A hybrid diploid potato population (Solanum phureja x Solanum stenotomum) with superior horticultural traits has been developed for breeding purposes. Because this highly advanced population has no resistance to potato virus Y (PVY), and because PVY pressure is very high in North Carolina, glandular trichomes from another diploid species, Solanum berthaultii, were introduced as a defense against aphids, the PVY vector. Two types of glandular trichomes, A and B, were transferred. Preliminary results show that genetic diversity and broad-sense heritability are adequate to transfer these traits; however, non-additive gene action was indicated in narrow-sense heritability estimates. Spearman's coefficients of rank correlation showed a low level of association between A and B trichomes; density of both types is postulated to be under the control of a Small number of genes.
Roger L. Vallejo, Wanda W. Collins, and Rocco Schiavone
Roger L. Vallejo, Wanda W. Collins, and Robert H. Moll
Glandular trichomes from some Solanum species have suppressed infestation by insects including green peach aphid, which is a main vector of potato virus Y (PVY) and potato leaf roll virus (PLRV), both of which contribute to a serious loss in potato production. Eight Solanum phureja Juz. et Buk.-S. stenotomum Juz. (Phu-Stn), three S. berthaultii Hawkes (Ber), nine F1 [(Phu-Stn) × Ber], fifteen backcross (BC) [(Phu-Stn) × F1], and seventeen reciprocal BC (BCR) [F1 × (Phu-Stn)] families were evaluated to determine the genetic variability and heritability of A and B glandular trichome density and polyphenol oxidase (PPO) activity. Experiments were carried out in completely randomized and randomized complete-block designs in the greenhouse. Genetic analysis was done using half-sib family and parent-offspring regression analysis. Phu-Stn showed a higher density of A trichomes than Ber and F1, while the BC and BCR had densities of A trichomes similar to Phu-Stn. B trichomes were not observed in Phu-Stn. Ber showed a high B trichome density, which was transmitted to the F1. In the BC, B trichomes were almost absent, but, in the BCR, the density of B trichomes was higher than that of BC. Ber and F1 had similar or higher PPO activity than Phu-Stn. PPO activity decreased in the BC, but, in the BCR, it was high and similar to Ber and F1. Broad-sense heritability estimates for A and B trichome density and PPO activity were from medium to high (0.48 to 0.77) in Phu-Stn, Ber, and F1. Narrow-sense heritability estimates for A and B trichome density and PPO activity were very low (0.04 to 0.24) in BC and BCR. In the BC families, additive genetic variance was very low for A and B trichome density and PPO activity. Half-sib family selection based on progeny testing and combined with BCs to Phu-Stn in subsequent generations would be a suggested breeding procedure to improve these traits. Phenotypic correlations between A and B trichome densities were 0.26 (F1) and 0.44 (BCR), between A trichome density and PPO activity 0.20 (F1) and 0.31 (BCR), and between B trichome density and PPO activity 0.04 (F1) and 0.27(BCR. Positive associations found between traits might facilitate simultaneous improvement for high levels of A and B trichome density and PPO activity.
Roger L. Vallejo, Wanda W. Collins, and Rocco D. Schiavone
Preliminary data on the genetics of glandular trichomes and the feasibility of incorporating A and B glandular trichomes and polyphenol oxidase (PPO) activity from Solanum berthaultii Hawkes (Ber) into an advanced S. phureja Juz. et Buk.-S. stenotomum Juz. (Phu-Stn) diploid potato population are presented. A random sample of four Phu-Stn clones was intercrossed with a sample of three Ber progenies (pollen bulk per progeny) segregating for high density of A and B glandular trichomes. Nine F1 families [(Phu-Stn) × Ber] were evaluated over two experiments in the greenhouse for A and B trichome density (5 mm2 of abaxial surface) and PPO activity 45 days after planting. A completely randomized design in an experimental hierarchical classification arrangement was used to quantify the genetic variability and to estimate broad-sense heritabilities (defined on an individual basis) for the attributes under study. To characterize the inheritance of these attributes further and to estimate narrow-sense heritabilities, fifteen backcross (BC) families were evaluated in a randomized complete-block design with three replications and analyzed by half-sib family analysis. Phenotypic and genotypic correlations between these attributes were also estimated. In the F1 families, broad-sense heritabilities were 0.59 and 0.41 for density of A and B trichomes, respectively. In the BC population, narrow-sense heritabilities were 0.15 and 0.08 for A trichome density and PPO activity, respectively; expression of B trichomes was almost absent in this generation. Genotype × environment interaction was significant for density of A trichomes and highly significant for density of B trichomes in F1 families. There was some level of positive association between density of A and B trichomes and PPO activity. Additive genetic variance of these attributes was very low in this study; therefore, selection schemes based on family selection and progeny testing would be an appropriate breeding strategy for improving these traits.
