Ray F. Severson

Leaf surface chemicals fromNicotiana affecting germination ofPeronospora tabacina (adam) sporangia

A bioassay was used to evaluate the effects of cuticular leaf components, isolated fromN. tabacum, N. glutinosa (accessions 24 and 24a), and 23other Nicotiana species, on germinationof P. tabacina (blue mold). The leaf surface compounds includedα- andβ-4,8,13,-duvatriene-l,3-diols (DVT-diols), (13-E)-labda-13-ene-8α-,15-diol (labdenediol), (12-Z)-labda-12,14-diene-8α-ol (cis-abienol), (13-R)-labda-8,14-diene-13-ol (manool), 2-hydroxymanool, a mixture of (13-R)-labda-14-ene-8α,13-diol (sclareol), and (13-S)-labda-14-ene-8α,13-diol (episclareol), and various glucose and/or sucrose ester isolates. The above in acetone were applied onto leaf disks of the blue moldsusceptibleN. tabacum cv. TI 1406, which was then inoculated with blue mold sporangia. Estimated IC50 values (inhibitory concentration) were 3.0μg/cm2 forα-DVT-diol, 2.9μ/cm2 forβ-DVT-diol, 0.4μg/cm2 for labdenediol and 4.7μg/cm2 for the sclareol mixture. Manool, 2-hydroxymanool, andcis-abienol at application rates up to 30μg/cm2 had little or no effect on sporangium germination. Glucose and/or sucrose ester isolates from the cuticular leaf extracts of 23Nicotiana species and three different fractions fromN. bigelovii were also evaluated for antimicrobial activity at a concentration of 30μg/cm2. Germination was inhibited by >20% when exposed to sugar esters isolated fromN. acuminata, N. benthamiana, N. attenuata, N. clevelandii, andN. miersii, and accessions 10 and 12 ofN. bigelovii. These results imply that a number of compounds may influence resistance to blue mold in tobacco.

Effects of cuticular duvane diterpenes from green tobacco leaves on tobacco budworm (Lepidoptera: Noctuidae) oviposition

Five cuticular chemical components isolated from the green leaves of tobacco introductions (TIs) and a commercial tobacco cultivar were tested for their effects on tobacco budworm,Heliothis virescens (F), oviposition in cage bioassays, and field experiments. These chemicals were sprayed onto budworm-resistant TI 1112 tobacco which produces low levels of most cuticular components. Individual duvane diterpenes (α- and β-4,8,13-duvatrien-1-ols and α- and β-4,8,13-duvatriene-1,3-diols) increased tobacco budworm egg laying on sprayed TI 1112 plants.cis-Abienol, docosanol, and docosanyl myristate were inactive.

Biorationals fromNicotiana protect cucumbers againstColletotrichum lagenarium (Pass.) ell. & halst disease development

Trichome exudate compounds isolated from leaves ofNicotiana tabacum, N. glutinosa (accessions 24 and 24a), and 21 otherNicotiana species were evaluated for biorational activity againstC. lagenarium, the anthracnose pathogen of cucumber. Exudate compounds tested were the cembrane diterpenes α- and β-4,8,13-duvatriene-1,3-diols (DVT diols); the labdane diterpenes (13-E)-labda-13-ene-8α,15-diol (labdenediol) and (13-R)-labda-14-ene-8α, 13-diol (sclareol); extracts fromN. gossei; and various sugar ester isolates. In dose-response experiments test compounds were applied to a water agar surface that was then inoculated with a conidial suspension. Low levels of most test compounds reduced or completely inhibited germination ofC. lagenarium conidia. IC50 values, concentrations (micrograms per square centimeter) at which conidium germination was reduced 50%, were 6.3 for DVT diols, 19.3 for sclareol, 1.0 for labdenediol, 2.8 for a mixture of sclareol and labdenediol, 1.2 for anN. gossei sucrose ester and 4.1 for theN. gossei crude extract. Higher levels of DVT diols and the sclareol-labdenediol mixture were required to reduce lesion size and number on inoculated cucumber leaves. At the highest concentration tested, 100 µg/cm2, the DVT diols and sclareol-labdenediol mixtures protected cucumbers against lesion development by 93 and 98%, respectively. Sugar ester mixtures from 20Nicotiana species and three different sugar ester fractions fromN. bideglovii had in vitro antifungal activity at a concentration of 48 µg/cm2. Sugar esters from nine of the species includingN. acuminata, N. attenuata, N. clevelandii, N. maritima, N. miersii, N. noctiflora, N. occidentalis, N. rustica, and fractions 10, 12, and 13 fromN. bideglovii completely inhibitedC. lagenarium conidium from germinating. Sugar ester mixtures from only four species,N. plumbaginifolia, N. bonariensis, N. simulans, andN. palmerii, had no significant effect onC. lagenarium conidium germination. These results suggest the potential ofNicotiana exudates as biorationals in reducing disease development.

