3-N-oxalyl-L-2,3-diaminopropanoic acid, a multifunctional plant metabolite of toxic reputation

3-N-oxalyl-L-2,3-diaminopropanoic acid (beta-ODAP) is a neuroexcitatory non- protein amino acid identified in grass pea (Lathyrus sativus L.) and in ginseng (Panax spp). Its presence in the drought tolerant grass pea is thought to be responsible for the crippling disease neurolathyrism, while in the longevity promoting ginseng it is named dencichine and praised for its haemostatic properties. Some properties of beta-ODAP are put in perspective

Grain legumes and human health

Since early civilisation in different continents, grain legumes have been a part of balanced diets together with cereals. In general the seeds from those two plant families give together a good source of essential amino acids and may have given an evolutionary advantage. A sustainable diet for vegetarians may not be possible without the protein-rich legumes. Consumed alone, legumes can be a mixed blessing because of their deficiency of some essential amino acids. This deficiency can be balanced in a varied diet containing components richer in tryptophan and the sulphur amino acid methionine and cysteine. Many legume species are exceptionally rich in secondary metabolites, some of which are beneficial to human health while without adequate processing through heating or leaching many can be detrimental to the health of the consumer. Soybean (Glycine max) is one of the most popular and also one of the more healthy species. The presence of genistein and other isoflavones makes it famous as an anti-cancer nutraceutical. Isoflavones and other phytoestrogens, occurring mainly in legumes, might be a factor promoting longevity. Other beneficial secondary metabolites are found among the non-protein amino acids. Legumes are very rich in an enormous variety of non-protein amino acids that can be beneficial or toxic. A beneficial non-protein amino acid occurs in high concentration in fenugreek seeds (Trigonella foenum-graecum), a component of curry, while a variety of toxic amino acids occurs in the genera Lathyrus and Vicia. Legume seeds also contain a variety of anti-nutritional factors that can be reduced by post-harvest processing such as fermentation or germination. Fermentation also improves the balance of essential amino acids. Numerous dietary products such as soy sauce or tempeh are traditional food ingredients derived from legumes

Plant toxin β-ODAP activates integrin β1 and focal adhesion : a critical pathway to cause neurolathyrism

Neurolathyrism is a unique neurodegeneration disease caused by beta-N-oxalyl-L-alpha, beta-diaminopropionic (beta-ODAP) present in grass pea seed (Lathyrus stativus L.) and its pathogenetic mechanism is unclear. This issue has become a critical restriction to take full advantage of drought-tolerant grass pea as an elite germplasm resource under climate change. We found that, in a human glioma cell line, beta-ODAP treatment decreased mitochondrial membrane potential, leading to outside release and overfall of Ca2+ from mitochondria to cellular matrix. Increased Ca2+ in cellular matrix activated the pathway of ECM, and brought about the overexpression of beta 1 integrin on cytomembrane surface and the phosphorylation of focal adhesion kinase (FAK). The formation of high concentration of FA units on the cell microfilaments further induced overexpression of paxillin, and then inhibited cytoskeleton polymerization. This phenomenon turned to cause serious cell microfilaments distortion and ultimately cytoskeleton collapse. We also conducted qRT-PCR verification on RNA-sequence data using 8 randomly chosen genes of pathway enrichment, and confirmed that the data was statistically reliable. For the first time, we proposed a relatively complete signal pathway to neurolathyrism. This work would help open a new window to cure neurolathyrism, and fully utilize grass pea germplasm resource under climate change

Evaluación del contenido del aminoácido no proteico ácido beta-3-N-oxalil-L-2,3 diaminopropiónico (beta-ODAP) en almortas (Lathyrus sativus L.) conservadas en el Banco de Germoplasma Hortícola del CITA

