Influence of alternating temperature preculture on cryopreservation results for potato shoot tips

Cryopreservation is the most suitable long-term storage method for genetic resources of vegetatively maintained crops like potato. In the Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) the DMSO droplet method is applied, and so far more than 1000 accessions are cryopreserved with an average regeneration rate of 58%. New experiments with four potato accessions using alternating temperatures (22/8 degrees C day/night temperature, 8 h photoperiod, 7 d) prior to cryopreservation showed improved regeneration. The influence of this preculture on the shoot tips was studied for two wild, frost resistant species Solanum acaule and S. demissum and for two cultivated, frost sensitive potatoes S. tuberosum 'Désirée' and 'King Edward'. Comparison of liquid and solid media after cryopreservation showed improved regeneration on solid media with higher regeneration percentages, less callus formation and better plantlet structure. In comparative analyses biochemical factors like soluble sugars, starch, and amino acid concentrations were measured. Shoot tips after constant and after alternating temperature preculture were analyzed. Total concentrations of soluble sugars (glucose, fructose, and sucrose) were higher for all accessions after the alternating temperature preculture, which could be the reason for improved cryopreservation results.     

Effect of mycorrhization on the accumulation of rishitin and solavetivone in potato plantlets challenged with<Emphasis Type="Italic"> Rhizoctonia solani</Emphasis>

The effect of colonization with the vesicular-arbuscular mycorrhizal fungus Glomus etunicatum on the content of rishitin and solavetivone was determined in potato plants cv. Goldrush challenged with Rhizoctonia solani. Mycorrhization stimulated significantly the accumulation of both phytoalexins in roots of plantlets challenged with R. solani but did not influence phytoalexin levels in non-challenged plantlet roots. No accumulation of solavetivone or rishitin was detected in shoots. In Petri dish bioassays, rishitin and solavetivone inhibited mycelial growth of R. solani.     

Influence of the nitrogen level on root growth and morphology of two potato varieties differing in nitrogen acquisition

Two potato (Solanum tuberosum L.) cultivars (Astrid and Bodenkraft) differing in their nitrogen acquisition from the soil (Hunnius, 1981) were used in nutrient solutions to study the effect of increasing concentrations of nitrate (0.05; 0.5; 5.0 mol m^-3) particularly on root growth and morphology. In each variety increasing nitrogen concentrations stimulated shoot growth more than root growth. At all nitrate concentrations, the variety with higher nitrogen acquisition (Astrid) had a significantly larger root system. The larger root system of Astrid compared to Bodenkraft was particularly evident when surface area and total length of the roots, instead of root dry weight were used as parameters. The results stress the importance of root length and surface area for nitrogen acquisition from soils.     

Examination of the hybrid origin of the wild potato Solanum ruiz-lealii Brücher

The common potato, tetraploid Solanum tuberosum spp. tuberosum L. (tbr), has a narrow genetic base but a large number of related tuber-bearing species that harbor genetic diversity for agronomic characters. The taxonomic status of Solanum ruiz-lealii Brücher (rzl), a diploid species endemic to Mendoza province, Argentina, is controversial. It has been described as a new species of non-hybrid origin and as a natural hybrid between S. kurtzianum Bitt. & Wittm. (ktz) and S. chacoense Bitt. (chc). The hypothesis of the hybrid origin of rzl is examined systematically for the first time by phenetic analyses of morphological and molecular (SSR markers) data, and cytological analyses of interspecific hybrids. The morphological, cytological and molecular data are congruent, and suggest that rzl is not a recent natural hybrid between the ktz and the chc populations studied but has probably originated by divergence of chc, or by hybridization between chc and another taxon.     

Effect of temperature shock and elevated incubation temperature on androgenic embryo yield of diploid potato

Using three diploid tuber-bearing Solanum clones as anther donors, experiments were conducted on the effect of high temperature shock and elevated incubation temperature during anther culture on androgenic embryo production. Five incubation treatments were tested on two clones and three treatments were repeated in a second experiment on one of the same clones and an additional one. In the first experiment, temperature treatment, genotype, date of culture initiation, and their interactions were all significant sources of variation. A treatment combining a high temperature shock (35 °C for 12 h) with elevated incubation temperature (30/20 °C) yielded 11 times as many embryos (44 per flask) as the control 20 °C (4 per flask). By conducting several replications per day of bud collection, the significant variation due to experimental dates was separated from experimental error to provide a more sensitive test of treatment effects. Temperature shock (35 °C 12h) during anther culture did not appear to influence the subsequent conversion rate of androgenic embryos.     

