Frost injury and heterogeneous ice nucleation in leaves of tuber-bearing solanum species : ice nucleation activity of external source of nucleants

The heterogeneous ice nucleation characteristics and frost injury in supercooled leaves upon ice formation were studied in nonhardened and cold-hardened species and crosses of tuber-bearing Solanum. The ice nucleation activity of the leaves was low at temperatures just below 0°C and further decreased as a result of cold acclimation. In the absence of supercooling, the nonhardened and cold-hardened leaves tolerated extracellular freezing between −3.5° and −8.5°C. However, if ice initiation in the supercooled leaves occurred at any temperature below −2.6°C, the leaves were lethally injured.

To prevent supercooling in these leaves, various nucleants were tested for their ice nucleating ability. One% aqueous suspensions of fluorophlogopite and acetoacetanilide were found to be effective in ice nucleation of the Solanum leaves above −1°C. They had threshold temperatures of −0.7° and −0.8°C, respectively, for freezing in distilled H₂O. Although freezing could be initiated in the Solanum leaves above −1°C with both the nucleants, 1% aqueous fluorophlogopite suspension showed overall higher ice nucleation activity than acetoacetanilide and was nontoxic to the leaves. The cold-hardened leaves survived between −2.5° and −6.5° using 1% aqueous fluorophlogopite suspension as a nucleant. The killing temperatures in the cold-hardened leaves were similar to those determined using ice as a nucleant. However, in the nonhardened leaves, use of fluorophlogopite as a nucleant resulted in lethal injury at higher temperatures than those estimated using ice as a nucleant.

     

Growth, partitioning, and harvest index of tuber-bearing solanum genotypes grown in two contrasting peruvian environments

Ten Solanum potato genotypes, including four primitive species and six hybrids, were grown to maturity near 230 and 3273 meters in elevation at two sites, Coast and Sierra, in Peru. Growth data, with emphasis upon tubers and leaves, were collected periodically to analyze the plant components which differed in these contrasting environments. Nine of the Solanum species/cultivars effectively partitioned dry matter into tubers with values reaching 73 to 85% of the total plant at mature harvest in the Sierra but dropping to 33 to 75% on the Coast. These harvest index differences were, however, accompanied by no consistent changes in total leaf area, specific leaf area, nor number of tuber initiated. Consistent differences did occur in having shorter plants in the Sierra and an increased tuber dry matter percentage, 20 to 28%, in the Sierra compared to 14 to 21% on the Coast. Linear relationships exist between plant tuber harvest index versus tuber yield and versus total plant dry matter on both the Coast and in the Sierra.     

Allelism of Endosperm Balance Number (EBN) in Mexican tuber-bearing Solanum species

Summary Endosperm Balance Number (EBN) is a genetic, dose-dependent crossability system functioning in tuber-bearing Solanum species. Each species has been assigned 1EBN, 2EBN, or 4EBN. Species thus designated cross only within their EBN group. Doubling of chromosome number also doubles the EBN. The ploidy: EBN ratio is not consistent among Solanum species. Some diploids are 2EBN while others are 1EBN. Some tetraploids are 4EBN while others are 2EBN. Species from Mexico typically have EBNs that are one-half of their ploidy [e.g. 2x(1EBN), 4x(2EBN)]. Hybrids of Mexican species and a South American species, 2x(1EBN) S. Commersonii, and its 4x(2EBN) colchicine derivative were made and crossed to 1, 2, and 4EBN standard testers to determine the relationship of the genetic organization of EBN among and within these species. Diploid hybrids crossed only to 1EBN standard testers. Hybrids of 4x(2EBN) S. commersonii and 4x(2EBN) Mexican species crossed almost exclusively to 2EBN standard testers. Complex tetraploid hybrids involving S. commersonii, S. stenophyllidium (a Mexican diploid), and Mexican tetraploids of series Longipedicellata also crossed only to 2EBN testers. The apparent lack of recombination and segregation for EBN in these hybrids indicates that the genomes of the Mexican diploid and tetraploid species carry EBN in a way genetically similar to that of the South American species S. Commersonii.Cooperative investigation of the U.S. Department of Agriculture, Agricultural Research Service and the Wisconsin Experiment Station. Supported in part by the USDA/Cooperative States Research Service Competitive Grant No. 83-CRCR-1-1253     

