Monitoring Indian "Superfood" Moringa oleifera Lam. - species-specific PCR-fingerprint-based authentication for more consumer safety.

Moringa oleifera Lam. has become one of the major new superfoods commonly available in the aisles of bio-shops and health-food sections in supermarkets of North America and Europe. While most of these products appear under the generic and scientifically inconclusive term "Moringa", the European Union, so far, has allowed commercialisation for the use in food and feed for M. oleifera only. M. oleifera is indigenous to India and South Asia, but large-scale cultivation of this species has spread to the tropical regions on all continents, with a strong focus on Africa, leading to a high risk of admixture with species like M. stenopetala (Baker f.) Cufod. that is native to Africa. In the present study, we have characterised six species of Moringa in order to develop a simple and robust authentication method for commercial products. While the plants can be discriminated based on the pinnation of the leaves, this does not work for processed samples. As alternative, we use the plastidic markers psbA-trnH igs and ycf1b to discern different species of Moringa and develop a diagnostic duplex-PCR that clearly differentiates M. oleifera from other Moringa species. This DNA-based diagnostic assay that does not rely on sequencing was validated with commercial products of "Moringa" (including teas, powders, or capsules). Our method provides a robust assay to detect adulterations, which are economically profitable for costly superfood products such as "Moringa".

Spiropyran-Based Photoisomerizable α-Amino Acid for Membrane-Active Peptide Modification.

Photoisomerizable peptides are promising drug candidates in photopharmacology. While azobenzene- and diarylethene-containing photoisomerizable peptides have already demonstrated their potential in this regard, reports on the use of spiropyrans to photoregulate bioactive peptides are still scarce. This work focuses on the design and synthesis of a spiropyran-derived amino acid, (S)-2-amino-3-(6'-methoxy-1',3',3'-trimethylspiro-[2H-1-benzopyran-2,2'-indolin-6-yl])propanoic acid, which is suitable for the preparation of photoisomerizable peptides. The utility of this amino acid is demonstrated by incorporating it into the backbone of BP100, a known membrane-active peptide, and by examining the photoregulation of the membrane perturbation by the spiropyran-containing peptides. The toxicity of the peptides (against the plant cell line BY-2), their bacteriotoxicity (E. coli), and actin-auxin oscillator modulation ability were shown to be significantly dependent on the photoisomeric state of the spiropyran unit.

Deciphering the regulatory networks involved in mild and severe salt stress responses in the roots of wild grapevine Vitis vinifera spp. sylvestris.

Transcriptional regulatory networks are pivotal components of plant's response to salt stress. However, plant adaptation strategies varied as a function of stress intensity, which is mainly modulated by climate change. Here, we determined the gene regulatory networks based on transcription factor (TF) TF_gene co-expression, using two transcriptomic data sets generated from the salt-tolerant "Tebaba" roots either treated with 50 mM NaCl (mild stress) or 150 mM NaCl (severe stress). The analysis of these regulatory networks identified specific TFs as key regulatory hubs as evidenced by their multiple interactions with different target genes related to stress response. Indeed, under mild stress, NAC and bHLH TFs were identified as central hubs regulating nitrogen storage process. Moreover, HSF TFs were revealed as a regulatory hub regulating various aspects of cellular metabolism including flavonoid biosynthesis, protein processing, phenylpropanoid metabolism, galactose metabolism, and heat shock proteins. These processes are essentially linked to short-term acclimatization under mild salt stress. This was further consolidated by the protein-protein interaction (PPI) network analysis showing structural and plant growth adjustment. Conversely, under severe salt stress, dramatic metabolic changes were observed leading to novel TF members including MYB family as regulatory hubs controlling isoflavonoid biosynthesis, oxidative stress response, abscisic acid signaling pathway, and proteolysis. The PPI network analysis also revealed deeper stress defense changes aiming to restore plant metabolic homeostasis when facing severe salt stress. Overall, both the gene co-expression and PPI network provided valuable insights on key transcription factor hubs that can be employed as candidates for future genetic crop engineering programs.

DNA barcoding as new diagnostic tool to lethal plant poisoning in herbivorous mammals.

Reliable identification of plant species in the digestive tract of a deceased animal often represents the major key to diagnose a lethal intoxication with poisonous plants in veterinary pathology. In many cases, identification of the species is challenging or even impossible because the diagnostic morphological features have been degraded, and because the interpretation of such features requires a considerable expertise in plant anatomy and biodiversity. The use of DNA barcoding markers can support or even replace classical morphological assessment. While these markers have been widely used for plant taxonomy, their forensic application to clarify causes of animal poisoning is novel. In addition, we use specific single-nucleotide polymorphisms as fingerprints. This allows for a clear decision even in cases, where the conventionally used statistical e-values remain ambiguous. In the current work, we explore the feasibility of this strategy in a couple of exemplary cases, either in concert with anatomical diagnostics, or in cases where visual species identification is not possible, or where chemical toxin detection methods are not well established, complex, time consuming and expensive.

