Plasma Membrane Alterations in Callus Tissues of Tuber-bearing Solanum Species during Cold Acclimation

Plasma membrane alterations in two tuber-bearing potato species during a 20-day cold acclimation period were investigated. Leaf-callus tissues of the frost-resistant Solanum acaule Hawkes `Oka 3878' and the frost-susceptible, commonly grown Solanum tuberosum `Red Pontiac,' were used. The former is a species that can be hardened after subjecting to the low temperature, and the latter does not harden. Samples for the electron microscopy were prepared from callus cultures after hardening at 2 C in the dark for 0, 5, 10, 15, and 20 days. After 20 days acclimation, S. acaule increased in frost hardiness from −6 to − 9 C (killing temperature), whereas frost hardiness of S. tuberosum remained unchanged (killed at −3 C). Actually, after 15 days acclimation, a −9 C frost hardiness level in S. acaule callus cultures had been achieved.     

Physiological responses to NaCl stress in three wild species of potato in vitro

In this study, responses of wild species of potato to NaCl stress were investigated in vitro. In S. stoloniferum and S. bulbosum, length of the shoot, fresh and dry weight, photosynthetic pigments, K⁺ concentration, K⁺/Na⁺ ratio, ascorbate pool, anthocyanin, and phenolic and flavonoid compounds were decreased in response to salinity. In these species, salinity increased the level of Na⁺, lipid peroxidation, proline and ion leakage percentage. In S. acaule, the length of the shoot, and fresh and dry weight were not affected by salinity. Photosynthetic pigments, Na⁺ concentration, proline, flavonoid and phenolic compounds quantities were increased and K⁺/Na⁺ ratio were decreased. K⁺ concentration, lipid peroxidation, ascorbate pool, anthocyanin and ion leakage were not changed by NaCl stress. Superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase and catalase activities were increased in all species. The results suggest that the non-enzymatic antioxidant capacity in S. acaule (salt tolerant) is more important than the enzymatic antioxidant capacity in comparison with the other species.     

Potato germ plasm enhancement with disomic tetraploid Solanum acaule. II. Assessment of breeding value of tetraploid F1 hybrids between tetrasomic tetraploid S. tuberosum and S. acaule

The breeding value of tetraploid F₁ hybrids between tetrasomic tetraploid S. tuberosum and the disomic tetraploid wild species S. acaule was examined. The F₁ hybrids showed a tuber yield and appearance comparable to those of their cultivated parent, indicating a potential as acceptable breeding stocks despite the 50% contribution to their pedigree from wild S. acaule. The cytological behavior of the tetraploid F₁ hybrids was examined to determine the probability of recombination for the introgression of S. acaule genes. The majority of the meiotic configurations at metaphase I was bivalents and univalents with mean frequencies of 17.6 and 9.9, respectively. Further, a low frequency of trivalents and quadrivalents was observed. An acceptable low level of meiotic irregularities were observed at the later stages of microsporogenesis, and a reasonable level of pollen stainability was obtained. Therefore, these hybrids could likely be employed for further introgression. From the cytological observations, the following speculations were drawn: (1) some genomic differentiation exists between the S. acaule genomes, (2) at least one of the S. acaule genomes may be homoeologous to the S. tuberosum genomes, (3) intergenomic recombination would likely occur due to the nature of the genomic constitution of the hybrids, and (4) the nature of sesquiploidy of the hybrids may facilitate efficient introgression and establishment of unique aneuploid and euploid recombinant genetic stocks.     

