La gesse commune (Lathyrus sativus L.) est une légumineuse cultivée principalement en Inde, au Bangladesh et en Ethiopie qui présente des niveaux de résistance élevés pour de nombreuses contraintes abiotiques, telles que la sécheresse et l'inondation. Dans ce travail, les capacités de tolérance du Lathyrus (lignées locales "Raipur" et "Bangladesh") à une autre contrainte abiotique, la présence de plomb, ont été déterminées des points de vue physiologique et moléculaire. Un système expérimental de culture hydroponique a été mis au point pour ces plantes. Le plomb y est introduit sous forme de nitrate de plomb (Pb(NO3)2). Les teneurs en plomb des différents organes des plantes (racines, tiges, feuilles) ont été déterminées par ICP-OES. Les réponses cellulaires dans ces organes ont été étudiées par RTPCR quantitative (PCR en temps réel). Des amorces spécifiques du Lathyrus ont été dessinées à partir des 11 séquences d'ADN complémentaires (ADNc) isolées pour la première fois et séquencées. L'un des ADNc isolé est complet et code une cystéine protéase (LsCP, 427 aa). Les autres sont des ADNc partiels et correspondent à une aspartique protéase (LsAP, 270 aa), deux ascorbate peroxidases cytosolique (LsAPXc, 195 aa) et peroxisomale (LsAPXp, 226 aa), une protéine de choc thermique ("Heat Shock Protein 70" ; LsHSP70, 287), une homoglutathion synthétase (LshGSHS, 329 aa), une glutathion S-transférase (LsGST, 66 aa), une glutathion réductase (LsGR, 336 aa), une phytochélatine synthétase (LsPCS, 64 aa), une phospholipase D a (LsPLDa, 288 aa), un transporteur membranaire spécifique du Pb (LsCNGC, 136) et une protéine soluble (LsABCt, 331 aa). Les plantes exposées au nitrate de plomb accumulent de grandes quantités de métal dans leurs racines sous forme de plomb fortement lié aux tissus. L'accumulation d'ARN messagers de la LsPLDa suggère que les racines subissent une contrainte cellulaire et s'y adaptent. La stimulation de l'expression des gènes LsGST, LsGR et LsAPX correspondrait à la formation de complexes Pb-glutathion et à une activation du cycle ascorbate-glutathion pour la neutralisation des espaces activées de l'oxygène (ROS) délétères. Dans les feuilles exemptes de plomb, l'augmentation de l'expression des gènes LsCP, LsAP, LsAPXc, LsAPXp, LsHSP70, LsGR et LsPCS semble indiquer l'émission d'un signal racinaire transmis de manière systémique vers le reste de la plante où il déclenche la sur-expression de ces gènes. Ceci pourrait permettre aux tissus épargnés par le polluant de se préparer à son éventuelle arrivée. La chélation du Pb avec de l'EDTA dans le milieu de culture conduit à son transport vers les parties aériennes et à son accumulation dans les feuilles. L'expression du gène LsCNGC chez ces plantes est activée dans les feuilles, suggérant une participation de ce transporteur à l'entrée des complexes Pb-EDTA dans le symplasme. Comme observé chez d'autres espèces végétales, le plomb affecte le métabolisme du glutathion chez la gesse commune. Cependant, la mise en évidence d'une réaction systémique en réponse au plomb à partir des racines ainsi que la surexpression de gènes d'autolyse sont réalisées pour la première fois et pourraient être des éléments contribuant fortement à la tolérance au plomb chez cette espace végétale sous-utilisée. / Grass pea (Lathyrus sativus L.) is a legume plant cultivated mostly in India, Bangladesh and Ethiopia. It possesses high tolerance levels to abiotic stresses like drought and flooding. In the present work, the tolerance of Lathyrus sativus L. (local lines "Raipur" and "Bangladesh") to another abiotic stress: lead, has been determined, using a physiological and a molecular approach. A hydroponic culture system has been set up. Lead was introduced as lead nitrate (Pb(NO3)2). Lead contents in various plant organs (roots, stems, leaves) were determined using ICP-OES. Cell responses in these organs were studied using real-time RT-PCR. Grass pea-specific primers were designed from the 11 complementary DNA sequences that were isolated and sequenced