La almorta (Lathyrus sativus L.) es una especie leguminosa con un alto contenido en proteína. Se trata de un cultivo muy rústico, adaptado a escenarios de sequía y suelos pobres, que es considerado como una planta modelo para sistemas de agricultura sostenible. Por ello, desde el proyecto “Siembra Teruel” se está impulsando su cultivo. Según la AECOSAN, el consumo de almorta para alimentación humana se considera seguro cuando el contenido en el aminoácido no proteico beta-ODAP reducido es inferior a un 1 %, en el marco de una dieta variada. En el presente trabajo se analiza el contenido en beta-ODAP en el grano seco de 5 muestras procedentes de la provincia de Teruel, y se compara con muestras de otros orígenes (Cuenca, Italia y Portugal). Aunque mayoritariamente la almorta se consume utilizando la harina obtenida del grano seco, como gachas o farinetas, también se pueden realizar otras recetas. Por ello, en el trabajo se analizan adicionalmente muestras susceptibles de ser utilizadas para el consumo (vaina verde, grano verde, grano precocido y seco). Los resultados muestran un contenido de beta-ODAP inferior al 1% sobre materia seca (sms) en todas las muestras analizadas, excepto para la vaina verde. Los contenidos de las muestras de Teruel varían entre 0,297% y 0,446%, valores superiores a los de las almortas de Italia y Portugal (0,190% y 0,261%, respectivamente) y similares a los de Cuenca (0,314%). En comparación con el grano seco (0,389%), los contenidos de la vaina y el grano en verde son superiores (1,054% y 0,625%, respectivamente) mientras que el de grano precocido y seco (0,217%) y el de la harina (0,256%) son inferiores.El trabajo se enmarca en el proyecto “Siembra Teruel”, del Fondo de Inversiones de Teruel (FITE), financiado por Gobierno de Aragón y Gobierno de España. Las accesiones han sido cultivadas en una parcela experimental de Muniesa en colaboración con los agricultores Félix Yus y Víctor Yus. La muestra de grano precocido y seco ha sido obtenida por Belén Soler (La Ojinegra, Alloza, Teruel).Publishe

Determination of the Neurotoxin 3-N-Oxalyl-2,3-Diaminopropionic Acid and Other Free Amino Acids in Lathyrus cicera and L. sativus Seeds by Reversed-Phase High-Performance Liquid Chromatography

© 2015, Springer Science+Business Media New York. A method for determination of the neurotoxic non-protein amino acid 3-N-oxalyl-2,3-diaminopropionic acid (ODAP) and other free amino acids in Lathyrus cicera and Lahtyrus sativus is presented. Seed extracts were derivatised by reaction with diethyl ethoxymethylenemalonate (DEEMM) and analysed by reversed-phase high-performance liquid chromatography (RP-HPLC). Calibration curves showed very good linearity of the response. The limits of detection (LOD) and quantification (LOQ) were 0.15 and 0.50 μM, respectively. The method has a high intra- [relative standard deviation (RSD) < 0.42 %] and inter-repeatability (RSD = 2.01–2.33 %) and a remarkable accuracy with a 99 % recovery in spiked samples. The method yielded similar results in comparison with a previously established colorimetric method. The method is very easy to carry out and allows for ready analysis of large number of samples using very basic HPLC equipment because the derivatised samples are very stable and have very good chromatographic properties.This work was carried out with the financial support of Junta de Andalucía (Spain) to the Laboratory of Bioactive and Functional Components of Plant Products (Instituto de la Grasa, C.S.I.C.) and the project PAI09-0015-7008 (Consejería de Educación y Ciencia, Junta de Comunidades de Castilla—La Mancha). Cristina Megias and Isabel Cortés-Giraldo are, respectively, recipients of a JAE-Doc (C.S.I.C.) contract and a JAE-Pre (C.S.I.C) fellowship from the “Junta para la Ampliación de Estudios” program (cofinanced by the European Social Fund). Thanks are due to María Dolores García Contreras for technical assistance. The authors are indebted to the Banco de Germoplasma Vegetal de Cuenca, BGV-CU (Cuenca, Spain) for supplying the plant material. We are grateful to the European Social Fund and to “Fundación Parque Científico y Tecnológico de Albacete” for additional financing.Peer Reviewe

Comportamiento fitotécnico y análisis químico proximal de grano de líneas promisoras de chícharo (Lathyrus sativus L.) en condiciones de la E.E.A. El Mantaro

