Evolutionary pathway of T-type Chloroplast DNA in potato

Potato was domesticated in the Andes of South America. However, the presently worldwide-grown potato (Solanum tuberosum L. ssp.tuberosum, 2n=4x=48) has characteristic T-type chloroplast DNA that was introduced after late blight epidemics in the mid-19th century from the Chilean potato (2n=4x=48) grown in the southern coastal regions in Chile. Among many wild potato species, the same chloroplast DNA was found only in some populations of a diploid speciesS. tarijense Hawkes (2n=2x=24), which ranges from central Bolivia to northwest Argentina. To elucidate an evolutionary pathway of T-type chloroplast DNA fromS. tarijense to Chilean potato, 215 accessions ofS. stenotomum Juz. et Buk., considered to be the most primitive, diploid cultivated potato species, from which all the Andean cultivated species evolved, and 286 accessions of the most widely grown, Andean tetraploid cultivated speciesS. tuberosum L. ssp.andigena Hawkes (2n=4x=48) were examined in this study. No accession ofS. stenotomum had T-type chloroplast DNA, while nine accessions, mostly from northwest Argentina, ofS. tuberosum ssp.andigena had T-type chloroplast DNA. Therefore, I conclude that some populations ofS. tarijense having T-type chloroplast DNA were naturally crossed as female withS. tuberosum ssp.andigena from which the Chilean potato was selected.     

Distribution of the 241 bp deletion of chloroplast DNA in wild potato species

The common potato (Solanum tuberosum L. ssp.tuberosum) has T-type chloroplast DNA characterized by a 241 bp deletion. To explore the maternal ancestry of the common potato, a total of 566 accessions of 35 wild species, collected mostly from central Bolivia to northern Argentina, were determined for presence or absence of the deletion by a simple PCR assay using primers flanking the deleted region of chloroplast DNA. Sixteen out of 80 accessions ofS. tarijense, S. berthaultii, andS. neorossii showed a shorter PCR amplified fragment. Sequencing of these fragments revealed that the same 241 bp was deleted at the same position in these accessions. This strongly suggests that the deletion event had occurred in wild species.     

Reaction ofSolanum tuberosum andSolanum species toMeloidogyne hapla

The reactions of 218 clones ofSolanum tuberosum and 238 accessions of 55 tuber-bearingSolanum species toMeloidogyne hapla were determined by planting them in soil naturally infested withM. hapla and the microsclerotial form ofVerticillium alboatrum. Degree of root galling was rated from none (0) to severe (5). Incidence of root galling on clones was slight as evidenced by ratings of 0, 0.5 and 1.0 for 38, 32 and 18 percent of them, respectively. As incidence of root galling increased there was a tendency for percentage of galled tubers and internal tuber infection byM. hapla to increase. Of many family progenies ofS. tuberosum tested, those from family B7147 (Penobscot x W39-1) were the most resistant toM. hapla. Incidence ofM. hapla root galls andVerticillium alboatrum wilt symptoms onS. tuberosum clones indicated there was no apparent interaction between these two pathogens. Eighty-one percent of the 238 accessions remained free from root galls.Solanum species free from root galls and represented by three or more accessions werebrachycarpum, bulbocastanum, cardiophyllum, hjertingii, hougasii, infundibuliforme, kurtzianum, martinasense, multidissectum, multiinterruptum, papita, sparsipilum, spegazzinii, stoloniferum, tarijense, andvernei. S. microdentum (PI320312) andS. tuberosum ssp.andigena (PI245926) had the most extensive root systems free ofM. hapla galls.     

Glandular pubescence,glycoalkaloid composition,and resistance to the green peach aphid,potato leafhopper,and potato fleabeetle inSolanum berthaultii

Fifteen accessions ofSolarium berthaultii Hawkes andS. berthaultii xS. tarijense Hawkes were assessed for resistance to field infestations of the green peach aphid,Myzus persicae (Sulzer), potato leafhopper,Empoasca fabae (Harris), and potato fleabeetle,Epitrix cucumeris (Harris). Accessions bearing both Type A and Type B glandular trichomes were much more resistant to the green peach aphid and potato leafhopper than accessions bearing Type A hairs alone. All accessions had significantly smaller populations of these 3 pests than S.tuberosum cultivars. Total glycoalkaloid (TGA) content of foliage and tubers was not correlated with insect populations. Foliar TGA levels of field-grown plants varied among accessions, ranging from < 2–240 mg/100 g fresh wt. Solasonine and solamargine were the major foliar glycoalkaloids while solamarines predominated in tubers.     

