Adapting Agriculture to Climate Change: A Synopsis of Coordinated National Crop Wild Relative Seed Collecting Programs across Five Continents.

The Adapting Agriculture to Climate Change Project set out to improve the diversity, quantity, and accessibility of germplasm collections of crop wild relatives (CWR). Between 2013 and 2018, partners in 25 countries, heirs to the globetrotting legacy of Nikolai Vavilov, undertook seed collecting expeditions targeting CWR of 28 crops of global significance for agriculture. Here, we describe the implementation of the 25 national collecting programs and present the key results. A total of 4587 unique seed samples from at least 355 CWR taxa were collected, conserved ex situ, safety duplicated in national and international genebanks, and made available through the Multilateral System (MLS) of the International Treaty on Plant Genetic Resources for Food and Agriculture (Plant Treaty). Collections of CWR were made for all 28 targeted crops. Potato and eggplant were the most collected genepools, although the greatest number of primary genepool collections were made for rice. Overall, alfalfa, Bambara groundnut, grass pea and wheat were the genepools for which targets were best achieved. Several of the newly collected samples have already been used in pre-breeding programs to adapt crops to future challenges.

Molecular networking-driven isolation of 8'-Glycosylated biscoumarins from Cruciata articulata.

A molecular networking-guided phytochemical investigation of Cruciata articulata led to the isolation of five unreported biscoumarins, four of which were characterized by a shared 6-methoxy-7,8'-dihydroxy-3,7'-biscoumarin aglycone. These were isolated alongside two known coumarin glycosides, daphnetin-8-O-ß-D-glucoside and 6'-acetoxy-daphnetin-8-O-ß-D-glucoside. Their structures were elucidated by extensive 1D and 2D NMR experiments, in combination with chemical transformation and MS/MS fragmentation analysis. Four of the biscoumarins were glycosylated at the 8' position: these are the first examples of this substitution pattern to be described in nature. All compounds were tested for cytotoxic, antimicrobial, anti-inflammatory, and α-glucosidase inhibitory properties, but did not display significant activity.

Cruciasides C-G, monoterpenoid glycosides from Cruciata articulata.

Cruciata articulata (L.) Ehrend. is a herbaceous species distributed in parts of Western Asia and the Mediterranean region. While research on other species in the Cruciata genus has revealed the presence of a range of flavonoids and terpenoids, few such studies have been conducted on C. articulata. Thus, in the current study, a phytochemical investigation of C. articulata was carried out. Molecular networking identified a large cluster of compounds sharing distinctive MS-MS fragmentation patterns that were targeted for isolation, leading to the isolation of five undescribed monoterpenoid glycosides, cruciasides C-G, along with two known monoterpenoid glycosides. The structures of these compounds were elucidated by using chemical and spectroscopic analyses, including 1D and 2D NMR, and MS-MS fragmentation. Structures for the ions observed in the MS-MS were proposed, and based on these fragmentation patterns, structures for several of the minor components observed in the molecular network were also proposed. All isolated compounds were tested for cytotoxic, anti-inflammatory, antimicrobial, and α-glucosidase inhibitory properties, but did not display any activity.

Unity in diversity-food plants and fungi of Sakartvelo (Republic of Georgia), Caucasus.

BACKGROUND: The Republic of Georgia is part of the Caucasus biodiversity hotspot, and human agricultural plant use dates back at least 6000 years. Over the last years, lots of ethnobotanical research on the area has been published. In this paper, we analyze the use of food plants in the 80% of Georgia not occupied by Russian forces. We hypothesized that (1) given the long tradition of plant use, and the isolation under Soviet rule, plant use both based on home gardens and wild harvesting would be more pronounced in Georgia than in the wider region, (2) food plant use knowledge would be widely and equally spread in most of Georgia, (3) there would still be incidence of knowledge loss despite wide plant use, especially in climatically favored agricultural regions in Western and Eastern Georgia. METHODS: From 2013 to 2019, we interviewed over 380 participants in all regions of Georgia not occupied by Russian forces and recorded over 19,800 mentions of food plants. All interviews were carried out in the participants' homes and gardens by native speakers of Georgian and its dialects (Imeretian, Rachian, Lechkhumian, Tush, Khevsurian, Psavian, Kakhetian), other Kartvelian languages (Megrelian, Svan) and minority languages (Ossetian, Ude, Azeri, Armenian, Greek). RESULTS: The regional division was based primarily on historic provinces of Georgia, which often coincides with the current administrative borders. The total number of taxa, mostly identified to species, including their varieties, was 527. Taxonomically, the difference between two food plant groups-garden versus wild-was strongly pronounced even at family level. The richness of plant families was 65 versus 97 families in garden versus wild plants, respectively, and the difference was highly significant. Other diversity indices also unequivocally pointed to considerably more diverse family composition of wild collected versus garden plants as the differences between all the tested diversity indices appeared to be highly significant. The wide use of leaves for herb pies and lactofermented is of particular interest. Some of the ingredients are toxic in larger quantities, and the participants pointed out that careful preparation was needed. The authors explicitly decided to not give any recipes, given that many of the species are widespread, and compound composition-and with it possible toxic effects-might vary across the distribution range, so that a preparation method that sufficiently reduces toxicity in the Caucasus might not necessary be applicable in other areas. CONCLUSIONS: Relationships among the regions in the case of wild food plants show a different and clearer pattern. Adjacent regions cluster together (Kvemo Zemo Racha, and Zemo Imereti; Samegrelo, Guria, Adjara, Lechkhumi and Kvemo and Zemo Svaneti; Meskheti, Javakheti, Kvemo Kartli; Mtianeti, Kakheti, Khevsureti, Tusheti. Like in the case of the garden food plants, species diversity of wild food plants mentioned varied strongly. Climate severity and traditions of the use of wild food plants might play role in this variation. Overall food plant knowledge is widely spread all-across Georgia, and broadly maintained.