Richard Grazzini, David Hesk, Ellen Yerger, Diana Cox-Foster, June Medford, Richard Craig, and Ralph O. Mumma
Biochemical and morphological components of 16 Pelargonium species and the P. ×hortorum interspecific complex were examined. Inflorescences and leaves of each species were analyzed for anacardic acids and the presence of glandular trichomes. Three species of the section Ciconium, P. acetosum, P. frutetorum, and P. inquinans, produced anacardic acids in association with glandular trichomes. only P. inquinans and P. frutetorum contained ω5- anacardic acids. An evolutionary model for the origin of anacardic acids and ω5- desaturation is proposed.
D.S. Douches, T.J. Kisha, J.J. Coombs, W. Li, W.L. Pett, and E.J. Grafius
The Colorado potato beetle, Leptinotarsa decemlineata (Say), is the most serious insect pest of potatoes throughout the eastern and north central United States. Host plant resistance to the Colorado potato beetle has been identified in wild Solanum species and Bt-transgenic potato lines. Detached-leaf bioassays (72 h) were conducted on insecticide-resistant, first instar Colorado potato beetles to study the effectiveness of individual and combined host plant resistance traits in potato. Potato lines tested include non-transgenic cultivars (`Russet Burbank', `Lemhi Russet', and `Spunta'), a line with glandular trichomes (NYL235-4), a line with high foliar leptines (USDA8380-1), and transgenic lines expressing either codon-modified Bt-cry3A or Bt-cry5 (Bt-cry1Ia1). Bt-cry3A transgenic lines, foliar leptine line, and foliar leptine lines with Bt-cry5 had reduced feeding compared to non-transgenic cultivars. Glandular trichome lines and glandular trichome lines with Bt-cry5 did not reduce feeding in this no-choice feeding study. Some Bt-cry5 transgenic lines, using either the constitutive promoters CaMV35s or (ocs)3mas (Gelvin super promoter), were moderately effective in reducing larval feeding. Feeding on Bt-cry5 transgenic lines with the tuber-specific patatin promoter was not significantly different than or greater than feeding on the susceptible cultivars. Mortality of first instars was highest when fed on the Bt-cry3A lines (68% to 70%) and intermediate (38%) on the Bt-cry5 `Spunta' line SPG3 where the gus reporter gene was not included in the gene construct. Host plant resistance from foliar leptines is a candidate mechanism to pyramid with either Bt-cry3A or Bt-cry5 expression in potato foliage against Colorado potato beetle. Without multiple sources of host plant resistance, long-term sustainability is questionable for a highly adaptable insect like the Colorado potato beetle.