Entomopathogenic fungi (Hypocreales) for control of potato psyllid, Bactericera cockerelli (Šulc) (Hemiptera: Triozidae) in an area endemic for zebra chip disease of potato

Potato psyllid, Bactericera cockerelli, is a serious pest of potato and other solanaceous vegetables in the United States, Mexico, Central America, and New Zealand and is responsible for transmission of Candidatus Liberibacter solanacearum which causes a disease known as “zebra chip” (ZC). Entomopathogenic fungi could provide a viable component for an integrated pest management strategy for control of B. cockerelli and other potato pest insects. Three field trials of commercial formulations of Metarhizium anisopliae (F 52®, Novozymes Biologicals) and Isaria fumosorosea (Pfr 97®, Certis USA) and abamectin (Agri-Mek®, Syngenta, USA) were conducted in Weslaco, Texas. Rates are expressed in quantity of product delivered in 375–470 l of water/ha. F 52 applied at 0.51, 1.1, and 2.2 l/ha and Agri-Mek applied at 584 ml/ha produced reductions of B. cockerelli eggs and nymphs of 45%, 59%, 67%, and 63%, respectively. Only Agri-Mek significantly reduced plant damage. Pfr 97 at 1.1 kg/ha with and without 1% Trilogy® (neem oil, Certis, USA), and Agri-Mek at 584 ml/ha resulted in psyllid reductions of 78%, 76%, and 84%, respectively. Significantly decreased plant damage and ZC symptoms were observed for all treatments. Tuber yields for Pfr plus Trilogy and Agri-Mek were significantly higher than the control. F 52 applied at 1.1 and 2.2 l/ha and Pfr 97 at 1.1 and 2.2 kg/ha produced 62%, 62%, 66%, and 65% reduction, respectively. Tuber yield for both rates of Pfr and the high rate of F 52 were significantly higher than the control. All fungal treatments significantly reduced plant damage and ZC symptoms.     

Stimulation of in vitro root and shoot growth of potato by increasing sucrose concentration in the presence of fluridone,an inhibitor of abscisic acid synthesis

Fluridone, an inhibitor of abscisic acid (ABA) biosynthesis, strongly stimulated rooting of nodal stem segments of potato (Solanum tuberosum L.) cultivar Arran Banner cultured in darkness on tuberisation medium. Inclusion of 10^-6 M ABA in the culture medium prevented this rooting response, indicating that root proliferation in the presence of fluridone could be due to inhibition of ABA synthesis. The rooting response to fluridone (increased total root number and root fresh weight) was obtained only at high sucrose concentrations (0.175 and 0.234 M) and was demonstrated with two potato cultivars and two culture media; one which favoured tuberisation and one which did not. Shoot numbers were also increased, but to a lesser extent than root numbers, and total fresh weight of plant material per culture was greatly increased by inclusion of both fluridone (10^-6 or 10^-5 M) and 0.234 M sucrose in the culture medium. The role of sucrose was not simply osmotic because when the osmolarity of fluridone medium was increased using mixtures of mannitol and sucrose, no root proliferation occurred unless sucrose predominated in the mixture.     

The effect of NaCl on proline accumulation in potato seedlings and calli

The effects of salt stress were studied on the accumulation and metabolism of proline and its correlation with Na⁺ and K⁺ content in shoots and callus tissue of four potato cultivars, viz., Agria, Kennebec (relatively salt tolerant), Diamant and Ajax (relatively salt sensitive). Na⁺ and proline contents increased in all cultivars under salt stress. However, K⁺ and protein contents decreased in response to NaCl treatments. The activities of enzymes involved in proline metabolism, Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (ProDH) increased and decreased, respectively, in response to elevated NaCl concentrations. The changes of P5CS and ProDH activities in more salt sensitive cultivars (Diamant, Ajax) were more than those in the tolerant ones. Then the stimulation of synthesis in combination with a partially increase of protein proteolysis, a decrease in proline utilization and inhibition of oxidation resulted in high proline contents in seedlings and calli under salt stress. In callus tissue, reduced growth and cell size may be partially responsible for high proline accumulation in response to high NaCl levels. However, although the basic proline contents in the seedlings of more salt tolerant cultivars were higher than the sensitive ones, a clear relationship was not generally observed between accumulation of proline and salt tolerance in potato.     