Physiological Comparison of Two Salt-Excluder Hybrid Grapevine Rootstocks under Salinity Reveals Different Adaptation Qualities.

Like other plant stresses, salinity is a central agricultural problem, mainly in arid or semi-arid regions. Therefore, salt-adapted plants have evolved several adaptation strategies to counteract salt-related events, such as photosynthesis inhibition, metabolic toxicity, and reactive oxygen species (ROS) formation. European grapes are usually grafted onto salt-tolerant rootstocks as a cultivation practice to alleviate salinity-dependent damage. In the current study, two grape rootstocks, 140 Ruggeri (RUG) and Millardet et de Grasset 420A (MGT), were utilized to evaluate the diversity of their salinity adaptation strategies. The results showed that RUG is able to maintain higher levels of the photosynthetic pigments (Chl-T, Chl-a, and Chl-b) under salt stress, and hence accumulates higher levels of total soluble sugars (TSS), monosaccharides, and disaccharides compared with the MGT rootstock. Moreover, it was revealed that the RUG rootstock maintains and/or increases the enzymatic activities of catalase, GPX, and SOD under salinity, giving it a more efficient ROS detoxification machinery under stress.

The fungal metabolite 4-hydroxyphenylacetic acid from Neofusicoccum parvum modulates defence responses in grapevine.

In a consequence of global warming, grapevine trunk diseases (GTDs) have become a pertinent problem to viticulture, because endophytic fungi can turn necrotrophic upon host stress killing the plant. In Neofusicoccum parvum Bt-67, plant-derived ferulic acid makes the fungus release Fusicoccin aglycone triggering plant cell death. Now, we show that the absence of ferulic acid lets the fungus secrete 4-hydroxyphenylacetic acid (4-HPA), mimicking the effect of auxins on grapevine defence and facilitating fungal spread. Using Vitis suspension cells, we dissected the mode of action of 4-HPA during defence triggered by the bacterial cell-death elicitor, harpin. Early responses (cytoskeletal remodelling and calcium influx) are inhibited, as well as the expression of Stilbene Synthase 27 and phytoalexin accumulation. In contrast to other auxins, 4-HPA quells transcripts for the auxin conjugating GRETCHEN HAGEN 3. We suggest that 4-HPA is a key component of the endophytic phase of N. parvum Bt-67 preventing host cell death. Therefore, our study paves the way to understand how GTDs regulate their latent phase for successful colonisation, before turning necrotrophic and killing the vines.

Surface wax in the ancestral grapevine Vitis sylvestris correlate with partial resistance to Powdery Mildew.

BACKGROUND: Powdery Mildew of Grapevine belongs to the major diseases in viticulture and requires intensive use of fungicides. Genetic introgression of resistance factors from wild grapes from North America and, recently, China, has been successful, but wine made from those varieties is still confronted with low consumer acceptance, due to differences in taste. RESULTS: The current work explores the potential of Vitis vinifera sylvestris, the wild ancestor of domesticated Grapevine, with respect to containing Erysiphe necator, the causative agent of Powdery Mildew. Making use of a germplasm collection comprising the entire genetic variability remaining in Germany, we show that there is considerable genetic variation in the formation of leaf surface waxes exceeding wax formation in commercial varieties. CONCLUSIONS: High wax formation correlates with reduced susceptibility to controlled infection with E. necator linked with perturbations of appressoria formation. We propose V. vinifera sylvestris as novel source for resistance breeding since it is genetically much closer to domesticated grapevine than the hitherto used sources from beyond the species barrier.

A Tunisian wild grape leads to metabolic fingerprints of salt tolerance.

Soil salinity is progressively impacting agriculture, including viticulture. Identification of genetic factors rendering grapevine (Vitis vinifera L.) resilience that can be introgressed into commercial varieties is necessary for safeguarding viticulture against the consequences of global climate change. To gain insight into the physiological and metabolic responses enabling salt tolerance, we compared a salt-tolerant accession of Vitis sylvestris from Tunisia, "Tebaba", with "1103 Paulsen" rootstock widely used in the Mediterranean. Salt stress was slowly increased, simulating the situation of an irrigated vineyard. We determined that "Tebaba" does not sequester sodium in the root but can cope with salinity through robust redox homeostasis. This is linked with rechanneling of metabolic pathways toward antioxidants and compatible osmolytes, buffering photosynthesis, such that cell-wall breakdown can be avoided. We propose that salt tolerance of this wild grapevine cannot be attributed to a single genetic factor but emerges from favorable metabolic fluxes that are mutually supportive. We suggest that introgression of "Tebaba" into commercial varieties is preferred over the use of "Tebaba" as a rootstock for improving salt tolerance in grapevine.