Allelism of endosperm balance number (EBN) in Solanum acaule Bitt. and other wild potato species

The inheritance of endosperm balance number (EBN), a genetic, dose-dependent crossability system functioning in tuber-bearing Solanum (potato) species, was investigated for certain wild potato species having an EBN equal to one half of their ploidy. The EBN of Solanum acaule, a disomic 4(2EBN) South American species, was investigated by producing F₁ and F₂ hybrids with artificial 4x(2EBN) S. commersonii. This allowed assessment of recombination among the two genomes of disomic S. acaule and that of S. commersonii. When crossability of the hybrids with 1EBN, 2EBN and 4EBN standards was tested, no variation for EBN was detected. The apparent lack of recombination and segregation for EBN in these hybrids indicates that the genomes of S. acaule and S. commersonii carry EBN in a genetically-similar way. Combined with previous reports, these data indicate that the inheritance of EBN is similar in widely-separated taxa from South America and Mexico.     

Diploid hybrids between allotetraploid wild potato species <Emphasis Type="Italic">Solanum acaule</Emphasis> Bitt., <Emphasis Type="Italic">S. stoloniferum</Emphasis> Schltdl. and dihaploids of <Emphasis Type="Italic">S. tuberosum</Emphasis> L.

The possibility to obtain diploid hybrids by pollination of allotetraploid wild potato species Solanum acaule and S. stoloniferum plants with fertile pollen of S. tuberosum dihaploids was demonstrated for the first time. Dihaploid hybrids have arisen with comparatively high frequency (from 12.5 to 33.3%). They were characterized by high regularity of meiosis and high fertility. They easily crossed with S. tuberosum dihaploids, forming viable progeny. This seems prospective for effective introgression of valuable genetic gene pool of wild allotetraploid potato species in breeding material of S. tuberosum on the diploid level.     

Relative sensitivity of photosynthesis and respiration to freeze-thaw stress in herbaceous species : importance of realistic freeze-thaw protocols

The relative effect of a freeze-thaw cycle on photosynthesis, respiration, and ion leakage of potato leaf tissue was examined in two potato species, Solanum acaule Bitt. and Solanum commersonii Dun. Photosynthesis was found to be much more sensitive to freezing stress than was respiration, and demonstrated more than a 60% inhibition before any impairment of respiratory function was observed. Photosynthesis showed a slight to moderate inhibition when only 5 to 10% of the total electrolytes had leaked from the tissue (reversible injury). This was in contrast to respiration which showed no impairment until temperatures at which about 50% ion leakage (irreversible injury) had occurred. The influence of freeze-thaw protocol was further examined in S. acaule and S. commersonii, in order to explore discrepancies in the literature as to the relative sensitivities of photosynthesis and respiration. As bath cooling rates increased from 1°C/hour to about 3 or 6°C/hour, there was a dramatic increase in the level of damage to all measured cellular functions. The initiation of ice formation in deeply supercooled tissue caused even greater damage. As the cooling rates used in stress treatments increased, the differential sensitivity between photosynthesis and respiration nearly disappeared. Examination of agriculturally relevant, climatological data from an 11 year period confirmed that air cooling rates in the freezing range do not exceed 2°C/hour. It was demonstrated, in the studies presented here, that simply increasing the actual cooling rate from 1.0 to 2.9°C/hour, in frozen tissue from paired leaflet halves, meant the difference between cell survival and cell death.     

