing and recombination in a hexaploid somatic hybrid, A206, the result of protoplast fusion between Solanum tuberosum (PI 203900, a tetraploid cultivated potato) and Solanum brevidens (PI 218228), a diploid, sexually incompatible, distant relative harboring several traits for disease resistance. Somatic hybrid A206 was crossed to Katahdin, a tetraploid potato cultivar, to generate a segregating population of pentaploid progeny. Although the clones of the tetraploid S. tuberosum lines PI 203900 and Katahdin were highly polymorphic, the diploid S. brevidens clone was homozygous at all but two of the tested RFLP loci. Thus, homeologous recombination could be detected only when S. tuberosum and S. brevidens chromosomes paired and the S. brevidens homologs then segregated into separate gametes. A bias toward homologous pairing was observed for all 12 chromosomes. At least four and perhaps six chromosomes participated in homologous pairing only; each of 24 progeny contained all S. brevidens-derived RFLP markers for chromosomes 4, 8, 9 and 10. The remaining six chromosomes paired with their homolog(s) about twice as often as expected if hexaploid pairings were completely random. Where detectable with RFLPs, homeologous recombinations (both single and double) occurred at a frequency of 1.31 per chromosome. Cytological observations of meiosis I in the somatic hybrid indicated that homeologous pairing had occurred. Enhanced recombinational activity was observed for chromosome 2. A specific small deletion from chromosome 4 was detected in A206 and 11 other somatic hybrids out of 14 screened. These hybrids represent 13 independent fusion events between the same clones of S. brevidens and S. tuberosum. In one instance, this deletion occurred in one of two plants resulting from the same callus, indicating that the loss occurred in culture after fusion had taken place. It is possible that this deletion contributes to somaclonal variation.