Crossability of some green-peach-aphid-resistant tuber-bearing solanums,potential bridging species,andSolanum tuberosum ssptuberosum

Blue light fluorescence microscopy was found to be a quick and easy procedure for assessing pollen tube development in interspecific pollinations ofSolanum. Based on penetration of pollen tubes to the base of the style, several combinations of green-peach-aphid-resistant species and potential bridging species to S.tuberosum ssp.tuberosum should be considered for embryo culture. Sterile or highly infertile hybrids were obtained between green-peach-aphid-resistantS. tuquerrense andS. tuberosum ssp.tuberosum and are described. Slightly fertile hybrids were obtained betweenS. tuquerrense andS. polytrichon, a species compatible withS. tuberosum ssp.tuberosum.     

Exploitation of Exotic, Cultivated Solanum Germplasm for Breeding and Commercial Purposes

Native potato species (NPS) are cultivated potatoes which do not belong to S. tuberosum ssp. tuberosum. They are maintained by the farmers of the Andes for subsistence under harsh environmental conditions where other potatoes cannot compete. Within the huge phenotypic variability between and within these species certain accessions possess resistances to different pests and diseases, remarkable culinary and processing quality and favourable nutritional properties. We present first evaluation results of a set of NPS for these characters and discuss prospects for exploiting this valuable germplasm for breeding and for developing novel products for niche markets.     

Identification of stem soft rot (Erwinia carotovora subspeciesatroseptica) resistance in potato

Development of potato cultivars resistant to Erwinia species (Erwinia carotovora subspeciesatroseptica, E. carotovora subspeciescarotovora, andErwinia chrysanthemi) causing soft rot of stems and tubers involves identification and transfer of the resistance to breeding lines possessing desirable agronomic traits. In total, 363 accessions of six wildSolanum species, three somatic hybrids and nine sexual progeny of somatic hybrids were screened for stem soft rot resistance. Thirty-one, 50, and 67 percent of accessions ofSolanum boliviense, Solanum chacoense, and Solanumsanctarosae, respectively, were resistant or highly resistant, whereas, 100, 97, and 95 percent of accessions ofSolanum canasense, Solanum tarijense, andSolanum spegazzinii were susceptible or highly susceptible, respectively. Two tuber soft-rot resistant somatic hybrids, A937 and T355-11 produced earlier by the fusion ofSolanum brevidens (PI 218228) andSolanum tuberosum (PI203900) orS. tuberosum cv. Russet Burbank, respectively, and the sexual progeny of A937 andS. tuberosum cv. Katahdin were also highly resistant. Transfer of resistance fromS. brevidens to somatic hybrid (A937) and subsequently to the progeny suggests that the resistance is simply inherited. In addition to the identification of 65 resistant or highly resistant clones of wildSolanum species, this study has identified three tuber soft rot-resistant clones that are also resistant to stem soft rot. Potato cultivars showed a range of responses to the pathogen.     

Seed protein variation within accessions of wild and cultivated potato species and inbredSolanum chacoense

Summary Soluble proteins in seed potato were analyzed by electrophoresis (PAGE). The Index of Variation (IV) was calculated in order to estimate variation within each accession of wild and cultivated potato species andS. chacoense selfed lines. Seeds of 34 accessions representing 14 species from six taxonomic series were analyzed. The IV ofS. chacoense selfed lines decreased from 47.7 in the S₂ to 12.9 in the S₇ generation. The average IV for the self-incompatible diploid species was 41.2, whereas that of the selfcompatible diploid species,S. polyadenium, was 21.3. TetraploidS. tuberosum ssp.andigena andS. sucrense had an average IV of 62.2, which is much higher than that observed in any other diploid species. The naturally selfing speciesS. acaule (4x), and its colchicine-doubled derivative (8x), showed very low variation (IV=7.5). The IV appears to reflect heterozygosity and is affected by the breeding mode and the genetic constitution of diploid parents for derived polyploids.     

Comparison of polyphenol oxidase in tubers ofSolanum tuberosum and the non-browning tubers ofS. hjertingii

Solarium hjertingii, a wild potato species, does not exhibit enzymatic browning nor blackspot. Blackspot and enzymatic browning are physiological disorders that are caused by the reaction of the enzyme polyphenol oxidase with phenolic substrates, resulting in discoloration of tuber cortex. We determined thatS. hjertingii has a low amount of at least one of the polyphenol oxidases in its tubers compared toS. tuberosum. Less than 5 μg ofS. tuberosum polyphenol oxidase protein, as determined from Western blot staining, had measurable enzymatic activity. In contrast,S. hjertingii protein with comparable staining had no detectable activity.     