Endosomes differ from plasma membranes in the phospholipid molecular species composition

125I-Labeled epidermal growth factor was incorporated into and highly concentrated in endosomes of Chinese hamster V79-UF cells during incubation at 37 degrees C for 8 min after binding to its receptors on the cell surface at 4 degrees C. From the labeled cells, endosomes were isolated by isopycnic centrifugation on a Percoll density gradient and then sucrose density gradients. The isolated endosomes were mostly free from contamination by Golgi, endoplasmic reticulum, lysosome, plasma membrane and mitochondria. Endosome membranes were found to differ from plasma membranes in the phospholipid composition. Sphingomyelin and phosphatidylserine were enriched in endosomes, compared with plasma membranes. Diacylphosphatidylcholine and diacylphosphatidylethanolamine were major phospholipids of the membranes in both organelles. The contents of molecular species of diacylphosphatidylcholine and diacylphosphatidylethanolamine with two monoenoic fatty acids were lower in endosomes than in plasma membranes. The differences in the polar head group and molecular species compositions of phospholipids between endosomes and plasma membranes did not change, regardless of whether or not the proportions of phospholipid molecular species in plasma membranes changed. The significance of the lipids in endosomes is discussed.     

The variation and evolution of selected species of solanum section basarthrum

A study of greenhouse-grown and field populations ofSolarium caripense Humboldt & Bonpland ex Dunal,S. tabanoense Correll, andS. trachycarpum Bitter & Sodiro, all diploid species (n = 12) of high altitudes of southern Central America and northern South America, revealed great morphological variation. Polygonographs utilizing seven characters (number of leaflets, leaf index, length of pubescence, number of flowers, anther length, corolla index, and pollen diameter) showed a wide range of variation and led to recognition of six morphologically distinct groups. Hybridizations with greenhouse populations showed that five of the morphological groups are reproductively isolated as well. A complex pattern of genetic variation involving various degrees and combinations of low crossing success, low seed set, lowered F₁ pollen fertility, and nonreciprocal crossability was found. Examinations of meiotic figures from hybrids revealed no gross chromosomal structural differences. Evidence indicates that genetic differences, including gene-cytoplasm interactions, are significant isolating barriers. A key to the species studied plus appropriate taxonomic notes are provided.Solanum heiseri is described as new, andS. trachycarpum is placed inS. sect.Basarthrum seriesCaripensia.     

Comparison of polyphenol oxidase expression in glandular trichomes of solanum and lycopersicon species

Tetralobulate glandular trichomes are present on the foliage of many solanaceous species. Resistance of many of these species to insects is conditioned by the ability of trichomes to rupture upon contact and to rapidly polymerize their contents, resulting in entrapment of insects in hardened trichome exudate. In the wild potato, Solanum berthaultii, polymerization of trichome exudate is initiated by a soluble M_r 59,000 polyphenol oxidase (PPO), which is a dominant protein constituent of the organ. PPOs, although ubiquitous in angiosperms, typically display great heterogeneity in molecular weight and are found at low levels in plant cells. Because of the unusually high accumulation and tissue-specific expression of the M_r 59,000 PPO in S. berthaultii glandular trichomes, we analyzed trichome proteins of a number of Lycopersicon and Solanum species to assess the extent to which possession of the M_r 59,000 PPO is conserved. Trichomes were collected manually and examined for PPO activity, immuno-cross-reactivity with S. berthaultiiM_r 59,000 PPO, and protein content. In addition, N-terminal amino acid sequences were obtained for five trichome PPOs. All species analyzed possessed trichome PPOs similar in structure and level of expression to that of S. berthaultii. The relationship between sequences and structures of these conserved PPOs and the variable PPOs of leaf is discussed.     

Antibodies to ovine adipocyte plasma membranes recognize tissue and species specific plasma membrane components

1. Ovine adipocyte plasma membrane (PM) contains three unique proteins that have relative molecular mass of 70, 106, and 110 kD which are lacking in PM from liver, kidney, heart, and red blood cells. 2. Two major proteins on ovine adipocyte PM having molecular mass of 44 and 46 kD which were also present on porcine adipocyte PM. 3. These ovine proteins could not be detected on either rat or chicken adipocyte PM.     

Acidic phospholipid species inhibit adenylate cyclase activity in rat liver plasma membranes.