Joseph J. Coombs, David S. Douches, Susannah G. Cooper, Edward J. Grafius, Walter L. Pett, and Dale D. Moyer
Colorado potato beetle (Leptinotarsa decemlineata Say) is the leading insect pest of potato (Solanum tuberosum L.) in northern latitudes. Host plant resistance is an important tool in an integrated pest management program for controlling insect pests. Field studies were conducted to compare natural host plant resistance mechanisms (glandular trichomes and Solanum chacoense Bitter-derived resistance), engineered [Bacillus thuringiensis (Bt) Berliner Bt-cry3A], and combined (glandular trichomes + Bt-cry3A and S. chacoense-derived resistance + Bt-cry3A transgenic potato lines) sources of resistance for control of colorado potato beetle. Six different potato clones representing five different host plant resistance mechanisms were evaluated for 2 years in a field situation under natural colorado potato beetle pressure in Michigan and New York, and in a no-choice field cage study in Michigan. In the field studies, the S. chacoense-derived resistance line, Bt-cry3A transgenic, and combined resistance lines were effective in controlling defoliation by colorado potato beetle adults and larvae. Effectively no feeding was observed in the Bt-cry3A transgenic lines. The glandular trichome line suffered less defoliation than the susceptible control, but had greater defoliation than the Bt-cry3A transgenic lines and the S. chacoense-derived resistance line. In the no-choice cage study, the Bt-cry3A transgenic lines and the combined resistance lines were effective in controlling feeding by colorado potato beetle adults and larvae with no defoliation observed. The S. chacoense-derived resistance line and the glandular trichome line suffered less defoliation than the susceptible control. Based on the results of the field trials and no-choice field cage studies, these host plant resistance mechanisms could be used to develop potato varieties for use in a resistance management program for control of colorado potato beetle.
Joseph J. Coombs, David S. Douches, Wenbin Li, Edward J. Grafius, and Walter L. Pett
The Colorado potato beetle [Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae)] is a destructive pest of the cultivated potato (Solanum tuberosum L.) in northern latitudes. Combining resistance mechanisms of leptine glycoalkaloids and glandular trichomes with the synthetic Bacillus thuringiensis Berliner (Bt) cry3A gene in potato may be an effective strategy for controlling the Colorado potato beetle. Bt-cry3A transgenic plants were developed for three potato lines with differing levels of resistance to Colorado potato beetle ['Yukon Gold' (susceptible control), USDA8380-1 (leptine glycoalkaloids), and NYL235-4 (glandular trichomes)]. Polymerase chain reaction, and Southern and northern blot analyses confirmed integration and transcription of the cry3A gene in the transgenic lines. Detached-leaf bioassays of the cry3A engineered transgenic lines demonstrated that resistance effectively controlled feeding by first instar Colorado potato beetles. The susceptible `Yukon Gold' control suffered 32.3% defoliation, the nontransformed high foliar leptine line (USDA8380-1) had 3.0% defoliation, and the nontransformed glandular trichome line (NYL235-4) had 32.9% defoliation. Mean percentage defoliation for all transgenic lines ranged between 0.1% and 1.9%. Mean mortality ranged from 0.0% to 98.9% among the Bt-cry3A transgenic lines, compared to 20% for the susceptible `Yukon Gold' control, 32.2% for USDA8380-1, and 16.4% for NYL235-4. Results indicate that genetic engineering and the availability of natural resistance mechanisms of potato provide the ability to readily combine host plant resistance factors with different mechanisms in potato.
Richard Grazzini, Donald Walters, Jody Harmon, David J. Hesk, Diana Cox-Foster, June Medford, Richard Craig, and Ralph O. Mumma
Diploid zonal geraniums (Pelargonium ×hortorum) are able to resist attack by small arthropod pests such as the two-spotted spider mite (Tetranychus urticae Koch) when exudate produced by tall glandular trichomes contains a high percentage of ω5-unsaturated anacardic acids. Trichomes of susceptible plants exude primarily saturated anacardic acids. Inbred mite-resistant and -susceptible geraniums were reciprocally crossed and the F1, F2, and backcross generations were examined for anacardic acid composition and trichome density. Selected F2 plants were bioassayed for resistance to two-spotted spider mites. High concentrations of ω5-unsaturated anacardic acids in resistant plants are conditioned by a single dominant allele. We propose that inheritance of tall glandular trichome density can be controlled by a small number of loci (possibly as few as one) exhibiting codominance. F2, with low densities of tall glandular trichomes and producing ω5-unsaturated anacardic acids, displayed effective resistance to two-spotted spider mites as measured by mite mortality and fecundity. A genetic model for the biosynthesis of anacardic acids is proposed.