Attraction of adult sweet potato weevils, Cylas formicarius elegantulus (Summers), (Coleoptera: Curculionidae), to sweet potato leaf and root volatiles.

A dual-choice olfactometer was developed to study the responses of sweet potato weevils,Cylas formicarius elegantulus (Summers), to volatiles from the sweet potato,Ipomoea batatas (L.) Lam. Both males and females were attracted by volatiles from sweet potato leaves and a methylene chloride leaf extract. Females, but not males, responded to volatiles from storage roots and a methylene chloride root extract. Leaves and storage roots from four sweet potato cultivars (Centennial, Jewel, Resisto, and Regal) were attractive to female weevils; however, the attractant response varied with cultivar. GC profiles from leaf and root extracts, and GC-MS analysis of leaf extract, for Jewel cultivar enabled the volatile peaks to be identified as sesquiterpenes.

Feeding and oviposition preferences of the Sweet Potato Weevil, Cylas formicarius elegantulus, on the outer periderm and exposed inner core of storage roots of selected Sweet Potato cultivars

Cores from sweet potato (Ipomoea batatas L.) storage roots, with either the outer periderm or inner core exposed, were presented to female sweet potato weevils, Cylas formicarius elegantulus (Summers) (Coleoptera: Curculionidae) in cage preference tests. Tests with weevils of different ages showed that feeding was constant 21 to 76 days after emergence, while oviposition showed a slight peak between 21 and 45 days. Differences in the levels of feeding and oviposition were noted for the periderms of four sweet potato cultivars, but these differences were not evident for the inner cores. Oviposition was reduced to low levels on the inner core and, therefore, the oviposition stimulant appears to reside in the root periderm. Feeding did occur on the inner cores, but at a reduced level compared to the root periderm in preference tests where both were available. Inner root cores have a potential use in the elucidation of the oviposition stimulant of the sweet potato weevil.

Feeding and oviposition preferences of sweet potato weevil,Cylas formicarius elegantulus (Summers), on storage roots of sweet potato cultivars with differing surface chemistries

Cores from sweet potato [Ipomoea batatas (L.) Lam.] storage roots (Centennial, Jewel, Resisto, and Regal cultivars) were presented to sweet potato weevils [Cylas formicarius elegantulus (Summers) (Coleoptera; Curculionidae)] in multiple-choice, limited-choice, and no-choice bioassays. Centennial, a susceptible cultivar in field-plot experiments, was preferred for feeding and oviposition by female weevils in choice bioassays, and for ovi-position in no-choice bioassays, compared to three other cultivars. Analysis of root surface chemistry showed a tentatively identified triterpenol acetate in Centennial, which was not found in the more resistant cultivars; another root surface component was found in higher concentrations in the more resistant cultivars.