Incubation of rat liver plasma membranes with liposomes of dioleoyl phosphatidic acid (dioleoyl-PA) led to an inhibition of adenylate cyclase activity which was more pronounced when fluoride-stimulated activity was followed than when glucagon-stimulated activity was followed. If Mn2+ (5 mM) replaced low (5 mM) [Mg2+] in adenylate cyclase assays, or if high (20 mM) [Mg2+] were employed, then the perceived inhibitory effect of phosphatidic acid was markedly reduced when the fluoride-stimulated activity was followed but was enhanced for the glucagon-stimulated activity. The inhibition of adenylate cyclase activity observed correlated with the association of dioleoyl-PA with the plasma membranes. Adenylate cyclase activity in dioleoyl-PA-treated membranes, however, responded differently to changes in [Mg2+] than did the enzyme in native liver plasma membranes. Benzyl alcohol, which increases membrane fluidity, had similar stimulatory effects on the fluoride- and glucagon-stimulated adenylate cyclase activities in both native and dioleoyl-PA-treated membranes. Incubation of the plasma membranes with phosphatidylserine also led to similar inhibitory effects on adenylate cyclase and responses to Mg2+. Arrhenius plots of both glucagon- and fluoride-stimulated adenylate cyclase activity were different in dioleoyl-PA-treated plasma membranes, compared with native membranes, with a new 'break' occurring at around 16 degrees C, indicating that dioleoyl-PA had become incorporated into the bilayer. E.s.r. analysis of dioleoyl-PA-treated plasma membranes with a nitroxide-labelled fatty acid spin probe identified a new lipid phase separation occurring at around 16 degrees C with also a lipid phase separation occurring at around 28 degrees C as in native liver plasma membranes. It is suggested that acidic phospholipids inhibit adenylate cyclase by virtue of a direct headgroup specific interaction and that this perturbation may be centred at the level of regulation of this enzyme by the stimulatory guanine nucleotide regulatory protein NS.     

A new species of solanum section geminata (Solanaceae) from western Mexico

Solanum malacothrix S. Knapp is described from the slopes of the Sierra Madre del Sur in the Río Balsas drainage in the state of Guerrero. It is apparently rare, and has been collected only twice. Its relationship to other species in Mexico and northern South America is discussed.     

The role of reactive oxygen species in copper-induced permeability of plasma membranes in Escherichia coli

The role of active oxygen species in the induction of nonselective cationic permeability of the plasma membrane of bacteria Escherichia coli B by the action of Cu2+ ions was studied. It was found that the increase in the amount of active oxygen species in the suspension after treating cells with copper occurred synchronously with the leakage of K+ cations from them. Evidence is presented that active oxygen species formed during the interaction of copper ions with bacteria under aerobic conditions are not involved in the induction of channel conductivity in the membrane. Moreover, the ability of oxygen to protect the membrane from the toxic action of copper was shown, and the activation of membrane damage by external reductants was confirmed. These data suggest that the barrier properties of the membrane are disturbed during the interaction of Cu+ ions with critical targets on the surface, the concentration of Cu+ being determined by all redox processes in the near-membrane space.     

Temperature effect of cholesterol association with synaptosomal plasma membranes of rabbit brain.

Association of exogenous cholesterol with rabbit brain synaptosomal plasma membranes follows an exponential path described by the general formula y = a X ebx. The co-operative nature of this association was shown when increasing amounts of unlabelled cholesterol glucoside (up to 0.5 mM) were added to a fixed amount (5 microM) of [14C]cholesterol, when a biphasic curve of the binding of [14C]cholesterol into the membranes was obtained. Arrhenius plots of this association revealed two break points which occur at 25 degrees C and 42 degrees C. The first break apparently corresponds to the transition from the crystalline to the gel phase. The second break may be due to the (continuously) increasing entropy of the system which creates at a certain point difficulties in the binding of cholesterol into the lipid bilayer.     

A new species of solanum section petota (Solanaceae) from northern Peru

Solanum guzmanguense. is a new species, of the wild potato alliance endemic in northern Peru. Its nearest relationships are with a small cluster of species restricted to southern Bolivia and northern Argentina.     

