Flowering of strict photoperiodic Nicotiana varieties in non-inductive conditions by transgenic approaches

The genus Nicotiana contains species and varieties that respond differently to photoperiod for flowering time control as day-neutral, short-day and long-day plants. In classical photoperiodism studies, these varieties have been widely used to analyse the physiological nature for floral induction by day length. Since key regulators for flowering time control by day length have been identified in Arabidopsis thaliana by molecular genetic studies, it was intriguing to analyse how closely related plants in the Nicotiana genus with opposite photoperiodic requirements respond to certain flowering time regulators. SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) and FRUITFULL (FUL) are two MADS box genes that are involved in the regulation of flowering time in Arabidopsis. SOC1 is a central flowering time pathway integrator, whereas the exact role of FUL for floral induction has not been established yet. The putative Nicotiana orthologs of SOC1 and FUL, NtSOC1 and NtFUL, were studied in day-neutral tobacco Nicotiana tabacum cv Hicks, in short-day tobacco N. tabacum cv Hicks Maryland Mammoth (MM) and long-day N. sylvestris plants. Both genes were similarly expressed under short- and long-day conditions in day-neutral and short-day tobaccos, but showed a different expression pattern in N. sylvestris. Overexpression of NtSOC1 and NtFUL caused flowering either in strict short-day (NtSOC1) or long-day (NtFUL) Nicotiana varieties under non-inductive photoperiods, indicating that these genes might be limiting for floral induction under non-inductive conditions in different Nicotiana varietie

Importance of juvenile hormone signaling arises with competence of insect larvae to metamorphose

AbstractJuvenile hormone (JH) postpones metamorphosis of insect larvae until they have attained an appropriate stage and size. Then, during the final larval instar, a drop in JH secretion permits a metamorphic molt that transforms larvae to adults either directly (hemimetaboly) or via a pupal stage (holometaboly). In both scenarios, JH precludes metamorphosis by activating the Kr-h1 gene through a JH receptor, Methoprene-tolerant (Met). Removal of Met, Kr-h1, or JH itself triggers deleterious precocious metamorphosis. Although JH is thought to maintain the juvenile status throughout larval life, various methods of depleting JH failed to induce metamorphosis in early-instar larvae. To determine when does JH signaling become important for the prevention of precocious metamorphosis, we chose the hemimetabolous bug, Pyrrhocoris apterus, and the holometabolous silkworm, Bombyx mori. Both species undergo a fixed number of five larval instars. Pyrrhocoris larvae subjected to RNAi-mediated knockdown of Met or Kr-h1 underwent precocious adult development when treated during the fourth (penultimate) instar, but younger larvae proved increasingly resistant to loss of either gene. The earliest instar developing minor signs of precocious metamorphosis was the third. Therefore, the JH-response genes may not be required to maintain the larval program during the first two larval instars. Next, we examined Bombyx mod mutants that cannot synthesize authentic, epoxidized forms of JH. Although mod larvae expressed Kr-h1 mRNA at severely reduced levels since hatching, they only entered metamorphosis by pupating after four, rarely three instars. Based on findings in Pyrrhocoris and Bombyx, we propose that insect postembryonic development is initially independent of JH. Only later, when larvae gain competence to enter metamorphosis, JH signaling becomes necessary to prevent precocious metamorphosis and to optimize growth

Climate Change Dependence in Ex Situ Conservation of Wild Medicinal Plants in Crete, Greece

Over 80% of the global population addresses their primary healthcare needs using traditional medicine based on medicinal plants. Consequently, there’s a rising demand for these plants for both household and industrial use at local, regional, national, and international levels. However, wild harvesting has negatively impacted natural ecosystems. Cultivating medicinal species has been proposed as a conservation strategy to alleviate this pressure. Yet, in this age of global climate change concerns, smallholder farmers’ views on the benefits of such cultivation clash with the uncertainties of climate change impacts, amplifying their anxieties. In this context, the climate change dependence of ex situ cultivation of ten wild medicinal taxa with significant ethnopharmacological interest in Crete, Greece, were studied, projecting their potential habitat suitability under various future climate scenarios. The results demonstrated species-specific effects. Based on the potential cultivation area gains and losses, these effects can be categorized into three groups. We also outlined the spatial patterns of these gains and losses, offering valuable insights for regional management strategies benefiting individual practitioners

A combined archaeobotanical and palaeogenetic analysis of charred pea (Pisum sativum) seeds from an Early Iron Age storage pit at the hill fort settlement Hissar, Leskovac, southeast Serbia