Sucrose and light effects on in vitro cultures of potato,chokecherry and saskatoon berry during low temperature storage

Cultures of potato (Solanum tuberosum) cv. Atlantic, chokecherry (Prunus virginiana L.) cv. Garrington and saskatoon berry (Amelancher alnifolia Nutt.) cv. Northline grown in vitro for 3 weeks at 24/22 °C, 16-h photoperiod, 150 μmol m⁻² s⁻¹ photosynthetic photon flux density (PPFD) mixed fluorescent/incandescent light were stored for 6, 9 and 12 weeks at 4 °C under 0 (darkness) and 3 μmol m⁻² s⁻¹ PPFD (690 nm red light continuous illumination). Growth regulators free MSMO medium either with or without 30 g l⁻¹ sucrose was used to store the cultures. All cultures retained capacity to re-grow after storage. Tested factors, sucrose, light and the length of the storage period had an impact on shoot quality and re-growth capacity of the cultures. For either light treatment sucrose was essential for the low temperature maintenance of vigorous stock plants of potato, if stored for over 6 weeks. Chokecherry and saskatoon cultures stored well without sucrose; although chokecherry benefited from sucrose in the storage medium when the stock cultures were kept at the low temperature for 12 weeks. Low light significantly improved quality of the stored potato cultures, but had very little effect on both chokecherry and saskatoon berry cultures. The woody plant cultures grew during storage, and the longer the stock plants were stored, the more vigorous cultures they generated. The results indicate that growers can successfully use their existing facilities, small refrigerators and coolers with low light intensity, set at 4 °C, for short term storage of potato, chokecherry and saskatoon berry cultures. The potato cultures, which are known to be sensitive to prolonged low temperature storage, should be frequently monitored and subcultured as required. On the other hand, the woody plant stock cultures do not require any special attention when kept at 4 °C and re-grow the most vigorous shoots if stored for at least 12 weeks. This revised version was published online in August 2006 with corrections to the Cover Date.     

QTL analysis of potato tuberization

Quantitative trait loci (QTLs) affecting tuberization were detected in reciprocal backcrosses between Solanum tuberosum and S. berthaultii. Linkage analyses were performed between traits and RFLP alleles segregating from both the hybrid and the recurrent parent using a set of framework markers from the potato map. Eleven distinct loci on seven chromosomes were associated with variation in tuberization. Most of the loci had small effects, but a QTL explaining 27% of the variance was found on chromosome 5. More QTLs were detected while following alleles segregating from the recurrent S. tuberosum parent used to make the backcross than were detected by following alleles segregating from the hybrid parent. More than half of the alleles favoring tuberization were at least partly dominant. Tuberization was favored by an allele from S. berthaultii at 3 of the 5 QTLs detected by segregation from the hybrid parent. The additive effects of the QTLs for tuberization explained up to 53% of the phenotypic variance, and inclusion of epistatic effects increased this figure to 60%. The most common form of epistasis was that in which presence of an allele at each of 2 loci favoring tuberization was no more effective than the presence of a favorable allele at 1 of the 2 loci. The QTLs detected for tuberization traits are discussed in relationship to those previously detected for trichome-mediated insect resistance derived from the unadapted wild species.Paper number 54 of the Department of Fruit and Vegetable Science, Cornell University     

