Green peach aphid resistance by glandular trichomes inSolanum tuberosum xS. berthaultii hybrids

Three interspecific F₃ hybrid clones ofSolarium tuberosum L. x 5.berthaultii Hawkes bearing both Type A and Type B glandular trichomes were assessed for resistance to the green peach aphid,Myzus persicae (Sulzer). In glasshouse trials, aphid mortality and population development on hybrid clones were similar to those on the aphid-resistant parent,S. berthaultii. Trichome exudate of the hybrid, B227-63, was as effective in aphid immobilization as that ofS. berthaultii. Field populations of the green peach aphid on hybrids were intermediate between those onS. berthaultii and susceptibleS. tuberosum cultivars.     

Phenolic constituents of glandular trichomes onSolanum berthaultii andS. polyadenium

Phenolic compounds in Type A glandular trichomes of two insectresistant potato species were separated and identified using a combination of high pressure liquid chromatography (HPLC), UV-visible spectrometry, and¹H proton nuclear magnetic resonance (NMR) procedures. Type A trichomes ofSolanum berthaultii andS. polyadenium contained a phenolic compound tentatively identified as the glucose ester of p-hydroxyphenylpropionic acid. Its presence in trichomes of both species suggests that this compound may play a major role in formation of the brownish, insect entrapping exudate characteristic of trichome-mediated insect resistance in these species. Chlorogenic acid, a major phenolic constituent of Type A trichomes ofSolanum polyadenium was not detected in trichomes of 5.berthaultii. The exclusive presence of chlorogenic acid in trichome exudates ofS. polyadenium may be responsible, at least in part, for the greater levels of insect entrapment by this species, compared with that ofS. berthaultii.     

Inheritance of tuber initiation in tuber bearingSolanum as influenced by photoperiod

Eight potato clones (three day-neutral (DN) Tuberosum, one dayneutral and four short-day (SD) Andigena) were used as parents. Eight genetic sets bearing P₁, P₂, P_1?, P_2?, F₁, F₂, BC₁, and BC₂ generations were developed. Two sets were DN×DN crosses, four SD×DN, and two SD×SD. Tuber initiation (TI) was evaluated in growth chambers with daylengths of 11, 13, and 15 hours. The stolons of each genotype were examined every eight days between the 38th and 86th day after planting. F₁ generation means from SD×DN crosses showed complete dominance of SD reaction (late TI) over DN reaction (early TI). F₁ generation means from DN × DN and SD× SD crosses showed either no dominance or a slight tendency towards the late TI parent. Within plot variances of the selfed generations suggested a variable degree of parental heterozygosity for the loci controlling TI. Andigena clones showed the greatest heterozygosity. Bell shaped frequency distributions in the segregating generations indicated that TI was under the control of several genes. It appears also that major and minor genes were involved. Broad sense heritability estimates of about 90% at 11 hours and 55% at 15 hours of daylength were computed.     

Glandular trichomes: Enzymic browning assays for improved selection of resistance to the green peach aphid

Polyphenoloxidase (PPO) activity in the glandular trichomes of 32Solanum tuberosum ×S. berthaultii F₃ hybrid clones differentially resistant to the green peach aphid,Myzus persicae (Sulzer) was determined. Within this population of segregating clones, aphid infestation levels varied 8-fold, while a 5-fold difference in glandular trichome PPO activity was observed. PPO activity and aphid resistance at three sample dates were correlated as follows: r = 0.38 (P = 0.05), r = 0.31 (P = 0.10), and r = 0.55 (P = 0.01), respectively. The total browning potential test (TBP), a modification of the PPO assay, was developed for use in screening for aphid resistance. The TBP procedure is rapid, inexpensive, and highly effective in identifying aphid resistant clones.     

A modified enzymic browning assay for potato glandular trichomes

Tetralobulate (Type A) glandular trichomes are present on foliage of many wild as well as cultivated species of potatoes. These trichomes confer resistance to insect pests if their density, gland size and biochemical products are similar as those ofSolanum berthaultti Hawkes andS. polyadenium Greenm. Hybrids ofS. tuberosum L. ×S. berthaultii have been screened for resistance by measurement of phenolic oxidation activity in their Type A trichomes by use of an enzymic browning assay. However, variability in the procedure has prompted the development of an improved assay.     