Kritsche Betrachtungen zur Nomenklatur argentinischer Wildkartoffeln

Summary Continuing the critical publications on the nomenclature of the Argentine wild potatoes, set on in 1956, now the results of 5 years' studies of the seriesAcaulia are discussed. This series is to be found in the highlands of the Cordilleras and its vegetation limit is at about 4700 m above sea-level. The frost-resistance ofS. acaule andS. aemulans is remarkably high (up to minus 10°). ForS. acaule cytological determinations have proved again the already known number 2n=48, besides this in the province of Jujuy a species with 2n=36 has been found, being probably a hybrid ofS. acaule andS. alticolum. This species with 36 chromosomes is increasingly spreading in certain arid areas in an altitude of 4000 m. ForS. aemulans the chromosome number 2n=24 is published for the first time. Reciprocal crosses under greenhouse conditions betweenS. aemulans (2n=24) andS. acaule (2n=48) showed no success. In opposition to the view ofHawkes the author insists onS. aemulans being a good species, endemical for the arid mountains of Famatina and Ambato in the provinces of La Rioja and Catamarca. Having been collected in 1907 for the last time, nowS. aemulans has anew been found by the author in the Famatina mountains.As toS. acaule and its numerous varieties and related species the author holds the following opinion: The termsS. depexum, Schreiteri, punae, subexinterruptum, checcae, caulescens, chorruense established byBitter, Bukasov, Juzepzuk, andHawkes, are superfluous, the real range of variability is moreover much greater. To illustrate this point of view some examples were set.By its valuable resistance-genes againstRhizoctonia, frost, and Virus,S. acaule, at present, has gained considerable importance for plant breeding. For this reason we think it the more necessary to elucidate the systematical and cytological problems of the whole SeriesAcaulia.

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Mit 8 Abbildungen     

Transfer of resistance to PLRV titer buildup from Solanum etuberosum to a tuber-bearing Solanum gene pool

Summary Hybrids between Solanum etuberosum and S. pinnatisectum harboring resistance to titer buildup of potato leafroll virus (PLRV) were reciprocally crossed with tuber-bearing wild species S. acaule and S. verrucosum. A total of 47 hybrids with acaule were obtained with the aid of embryo rescue and sterile culturing of embryos from imbibing seeds. All but two hybrids with acaule had low pollen stainabilities or were pollen sterile. Hybrid seeds from crosses with verrucosum were easily obtained, and the triploid progenies were sterile. Hybrid progeny were screened for resistance to PLRV infection by viruliferous green peach aphid and for resistance to titer buildup. Although hybrids did not exhibit resistance to infection, PLRV was not detectable using ELISA. Virus was detected, however, by graft transmission to Datura stramonium. Crosses of fertile acaule-etuberosum-pinnatisectum hybrids with S. phureja, a cultivated diploid, using the latter as pollen parent, produced berries but seed did not complete development and was aborted. Rescue of immature embryos 25 days after pollination by excision from berries and sterile culture produced vigorous, pot-cultured plants. Segregation of susceptible (virus detected) and resistant (virus undetected) progenies suggests simple inheritance.     

Isolation of polymorphic microsatellites in the stemless thistle (Cirsium acaule) and their utility in other Cirsium species

The genus Cirsium includes species with both widespread and restricted geographical distributions, several of which are serious weeds. Nine polymorphic microsatellite loci were isolated from the stemless thistle Cirsium acaule. Eight were polymorphic in C. acaule, six in C. arvense and seven in C. heterophyllum. One locus monomorphic in C. acaule showed polymorphism in C. heterophyllum. The mean number of alleles per locus was 4.1 in C. acaule, 6.2 in C. arvense and 2.9 in C. heterophyllum. These nine loci were also amplified in C. eriophorum and C. vulgare, suggesting that these markers may be of use throughout the genus.     

Causes of Flowering of Long-Day Potato Species under Short-Day and Cold-Night Conditions

Flowering and tuber formation in high-mountain potato species Solanum sparsipilum Bitt., S. acaule Bitt., S. punae Juz., S. demissum Lindl., and a tuber crop Ullucus tuberosus Caldas. were investigated. All these species are characterized by absolute requirement of long day-length for flowering and short day-length for tuberization. Plants were grown under the following conditions: natural day-length with a photoperiod of 17 h or longer (treatment 1), short days with a photoperiod of 12 h and warm nights (15–20°C) (treatment 2), and short days with cold nights (5–6°C) (treatment 3). In the first treatment, plants produced flowers but no tubers. In the second treatment, plants produced tubers but no flowers. In the third treatment, plants produced both flowers and tubers. In leaves of S. acaule and U. tuberosus, the levels of gibberellins and ABA were determined. A high activity of gibberellins in the third treatment was similar to that in the first treatment, whereas high ABA activity in the third treatment was similar to that in the second treatment. It is supposed that cold nights retard the destruction of GA in plants during the dark period of diurnal cycle and ensure a permanently high level of gibberellins, which facilitates flowering of long-day species under short-day conditions. The high level of ABA is considered a plant response to short-day conditions, which is favorable for tuberization.     