The effect of starch and ineubation temperature in anther culture of potato

Three Andean tetraploid potato genotypes (2n=48) and 7 anther-derived dihaploids (2n=24) originating from two of the tetraploids were used in anther culture. Relative number of embryos/vial was significantly higher when the anther culture media was gelatinized with 3% potato starch than when Gelrite or wheat starch (3%) were used as gelatinizing agents. The degree of anther culture response varied between tetraploids but also within a group of related dihaploids. Additionally, the embryo production of individual genotypes, tetraploids as well as dihaploids, was dependent on the incubation temperature (10, 15, 20, 25, 30°C) of the anther culture. The incubation temperature of the anther culture was also important for the regeneration rate. Direct regeneration was mostly stimulated when the anther culture was incubated at 20°C.Abbreviations BAP 6-benzylaminopurine - IAA indole-3-acetic acid     

Nutrient uptake by potato crops grown on two soils with contrasting physical properties

Potatoes were grown on two contrasting soils but in adjacent sites to investigate the effect of soil type on tuber production, nutrient uptake and nutrient inflow rates (uptake rate per unit length of root). The year of the study was wetter than normal. Tuber growth, root growth and nutrient uptake were all greater on the coarse rather than the fine-textured soil. However there was no difference in nutrient inflow rates between plants growing in the two soils. Therefore, it was concluded that the crop on the finer textured soil did not have an adequate nutrient supply, particularly of N, relative to the crop on the coarser-textured soil. The reasons for the low supply of nitrogen in the fine textured soil are not clear, but it might have been due to the smaller root system or to enhanced losses of nitrogen by denitrification caused by the combination of soil physical properties and poor drainage in a wet year.     

QTL analysis of potato tuber dormancy

The potential loss of chemical sprout inhibitors because of public concern over the use of pesticides underscores the desirability of breeding for long dormancy of potato (Solanum tuberosum L.) tubers. Quantitative trait locus (QTL) analyses were performed in reciprocal backcrosses between S. tuberosum and S. berthaultii toward defining the complexity of dormancy. S. berthaultii is a wild Bolivian species characterized by a short-day requirement for tuberization, long tuber dormancy, and resistance to several insect pests. RFLP alleles segregating from the recurrent parents as well as from the interspecific hybrid were monitored in two segregating progenies. We detected QTLs on nine chromosomes that affected tuber dormancy, either alone or through epistatic interactions. Alleles from the wild parent promoted dormancy, with the largest effect at a QTL on chromosome 2. Long dormancy appeared to be recessive in the backcross to S. berthaultii (BCB). In BCB the additive effects of dormancy QTLs accounted for 48% of the measured phenotypic variance, and adding epistatic effects to the model explained only 4% more. In contrast, additive effects explained only 16% of the variance in the backcross to S. tuberosum (BCT), and an additional 24% was explained by the inclusion of epistatic effects. In BCB variation at all QTLs detected was associated with RFLP alleles segregating from the hybrid parent; in BCT all QTLs except for two found through epistasis were detected through RFLP alleles segregating from the recurrent parent. At least three dormancy QTLs mapped to markers previously found to be associated with tuberization in these crosses.Paper number 55 of the Department of Fruit and Vegetable Science, Cornell University     

Influence of culture temperature on the growth,biochemical composition and fatty acid profiles of six Antarctic microalgae

The growth, biochemical composition and fatty acid profiles of six Antarctic microalgae cultured at different temperatures, ranging from 4, 6, 9, 14, 20 to 30 ^∘C, were compared. The algae were isolated from seawater, freshwater, soil and snow samples collected during our recent expeditions to Casey, Antarctica, and are currently deposited in the University of Malaya Algae Culture Collection (UMACC). The algae chosen for the study were Chlamydomonas UMACC 229, Chlorella UMACC 234, Chlorella UMACC 237, Klebsormidium UMACC 227, Navicula UMACC 231 and Stichococcus UMACC 238. All the isolates could grow at temperatures up to 20 ^∘C; three isolates, namely Navicula UMACC 231 and the two Chlorella isolates (UMACC 234 and UMACC 237) grew even at 30 ^∘C. Both Chlorella UMACC 234 and Stichococcus UMACC 238 had broad optimal temperatures for growth, ranging from 6 to 20 ^∘C (μ = 0.19 – 0.22 day^–1) and 4 to 14 ^∘C (μ = 0.13 – 0.16 day^–1), respectively. In contrast, optimal growth temperatures for NaviculaUMACC 231 and Chlamydomonas UMACC 229 were 4 ^∘C (μ = 0.34 day^–1) and 6–9 ^∘C (μ = 0.39 – 0.40 day^–1), respectively. The protein content of the Antarctic algae was markedly affected by culture temperature. All except Navicula UMACC 231 and Stichococcus UMACC 238 contained higher amount of proteins when grown at low temperatures (6–9 ^∘C). The percentage of PUFA, especially 20:5 in Navicula UMACC 231 decreased with increasing culture temperature. However, the percentages of unsaturated fatty acids did not show consistent trend with culture temperature for the other algae studied.     