The germplasm release of NYL 235–4, a clone with resistance to the Colorado potato beetle

Clones selected from the wild diploid species,Solanum berthaultii, have been shown to possess valuable levels of resistance to the Colorado potato beetle (Leptinotarsa decemlineata), as well as to insects such as aphids, flea beetles, leafhoppers and the potato tuber moth. Resistance is associated with the presence of glandular trichomes on the foliage ofS. berthaultii (2). Six generations after producing a hybrid between the diploid species and a tetraploid variety, a clone with good beetle and leafhopper resistance, as well as reasonable agronomic characteristics has been produced.     

Inheritance of resistance to sclerotinia stem rot (Sclerotinia trifoliorum) in faba beans (Vicia faba L.)

Stem rot, a fungal disease caused by Sclerotinia trifoliorum Eriks., is often a serious problem in many important forage legumes including faba beans (Vicia faba L.). Understanding the inheritance of resistance to the disease is essential for effective breeding of resistant cultivars. Experiments were conducted to study the inheritance of resistance to stem rot of faba beans. The F₁, F₂, and the backcross generations of five crosses between four resistant and four susceptible populations (Alto × Polycarpe, A-90 × Polycarpe, ILB-1814 × A-247, A-90 × A-244, VT × Tanagra) were used. The eight populations were crossed properly in the field, and progenies of F₁ and F₂, as well as backcross progenies of F₁ with each of their parents, were evaluated for resistance to stem rot disease under controlled conditions after artificial inoculation of the plants with carrot root pieces colonized by the fungus. On the assumption that inheritance of stem rot resistance is governed by a single dominant gene, no significant differences were found between the observed and the expected frequencies of resistance for progenies, except for one cross. As the expression of resistance to the disease fits the expected ratios for a single dominant gene model, it is concluded that the inheritance of resistance to sclerotinia stem rot in the evaluated faba bean populations is controlled by a single dominant gene.     

Glycoalkaloid content in tubers of hybrid and backcross populations from aSolanum tuberosumx s. chacoense cross

The glycoalkaloid contents of tubers were measured in hybrid and back-cross populations derived from a cross ofSolarium chacoense Bitter (PI 472810, 2n=2x=24) withSolanum tuberosum L. (2n=4x=48). In tubers sampled from near the median size in each population and parent, the mean total glycoalkaloid (TGA) contents of theS. chacoense andS. tuberosum parent genotypes, and the F₂ and F₄ hybrid genotypes were 245, 4, 39, and 35 mg/ 100 g fresh weight (fw), respectively. Backcrossing the F₂ genotypes toS. tuberosum reduced the mean TGA content to 15 mg/100 g fw. A second backcross further reduced the content to 9 mg/100 g fw. However, some genotypes in both backcross populations (25% and 10%) had TGA contents above the generally accepted upper limit of 20 mg/100 g fw. Tuber size was significantly smaller in theS. chacoense parent and the hybrid populations compared with the size in theS. tuberosum parent and backcross populations. In a second sampling of tubers from each population, intentionally selected to equalize tuber size among populations, the mean tuber size for populations was reduced by 79%. The mean population TGA content increased by 44%. However, the differences and rankings among parent, hybrid, and backcross populations were the same for both tuber samplings.     

Leptines and other glycoalkaloids in tetraploidSolanum tuberosum xSolanum chacoense F2 Hybrid and Backcross Families

The leptine glycoalkaloids fromSolanum chacoense Bitter are potential resistance factors to Colorado potato beetle,Leptinotarsa decemlineata Say. A selected sample of 17 F₁ hybrids (4x) from crosses of a 4xS. chacoense clone that synthesizes leptines (8380-1) with 4xS. tuberosum L. lines were selfed to generate F₂ hybrids (4x) and were also backcrossed toS. tuberosum. Glycoalkaloid (GA) content in foliage and tubers was measured for 786 sibling genotypes from the two generations in the field at Beltsville, Maryland. Leptines were found in the foliage of all F₂ hybrids and in 98% of the backcross genotypes. Leptines were not detected in tubers from either generation. Foliage concentrations of leptines for the F₂ hybrids averaged 156 mg/100g fresh weight (FW). The proportion of the total GA content consisting of leptines averaged 42%. By backcrossing the F₁ hybrids toS. tuberosum the average concentration of leptines was reduced to 44 mg/100g FW. The proportion of leptines averaged 16%. The tuber contents of solanine plus chaconine glycoalkaloids averaged 52 mg/100g FW for the F₂ hybrids and 27 mg/100g for the backcross genotypes.     