A unique example of 2,572 carbonized pea seeds was recovered from the 11th century B.C. deposits at Hissar, a multilevel settlement of the Brnjica cultural group (1,350-1,000 B.C.) in southeast Serbia. Five hundred seeds maintained pea-like hilum, only few had preserved intact smooth-surfaced testa and the majority of seeds had no seed coat. Fortunately, only few seeds of other pulses were observed in the sample and their identity was clearly determined. Applying the exclusion principle, all the “naked”, 3-4 mm large seeds, mainly broad ellipsoid and less often globose, with flattenings or concavities, were determined to correspond to cultivated pea (Pisum sativum L.). Additionally, a high thousand seed weight of charred seeds (24.4 g) suggested cultivated status. To confirm our finding, we processed two samples with molecular tools. A sequence analysis of four chloroplast DNA loci (trnSG, trnK, matK and rbcL) in total length of 1329bp, showed intermediate position to cultivated P. sativum and wild P. sativum subsp. elatius

Evolution of casein kinase 1 and functional analysis of new doubletime mutants in Drosophila

Circadian clocks are timing devices that rhythmically adjust organism’s behavior, physiology, and metabolism to the 24-h day-night cycle. Eukaryotic circadian clocks rely on several interlocked transcription-translation feedback loops, where protein stability is the key part of the delay between transcription and the appearance of the mature proteins within the feedback loops. In bilaterian animals, including mammals and insects, the circadian clock depends on a homologous set of proteins. Despite mostly conserved clock components among the fruit fly Drosophila and mammals, several lineage-specific differences exist. Here we have systematically explored the evolution and sequence variability of insect DBT proteins and their vertebrate homologs casein kinase 1 delta (CKIδ) and epsilon (CKIε), dated the origin and separation of CKIδ from CKIε, and identified at least three additional independent duplications of the CKIδ/ε gene in Petromyzon, Danio, and Xenopus. We determined conserved regions in DBT specific to Diptera, and functionally tested a subset of those in D. melanogaster. Replacement of Lysine K224 with acidic residues strongly impacts the free-running period even in heterozygous flies, whereas homozygous mutants are not viable. K224D mutants have a temperature compensation defect with longer free-running periods at higher temperatures, which is exactly the opposite trend of what was reported for corresponding mammalian mutants. All DBTs of dipteran insects contain the NKRQK motif at positions 220–224. The occurrence of this motif perfectly correlates with the presence of BRIDE OF DOUBLETIME, BDBT, in Diptera. BDBT is a non-canonical FK506-binding protein that physically interacts with Drosophila DBT. The phylogeny of FK506-binding proteins suggests that BDBT is either absent or highly modified in non-dipteran insects. In addition to in silico analysis of DBT/CKIδ/ε evolution and diversity, we have identified four novel casein kinase 1 genes specific to the Drosophila genus

Functional analyses of AGAMOUS family members in Nicotiana benthamiana clarify the evolution of early and late roles of C-function genes in eudicots

[EN] The C-function, according to the ABC model of floral organ identity, is required for stamen and carpel development and to provide floral meristem determinacy. Members of the AG lineage of the large MADS box gene family specify the C-function in a broadly conserved manner in angiosperms. In core eudicots, two sub-lineages co-exist, euAG and PLE, which have been extensively characterized in Antirrhinum majus and Arabidopsis thaliana, where strong sub-functionalization has led to highly divergent contributions of the respective paralogs to the C-function. Various scenarios have been proposed to reconstruct the evolutionary history of the euAG and PLE lineages in eudicots, but detailed functional analyses of the roles of these genes in additional representative species to validate evolutionary hypotheses are scarce. Here, we report functional characterization of euAG- and PLE-like genes in Nicotiana benthamiana through expression analyses and phenotypic characterization of the defects caused by their specific down-regulation. We show that both paralogs redundantly contribute to the C-function in this species, providing insights on the likely evolution of these gene lineages following divergence of the major groups within the eudicots (rosids and asterids). Moreover, we have demonstrated a conserved role for the PLE-like genes in controlling fruit dehiscence, which strongly supports the ancestral role of PLE-like genes in late fruit development and suggests a common evolutionary origin of late developmental processes in dry (dehiscent) and fleshy (ripening) fruits.We thank Rafael Martinez-Pardo and Eugenio Grau for technical support, Amy Litt (New York Botanical Gradens, Bronx, NY, USA) for providing VIGS plasmids, Eugenio Minguet for help with the LFY matrix and Morpheus tool, and Francisco Madueno, Monica Colombo and Barbara Ambrose (NYBG) for critical reading of the manuscript. The work was funded by grant BIO2009-09920 from the Spanish Ministerio de Ciencia e Innovacion to C.F.Fourquin, C.; Ferrandiz Maestre, C. (2012). Functional analyses of AGAMOUS family members in Nicotiana benthamiana clarify the evolution of early and late roles of C-function genes in eudicots. Plant Journal. 71(6):990-1001. https://doi.org/10.1111/j.1365-313X.2012.05046.xS990100171