Cloning and Functional Characterization of SAD Genes in Potato

Stearoyl-acyl carrier protein desaturase (SAD), locating in the plastid stroma, is an important fatty acid biosynthetic enzyme in higher plants. SAD catalyzes desaturation of stearoyl-ACP to oleyl-ACP and plays a key role in determining the homeostasis between saturated fatty acids and unsaturated fatty acids, which is an important player in cold acclimation in plants. Here, four new full-length cDNA of SADs (ScoSAD, SaSAD, ScaSAD and StSAD) were cloned from four Solanum species, Solanum commersonii, S. acaule, S. cardiophyllum and S. tuberosum, respectively. The ORF of the four SADs were 1182 bp in length, encoding 393 amino acids. A sequence alignment indicated 13 amino acids varied among the SADs of three wild species. Further analysis showed that the freezing tolerance and cold acclimation capacity of S. commersonii are similar to S. acaule and their SAD amino acid sequences were identical but differed from that of S. cardiophyllum, which is sensitive to freezing. Furthermore, the sequence alignments between StSAD and ScoSAD indicated that only 7 different amino acids at residues were found in SAD of S. tuberosum (Zhongshu8) against the protein sequence of ScoSAD. A phylogenetic analysis showed the three wild potato species had the closest genetic relationship with the SAD of S. lycopersicum and Nicotiana tomentosiformis but not S. tuberosum. The SAD gene from S. commersonii (ScoSAD) was cloned into multiple sites of the pBI121 plant binary vector and transformed into the cultivated potato variety Zhongshu 8. A freeze tolerance analysis showed overexpression of the ScoSAD gene in transgenic plants significantly enhanced freeze tolerance in cv. Zhongshu 8 and increased their linoleic acid content, suggesting that linoleic acid likely plays a key role in improving freeze tolerance in potato plants. This study provided some new insights into how SAD regulates in the freezing tolerance and cold acclimation in potato.     

Interspecific somatic hybrids between wild potato Solanum acaule Bitt. and anther-derived dihaploid potato (Solanum tuberosum L.)

Solanum acaule Bitt. is a disomic tetraploid (4x) wild potato species which is resistant to several potato diseases. Introgression of disease resistance and abiotic stress tolerance to the tetrasomic tetraploid (4x) cultivated potato (S. tuberosum L.) gene pool via crossing has been limited due to the difference in the endosperm balance number. In the present study, protoplast fusion was applied to produce hexaploid (6x) somatic hybrids between the parental lines, tetraploid (4x) S. acaule and two anther-derived dihaploid (2x) lines of S. tuberosum cv. White Lady. One callus (0.4%) of a total of 229 calli obtained regenerated into shoots in the fusion combination S. acaule (+) White Lady 15.dh.8.2.2. All the regenerated shoots were confirmed to be interspecific somatic hybrids using species-specific RAPD markers. In another fusion combination, S. acaule (+) White Lady 7.dh.23.1.1, fifteen calli (5%) regenerated into a total of sixteen shoots from 289 calli. All the analysed somatic hybrids between S. acaule and S. tuberosum were hexaploid. The mean DNA content (2C value) of the combination S. acaule (+) White Lady 15.dh.8.2.2 somatic hybrids (4.55 pg), was approximately the sum (4.69 pg) of the DNA contents of the parental lines, S. acaule (2.95 pg) and S. tuberosum (1.74 pg). In the greenhouse, the two somatic hybrids analysed were normal in their morphological characteristics and more vigorous than their parental lines. Most of the morphological characteristics were closer to the tetraploid S. acaule than to the dihaploid S. tuberosum. The interspecific somatic hybrids are currently being tested for frost tolerance and glycoalkaloid composition. Received: 19 January 1998 / Revision received: 27 March 1998 / Accepted: 20 April 1998     

