Reconsideration of the term ‘vitrification’ as used in micropropagation

The term vitrification is currently used to describe two types of processes related to tissue-cultured plant material. The first is used to describe organs and tissues having an abnormal morphological appearance and physiological function. The second is used to describe the transition from liquid to solid state, i.e. the formation of ice during low temperature storage of in vitro cultured cells, tissues and organs. Use of the same term to define two greatly different processes in the same research area can only lead to confusion, especially for key words. Thus it is appropriate to reconsider the usage of vitrification in the first sense mentioned above. It is recommended that the term vitrification should no longer be used to indicate plant material with an abnormal morphological appearance and physiological function, and should be substituted by the term hyperhydricity.This paper was written as a follow up of the discussions held during the workshop on Vitrification at the IAPTC-Congress in Amsterdam (1990). Different eminent research workers in the field of vitrification were invited to formulate their preference for a new terminology, and they came to a consensus.     

Importance of the iron chelate formula for micropropagation of Rosa hybrida L. ‘Moneyway’

In vitro propagation of the rose rootstock Moneyway was investigated on the following media: Murashige and Skoog (MS), Quoirin and Lepoivre (QL) and Woody Plant (WP). Growth, which was measured as length of shoots after a 6-week period, was faster on MS and QL than on WP. In spite of the better growth, chlorosis of newly formed leaves occurred from the third week on and was correlated with a lower chlorophyll content of shoots. Replacement of FeEDTA by FeEDDHA in QL and MS resulted in the development of green shoots for more than 3 months. The occurrence of chlorosis was not pH directed since the pH of QL with FeEDTA or FeEDDHA had not changed after 6 weeks of growth. Addition of the light absorbing dye fast yellow 9 to QL with FeEDTA also resulted in green shoots with a higher chlorophyll content. It is suggested that FeEDDHA is a more photostable chelate than FeEDTA, resulting in a higher availability of iron for the rose shoots. The impact of the iron chelate formula on the micropropagation of plant species that are susceptible to iron deficiency is discussed.Abbreviations BA 6-benzyladenine, fast yellow 9-4-amino-1,1-azobenzene-3,4-disulfonic acid - FeEDTA ferric ethylenediamine tetraacetate - FeEDDHA ferric ethylenediamine di(o-hydroxyphenylacetate) - IAA indole-3-acetic acid - IBA indole-3-butyric acid - LSD least significant difference - NAA -naphthaleneacetic acid - P probability     

Imitation of lever-pressing ‘strategies’ during the operant conditioning of albino rats

This experiment was carried out on ninety-one male Wistar rats in order to study the ability to learn by observation in an operant conditioning situation. All rats were trained on a ‘multiple’ schedule over twelve daily sessions. Four observer groups were constituted: One control group and three groups paired with demonstrator rats which used well-known and distinct lever-pressing ‘strategies’. During each session, 15 min of demonstration or simple presence of a naïve companion were followed by 30 min of training. It was shown that the rats which were paired with a demonstrator have a higher response rate than controls, but only during the first session. During the eleven following sessions, no significant differences could be observed between the four observer groups. Nevertheless, if we consider the percentage of utilization of the various ‘strategies’ by each rat, we can notice significant correlations between demonstrators and observers, during the last three sessions.     

Genetic variation in the ‘Japanese apogamous form’ of the fernAsplenium unilaterale Lam.

Genetic variation was examined in the Japanese apogamous and sexual forms ofAsplenium unilaterale by electrophoretic analysis of eight enzyme systems. The apogamous form consisted of four biotypes (A, B, C, and D) which show different band patterns. Biotypes C and D were distinguished from biotype A by a difference at only one locus each:Mdh-1 andPgi-2, respectively. Biotypes A, C, and D were sufficiently differentiated from the sexual form (Nei's genetic distance (D)=0.50). This suggests that the apogamous form is not directly derived from the sexual form. However, biotype B expressed the combined band pattern of biotype C and the sexual form, leading to the conclusion that biotype B probably originated by hybridization between those two.     