Aleksandar Mikić, the legume (re)searcher

Aleksandar Mikić was born in Pančevo, Serbia, on 9 January 1974, and died on 5 September 2021, Novi Sad, Serbia. He received BSc in 1998, MSc in 2008, and PhD in 2014 in Plant Genetics and Breeding at the University of Novi Sad, Faculty of Agriculture. He worked as Research Associate and annual legume breeder at the Institute of Field and Vegetable Crops in Novi Sad, Serbia, from 2000 to 2018. His fields of interest encompassed conventional and molecular genetics, genetic resources, breeding, agronomy, agroecology, and crop history of annual legumes. Dr. Mikić co-authored more than 350 journal papers, about 120 international conference papers, over 10 book chapters and one book. He has also authored more than 30 cultivars of annual legumes and various forage crops registered in Serbia and abroad. Aleksandar's scientific horizons were endless, including archaeobotany, linguistics, plant biology, breeding, and farmers communities

Reports on establishing an ex situ site for ‘beautiful’ vavilovia (Vavilovia formosa) in Armenia

Vavilovia (Vavilovia Fed.) is one of the five genera in tribe Fabeae and consists of only one species, ‘beautiful’ vavilovia (Vavilovia formosa (Stev.) Fed.). The main centre of distribution is the Central and Eastern Caucasus, with a disjunct distribution among high alpine areas in the region, extending as far as West Turkey, Lebanon and Iran. In Armenia, in situ studies on Vavilovia started in the late 1930s. In July and August 2009, three expeditions were conducted to two locations: two to the Ughtasar Mountain and one to the Geghama Mountains. The first expedition to Ughtasar resulted in fresh plant collections and soil analysis for one of the sites. The expedition to Geghama established the existence of Vavilovia in the region of Lake Aknalitch. The second expedition to Ughtasar provided immature fruits and seeds. Collected plant material was transplanted into the Flora and Vegetation of Armenia plot of the Yerevan Botanic Garden established in 1940. Today, along with other plants the plot contains more than 200 species of wild relatives of cultural plants from 130 genera, including indiginous species of tribe Fabeae such as Vavilovia. The transplanted plants will continue to be monitored to see if the plants go on to successfully flower and set seed or whether further sites, possibly at higher altitudes might need to be tested to meet the long term conservation requirements of this iconic legume. These co-ordinated efforts provide a good example of an ex situ conservation strategy for Vavilovia formosa, which, if successful will improve access and utility for the whole legume research community

The bicentenary of the research on 'beautiful' vavilovia (Vavilovia formosa), a legume crop wild relative with taxonomic and agronomic potential

Vavilovia formosa is a relict, endangered species from the highlands of the Caucasus and the Near East. Described in 1812, it has had an uncertain status and was finally recognized as a separate genus of tribe Fabeae (Fabaceae). Our informal international group was established in 2007 to revive the interest in this species as it had been seriously neglected for decades. Here, we provide an overview of the accumulated knowledge on V.formosa and present the results of the most recent multidisciplinary research. Three expeditions were made to two locations in Armenia in 2009, providing the material for anatomical, morphological, chemical and molecular analysis. Unlike previous attempts, ex situ conservation in Yerevan and in vitro propagation, important for potential interspecific hybridization, were successful. Molecular tools were used to clarify the taxonomic position of V.formosa, often considered the closest to the extinct ancestor of the whole tribe. The analysis of four informative regions of plastid and nuclear DNA showed that V.formosa belongs to the same clade as Lathyrus and Pisum, with a distinct status. Preservation and maintenance of V.formosa remains the only basis for further development of all other scientific aspects, especially breeding and uses in agronomy.

Spatial patterns and intraspecific diversity of the glacial relict legume species Vavilovia formosa (Stev.) Fed. in Eurasia

Vavilovia formosa is one of five genera in tribe Fabeae, (Fabaceae, Leguminosae) with close phylogenetic relationships to Pisum. It grows in subalpine and alpine levels in Armenia, Azerbaijan, Georgia, Iran, Iraq, Lebanon, Russia and Turkey and is recognized as an endangered and protected plant. This study was conducted to reveal its intraspecific variability, as well as to predict the past, extant and future species distribution range. We analysed 51 accessions with common phylogenetic markers (trnF-trnL, trnS-trnG, matK, rbcL, psbA-trnH and ITS). These represent in total up to 2551 bp of chloroplast and 664 bp of nuclear sequences per sample. Two populations from Turkey and Armenia were analysed for genetic diversity by AFLP