Joseph J. Coombs, David S Douches, Wenbin Li, Edward J. Grafius, and Walter L. Pett
The colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), is the leading insect pest of potato (Solanum tuberosum L.) in northern latitudes. Host plant resistance has the potential use in an integrated pest management program for control of colorado potato beetle. During the 1998 and 1999 seasons, field studies were conducted to compare natural (leptine glycoalkaloids and glandular trichomes), engineered (Bt-cry3A and Bt-cry5 transgenic potato lines), and combined (Bt-cry5+glandular trichomes) plant resistance mechanisms of potato for control of colorado potato beetle. Nine different potato clones representing five different host plant resistance mechanisms were evaluated under natural colorado potato beetle infestation at the Montcalm Research Farm in Entrican, Michigan. The Bt-cry3A transgenic lines, the high leptine line (USDA8380-1), and the high foliar glycoalkaloid line (ND5873-15) were most effective for controlling defoliation by colorado potato beetle adults and larvae. The Bt-cry5 line (SPc5-G2) was not as effective as the Bt-cry3A transgenic lines ('Russet Burbank Newleaf,' RBN15, and YGc3.1). The glandular trichome (NYL235-4) and Bt-cry5+glandular trichome lines proved to be ineffective. Significant rank correlations for the potato lines between the two years were observed for egg masses, second and third instar, and fourth instar seasonal cumulative mean number of individuals per plant, and defoliation. Egg mass and first instar seasonal cumulative mean number of individuals per plant were not strong indicators of host plant resistance in contrast to second and third instars or adults. Based on these results, the Bt-cry3A transgenic lines, the high leptine line, and the high total glycoalkaloid line are effective host plant resistance mechanisms for control of colorado potato beetle.
Richard Grazzini, David Hesk, Ellen Yerger, Diana Cox-Foster, June Medford, Richard Craig, and Ralph O. Mumma
Composition of anacardic acids (phenolic acids known to be associated with small pest resistance in Pelargonium ×hortorum) was examined in 13 diploid and 25 tetraploid cultivars by high-performance liquid chromatography (HPLC). The presence of an unusual desaturation (omega (ω)-5) in the alkyl tail of anacardic acids present only in glandular trichome exudate of pest-resistant diploid inbred lines had previously been associated with a sticky-trap pest-resistance phenomenon. In this study, we examine Pelargonium cultivars for variability in anacardic acid composition to assess the distribution of ω5 desaturation among commercial cultivars, to determine possible interactions between ω5 desaturation and other plant desaturation mechanisms, and to examine the possible impact of ploidy on ω5 desaturation. An unsaturation index (UI) is derived to compare exudates differing widely in composition yet which may provide a similarly effective sticky-trap pest-resistance mechanism based on exudate viscosity. ω-5 Anacardic acids were observed in the glandular trichome exudate of all 38 commercial cultivars examined. No diploid cultivar produced ω5- and ω9- anacardic acids, although the simultaneous production of ω5 and ω9- anacardic acids was observed in three tetraploid cultivars. Total ω5- anacardic acids comprised from 42.4% (tetraploid cultivar Perlenkette-syn. Snowhite, Weiss) to 86.8% (tetraploid cultivar Amanda). Commercial P. ×domesticum cultivars had no ω5 anacardic acids. UIs ranged from 60.9 (tetraploid cultivar Dixieland) to 103.4 (diploid cultivar Pinto White). In contrast, anacardic acids collected from a pest-susceptible inbred line contained no ω5- anacardic acids and had a UI of 38.7. No significant differences among ploidy levels were observed for UIs or for most specific anacardic acid components, with the exception of 24:1 ω5- anacardic acid, in which the mean diploid value (32.1%) was significantly higher than that of the mean tetraploid value (27.6%). We conclude that ω5- anacardic acid production occurs in all Pelargonium cultivars observed and that these cultivars are predicted to exhibit resistance to small arthropod pests. Significant genetic variability in specific anacardic acid composition appears to exist among Pelargonium cultivars, suggesting that breeding for pest resistance can be readily monitored by HPLC of anacardic acids.