Respiratory enzyme activities correlate with anoxia tolerance in salt marsh grasses

Salt marsh communities are known for well-defined species zonation patterns. Lower limits of plant growth are thought to be set by an ability to tolerate anoxic sediments, but the physiological differences between species have not previously been examined. To investigate responses to anoxic sediments, several estuarine species were grown in greenhouse experiments to compare how respiratory processes were affected by flooding. Metabolic characteristics related to respiration and anoxia tolerance were studied in the emergent estuarine species Spartina alterniflora, S. anglica, S. densiflora, S. foliosa, S. alterniflora × S. foliosa hybrids, S. patens, and Distichlis spicata and compared to the inland species maize (Zea mays). All species showed a strong ability to respire anaerobically, indicating flooding tolerance. High intertidal marsh species had significantly higher root aerobic respiration enzyme activities compared to low intertidal species that may suggest lower aerobic demand in low marsh species. Some higher marsh species showed an apparent high sensitivity to sulfide that may be related to high cytochrome c oxidase activities. In contrast, the low marsh species S. alterniflora and S. anglica had lower aerobic respiration enzyme activities and a lower sensitivity to sulfide. Thus differences in aerobic demand and sulfide sensitivity may influence estuarine species zonation.     

Changes in stomatal characteristics and photochemical efficiency during leaf development in six species of Sorbus

Measurements of Sorbus stomata size and density, maximal photochemical efficiency of photosystem II (F_v/F_m), variable-to-initial fluorescence ratio (F_v/F₀) and potential electron acceptor capacity (‘area’) were performed during leaf development in four parental diploid species, S. aria, S. aucuparia, S. chamaemespilus, S. torminalis, and two hybrid species, S. hazslinszkyana and S. intermedia. In fully expanded mature leaves, stomata lengths and densities were significantly larger in the shrub S. chamaemespilus than in the five tree species. The best performance of both the F_v/F_m and the F_v/F₀ ratio was recorded in S. intermedia, whereas S. chamaemespilus had the highest value of ‘area’. From a physiological point of view, the results of this study showed that the photosystem II reaction centers remained intact functionally through all phenological stages of leaf expansion for all examined species of Sorbus.     

An interpretation of genomic balance in seed formation in interspecific hybrids of Solanum

Summary To investigate the mechanisms of seed failure in intraspecific and interspecific crosses of Solanum two diploid, S. commersonii and Group Phureja, and one tetraploid species, S. acaule, species were crossed and the seeds were analyzed for embryo and endosperm development. Many seeds of certain crosses observed seven days after pollinations were found to contain abnormal embryos and degenerating endosperms. In some cases seeds contained an embryo with no endosperm, or an endosperm with no embryo. Other interspecific crosses which were predicted to fail actually produced seeds with normally developed embryos and endosperms. To further characterize the intra- and interspecific embryos and endosperms the nuclear DNA was measured. There are several ways to explain the ploidy levels of embryos and endosperms among the crosses, the occurrence of unreduced gametes in some cases and genomic instability in other cases. The latter resulted in chromosome loss at meiotic and mitotic divisions. Genomic balance in interspecific seeds is critical to both embryo and endosperm development.Scientific Journal Series Article No. 14636 of the Minnesota Experiment Station     

Molecular identification of seven Sarcocystis species in red deer (Cervus elaphus) from Lithuania