Potential anti-neuroblastoma agents from Juniperus oblonga.

Neuroblastoma (NB) is a neuroendocrine tumor derived from neural crest cells. Approximately 90% of cases occur in children less than 5 years old. The amplification of MYCN correlates with high-risk neuroblastoma and patients with MYCN amplified showed poorer prognosis than those without MYCN amplification. In this study, three compounds isolated from Juniperus oblonga showed anti-proliferative activity against NB cell lines with and without tetracycline inducible MYCN over-expression which were identified as (-)-deoxypodophyllotoxin (1), (-)-matairesinol (2) and (+)-isocupressic acid (3). The effects of compounds 2 and 3 in NB cells included a decrease in NB cell viability and induction of apoptosis. Compound 1 was more effective in NB cells over-expressing MycN. Compound 1 also showed almost 2-fold induction of intracellular free calcium levels in M2(+) cells, which may indicate a different mechanism of action for this compound. Cytotoxicity studies against the human embryonic kidney cell (HEK-293) showed compounds 1, 2 and 3 were ineffective in the non-cancer cells at concentrations approximating their IC50 against the NB cell lines. These results may lead to safer and more effective treatment options for NB patients especially for those with high-risk NB.

Unusual Flavones from Primula macrocalyx as Inhibitors of OAT1 and OAT3 and as Antifungal Agents against Candida rugosa.

A bioactivity guided program exploring the interaction of phytochemicals in the entire plant Primula macrocalyx with the organic anion transporters (OAT1 and OAT3) and microorganisms led to the elucidation of ten known flavones (1-4, 6-10, 12) and two previously undescribed flavones (5, 11). The structures of the compounds were determined by extensive analysis of spectroscopic data, as well as by comparison with data from previous reports. Two known flavones (9, 12) are reported for the first time from the family Primulaceae. All compounds were evaluated for inhibition of OAT1 and OAT3. Six flavones (2, 3, 6-8, 12) showed potent inhibitory activity on OAT1, while seven flavones (2, 3, 6-9, 12) showed marked inhibitory activity on OAT3, with IC50 ≤ 10.0 µM. Antimicrobial activities of crude fractions against sixteen microorganisms were tested to give a target yeast strain Candida rugosa for further evaluation of MICs on the isolates. Three flavones (7, 8, 12) showed marked antifungal activity with MIC < 2.0 µM. To our knowledge, this study is the first to evaluate these flavones as inhibitors of the OAT1 and OAT3, and as antifungal agents.

Labdane and Abietane Diterpenoids from Juniperus oblonga and Their Cytotoxic Activity.

A phytochemical investigation of the whole plant of Juniperus oblonga led to the isolationof one previously undescribed labdane diterpenoid, (4R,5S,9S,10R)-13-des-ethyl-13-oxolabda-8(17),11E-dien-19-oic acid (1), together with nine known diterpenoids (2-3, 6-12), two lignans (4, 5),and a coumarin (13). The structures of all the compounds were elucidated on the basis ofspectrometric data, primarily one-dimensional (1D)- and two-dimensional (2D)-NMR and massspectrometry. Electronic circular dichroism (ECD) calculations determined the absoluteconfiguration of 1. In addition, the isolated compounds were evaluated for their cytotoxic activityagainst three human tumor cell lines (HepG2, MCF-7, and HeLa). 6,12-Dihydroxyabieta-5,8,11,13-tetraen-7-one (6) showed moderate cytotoxicity against all three cell lines with IC50 values rangingfrom 24.41 µM to 58.39 µM and trilobinone (10) showed weaker activity with IC50 values rangingfrom 56.93 µM to 79.98 µM. None of the isolated diterpenoids have been previously reported fromJuniperus oblonga, and five compounds are here reported from the genus Juniperus for the first time.

Biflavonoids from Juniperus oblonga inhibit organic anion transporter 3.