here for the first time. One of the cDNA is full-length and codes a cystein protease (LsCP). The others are partial cDNA and code an aspartic protease (LsAP, 270 aa), two ascorbate peroxidases, a cytosolic (LsAPXc, 195 aa) and a peroxisomal (LsAPXp, 226 aa), a heat shock protein 70 (LsHSP70, 287), a homoglutathione synthetase (LshGSHS, 329 aa), a glutathione-S-transferase (LsGST, 66 aa), a glutathione reductase (LsGR, 336 aa), a phytochelatin synthetase (LsPCS, 64 aa), a phospholipase D a (LsPLDa, 288 aa), a membrane transporter specific of lead (LsCNGC, 136) and a soluble protein (LsABCt, 331 aa). The plants exposed to lead accumulate large amounts of the metal in their roots only and the metal is tightly bound to the tissues. mRNA accumulation for LsPLDa suggest that root tissues are under stress and coping. The increases in LsGST, LsGR and LsAPX transcripts suggest that Pb-glutathione complexes are formed and that the ascorbate-glutathione cycle is stimulated to scavenge deleterious activated oxygen species, in the root tissues. In lead-free leaves, LsCP, LsAP, LsAPXc, LsAPXp, LsHSP70, LsGR et LsPCS genes are overexpressed. This indicates that a root signal is emitted by these leaded tissues and transported systemically to the rest of the plant where it stimulates the expression of the genes mentioned above. This could be a prevention mechanism to prepare leaf tissues for the possibility of lead spreading. Chelation of lead with EDTA in the growth medium facilitates the transport of the pollutant to the leaves where it accumulates. In these tissues, the LsCNGC is over-expressed suggesting that this transporter is involved in the translocation of the Pb-EDTA complexes into the leaf symplasm. As was observed previously in other plant species, lead affects glutathione metabolism in grass pea. However, the observation of a systemic signal originating from the leaves in response to lead and the over-expression of autolytic genes are reported here for the first time and could be contributing elements to lead tolerance in this under-utilized plant species.

Experiments were conducted to determine the outcrossing rate, the inheritance of markers and establish a basic linkage map in grasspea, <i> Lathyrus sativus </i>L. The outcrossing rate in a white-flowered line of grasspea ranged from 1.7 to 2.7% among eight combinations of gene frequency and location. The outcrossing rate in this study (2.2 ± 0.7%) suggests that individual lines of grasspea should be maintained in isolation to maintain their genetic integrity. Inheritance and linkage were determined for one morphological, 11 isozyme and 72 RAPD markers in five F₂ populations (all RAPD markers were in one F₂ population). The inheritance of flower colour was monogenic with colour dominant over white. The isozymes, ACO-1, ACO-2, AAT-1, AAT-2, EST-6, FDH, LAP-1, PGD-2, SKDH and TPI-1, were codominantly expressed with monogenic inheritance. The isozymes LAP-1 and PGD-2 segregated in a non-Mendelian ratios in the crosses PI 426891.1.3 x PI 283564c.3.2 and PI 426891.1 x PI 172930.4, respectively. The isozymeEST-3 was monogenically inherited and dominantly expressed. Most RAPD markers segregated in a 3:1 ratio. Marker UBC368_425/655 segregated in a co-dominant fashion. The RAPD markers UBC304₈₃₁, UBC304₉₆₄, UBC308₉₉₀, UBC322₁₄₃₂, UBC328₈₃₁, UBC332₁₁₁₈, UBC3321₁₅₈₁, UBC333₆₁₇, UBC349<sub>752<?sub>, UBC365₁₀₁₃ and UBC388₄₅₉ showed distorted segregation. In two F₂ populations, PI 283564c.3 x PI 426885.2 and PI 358601.5 x PI 173714.5, a linkage between AAT-2 and SKDH was reconfirmed. In the cross PI 426891.1.3 x PI 283564c.3.2, one morphological, three isozyme and 71 RAPD markers were mapped resulting in the delineation of 14 linkage groups including 69 markers (1 morphological, 3 isozyme and 65 RAPD markers). The total genome length covered by these 75 markers (69 linked and six unlinked) was about 864 cM. Considering cost, simplicity and abundance, RAPD analysis was more efficient than isozyme analysis in developing linkage map.