Influence of root temperature on growth of Pinus sylvestris,Fagus sylvatica,Tilia cordata and Quercus robur

One-year-old tree seedlings were incubated in a greenhouse from April to July, under natural daylight conditions, with their root systems at constant temperatures of 5, 10, 15, 20, 25, 30 and 35 °C and with the above ground parts kept at a constant air temperature of 18–20 °C. The course of height growth, total mass increment, root, shoot and leaf weight as well as leaf areas were measured. The results indicate that clear differences exist in the optimal root zone temperatures for various growth parameters in different tree species. Pinus sylvestris had a maximal height increment at about 5–10 °C and maximal total mass increment at 15 °C root temperature. In contrast, the optimum for Quercus robur was at 25 °C. Tilia cordata and Fagus sylvatica had their optima for most growth parameters at 20 °C. The root temperature apparently indirectly influenced photosynthesis (dry weight accumulation) and respiration loss. From the observed symptoms and indications in the literature it seems probable that a change in hormone levels is involved as the main factor in the described effects. Variation of root temperature had only an insignificant effect on bud burst and the time at which the shoots sprouted. Apparently species of northern origin seem to have lower root temperature optima than those of more southern origin. This is to be verified by investigation of other tree species.     

Phenotypic characterization and bulk segregant analysis of anther culture response in two backcross families of diploid potato

Two diploid (2n=2x=24) backcross potato populations (PBCp, and CBC) were characterized for anther culture response (ACR). PBCp (Solanum phureja Juz. & Buk. genotype 1-3 × CP2) and CBC (CP2 × S. chacoense Bitt. genotype 80-1) resulted from a cross between CP2 (intermediate ACR) and its parents, S. chacoense 80-1(low ACR) and S. phureja 1-3 (high ACR). Three components of ACR were initially investigated: embryos per anther (EPA), embryo regeneration rate and percent monoploids (2n=1x=12) among regenerants. EPA was selected for further characterization because of its relative stability. In a series of studies of EPA on a total of 44 genotypes within CBC, nine high (mean EPA=2.5) and ten low (mean EPA=0.02) selections were made. In PBCp, ten high (mean EPA= 4.7) and ten low (mean EPA= 0.05) selections were made from 67 genotypes. High and low selections were used for bulk segregant analysis to screen 214 RAPD primers as candidate markers linked to EPA. Bands amplified by OPQ-10 and OPZ-4 were associated in coupling and repulsion, respectively, to ACR in PBCp. A band amplified by OPW-14 primer was associated in coupling to ACR in CBC. One-way ANOVAs using presence/absence of each candidate band to classify additional genotypes in each population verified association of the markers with EPA.     

Mould growth on building materials under low water activities. Influence of humidity and temperature on fungal growth and secondary metabolism

The influence of relative humidity (RH) and temperature on growth and metabolism of eight microfungi on 21 different types of building material was investigated. The fungi were applied as a dry mixture to the materials, which were incubated at 5°C, 10°C, 20°C and 25°C at three humidity levels in the range 69–95% RH over 4–7 months. The lower limit for fungal growth on wood, wood composites and starch-containing materials was 78% RH at 20–25°C and increased to 90% RH at 5°C. An RH of 86% was necessary for growth on gypsum board. Ceramic materials supported growth at RH >90%, although 95% RH was needed to yield chemically detectable quantities of biomass. Almost exclusively only Penicillium, Aspergillus and Eurotium (contaminant) species grew on the materials. Production of secondary metabolites and mycotoxins decreased with humidity and the quantities of metabolites were insignificant compared with those produced at high RH (RH >95%), except in the case of Eurotium.