Two ways to die: Species dependent PCD modes in grapevine cells.

Programmed cell death (PCD) is considered as a hallmark of strain-specific immunity. In contrast, generic basal immunity is thought to act without PCD. This classical bifurcation has been questioned during recent years. Likewise, the role of jasmonate signalling for these two modes of innate immunity has remained ambiguous. We have addressed both questions using two closely related grapevine cell lines (V. rupestris, V. vinifera cv. 'Pinot Noir') that contrast in their cell-death response to the bacterial elicitor harpin and the hormonal trigger methyl jasmonate (MeJA). We follow different cellular (loss of membrane integrity, mortality), molecular (induction of transcripts for phytoalexin synthesis and for metacaspases), as well as metabolic (sphingolipid profiles) responses to the two triggers in the two cell lines. The role of NADPH oxidases and induction of transcripts for the class-II metacaspases MC5 differ qualitatively between the two cell lines. We tested a possible role of sphingolipid metabolism but can rule this out. We propose a model, where V. rupestris, originating from co-evolution with several biotrophic pathogens, readily activates a hypersensitive cell death in response to harpin, while the context of MeJA-induced cell death in 'Pinot Noir' might not be related to immunity at all. We propose that the underlying signalling is modular, recruiting metacaspases differently depending on upstream signalling.

Peruvian Amaranth (kiwicha) Accumulates Higher Levels of the Unsaturated Linoleic Acid.

Grain amaranth (Amaranthus spp.) is an emerging crop rich in proteins and other valuable nutrients. It was domesticated twice, in Mexico and Peru. Although global trade is dominated by Mexican species of amaranth, Peruvian amaranth (A. caudatus, kiwicha) has remained neglected, although it harbours valuable traits. In the current study, we investigate the accumulation of polyunsaturated fatty acids, comparing four genotypes of A. caudatus with K432, a commercial variety deriving from the Mexican species A. hypochondriacus under the temperate environment of Southwest Germany. We show that the A. caudatus genotypes flowered later (only in late autumn), such that they were taller as compared to the Mexican hybrid but yielded fewer grains. The oil of kiwicha showed a significantly higher content of unsaturated fatty acids, especially of linoleic acid and α-linolenic acid compared to early flowering genotype K432. To gain insight into the molecular mechanisms behind these differences, we sequenced the genomes of the A. hypochondriacus × hybridus variety K432 and the Peruvian kiwicha genotype 8300 and identified the homologues for genes involved in the ω3 fatty-acid pathway and concurrent oxylipin metabolism, as well as of key factors for jasmonate signalling and cold acclimation. We followed the expression of these transcripts over three stages of seed development in all five genotypes. We find that transcripts for Δ6 desaturases are elevated in kiwicha, whereas in the Mexican hybrid, the concurrent lipoxygenase is more active, which is followed by the activation of jasmonate biosynthesis and signalling. The early accumulation of transcripts involved in cold-stress signalling reports that the Mexican hybrid experiences cold stress already early in autumn, whereas the kiwicha genotypes do not display indications for cold stress, except for the very final phase, when there were already freezing temperatures. We interpret the higher content of unsaturated fatty acids in the context of the different climatic conditions shaping domestication (tropical conditions in the case of Mexican amaranth, sharp cold snaps in the case of kiwicha) and suggest that kiwicha oil has high potential as functional food which can be developed further by tailoring genetic backgrounds, agricultural practice, and processing.

Dual domestications and origin of traits in grapevine evolution.

We elucidate grapevine evolution and domestication histories with 3525 cultivated and wild accessions worldwide. In the Pleistocene, harsh climate drove the separation of wild grape ecotypes caused by continuous habitat fragmentation. Then, domestication occurred concurrently about 11,000 years ago in Western Asia and the Caucasus to yield table and wine grapevines. The Western Asia domesticates dispersed into Europe with early farmers, introgressed with ancient wild western ecotypes, and subsequently diversified along human migration trails into muscat and unique western wine grape ancestries by the late Neolithic. Analyses of domestication traits also reveal new insights into selection for berry palatability, hermaphroditism, muscat flavor, and berry skin color. These data demonstrate the role of the grapevines in the early inception of agriculture across Eurasia.