Fodder from lasting grass growths serves from 80% to fed ruminants. Therefore is setting high emphasis on quality fodder from trifolium, grass and trifolium-grass growths. On this account fit itrodukcion try in these growths imply at full blast useful kinds plants (trifolium), which have improving effects on fodder. Once of these improving products is also Lathyrus pratensis. His introduction increases substitution legumin in lasting meadow growths and share rhizobial nitrogen on production knows - nose fodder and nutritive valuable degestible nitrogen factory materials. Occurrence Lathyrus pratensis is impressed with type herbal growth and his differential management.

京都大学 / 0048 / 新制・課程博士 / 博士(人間・環境学) / 甲第19085号 / 人博第738号 / 新制||人||177(附属図書館) / 26||人博||738(吉田南総合図書館) / 32036 / 京都大学大学院人間・環境学研究科相関環境学専攻 / (主査)教授 瀬戸口 浩彰, 教授 加藤 眞, 教授 市岡 孝朗 / 学位規則第4条第1項該当 / Doctor of Human and Environmental Studies / Kyoto University / DGAM

El hombre maneja la mayor parte de la superficie terrestre y una gran proporción está dedicada a actividades agropecuarias. El modelo de producción agropecuaria vigente, fuertemente influenciado por la Revolución Verde, tiene problemas ambientales y sociales severos que ponen en duda su sustentabilidad. La agroecología y la restauración productiva son enfoques alternativos, que consideran las dimensiones ecológicas, económicas y sociales de los sistemas agroalimentarios. El sudoeste bonaerense tiene características marginales para la agricultura y aptitud principalmente ganadera. La restauración productiva de los pastizales naturales de esta región mediante la reincorporación de especies vegetales claves del ecosistema original para generar agroecosistemas más resilientes y sustentables es una prioridad que debe ser atendida. El objetivo de esta tesis fue aumentar el conocimiento sobre algunas especies de leguminosas herbáceas nativas de la región con potencial para fijar nitrógeno, mejorar la oferta forrajera y enriquecer los pastizales naturales degradados. En primer lugar se realizó una búsqueda bibliográfica para detectar y caracterizar las especies de leguminosas herbáceas nativas reportadas para el área de estudio; luego se localizaron y georreferenciaron sus poblaciones. Posteriormente se estudió la germinación, capacidad de fijación biológica de nitrógeno (FBN) y calidad nutritiva de algunas especies. Se detectaron 30 especies correspondientes a 10 géneros, 19 de ellas potencialmente útiles en agroecosistemas y 16 encontradas en alguno de los 43 sitios visitados, siendo Adesmia, Lathyrus y Vicia los géneros con más especies y A. incana y Rhynchosia senna las más frecuentemente halladas. Se estudió la germinación en laboratorio de las especies más promisorias de las que se colectó semilla: Adesmia filipes (Af), A. incana (Ai), A. muricata (Am), Lathyrus nervosus (Ln), L. pubescens (Lp), Rhynchosia diversifolia (Rd) y R. senna (Rs), evaluando el porcentaje de germinación (PG) y la respuesta a tratamientos pregerminativos aplicados para romper la dormición seminal. Las respuestas fueron variables según las especies; todas presentan un 45% o más de semillas dormantes y para la mayoría se lograron PG mayores al 50%. Los tratamientos más efectivos fueron los de escarificación física; la escarificación térmica por inmersión en agua caliente fue efectiva para Lp, Rd y Rs. Paralelamente, se aislaron cepas de rizobio nodulantes de Ai y Lp, y se evaluó la FBN en esta última utilizando inoculante propio y comercial. Lp solo estableció simbiosis efectivas con rizobios nativos y tuvo una eficiencia de fijación del 50%, sin diferencias en el contenido de nitrógeno entre tratamientos con nitrógeno mineral o simbiótico, pero con mayor producción de biomasa en el primero. Por último, se analizaron algunos parámetros de calidad nutritiva de las especies Ai, Ln, Lp, L. subulatus, Rd y Rs. En estado vegetativo, todas mostraron contenidos de proteína bruta semejantes a los de leguminosas cultivadas y algunas un