Changes in micro RNA expression in a wild tuber-bearing Solanum species induced by 5-Azacytidine treatment

Phenotypic plasticity is often postulated as a principal characteristic of tuber-bearing wild Solanum species. The hypotheses to explore this observation have been developed based on the presence of genetic variation. In this context, evolutionary changes and adaptation are impossible without genetic variation. However, epigenetic effects, which include DNA methylation and microRNAs expression control, could be another source of phenotypic variation in ecologically relevant traits. To achieve a detailed mechanistic understanding of these processes, it is necessary to separate epigenetic from DNA sequence-based effects and to evaluate their relative importance on phenotypic variability. We explored the potential relevance of epigenetic effects in individuals with the same genotype. For this purpose, a clone of the wild potato Solanum ruiz-lealii, a non-model species in which natural methylation variability has been demonstrated, was selected and its DNA methylation was manipulated applying 5-Azacytidine (AzaC), a demethylating agent. The AzaC treatment induced early flowering and changes in leaf morphology. Using quantitative real-time PCR, we identified four miRNAs up-regulated in the AzaC-treated plants. One of them, miRNA172, could play a role on the early flowering phenotype. In this work, we showed that the treatment with AzaC could provide meaningful results allowing to study both the phenotypic plasticity in tuber-bearing Solanum species and the inter-relation between DNA methylation and miRNA accumulations in a wide range of species.     

Simultaneous detection and functional response of testosterone and estradiol receptors in osteoblast plasma membranes

Osteoblasts derived from the periosteal surfaces of two-three-week-old male broiler chicken tibias were cultured for eight days. The cells were then loaded with fura-2/AM ester to detect surges in intracellular Ca(2+). Treatment with 10(-7) M testosterone (T) or 17beta-estradiol (E) elicited a rapid (within seconds) response that was substantially reduced by introducing the calcium chelating agent EGTA or the calcium-channel blocker verapamil. The hormones were equally effective when covalently linked to bovine serum albumin (BSA), a procedure that ensures the hormone does not enter the cells. The rapid response to surface-bound steroids indicates that the responses were invoked through plasma-membrane receptors. The source of Ca(2+) was shown to be through entry from external sources, as well as from intracellular stores. Flow cytometry of fluorescein-tagged T-BSA and E-BSA revealed that osteoblasts derived from male chickens had similar and substantial levels of both receptors.     

The utility of NBS profiling for plant systematics: a first study in tuber-bearing Solanum species

Systematic relationships are important criteria for researchers and breeders to select materials. We evaluated a novel molecular technique, nucleotide binding site (NBS) profiling, for its potential in phylogeny reconstruction. NBS profiling produces multiple markers in resistance genes and their analogs (RGAs). Potato (Solanum tuberosum L.) is a crop with a large secondary genepool, which contains many important traits that can be exploited in breeding programs. In this study we used a set of over 100 genebank accessions, representing 49 tuber-bearing wild and cultivated Solanum species. NBS profiling was compared to amplified fragment length polymorphism (AFLP). Cladistic and phenetic analyses showed that the two techniques had similar resolving power and delivered trees with a similar topology. However, the different statistical tests used to demonstrate congruency of the trees were inconclusive. Visual inspection of the trees showed that, especially at the lower level, many accessions grouped together in the same way in both trees; at the higher level, when looking at the more basal nodes, only a few groups were well supported. Again this was similar for both techniques. The observation that higher level groups were poorly supported might be due to the nature of the material and the way the species evolved. The similarity of the NBS and AFLP results indicate that the role of disease resistance in speciation is limited.     

Calcium transport activities of plasma membranes isolated from the livers of various animal species

1. Plasma membranes of comparable yield and purity were isolated from the livers of various animal species belonging to phylogenetic groups from Amphibia to Mammalia. 2. Calcium transport activity was observed in all liver plasma membranes examined. 3. No phylogenetic pattern of expression of the liver plasma membrane calcium transport system was observed, with the order of activity being: guinea pig greater than rabbit greater than frog greater than chicken = hamster greater than rat = budgerigar = turtle greater than beef cattle greater than mouse = duck. 4. Calcium transport activity was only 9.7 and 8.7% of adult frog levels in plasma membranes isolated from the livers of tadpoles without and with limbs, respectively. 5. Liver plasma membrane calcium transport activity was 25% higher in adult chickens than in day-old chicks. 6. A possible role for thyroid hormone in the development of the liver plasma membrane calcium transport system is discussed.