Production of hairy root cultures of lettuce (Lactuca sativa L.)

Hairy root cultures of lettuce (Lactuca sativa L.) were obtained by inoculation of cotyledonary leaves of in vitro lettuce seedlings (cvs. Nansen and Ljubljanska ledenka) with Agrobacterium rhizogenes A4M70GUS. Approximately in 96.7% cvs. Nansen and in 91.2% Ljubljanska ledenka inoculated explants produced hairy root when they were incubated on Murashige and Skoog (MS) half-strength medium without plant growth regulators. A total of 54% of all hairy root cultures expressed GUS activity. Every hairy root represented an independent transformation event. Line Ljubljanska ledenka 18 showed the highest biomass (5.5 times the biomass of control root). A PCR analysis of the genomic DNA confirmed the presence of marker and target genes in 15 hairy roots examined.     

Leaf anatomy of Carex humilis does not correlate with orographic,geological and bioclimatic habitat conditions in C&SE Europe

This paper presents the results of a multivariate morphometric study of leaf anatomical characters in different, geographically very distant populations of taxon Carex humilis from Hungary, Romania, Serbia, Bosnia & Herzegovina, Montenegro and Albania with the aim to reveal the trends of population differentiation. Analyses were performed on the cross-section of 173 leaves collected from 12 populations. In order to establish the overall morphological variation and relationships between individuals from all populations, principal component analyses (PCA) and canonical discriminant analysis (CDA) have been done. The UPGMA clustering analyses based on leaf anatomical characters and habitat climatic characteristics were carried out to explore whether the observed anatomical differences are a result of adaptive responses. Regression analysis (linear regression) was performed to identify the level of correlation between leaf anatomical characters and basic orographic, geological, and bioclimatic habitat characteristics. Quite unexpectedly, most of observed groups are formed of geographically very distant populations which are living in extremely different climatic and geological conditions, indicating that general anatomical differentiation in Carex humilis in C&SE Europe cannot be explained by the environmental impacts, and basically do not represent an adaptive response to different climatic or geological condition.     

Distribution and diversity of Arctic-Alpine species in the Balkans

The distributions of 77 Arctic-Alpine species in the Balkans are mapped and the centers of their richness and diversity presented. Within the Dinaric Alps these are Mts Vranica, Durmitor, and Prokletije; in the Scardo-Pindhic mountains, ?arplanina–Rudoka–Korab form a continuous chain; in the Rhodope-Rila mountain system there are Mts Vito?a, Rila, and Pirin; while in the Balkan mountain system there are the West and Central part of Stara planina. A comparison of floristic richness and distribution of Arctic-Alpine flora in relation to altitude, geographical location, and geological substrate is made. Correlations between floristic richness and geographical distance of the Balkan mountains from the two main centers of Arctic-Alpine flora in Central Europe are also provided.     

Gibberellic acid promotes in vitro regeneration and shoot multiplication in <Emphasis Type="Italic">Lotus</Emphasis><Emphasis Type="Italic">corniculatus</Emphasis> L.

Shoots of Lotus corniculatus L., previously transformed with Agrobacterium tumefaciens LBA4404/pTOK233, were grown in gibberellic acid (GA₃)-containing media in an attempt to improve their growth and multiplication. Nodal stem segments of four poorly multiplying clones, a clone with very good multiplication, and two non-transformed clones were incubated in media containing GA₃ for 3 weeks, and then returned to a hormone-free medium for a further 3–5 subculture periods. Gibberellic acid increased the number of axillary buds from one in controls to 2–13 shoots during the first 3 weeks of subculture in GA₃-containing media. The multiplied buds, which apparently derived from additional axillaries, continued to multiply in hormone-free medium, producing bunches of more than 50 shoots after 9 weeks. In addition, many nodal segments also produced callus tissues at the basal end, or along the internode below the node, in which abundant shoot meristems formed de novo. Histological examination confirmed their origin via organogenesis. About 25% of regenerated shoots rooted spontaneously. The application of auxins significantly improved rooting in more than 75% of all clones. Well-developed plantlets were produced upon the transfer of rooted shoots to pots. No differences in responses to hormones were observed between LBA4404/pTOK233-transformed clones and non-transformed clones, indicating that the reaction to GA₃ was not the consequence of transformation events. The results point to a possible involvement of GA₃ in lateral branching. They might also be recommended as a procedure suitable for increasing the production of transformed plants in L. corniculatus.     

Use of Chenopodium murale L. transgenic hairy root in vitro culture system as a new tool for allelopathic assays

We investigated Chenopodium murale transgenic hairy root in vitro culture system as a new tool for allelopathic assays. Transgenic hairy roots were induced by Agrobacterium rhizogenes A4M70GUS from roots, cotyledons, leaves, and internodes of C. murale seedlings. Roots were found to be the best target explants, providing transformation efficiency of up to 11.1%. Established hairy root clones differed in their morphology and growth potential. Molecular characterization of these clones was carried out by PCR, RT-PCR and histochemical GUS analyses. No differences in rol gene expression were observed. Liquid culture system of characterized hairy root clones was maintained for over 2 years. Six hairy root clones were selected for assaying the allelopathic effect of their growth medium against germination and seedling elongation of wheat and lettuce test plants. The inhibitory potential varied depending on the hairy root clone. Some transgenic clones showed significantly higher inhibition compared to wild-type roots. These results revealed that hairy roots as an independent system synthesize some bioactive substances with allelopathic activity and exude them into the growth medium. Concentrations of caffeic, ferulic and p-coumaric acids (0.07-2.85 μmol/L) identified by HPLC analysis in the growth media were at least 1000 times lower than the inhibitory active concentration (5 mmol/L) of pure grade phenolic acids, suggesting that they have a limited role in the allelopathic phenomena of C. murale. The presented hairy root system appears to be a suitable tool for further investigation of the potential and nature of root-mediated allelopathic interference of C. murale.     