The diaphragm muscles of 77 free-ranging red deer (Cervus elaphus) were examined for Sarcocystis species in Lithuania. Sarcocysts were detected in 61 out of 77 (79.2%) animals investigated. A total of 60 isolated sarcocysts were identified to species using subunit I of cytochrome c oxidase (cox1) sequence analysis. Overall, seven species, S. entzerothi, S. hjorti, S. iberica, S. linearis, S. pilosa, S. truncata and S. venatoria, were confirmed in Lithuanian red deer. Sarcocystis entzerothi was reported in red deer for the first time. Previously this species was shown to use sika deer as well as roe deer and fallow deer as an intermediate host. Based on cox1, with the addition of the current data, altogether 13 Sarcocystis species have so far been shown to use red deer as an intermediate host. Species detected in red deer demonstrated considerable differences in intraspecific genetic variation at cox1. Genetic distances between different samples of S. hjorti and S. linearis were calculated using principal coordinates analysis (PCoA), implying molecular divergence of same Sarcocystis species using different hosts in the same geographical area and divergence of those employing same intermediate host species from different areas.     

Scanning Electron Microscopy of Ascospores of Schwanniomyces

Ascospores of the four recognized species of Schwanniomyces were examined by scanning electron microscopy. Spores of S. alluvius, S. castellii, and S. occidentalis, which were essentially identical, had abundant, long protuberances and wide, thin equatorial rings. The two known strains of S. persoonii differed from the other species as well as from each other. One strain had spores with a wide ring but only a few short protuberances; spores from the second strain were covered with craterlike depressions and had a narrow ring. Also examined were spores of Schwanniomyces hominis (=Saccharomyces rosei) which lacked a ring and were covered with short irregularly shaped protuberances, a finding consistent with the morphology of spores from other strains of S. rosei.     

Foliar flavonoids of North American Solanum section Solanum

A total of ten flavonoids, all flavonols, were isolated from leaves of eleven species of Solanum sect. Solanum. Most species had relatively few compounds, primarily quercetin 3-O-glycosides. Little intraspecific variation in flavonoids was observed, except in S. americanum were it correlated with previously recognized races. Flavonoid data are of little help in determining ancestries of polyploid species, but do rule out S. sarrachoides as a progenitor of S. villosum and S. nigrum. Solanum sarrachoides is unique among species examined in having free flavonoid aglycones as well as extensive 3-O-methoxylation.     

Analysis of the wild potato germplasm of the series Acaulia with AFLPs: implications for ex situ conservation

The wild potato germplasm of the series Acaulia maintained at the Centre for Genetic Resources, The Netherlands, currently consists of 314 accessions. This collection comprises seed samples of the species Solanum acaule (ssp. acaule, ssp. aemulans, ssp. palmirense and ssp. punae) and Solanum albicans collected from South America. In order to validate taxonomic classification, to investigate the extent of redundancy and to study the distribution of genetic diversity across the collection area, the entire collection was analysed with two AFLP primer pairs on two plants per accession. Within the entire sample a total number of 130 polymorphic bands were scored for the two primer pairs. An UPGMA cluster analysis grouped the majority of plants according to the species and subspecies. A total number of 16 misclassifications were identified, including four cases that did not seem to belong to the series Acaulia. Two accessions were found to consist of plants of different AFLP clusters. AFLP data also allowed the taxonomic classification of the subspecies of 97 accessions that previously were described as S. acaule only. For 126 accessions the two individuals studied displayed identical AFLP profiles. Forty six of these 126 accessions shared their profiles with both or single plants of other accessions. These were all tested for identical profiles for a third primer pair, resulting in 15 duplication groups consisting of a total number of 22 accessions and 14 single plants. Analyses of molecular variance (AMOVA) were performed to examine the distribution of genetic variation. Comparison of geographic distances between the collection site of plants and the number of AFLP polymorphisms revealed no consistent relationship between geographic distance and genetic diversity. AFLP analysis appeared to be an efficient method to verify taxonomic classification and to identify redundancies in the wild germplasm of the series Acaulia. Implications of the results for the ex situ conservation of wild potato germplasm are discussed. Received: 6 November 2000 / Accepted: 20 April 2001