Leaf characteristics and diurnal variation of chlorophyll fluorescence in leaves of the ‘bana’ vegetation of the amazon region

In six dominant species of the Amazonian ‘Bana’ vegetation, leaf blade characteristics, pigment composition, and chlorophyll (Chl) fluorescence parameters were measured in young and mature leaves under field conditions. Leaf δ¹³C was comparable in the six species, which suggested that both expanding and expanded leaves contained organic matter fixed under similar intercellular and ambient CO₂ concentration (C _i/C _a). High leaf C/N and negative δ¹⁵N values found in this habitat were consistent with the extreme soil N-deficiency. Analysis of Chl and carotenoids showed that expanding leaves had an incomplete development of photosynthetic antenna when compared to adult leaves. Dynamic inactivation of photosystem 2 (PS2) at midday was observed at both leaf ages as F_v/F_m decreased compared to predawn values. Adult leaves reached overnight F_v/F_m ratios typical of healthy leaves. Overnight recovery of F_v/F_m in expanding leaves was incomplete. F₀ remained unchanged from midday to predawn and F_v tended to increase from midday to predawn. The recovery from midday depression observed in adult leaves suggested an acclimatory down-regulation associated with photo-protection and non-damage of PS2.     

A submerged culture system for rapid micropropagation of the commercially important aquarium plant, ‘Amazon sword’ (Echinodorus ‘Indian Red’)

Echinodorus ‘Indian Red’ is an underwater plant, used worldwide for aquarium ornamentation. An efficient method for in vitro propagation and plantlet acclimatization of this popular aquarium plant was standardized. Surface-disinfected shoot-tips were cultured in submerged conditions in a solid–liquid bilayer medium, consisting of an upper, liquid layer (sterile distilled water) and a lower, solid layer Murashige and Skoog (MS) basal medium supplemented with 3.0% (w/v) sucrose, 0.8% (w/v) agar-agar, and plant growth regulators (PGRs) in different combinations and concentrations. The combination of 2.5 mg L⁻¹ 6-benzylaminopurine and 1.0 mg L⁻¹ α-naphthaleneacetic acid improved the multiplication rate to a maximum of 26.8 ± 0.51 shoots per explant after 60 d of culture. The number of multiplied shoots increased with each regeneration cycle, thus from only 26.8 ± 0.51 shoots per explant (first regeneration cycle), this number increased to 33.5 ± 0.58 (second regeneration cycle), and to 38.3 ± 0.62 for the third regeneration cycle with the same medium composition. The highest number of roots (8.3 ± 0.28) per shoot was induced in the presence of 1.0 mg L⁻¹ indole-3-butyric acid, but further growth of these roots was stunted. The best rooting was achieved on PGR-free ½-strength MS medium, where 6.1 ± 0.21 roots per shoot were induced with 5.8 ± 0.35 cm length after 30 d of culture. The regenerated plantlets were successfully acclimatized to submerged underwater conditions, with 100% survival rate. The present protocol is suitable for the commercial propagation of Echinodorus ‘Indian Red’ for aquarium-industries.

     

Temperature- and seed-batch-related variation in the kinetic behaviour of phytochrome under ‘high-irradiance-response’ conditions

The characteristics of the high-irradiance response (HIR) of plant photomorphogenesis are thought to be the result of the interaction of both the light and dark reactions of phytochrome. Thus any variation in the rates of the dark reactions may be expected to lead to variation in the characteristics of the HIR. We report here substantial differences in the rates of the dark reactions between different seed batches of a single species (Sinapis alba L.), and also between different organs of seedlings from each of the batches of seed. Calculations of phytochrome dynamics from the measured dark-reaction rates show that the behaviour of Pfr under HIR conditions will vary considerably according to seed batch and seedling organ. Much larger differences in dark-reaction rates, and the resulting phytochrome dynamics, were found between 25° and 10° C. These lead to the prediction that the HIR will be much reduced at the lower temperature, and may be absent in some cases.Abbreviations and symbols HIR high-irradiance response - Pfr far-red-absorbing form of phytochrome - Pr red-absorbing form of phytochrome - Ptot total phytochrome, Pr+Pfr - _ss Pfr/Ptot ratio which immediately establishes the phytochrome steady state     