contenido mayor, y buena digestibilidad. Solo se detectaron taninos precipitantes de proteínas en Rd, Rs y Ai en estado reproductivo, siendo de moderados a altos solo en Rd. Por las características evaluadas en este trabajo y las reportadas en la bibliografía, muchas de estas especies merecen ser consideradas para la restauración productiva de pastizales semiáridos y subhúmedos del centro de Argentina; nuevos estudios orientados a analizar diferentes modos de obtener semillas para cultivo a mediana y gran escala contribuirían a concretar su utilización. / Human manages and handles most of the land surface and a large proportion of it is dedicated to agricultural activities. The current model of agricultural production, strongly influenced by the Green Revolution, has severe environmental and social problems that call into question its sustainability. Agroecology and productive restoration are alternative approaches, which consider the ecological, economic and social dimensions of agri-food systems. The southwest of Buenos Aires province has marginal characteristics for agriculture and its aptitude is mosty for cattle grazing. Productive restoration of natural grasslands of this area through reincorporation of key plant species from the original ecosystem to generate more resilient and sustainable agroecosystems is a priority that must be addressed. The aim of this thesis was to increase the knowledge on some species of herbaceous legumes native to the region with the potential to fix nitrogen, improve forage supply and enrich degraded natural grasslands. First, a bibliographic search was carried out to detect and characterize the native herbaceous legume species reported for the study area; then their populations were located and georeferenced. Subsequently, germination, biological nitrogen fixation capacity (BNF) and nutritional quality of some species were studied. 30 species were detected corresponding to 10 genera, 19 of them potentially useful in agroecosystems and 16 found in at least one of the 43 sites visited, with Adesmia, Lathyrus and Vicia being the genera with more species and A. incana and Rhynchosia senna the most frequently found. Germination in lab conditions of the most promising species from which seeds were collected was studied: Adesmia filipes (Af), A. incana (Ai), A. muricata (Am), Lathyrus nervosus (Ln), L. pubescens (Lp), Rhynchosia diversifolia (Rd) and R. senna (Rs), evaluating the germination percentage (GP) and the response to pregerminative treatments applied to break seed dormancy. Responses were variable according to the species; all present 45% or more dormant seeds and for most of the species a GP greater than 50% was achieved. The most effective treatments were physical scarification; thermal scarification by immersion in hot water was effective for Lp, Rd and Rs. Simultaneously, nodulant rhizobium strains of Ai and Lp were isolated, and BNF was evaluated in Lp using commercial and self-produced inoculant. Lp only established effective symbiosis with native rhizobia and had a fixing efficiency of 50%, without differences in nitrogen content between treatments with mineral or symbiotic nitrogen, but with higher biomass production in the former. Finally, some parameters of nutritional quality of the species Ai, Ln, Lp, L. subulatus, Rd and Rs were analyzed. In vegetative state all of them showed crude protein contents similar to those of cultivated legumes and some of them a higher content, and good digestibility. Protein precipitating tannins were detected only in Rd, Rs and Ai in reproductive status, being moderate to high only in Rd. For the characteristics evaluated in this work and those reported in the literature, many of these species deserve consideration for productive restoration of semi-arid and sub-humid grasslands of central Argentina; new studies focused on analyzing different ways to obtain seeds for medium and large-scale cultivation would help to make their use possible.