Potato germplasm collecting expedition to Chile, 1989, and utility of the Chilean species

A joint Chilean, United States potato (Solanum sect.Petota) germplasm collecting expedition was conducted in Chile between January 21–March 30, 1989. The trip resulted in collections ofSolanum brevidens (50 accessions),S. etuberosum (28 accessions),S. maglia (3 accessions) and indigenous primitive landraces ofS. tuberosum (24 accessions). Comments are presented on the habitats, distribution, and potential and realized breeding value of these species.     

Cytoplasmic substitution inSolanum. I. Seed production,germination, and sterilities of reciprocal backcross generations

Summary Genes ofSolanum tuberosum L. ssp.tuberosum were introduced into cytoplasm ofS. tuberosum ssp.andigena (Juz. & Buk.) Hawkes andS. phureja Juz. & Buk. to reduce cytoplasmic sterilities. Alleles that influenced berry set and seed content produced differences between reciprocal progenies. Their expressions were interpreted in terms of genes of the maternal and/or paternal parents. When the ssp.andigena genotype was introduced from the maternal parent if favoured high berry set and low seed number. Maternal ssp.tuberosum favoured lower berry set and higher seed number. The source ofS. phureja cytoplasm, aS. phureja×S. chacoense F₁, used as maternal parent favoured lower berry and seed set. During successive backcrosses progeny expressions approached those of the recurrent ssp.tuberosum pollen parents. A low level of pollen sterility occurred in BC₂ plants, unrelated to direction of cross or cytoplasmic factors. By BC₄, seed set and fertility were as good as ssp.tuberosum, and recurrent backcrossing could be terminated without loss of seed production or fertility. Pennsylvania Agricultural Experiment Station Journal Series No. 7493.     

Characterization ofSolanum tuberosum simple sequence repeats and application to potato culiwar identification

With the continued introduction of new potato cultivars, accurate identification is becoming difficult but is essential for maintaining cultivar integrity and Plant Breeders’ Rights. Hypervariable DNA sequences, referred to as simple sequence repeats (SSRs) or microsatellites, have been reported to be an excellent source of genetic markers. To determine the abundance, distribution, and composition of SSRs withinSolanium tuberosum, 252 sequences were searched for tetranucleotide and smaller SSRs with a minimum length of 20 nucleotides and a maximum discrepancy of two nucleotides. In total, 40 unique SSRs were observed in the 252S. tuberosum sequences examined and occurred at a frequency of one SSR every 8.1 kb. To assess the ability of site-specific amplified SSRs to identify potato cultivars, a simple (TCAC)_m and compound (TCAC)_m ? (CTT)_n SSR 5’ to the starch synthase gene and a compound (C)_p ? (CT)_q ? (AT)_r ? (G)_s SSR 5’ to the sequence encoding mature proteinase inhibitor I, were examined and shown to produce unique DNA profiles for 73 of 95 tetraploid cultivars. In total, 24 alleles were observed at these loci and the accurately sized amplified DNA products can be used to establish a database for cultivar identification. Site-specific amplified alleles were somatically stable and have been conserved in clonal variants of Russet Burbank independently maintained for almost seven decades, a characteristic essential for cultivar identification. As genetic markers, the abundant, informative, and easily examined site-specific amplified alleles of SSRs are ideal for quickly and accurately determining cultivar identity of S.tuberosum ssp.tuberosum.     

An analysis of the hybrid nature ofsolanum xedinense Berth, using molecular, cytological and crossability studies

Summary Solanum xedinense Berth., a pentaploid (2n=5x=60) interspecific hybrid between the cultivated potato and the wild speciesSolanum demissum Lindl., represents evidence of genetic exchange between wild and cultivated species. It has been found in Central Mexico at altitudes between 2000 and 3500 meters, growing in and along the edges of cultivated fields, roadside thickets, forest margins and along irrigation ditches. This study examined the hybrid nature of S.xedinense using molecular, cytological and crossability data. RAPD and AFLP markers were used to supportS. xedinense as a hybrid between the cultivated potato andS. demissum. Meiotic analysis and crossability studies were used to examine the potential ofS. xedinense to form offspring. Several accessions ofS. tuberosum L. subsp.tuberosum and subsp.andigena Hawkes,S. demissum and an accession ofS. xedinense were used in the study. Artificial hybrids betweenS. tuberosum subsp.tuberosum andS. demissum were included for comparison. Molecular analysis positioned the natural and artificial hybrids as an intermediate group between the parental populations, supporting their relationship with the parental populations. Meiosis was abnormal in both natural and artificial hybrids, and reduced crossability was noted forS. xedinense. Although gene flow appears to happen between wild and cultivated potato, the potential for the natural hybrids to become established may depend mostly on their ability to clonally propagate themselves.     