El trabajo de investigación se ejecutó en la E.E.A. “El Mantaro” – UNCP, distrito El Mantaro, provincia Jauja, región Junín; en la especie Lathyrus sativus L. originaria de zonas áridas del Mediterráneo, y considerada como una leguminosa con buena capacidad productiva por su tolerancia a sequias y heladas. El material genético utilizado fueron cuatro líneas promisorias seleccionadas en varios ciclos de selección de un total de 60 líneas por su aclimatación a los ambientes de sierra centro (El Mantaro) y norte del país (Callejón de Huaylas); teniendo como objetivos: a) Determinar el comportamiento fitotécnico del rendimiento de grano de líneas promisorias de chícharo (Lathyrus sativus L.) en condiciones de la E.E.A. El Mantaro, b) Analizar los componentes de rendimiento en dos tipos de siembra de las líneas y c) Realizar el análisis químico proximal de los granos. Fue sembrado bajo dos tipos de siembra a chorro continuo y localizado, mediante el Diseño de Bloques Completamente Randomizado con arreglo factorial 4×2 de 4 repeticiones y 4 tratamientos por los 2 tipos de siembra. Resultado: En días al 50% de floración las líneas a1(Mant. 01-3556) y a3 (Ancash M-012018) mostraron precocidad con 10.51, 10.54 días; en número de vainas/planta destacaron las interacciones a4b2 (Ancash Q-022018/ localizado) y a1b2 (Mant. 01-3556/ localizado) con promedio de 16.82 y 16.35; en peso de 100 granos destacaron las interacciones a1b1 (Mant. 01-3556/ chorro continuo) y a1b2 (Mant. 01-3556/localizado) con promedios de 29.03 y 27.88 g. y en peso por parcela la línea a1 (Mant. 01-3556) en interacción con el tipo de siembra b2 (localizado) tuvo como promedio 2.56 kg/parcela siendo superiores a las demás. En el análisis químico proximal las líneas en estudio presentan mayor contenido de proteína total (Nx6.25) que oscila entre 24.40 % y 23.61%, superando a otras leguminosas como arveja, haba y garbanzo

Genomics and biochemical analyses reveal a metabolon key to β-L-ODAP biosynthesis in Lathyrus sativus

Grass pea (Lathyrus sativus L.) is a rich source of protein cultivated as an insurance crop in Ethiopia, Eritrea, India, Bangladesh, and Nepal. Its resilience to both drought and flooding makes it a promising crop for ensuring food security in a changing climate. The lack of genetic resources and the crop’s association with the disease neurolathyrism have limited the cultivation of grass pea. Here, we present an annotated, long read-based assembly of the 6.5 Gbp L. sativus genome. Using this genome sequence, we have elucidated the biosynthetic pathway leading to the formation of the neurotoxin, β-L-oxalyl-2,3-diaminopropionic acid (β-L-ODAP). The final reaction of the pathway depends on an interaction between L. sativus acyl-activating enzyme 3 (LsAAE3) and a BAHD-acyltransferase (LsBOS) that form a metabolon activated by CoA to produce β-L-ODAP. This provides valuable insight into the best approaches for developing varieties which produce substantially less toxi

Spasticity Mechanisms – for the Clinician

Spasticity, a classical clinical manifestation of an upper motor neuron lesion, has been traditionally and physiologically defined as a velocity dependent increase in muscle tone caused by the increased excitability of the muscle stretch reflex. Clinically spasticity manifests as an increased resistance offered by muscles to passive stretching (lengthening) and is often associated with other commonly observed phenomenon like clasp-knife phenomenon, increased tendon reflexes, clonus, and flexor and extensor spasms. The key to the increased excitability of the muscle stretch reflex (muscle tone) is the abnormal activity of muscle spindles which have an intricate relation with the innervations of the extrafusal muscle fibers at the spinal level (feed-back and feed-forward circuits) which are under influence of the supraspinal pathways (inhibitory and facilitatory). The reflex hyperexcitability develops over variable period of time following the primary lesion (brain or spinal cord) and involves adaptation in spinal neuronal circuitries caudal to the lesion. It is highly likely that in humans, reduction of spinal inhibitory mechanisms (in particular that of disynaptic reciprocal inhibition) is involved. While simply speaking the increased muscle stretch reflex may be assumed to be due to an altered balance between the innervations of intra and extrafusal fibers in a muscle caused by loss of inhibitory supraspinal control, the delayed onset after lesion and the frequent reduction in reflex excitability over time, suggest plastic changes in the central nervous system following brain or spinal lesion. It seems highly likely that multiple mechanisms are operative in causation of human spasticity, many of which still remain to be fully elucidated. This will be apparent from the variable mechanisms of actions of anti-spasticity agents used in clinical practice