A₂bu composed 20-40% of total free amino acids in flatpea tissues and constituted 2-4% of the tissue dry weight. Higher concentrations of A₂bu were found in the leaves than in the roots. A₂bu concentration in leaves and stems increased slightly with plant age. Higher nitrogen availability increased the content of A₂bu in flatpea, a response accompanied by an increase in the contents of soluble protein and other nitrogenous compounds. When exogenous nitrogen supply was decreased, A₂bu levels decreased significantly. Rhizobium infection had no effect on the A₂bu production by flatpea. Ammonium was toxic to flatpea growth. Together with typical toxic symptoms, A₂bu elevation was observed in flatpea plants fed with ammonium. Water-deficit stress also elevated A₂bu content of flatpea. The elevation of A₂bu concentration in aerial tissues of flatpea under stress may not be high enough to decrease the value of flatpea as a forage. 4-Aminobutyric acid (Abu), homoserine (Hse), and asparagine (Asn) were the other major free amino acids in flatpea. As with A₂bu, levels of Hse were higher in the leaves than in the roots. The opposite was true for Abu and Asn. The concentration of Abu in the stems increased consistently with plant age. In response to stress conditions, Abu accumulated in flatpea, especially in stems and roots. Asn was the most prevalent free amino acid in the roots of flatpea. Asn levels in roots increased with plant age and accounted for the greatest portion of the increase in the free amino acid pool in the roots of plants subjected to the water stress or supplied with nitrogen in the form of ammonium ions. Levels of Hse in flatpea were changed little in response to the experimental treatments. Relative amounts of major amino acids in flatpea changed with respect to plant organs and experimental factors. If expressed as ratios, the resulting values suggest metabolic relationships. / Ph. D. / incomplete_metadata

The theme of the thesis are climbing legumes meadow vetchling (Lathyrus pratensis) and tufted vetch (Vicia cracca). The literature search defines watched kinds of legumes and their occurence in permanent grassland. There is summarized seed growing of legumes because of its problematic production. Legumes are valuable components of grassland especially thanks to fixation of nitrogen and high capacity of proteins. The practical part is focused on observation of floristic compilation of watched locations for three years. Particular locations were compared from the point of view of changes in land management and weather. Pods and seeds of observed kinds were collected and evaluated in single years. Experiments of seed's field germination were done after different period from picking and in case of various treatment.

In this study, morphology, anatomy and systematics of the Genus Lathyrus L. (Leguminosae) in Central Anatolia is presented. Comparative morphological characters and their variation in the Genus / Calyx, leaf, corolla and pollen grains micro-morphology of the species / Anatomy of the species / Ecology, endemism, phytogeography and IUCN threat categories of the species / Numerical analysis and Revision of the genus in Central Anatolia were conducted. For the first time the calyx, corolla and leaf micromorphology, and the anatomical characteristics of Lathyrus were examined. Infrageneric delimitation of the species is performed by using multivariate analysis. As a result of morphological and anatomical data, it is suggested that L. haussknechtii should be classified as different species not variety of L. brachypterus. v The collected specimens was crosschecked with neighboring floras and the existing type specimens known from Turkey ANK, GAZI, HUB, KNYA, CUFH, Erciyes, and Royal Botanic Garden Edinburgh (RBGE) herbaria. Morphological and micromorphological characters of pollen grains were revealed. It was determined that the pollen grains size and sculpturing were important diagnostic characters for the species. The statistical analysis was applied to compare P/E ratios of the pollen grains of studied taxa. According to this analysis, the P/E ratio is important diagnostic feature for most of the sections and the species. Sectional key and species key were prepared and updated. Expanded descriptions, GPS data, some photographs and some notes on the taxonomy of the species were given.