Biochemical investigations of antibiosis material in leaf exudate of wildNicotiana species and interspecific hybrids

A model system involving severalNicotiana species containing novel nicotine alkaloids was used to study heritability and expression of alkaloid production in leaf trichomes. The three species that comprise the section Repandae (N. repanda, N. stocktonii, andN. nesophila) were hybridized with eitherN. tabacum orN. sylvestris (neither of which producesN-acylnornicotine). The progeny of the hybrid with sylvestris produced theN-acylnornicotines at a level found in the Repandae parent.Nicotiana repanda was crossed toN. tabacum, and the F2 progeny produced the alkaloid at the same level as the original Repandae parent. Inheritance of the ability to acylate nornicotine in Repandae species is inherited in hybrids in a dominant manner These and other data obtained suggest that theN-acyltransferase that acylates nornicotine in Repandae species inherited in hybrids is in a dominant manner and that the regulatory sequence(s) for the gene is expressed in leaf trichomes when the gene is in a foreignNicotiana background.

Survival and development of tobacco hornworm larvae on tobacco plants grown under elevated levels of ozone.

Tobacco plants, Nicotiana tabacum were grown under different levels of ozone (O3) in open-top chambers. Ozone concentrations were established by charcoal filtration, which reduced O3 to approximately one-half ambient, or by the addition of O3 to unfiltered air to increase concentrations to approximately 1.4 or 1.7 times ambient O3. Survival of tobacco hornworm, Manduca sexta, larvae was increased when second instars were fed tobacco leaves grown in chambers with elevated levels of O3. Second instars also gained significantly more weight when they were fed for one week on plants exposed to elevated levels of O3 than when they were fed plants grown in charcoal-filtered air. Ozone-treated tobacco plants had higher levels of total nitrogen (primarily reduced nitrogen) and soluble carbohydrates (sugars), and lower levels of leaf-surface components, starch, nicotine, and rutin. Increased survival and growth response of hornworm larvae to elevated O3 levels in these experiments suggests that similar responses could occur in the southeastern US tobacco production areas where O3 levels can be high enough to injure tobacco plants.

Characterization of an oviposition stimulant from the surface of sweet potato Ipomoea batatas storage roots for the sweet potato weevil, Cylas formicarius elegantulus

An improved laboratory bioassay was used to characterize an oviposition stimulant from the surface of sweet potato Ipomoea batatas (L.) Lam. storage roots for the sweetpotato weevil, Cylas formicarius elegantulus (Summers). Filter paper discs impregnated with a methylene chloride surface extract of sweet potato storage roots induced significantly (p < 0.05) higher oviposition on root cores than those treated with solvent only. Significantly higher oviposition was also observed in the nonpolar fractions, especially one that contains a tentatively identified triterpenoid present in susceptible cultivars.

Ovipositional response of tobacco budworm moths (Lepidoptera: Noctuidae) to cuticular labdanes and sucrose esters from the green leaves ofNicotiana glutinosa L. (Solanaceae)

Field plots of three accessions ofNicotiana glutinosa L. (Nicotiana species accessions 24, 24A, and 24B) at Oxford, North Carolina and Tifton, Georgia were heavily damaged by natural populations of tobacco budworms,Heliothis virescens (F.), during 1985–1989. Experiments in outdoor screen cages demonstrated that all accessions ofN. glutinosa were as prone to oviposition byH. virescens moths as was NC 2326, a commercial cultivar of flue-cured tobacco,N. tabacum L. However, in greenhouse experiments, tobacco budworm larvae did not survive or grow as well when placed on plants ofN. glutinosa as they did when placed on plants of NC 2326. Four labdane diterpenes (manool, 2-hydroxymanool, a mixture of sclareols, and labda-13-ene-8α,15-diol [labdenediol]) and two sucrose ester fractions (2,3,4-tri-O-acyl-3′-O-acetyl-sucrose [G-SE-I] and 2,3,4,-tri-O-acyl-sucrose [G-SE-II]) were isolated from green leaves of the three accessions ofN. glutinosa. These components were bioassayed for their effects on the ovipositional behavior of tobacco budworm moths using small screen cages in a greenhouse at Oxford, North Carolina. Labdenediol, manool, and both sucrose ester fractions stimulated tobacco budworm moths to oviposit on a tobacco budworm-resistant Tobacco Introduction, TI 1112 (PI 124166), when these materials were sprayed onto a leaf.