Association between trichome characters and agronomic traits inSolanum tuberosum (L.)XS. berthaultii (hawkes) hybrids

In 1977, a program to develop potato cultivars with insect resistance based on the glandular trichomes found onSolanum berthaultii was initiated at Cornell University. Associations between agronomic and trichome characters were suspected due to difficulties in selecting hybrid plants with both insect resistance and acceptable agronomic characteristics. Plants from backcrosses and intercrosses were studied. A strong association between the presence of B trichome droplets and undesirable agronomic characteristics was established in backcrossed generations. The backcross plants bearing droplets produced significantly lower yields, fewer tubers, later maturing plants, and poorer foliage and tuber appearance than their sibs without the droplets. In intercrosses, no associations were found. Good MEBA scores were seldom found in backcrossed plants without B droplets. The associations observed are speculated to be due to linkage or structural genomic differentiation between the genomes of the species involved in these populations.     

Potato haploid performance in 2x × 4x crosses

Nineteen haploids (2n = 2x = 24) extracted from four tetraploids ofSolanum tuberosum (2n = 4x = 48) with high 2n egg frequencies and known mode of 2n egg formation were evaluated via 4x progeny from 2x × 4x crosses (haploid × cultivar) for total tuber yield, tuber appearance and tuber set. Ninety-eight, 4x families (15-20 genotypes each) were planted in the randomized complete block design with two replications at two locations. Analyses of variance were conducted for all haploids over three common testers, and each of the four groups of haploids individually over a variable number of testers. The performance of haploids, based on general combining ability effects, was consistent for all traits, when tested with either three or more testers. Diploid clones can be evaluated in 2x × 4x crosses using three unrelated adapted 4x testers. Nonorthogonal comparisons indicated no significant differences for all traits among four groups of haploids, and between modes of 2n egg formation. Significant variation within each group of haploids indicates that selection at the haploid level should be carried out among haploids from either the same or different cultivars. Superior haploids can be utilized in 2x × 4x and 2x × 2x breeding schemes to generate 4x clones.     

Immunodetection of Polyphenol oxidase in glandular trichomes ofS. berthaultii,S. tuberosum and their hybrids

Type A glandular trichomes confer insect resistance to the wild potato,S. berthaultii Hawkes by oxidatively polymerizing their contents upon breakage, resulting in insect entrapment. Polyphenol oxidase (PPO) is responsible for the oxidative polymerization of Type A glandular trichome exudate.S. tuberosum L. trichomes possess low amounts of PPO and lack the ability to either polymerize their contents or entrap insects. Polyclonal antibodies raised against purifiedS. berthaultii glandular trichome PPO were used to develop an enzyme-linked immunosorbent assay (ELISA) to quantify PPO content ofS. berthaultii, S tuberosum, and hybrid type A glandular trichomes. The PPO ELISA and immunoblots were used to quantify PPO expression as a function of leaf development and genetic background. PPO accumulates to its maximal levels in trichomes prior to completion of leaflet expansion. In advanced breeding generations, trichome PPO is expressed at levels varying from that found inS. tuberosum to that found inS. berthaultii. By serving as an initial screen for trichome PPO, this ELISA may be useful to assist rapid introgression of this insect resistance trait into adaptedS. tuberosum backgrounds.     

Characterization of a new gibberellin related dwarfing locus in potato (Solanum tuberosum L.)

Gibberellins have been shown to be involved in such important processes as seed germination, flowering, maturation and tuberization in potato. Parents fromSolanum tuberosum Groups Andigena and Tuberosum were found to produce progeny containing dark green, rosette dwarf individuals which could be completely restored to normal appearance by exogenous GA₃. Test crosses indicated that this phenomenon can be explained by the action of a single locus, the dwarfing characteristic being conferred by the nulliplex condition. The gene symbol “ga₁” is proposed. Group Andigena dwarfs demonstrated much greater tuberization compared to their normal full sibs or dwarfs restored to normal phenotype by exogenous GA₃. The possibility that such gibberellin mutants may be involved in the long day adaptation of Gp. Tuberosum is discussed. This character may also be a useful tool for investigating various aspects of the gibberellin status of the potato.     