A Phenotypic,Molecular and Biochemical Characterization of the First Cisgenic Scab-Resistant Apple Variety ‘Gala’

Scab resistance is one of the most important goals of apple breeding, typically achieved by time-consuming and expensive conventional breeding techniques. Cisgenesis, which is the genetic modification of a recipient organism with genes from a crossable—sexually compatible—organism, is a promising tool for plant breeding to develop disease resistance in a rapid way. A cisgenic, scab-resistant line of the apple variety ‘Gala’ expressing the native apple scab resistance gene Rvi6 (formerly HcrVf2) under control of its own regulatory sequences has been recently developed. In this paper, we present the results from a phenotypic, molecular and biochemical evaluation of clonal replicates of this line (C11.1.53). The phenotype (shoot length, shoot diameter, internode length, number of leaves, leaf length and leaf width) of C11.1.53 was compared to that of the Gala parental background over a period of 108 days. Only a few statistically significant differences were detected, which are probably due to small differences in the quality of the budwood used for grafting rather than effects related to the presence of the cisgene. As the expression of a resistance gene can affect the downstream cascade of plant defence responses, a selection of apple defence-related genes was analyzed by quantitative real-time PCR analysis. These genes are also known as major allergen genes in apple. Even if three out of ten apple allergen genes tested in the leaves differed in the cisgenic line compared to both Gala (background) and ‘Florina’ (the variety from which the Rvi6 gene was cloned), using 2D-PAGE, we were unable to find any significant difference in the expressed proteomes of the leaves of C11.1.53 compared to Gala. Results are discussed in the context of a possible use of cisgenic lines for fruit crop improvement.     

Phenotypic variation and ploidy level of plants regenerated from protoplasts of tetraploid potato (Solanum tuberosum L. cv. ‘Bintje’)

Summary A wide range of phenotypic variation occurred among protoplast — derived plants of tetraploid potato cultivar Bintje. The variant plants had alterations in growth and vigour, and in leaf and stem characteristics. The results suggest that the altered morphologies are caused predominantly by changes in ploidy levels. Some alterations could be attributed typically to octoploidy and aneuploidy. The occurrence of mixoploidy indicates that at least part of the observed variation arose during culture stage. The exogeneous cytokinin or auxin level and their combination during in vitro phase influenced the frequency of the variants observed. The origin of variation is discussed.     

Intra- and interspecific variation in trophic ecology of ‘predatory’ ants in the subfamily Ponerinae

1. The characterisation of energy flow through communities is a primary goal of ecology. Furthermore, predator–prey interactions can influence both species abundance and community composition. The ant subfamily Ponerinae includes many predatory species that range from generalist insectivores to highly specialised hunters that target a single prey type. Given their high diversity and ubiquity in tropical ecosystems, measuring intra- and interspecific variation in their trophic ecology is essential for understanding the role of ants as predators of insect communities. 2. The stable isotopic composition of nitrogen of 22 species from the ant subfamily Ponerinae was measured, relative to plants and other predatory and herbivorous insects at two Atlantic Forest sites in Argentina. The study tested the general assumption that ponerine ants are all predatory, and examined intra- and interspecific variation in trophic ecology relative to habitat, body size and cytochrome c oxidase subunit 1 sequences (DNA barcoding). 3. Stable isotope analysis revealed that most ponerines occupy high trophic levels (primary and secondary predators), but some species overlapped with known insect herbivores. Species residing at low trophic levels were primarily arboreal and may rely heavily on nectar or other plant-based resources in their diet. In addition, larger species tend to occupy lower trophic positions than smaller species. 4. Although some of the species were divided into two or more genetic clusters by DNA barcoding analysis, these clusters did not correspond to intraspecific variation in trophic position; therefore, colony dietary flexibility most probably explains species that inhabit more than one trophic level.     