La gesse commune (Lathyrus sativus L.) est une légumineuse cultivée principalement en Inde, au Bangladesh et en Ethiopie qui présente des niveaux de résistance élevés pour de nombreuses contraintes abiotiques, telles que la sécheresse et l'inondation. Dans ce travail, les capacités de tolérance du Lathyrus (lignées locales " Raipur " et " Bangladesh ") à une autre contrainte abiotique, la présence de plomb, ont été déterminées des points de vue physiologique et moléculaire. Un système expérimental de culture hydroponique a été mis au point pour ces plantes. Le plomb y est introduit sous forme de nitrate de plomb (Pb(NO3)2). Les teneurs en plomb des différents organes des plantes (racines, tiges, feuilles) ont été déterminées par ICP-OES. Les réponses cellulaires dans ces organes ont été étudiées par RTPCR quantitative (PCR en temps réel). Des amorces spécifiques du Lathyrus ont été dessinées à partir des 11 séquences d'ADN complémentaires (ADNc) isolées pour la première fois et séquencées. L'un des ADNc isolé est complet et code une cystéine protéase (LsCP, 427 aa). Les autres sont des ADNc partiels et correspondent à une aspartique protéase (LsAP, 270 aa), deux ascorbate peroxidases cytosolique (LsAPXc, 195 aa) et peroxisomale (LsAPXp, 226 aa), une protéine de choc thermique (" Heat Shock Protein 70 " ; LsHSP70, 287), une homoglutathion synthétase (LshGSHS, 329 aa), une glutathion S-transférase (LsGST, 66 aa), une glutathion réductase (LsGR, 336 aa), une phytochélatine synthétase (LsPCS, 64 aa), une phospholipase D a (LsPLDa, 288 aa), un transporteur membranaire spécifique du Pb (LsCNGC, 136) et une protéine soluble (LsABCt, 331 aa). Les plantes exposées au nitrate de plomb accumulent de grandes quantités de métal dans leurs racines sous forme de plomb fortement lié aux tissus. L'accumulation d'ARN messagers de la LsPLDa suggère que les racines subissent une contrainte cellulaire et s'y adaptent. La stimulation de l'expression des gènes LsGST, LsGR et LsAPX correspondrait à la formation de complexes Pb-glutathion et à une activation du cycle ascorbate-glutathion pour la neutralisation des espèces activées de l'oxygène (ROS) délétères. Dans les feuilles exemptes de plomb, l'augmentation de l'expression des gènes LsCP, LsAP, LsAPXc, LsAPXp, LsHSP70, LsGR et LsPCS semble indiquer l'émission d'un signal racinaire transmis de manière systémique vers le reste de la plante où il déclenche la sur-expression de ces gènes. Ceci pourrait permettre aux tissus épargnés par le polluant de se préparer à son éventuelle arrivée. La chélation du Pb avec de l'EDTA dans le milieu de culture conduit à son transport vers les parties aériennes et à son accumulation dans les feuilles. L'expression du gène LsCNGC chez ces plantes est activée dans les feuilles, suggérant une participation de ce transporteur à l'entrée des complexes Pb-EDTA dans le symplasme. Comme observé chez d'autres espèces végétales, le plomb affecte le métabolisme du glutathion chez la gesse commune. Cependant, la mise en évidence d'une réaction systémique en réponse au plomb à partir des racines ainsi que la surexpression de gènes d'autolyse sont réalisées pour la première fois et pourraient être des éléments contribuant fortement à la tolérance au plomb chez cette espèce végétale sous-utilisée.

Two experiments were conducted with sheep to determine the nutritional value of 'Lathco' flatpea (Lathyrus sylvestris) hay. In experiment 1, a finishing trial was conducted with 50 group fed crossbred lambs (25 ewes and 25 wethers) fed diets consisting of 70% chopped forage and 30% ground corn grain for 70 d. Five pens of five wethers and five pens of five ewes were selffed diets in which the forage consisted of 100:0, 75:25, 50:50, 25:75, and 0:100 proportions of alfalfa (Medicago sativa L.) and flatpea hay. Feed efficiencies decreased linearly (P<.05) as flatpea increased in the diet. Linear decreases (P<.05) were obtained in kidney and pelvic fat, backfat, leg conformation, and yield grade with increased flatpea in the diet. Blood urea-N (BUN), ruminal pH and NH3-N increased (P<.005) as proportion of flatpea increased. Total volatile fatty acid (VFA) and acetate concentration decreased linearly (P<.005) and propionate, isovalerate, and valerate levels increased linearly (P<.01) with increased proportions of flatpea hay. In experiment 2, 30 wether lambs were fed diets consisting of 100:0, 75:25, 50:50, 25:75 and 0:100 proportions of chopped alfalfa and flatpea hays in a digestion trial. Neutral detergent fiber (NOE), acid detergent fiber (ADE), cellulose, hemicellulose and energy digestibilities decreased linearly (P<.01) with increased levels of flatpea hay. Nitrogen retention was not different between sheep fed the different diets. Apparent absorbtion and retention of Ca decreased linearly (P<.05) with increased level of flatpea hay Ruminal pH and NHa-N, and BUN increased linearly (P<.OOl) as flatpea was increased in the diet. Ruminal total VEA, acetate and acetate to propionate ratio decreased linearly (P<.001) and propionate, isobutyrate, isovalerate, and valerate levels increased linearly (P<.001) as level of flatpea increased. / Master of Science