Mechanical interaction between larvae of the Colorado potato beetle and glandular trichomes ofSolanum berthaultii Hawkes

The rate of accumulation of glandular trichome exúdate ofSolanum berthaultii on the tarsi of Colorado potato beetle (Leptinotarsa decemlineata) larvae depended on larval size (instar), with large larvae bearing the hardened, darkened exudate on more of their tarsi than did small larvae. In greenhouse studies, third instar larvae accumulated large amounts of exudate regardless of which of 2S. berthaultii accessions they were exposed to, but exudate accumulation by first instars was influenced significantly by plant accession. Tarsal accumulation of exudate was lower in field plots of bothS. berthaultii accessions, but tended to increase with time of exposure. The significant differences in exudate accumulation among larvae exposed to the 2 accessions in the greenhouse were not evident in the field. The density of type A glandular trichomes on the abaxial leaflet surface differed significantly between the 2 accessions in both greenhouse and field. Despite these effects of host and environment on the accumulation of readily observed amounts of trichome exudate on larval tarsi, no larvae were trapped by the adhesive exudate.     

Improved procedures for the isolation and culture of protoplasts fromSolanum etuberosum

An efficient and convenient procedure for the isolation of protoplasts fromSolanum etuberosum is described. Excised leaves of plants grownin vitro were preconditioned for 3 days in the dark at 4 C. A 16 h enzyme digestion followed with purification by flotation in Babcock bottles produced a mean yield of 12.5 × 10⁶ protoplasts/g fresh weight. Protoplast viability as judged by morphology at the time of isolation was 89%. Liquid preculture for 1 day at 1 × 10⁶ protoplasts/ml was used prior to plating at 1 × 10⁵ protoplasts/ml and gave a mean survival of protoplasts of 43% after 8 days in culture.     

Utilizing the frost resistance of diploid solanum species

Introductions of 13 frost-resistant diploid (2n=2x=24)Solanum species (2 non-tuberous) were inter-crossed to produce F₁ hybrids involving two frost resistance sources. Successful F₁ combinations were: (i) inter-crossed to produce hybrids involving 3 to 4 frost resistance sources, (F₁×F₁); and (ii) crossed with haploids (2n=24) of frost-susceptible tetraploid (2n=4x=48)S. tuberosum subsp.tuberosum, (F₁×tbr). Thirty-five of the 75 crosses between the 13 species produced viable seed. The F₁ and F₁×F₁ progenies segregated to yield plants more resistant to freezing temperatures than the parental species. All attempts to cross these hybrids with 24-chromosometbr were successful. Although tuber production improved following the first cross totbr, the frequency of resistant plants decreased approximately 50%. Thus far, all first back-cross attempts, (F₁×tbr)×tbr, have succeeded. Utilizing hybrids synthesized from several sources of frost resistance as the donor parents, and clonal selections of 24-chromosometbr as the recurring parents, may provide a high order of resistance throughout the backcrossing program and implement selection of frost-resistant, long-day tuberizing parental stocks acceptable to the potato breeder.     

Performance of populations derived by selecting for resistance to potato leafhopper in A 4XSolanum Tuberosum X 2XSolanum Chacoense cross

Intercross and backcross populations derived from aSolanum tuberosum L. (2n = 2x = 48) ×Solanum chacoense Bitt. (2n = 2x = 48) cross by selecting for resistance to potato leafhopper (PLH) (Empoasca fabae Harris), were tested in the field. In F2 and F4 populations, the levels of nymphal infestation and leaf necrosis from PLH feeding were considerably lower and tuber yields were much lower compared with those measured on the susceptible cultivar Kennebec. No differences were found between the two populations. Backcrosses of the F2 and F4 populations toS. tuberosum decreased resistance to PLH. Resistance to infestation was still greater than that of Kennebec; severity of leaf necrosis equaled that of Kennebec. Tuber yields increased, but were less than Kennebec’s yield. Foliage types shifted closer to those ofS. tuberosum. Three generations of selection and intercrossing within the backcross populations did not change the level of resistance to PLH but did tend to lower tuber yields. Foliage types remained primarilyS. tuberosum in appearance although distinctive types segregated with increased frequency. A second backcross toS. tuberosum did not change the level of resistance to PLH or the level of tuber yield relative to those of the first backcross or Kennebec.     