Use of RAPD markers in potato genetics: Segregations in diploid and tetraploid families

The objective of the present study was to evaluate random amplified polymorphic DNA (RAPD) as a source of markers for use in investigations in potato genetics. Segregation of 18 loci in diploidSolanum goniocalyx xS. phureja and 12 loci in tetraploidS. tuberosum ssp.tuberosum ×S. tuberosum ssp.andigena families fitted Mendelian and tetrasomic ratios, respectively. Eight loci in the diploid progeny were arranged in three linkage groups. Segregations of these markers fitting expected ratios indicate that they can be effectively used in potato genetics, breeding and evolution.     

Fine screeningSolanum (potato) germplasm accessions for resistance to Colorado potato beetle

This study was conducted to investigate the utility of systematic screening at the genotype level withinSolanum accessions highly resistant to the Colorado potato beetle. Evaluations of clonally replicated genotypes showed that most accessions reported to be uniform when screened as populations contained small but significant variation among genotypes for resistance to oviposition, larvae, and defoliation, differences for numbers of larvae being most common. Adult counts and percentage defoliation were not as useful in evaluating among-genotype variability in beetle resistance. Genotypes ofS. pinnatisectum WRF 343 andS. tarijense PI 473227 were the most uniformly and highly resistant to Colorado potato beetle. Genotypes of S.berthaultii PI 473331,S. chacoense PI 473405, andS. tarijense PI 473336 were moderately to highly resistant, and genotypes ofS. bukasovii PI 473494 andS. canasense PI 230511 were uniformly susceptible to Colorado potato beetle. Nonparametric correlation analyses indicated that number of egg masses, small larvae, large larvae, and defoliation scores were positively correlated, negatively correlated, or not correlated, depending on the species. One generation of selection attempting to segregate resistance and susceptibility in nearly uniform and highly resistantS. pinnatisectum WRF 343 resulted in “divergent” populations that could not be distinguished from each other or the base population. Thus, through genotype (“fine”) screening and selection, we showed that some existing populations are virtually pure for extreme resistance. Use of individuals from such families would make screening breeding populations more efficient, and reduce the risk of losing resistance genes that have non-dominant effects. Fine screening, recurrent selection, and maintenance of such elite populations is recommended as an extension of population-based evaluation usually done by genebanks.     

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 forty-four potato varieties for resistance to common scab,black scurf and silver scurf

Common scab, black scurf and silver scurf belong to serious diseases of cultivated potato that especially impair tuber quality. Tuber infection in 44 potato (Solanum tuberosum ssp. tuberosum L.) varieties was evaluated using two measures – severity and incidence of infected tubers. Both measures were highly correlated. An important result derived from this study supports fungicide application against potato late blight secondarily acts on reducing tuber infection by all evaluated pathogens. Therefore, the conventional potato growing seems to be more effective compared to ecological growing. The pedigree analysis of 44 varieties indicated the presence of Solanum demissum genotypes in the variety genome may negatively affect variety response to Helminthosporium solani presence.     

Progress report: Resistance to the Colorado potato beetle and potato leafhopper inSolanum tuberosum subsp.andigena

An extensive collection of 1527 clones ofS. tuberosum subsp.andigena was screened in 1973 in unreplicated plots to test for resistance to the Colorado potato beetle and potato leafhopper. All clones were susceptible to the beetle. Of 17 clones selected in the 1973 tests for resistance to the potato leafhopper, 7 selections were significantly more resistant than the control (Katahdin) in a replicated experiment in 1974.     

Cytoplasmic substitution inSolanum. II. Tuber characteristics of reciprocal backcross progeny

Summary Recurrent backcrosses (BC) were used to introduce nuclear factors ofS. tuberosum L. ssp.tuberosum into the cytoplasm of Andean potatoesS. tuberosum ssp.andigena (Juz. and Buk.) Hawkes andS. phureja Juz. and Buk. Each generation BC was produced in reciprocal directions to test the effect of cytoplasmic substitution on yield. This report analyses tuber number, tuber weight, and tuber length of BC₂ and BC₃ substitution into ssp.andigena cytoplasm and BC₂ and BC₂ substitutions intoS. phureja cytoplasm. Direction of cross had no consistent significant effect on yield components of ssp.andigena substitution lines. InS. phureja reciprocal progenies direction of cross was responsible for significant differences of some reciprocal sets. When there were significant differences the higher yield usually occurred when ssp.tuberosum was the pistillate parent, but in some progenies when ssp.tuberosum was the pistillate parent the yield was significantly lower than in the reciprocals. It is suggested that differences between reciprocals resulted from chromosomally encoded gene action, resulting from maternal and/or paternal effects, rather than cytoplasmic factors. Authorized for publication as paper No. 7494 in the Journal Series of The Pennsylvania Agriculture Experiment Station.