High interannual variation in the hatching sex ratio of Tengmalm’s owl broods during a vole cycle

The sex ratio at hatching in broods of Tengmalms owl (Aegolius funereus) in northern Sweden was investigated for 3 years characterized by different phases of the vole and owl cycle. Previous work showed the sex ratio in this species to be male-biased for 1 year with a favourable food supply, and that in feeding experiments male nestlings (but not females) suffered higher mortality when food was limited, but not otherwise. Here we present data from a complete 3-year owl cycle, showing that mean brood sex ratio varied significantly among years, being male-biased (65% males) in the first year of high owl breeding density, unbiased (49%) in the second year of high owl breeding density, and female-biased (33% males) in the owls low year. Brood sex ratio did not vary significantly within years with laying date or parental age. Vole availability, and therefore the owls food supply, declined during the 3 years studied. Tengmalms owl parents thus appear to adaptively adjust the sex ratio of their broods according to the expected annual mortality risk of sons.     

Contrasting patterns of disparity suggest differing constraints on the evolution of trilobite cephalic structures during the Cambrian ‘explosion’

Trilobites are an abundant group of Palaeozoic marine euarthropods that appear abruptly in the fossil record c. 521 million years ago. Quantifying the development of morphological variation (or ‘disparity’) through time in fossil groups like trilobites is critical in understanding evolutionary radiations such as the Cambrian ‘explosion’. Here, I use geometric morphometrics to quantify ‘cumulative disparity’ in functionally-important structures within the trilobite cephalon across their initial radiation during Cambrian Series 2. Overall cephalic disparity increased rapidly and attained a maximum within several million years. This pattern is dominated by the cephalic outline (in particular the genal spines), reflecting rapid, convergent expansion to the extremes of morphospace in a few early families. In contrast, removing the outline and focusing on structures such as the glabella and eye ridges (associated with feeding and vision, respectively) showed a more gradual increase in disparity, closer in line with taxonomic diversity and supporting the hypothesis of a relatively accurate trilobite fossil record. These contrasting patterns suggest that disparity in different structures was constrained in different ways, with extrinsic (ecological) factors probably having the major impact on overall disparity. It also implies that patterns of disparity in isolated substructures cannot necessarily be taken individually as representative of overall morphologies.     

How Are ‘Barack Obama’ and ‘President Elect’ Differentially Stored in the Brain? An ERP Investigation on the Processing of Proper and Common Noun Pairs

Background

One of the most debated issues in the cognitive neuroscience of language is whether distinct semantic domains are differentially represented in the brain. Clinical studies described several anomic dissociations with no clear neuroanatomical correlate. Neuroimaging studies have shown that memory retrieval is more demanding for proper than common nouns in that the former are purely arbitrary referential expressions. In this study a semantic relatedness paradigm was devised to investigate neural processing of proper and common nouns.

Methodology/Principal Findings

780 words (arranged in pairs of Italian nouns/adjectives and the first/last names of well known persons) were presented. Half pairs were semantically related (“Woody Allen” or “social security”), while the others were not (“Sigmund Parodi” or “judicial cream”). All items were balanced for length, frequency, familiarity and semantic relatedness. Participants were to decide about the semantic relatedness of the two items in a pair. RTs and N400 data suggest that the task was more demanding for common nouns. The LORETA neural generators for the related-unrelated contrast (for proper names) included the left fusiform gyrus, right medial temporal gyrus, limbic and parahippocampal regions, inferior parietal and inferior frontal areas, which are thought to be involved in the conjoined processing a familiar face with the relevant episodic information. Person name was more emotional and sensory vivid than common noun semantic access.

Conclusions/Significance

When memory retrieval is not required, proper name access (conspecifics knowledge) is not more demanding. The neural generators of N400 to unrelated items (unknown persons and things) did not differ as a function of lexical class, thus suggesting that proper and common nouns are not treated differently as belonging to different grammatical classes.