Glycoalkaloid content in foliage of hybrid and backcross populations from aSolanum tuberosum X S. chacoense cross

The glycoalkaloid contents of foliage were measured in populations derived from a cross ofSolanum chacoense Bitter (PI 472810) with a composite sample of genotypes from a randomly matedSolanum tuberosum L. population. The mean total glycoalkaloid contents of the parent PI 472810 andtuberosum populations, and the F₂ and F₄ populations were 856, 121, 286, and 279 mg/100g fresh weight, respectively. The total glycoalkaloid content of the first backcross population, F₂ xtuberosum, was 290 mg/100 g, close to the content of the F₂. In a second backcross totuberosum, the mean total content was 148 mg/100 g, close to the content of the parenttuberosum population. Only the glycoalkaloids α-solanine and α-chaconine were found in PI 472810, whereas in the parenttuberosum, hybrid, and backcross populations, the glycoalkaloids α & β-solamarine also were found in a small number of genotypes. The ratios of chaconine to solanine contents were significantly (P≤.05) different in the parent PI 472810 andtuberosum populations, 2.55 and 2.12, respectively. The ratios in the F₂ and F₄ populations were similar to that of PI 472810, whereas the ratios in the backcross populations were closer to that of thetuberosum parent. The levels of foliage glycoalkaloids in the hybrid and backcross populations paralleled the levels of potato leafhopper,Empoasca fabae Harris, resistance measured in a previous study.     

Solanum berthaultii Hawkes affects the digestive system,fat body and ovaries of the Colorado potato beetle

Ovarian development and fecundity of Colorado potato beetles reared on the resistant potato species,Solanum berthaultii were reduced compared to females reared on the susceptible potato,S. tuberosum. Striking changes in the appearance of the digestive tract and large quantities of undigested plant material in the midgut of females reared onS. berthaultii suggest that ovarian development might be impaired by poor nutritional quality ofS. berthaultii foliage. Alternatively, enlargement of the midgut by undigested plant material might physically limit ovarian development and subsequent fecundity. These results suggest that a major impact ofS. berthaultii on CPB performance occurs in the physiology of digestion and reproduction.     

Seed set in 4x x 2x crosses as related to 2n pollen frequency

Seeds per fruit following 4x × 2x crosses in potato are roughly related to the frequency of 2n pollen. Our goal was to examine precisely the relationship between frequency of stainable 2n pollen and seeds per fruit (s/f) in 4x × 2x crosses. A 4x cultivar with high female fertility and 21 2n pollen producing 2x hybrids were used as seed and pollen parents, respectively. 2n pollen frequencies varied from 1 to 99% in the 2x hybrids and seed set ranged from 0 to 305 s/f in 4x×2x crosses. Mean seeds per fruit increased in response to increases from 1 to 50% 2n pollen, but did not increase in response to using a pollen sample with a 2n pollen frequency over 60%. Seed set averaged 11 s/f corresponding to pollen samples with 1–5% 2n pollen, 50 s/f at 10–20%, 125 s/f at 20–30%, and over 170 s/f at 2n pollen frequencies over 60%. Seed set in 210 control 4x × 4x crosses indicated that equivalent seed set occurs in 4x × 2x crosses when 2n pollen frequencies are over 20–30%. The 2x hybrids differed in mean and variation of 2n pollen frequencies and seed set in 4x × 2x crosses. One 2x hybrid with a meiotic aberration (sy-3) resulting in all univalents at Metaphase I (ordinarily leading to male sterility) and parallel spindles (ps) at Anaphase II had close to 100% 2n pollen (of the stainable pollen) and gave equivalent s/f in 4x × 2x crosses to s/f in 4x × 4x crosses. It is apparent that ample seed set occurs for using the 4x × 2x breeding scheme with 2n pollen frequencies over 10% and that even very low 2n pollen frequencies (1–10%) provide seed for the 4x×2x breeding approach.