Overexpressing AtCKX1 in Potato Plants grown In Vitro: The Effects on Cytokinin Composition and Tuberization

Journal of Plant Growth Regulation - Over the last years, cytokinin deficiency has been studied in a variety of plant species, using transgenic expression of cytokinin oxidase/dehydrogenase genes...     

Micromorphological and anatomical characteristics of Salvia amplexicaulis Lam., S. jurisicii Košanin and S. ringens Sibth. & Sm. (Lamiaceae)

Abstract

In this study, we examined Salvia amplexicaulis Lam., S. jurisicii Ko?anin and S. ringens Sibth. & Sm. collected in the Republic of North Macedonia, which are for the first time subjected to detailed micromorphological and structural analysis using light and scanning electron microscopy. The nutlets and mucilage were additionally subjected to spectroscopic analysis using Raman and Attenuated Total Reflectance Fourier Transform Infrared spectroscopy. The anatomical structure of stems and leaves is described and compared. The stems, leaves and calyces bear numerous one- and multi-cellular nonglandular trichomes, and various peltate, capitate and digitiform glandular trichomes. The nutlets differ in size and shape, as well as in myxocarpy. The nutlets predominantly contained α-linolenic and linoleic acid, whereas the mucilages are primarily formed of polysaccharides. The results obtained in this study confirmed the importance of micromorphological and anatomical analysis of Salvia L. spp. plant parts, particularly trichomes and nutlets, and hence contributing to the knowledge about the variety of micromorphological characteristics within the genus Salvia L.     

Radioprotectors – the Evergreen Topic

To protect organisms from ionizing radiation (IR), and to reduce morbidity or mortality, various agents, called radioprotectors, have been utilized. Because radiation‐induced cellular damage is attributed primarily to the harmful effects of free radicals, molecules with radical‐scavenging properties are particularly promising as radioprotectors. Early development of such agents focused on thiol synthetic compounds, known as WR protectors, but only amifostine (WR‐2721) has been used in clinical trials as an officially approved radioprotector. Besides thiol compounds, various compounds with different chemical structure were investigated, but an ideal radioprotector has not been found yet. Plants and natural products have been evaluated as promising sources of radioprotectors because of their low toxicity, although they exhibit an inferior protection level compared to synthetic thiol compounds. Active plant constituents seem to exert the radioprotection through antioxidant and free radical‐scavenging activities. Our research established that plants containing polyphenolic compounds (raspberry, blueberry, strawberry, grape, etc.) exhibit antioxidative activities and protect genetic material from IR.     

Future area expansion outweighs increasing drought risk for soybean in Europe

The European Union is highly dependent on soybean imports from overseas to meet its protein demands. Individual Member States have been quick to declare self-sufficiency targets for plant-based proteins, but detailed strategies are still lacking. Rising global temperatures have painted an image of a bright future for soybean production in Europe, but emerging climatic risks such as drought have so far not been included in any of those outlooks. Here, we present simulations of future soybean production and the most prominent risk factors across Europe using an ensemble of climate and soybean growth models. Projections suggest a substantial increase in potential soybean production area and productivity in Central Europe, while southern European production would become increasingly dependent on supplementary irrigation. Average productivity would rise by 8.3% (RCP 4.5) to 8.7% (RCP 8.5) as a result of improved growing conditions (plant physiology benefiting from rising temperature and CO₂ levels) and farmers adapting to them by using cultivars with longer phenological cycles. Suitable production area would rise by 31.4% (RCP 4.5) to 37.7% (RCP 8.5) by the mid-century, contributing considerably more than productivity increase to the production potential for closing the protein gap in Europe. While wet conditions at harvest and incidental cold spells are the current key challenges for extending soybean production, the models and climate data analysis anticipate that drought and heat will become the dominant limitations in the future. Breeding for heat-tolerant and water-efficient genotypes is needed to further improve soybean adaptation to changing climatic conditions.     

Pseudotsuga menziesii (Pinaceae): Volatile Profiles,Antimicrobial Activity and Toxicological Evaluation of Its Essential Oil

The present article investigates the chemical composition of volatiles of essential oil (EO) and headspace (HS) fraction, as well as biological activities of EO obtained from needles with twigs of Pseudotsuga menziesii var. menziesii cultivated in Serbia. The major class of compounds was monoterpene hydrocarbons with α-terpinolene, sabinene and β-pinene (EO), and sabinene, α-terpinolene and β-pinene (HS) as the dominant volatiles. Tested EO exhibited mostly low antimicrobial potential against investigated strains (ATCC and respiratory isolates), where MICs ranged 1.25–20.00 mg/mL. Nevertheless, based on presented results, where antimicrobial testing was done for the first time on human respiratory system isolates, there is a potential of this EO to be used as an adjuvant in the treatment of human respiratory infections, especially those caused by Pseudomonas aeruginosa or Candida albicans strains. Regarding toxicological evaluation, EO showed moderate toxicity in Artemia salina toxicity bioassay (LC₅₀=347.41, after 24 h) as well as week toxicity against Drosophila melanogaster with the ability only to moderately delay larval and pupal development.