Organic anion transporters (OATs in humans, Oats in rodents) play an important role in the distribution and excretion of numerous endogenous metabolic products and exogenous organic anions, including a host of widely prescribed drugs. Their ligand recognition is also important for drug therapy and development. In this study, the n-butanol and dichloromethane soluble fractions of Juniperus oblonga were found to inhibit OAT3 in vitro and three biflavonoids were found to be responsible for this activity. One of these compounds, amentoflavone exhibited stronger inhibition than probenecid, a known strong inhibitor of OAT3. Biological characterization of amentoflavone in vivo also showed inhibition of Oat3. Preliminary observations of structure-activity relationships suggest that the biflavonoids are more potent inhibitors of this transporter than their corresponding monomer, and that methylation of even a single hydroxyl group results in a substantial decrease in activity. This greater potency of the biflavonoids may indicate the need for a more in-depth investigation of the distribution of biflavonoids in plants used as foodstuffs and herbal medicines, due to their potential for causing interactions with OAT3 substrate drugs.

Isoflavones from Camphorosma lessingii Inhibit the Organic Anion Transporters OAT1 and OAT3.

Phytochemical investigation of Camphorosma lessingii has resulted in the isolation of four previously unreported isoflavones (1: -4: ) and eight known compounds (5: -12: ). Nine of these compounds (1: -6, 8: -10: ) are reported for the first time from members of the family Amaranthaceae. The structures of all isolated compounds were determined by spectroscopic methods, primarily one-dimensional and two-dimensional nuclear magnetic resonance and mass spectrometry. The absolute configuration of 6: was confirmed by circular dichroism. Inhibition of the organic anion transporters, OAT1 and OAT3, by the isolated compounds was evaluated. Among them, 7, 2'-dihydroxy- 6,8-dimethoxyisoflavone (1: ), 2'-hydroxy-6,7,8-trimethoxyisoflavone (2: ), 6,2'-dihydroxy-7,8-dimethoxyisoflavone (3: ), and 7-methoxyflavone (5: ) showed a significant inhibitory effect on 6-carboxyfluorescein uptake mediated by OAT1 and OAT3.

A comparative ethnobotany of Khevsureti, Samtskhe-Javakheti, Tusheti, Svaneti, and Racha-Lechkhumi, Republic of Georgia (Sakartvelo), Caucasus.

BACKGROUND: The Republic of Georgia (Sakartvelo in Georgian language) is part of the Caucasus biodiversity hotspot, and human agricultural plant use dates bat at least 6000 years. However, little ethnobiological research has been published from the region since the 1940s. Given the lack of recent research in the region, the present study we report on plant uses in Skartvelo (Republic of Georgia), Caucasus. We hypothesized that, (1) given the long tradition of plant use, and the isolation under Soviet rule, plant use both based on homegardens and wild harvesting would be more pronounced in Georgia than in the wiser region, (2) the Soviet occupation would have had broad influence on plant use, and (3) there would still be incidence of knowledge loss despite wide plant use. METHODS: Fieldwork was conducted in Khevsureti, Samtskhe-Javakheti, Tusheti, Svaneti, and Racha in July-August 2013, July-August 2014, and September-October 2015. Interviews using semi-structured questionnaires were conducted with 170 participants (80 women and 90 men) after obtaining their oral prior informed consent. All interviews were carried out in the participants' homes and gardens by native speakers of Georgian and its local dialects (Svan, Tush, Khevsur, Psav), or, where participants spoke these as their native language, Armenian, Russian, or Greek. RESULTS: In the present study we encountered 480 plant species belonging to 249 genera of 95 families being used in the research region. The highest number of species and of unique species were reported from the remote Tusheti-Khevsureti region. Informant consensus and number of use reports were highest for each region in the food and medicinal use categories. Of the 480 plants being used in the research region 282 species were exclusively wild-harvested, 103 were grown in homegardens, and 84 were both grown in gardens and sourced in the wild. CONCLUSIONS: Plant species, and uses, found in our study, both for Georgia in general, as well as for its regions, showed clear relations to the wider Caucasus - Asia Minor - Balkans cultural complex. However, plant use in Georgia was much more diverse than reported in other studies from Eurasia.

Scrophularia orientalis extract induces calcium signaling and apoptosis in neuroblastoma cells.

Effective neuroblastoma (NB) treatments are still limited despite treatment options available today. Therefore, this study attempted to identify novel plant extracts that have anticancer effects. Cytotoxicity and increased intracellular calcium levels were determined using the Sulforhodamine B (SRB) assay and Fluo4-AM (acetoxymethyl) staining and fluorescence microscopy in NB cells in order to screen a library of plant extracts. The current study examined the anticancer effects of a dichloromethane extract from Scrophularia orientalis L. (Scrophulariaceae), a plant that has been used in Traditional Chinese Medicine. This extract contained highly potent agents that significantly reduced cell survival and increased calcium levels in NB cells. Further analysis revealed that cell death induced by this extract was associated with intracellular calcium release, opening of the MPTP, caspase 3- and PARP-cleavage suggesting that this extract induced aberrant calcium signaling that resulted in apoptosis via the mitochondrial pathway. Therefore, agents from Scrophularia orientalis may have the potential to lead to new chemo-therapeutic anticancer drugs. Furthermore, targeting intracellular calcium signaling may be a novel strategy to develop more effective treatments for NB.