Discovery of two new species of Crotalaria (Leguminosae, Crotalarieae) from Western Ghats, India.

Two new species of Fabaceae-Papilionoideae are described and illustrated. Crotalaria suffruticosa from Karul Ghat region of Maharashtra is morphologically close to C. albida and C. epunctata. C. multibracteata from Panhala region of Maharashtra resembles C. vestita. C. suffruticosa differs from C. albida and C. epunctata in its habit, leaf, inflorescence, callosity, keel type, stigma, style morphology and number of seeds/pod. To test if the new species differ from their morphologically most similar species, we measured various traits and performed a Principal Component Analysis (PCA). This analysis shows that the new species differs from similar species in gross morphology for several diagnostic traits and showed correlations between the variables or distance among groups and estimated the contribution of each character. Phylogenetic analyses were also conducted based on nuclear (ITS) and plastid (matK) markers. The analyses revealed nucleotide differences between the new species and their close allies attributing to their distinctiveness. A map and key including all species of Crotalaria from Maharashtra state are provided. Conservation status of the two new species have also been assessed.

Clock-dated phylogeny for 48% of the 700 species of Crotalaria (Fabaceae-Papilionoideae) resolves sections worldwide and implies conserved flower and leaf traits throughout its pantropical range.

BACKGROUND: With some 700 species, the pantropical Crotalaria is among the angiosperm's largest genera. We sampled 48% of the species from all sections (and representatives of the 15 remaining Crotalarieae genera) for nuclear and plastid DNA markers to infer changes in climate niches, flower morphology, leaf type, and chromosome numbers. RESULTS: Crotalaria is monophyletic and most closely related to African Bolusia (five species) from which it diverged 23 to 30 Ma ago. Ancestral state reconstructions reveal that leaf and flower types are conserved in large clades and that leaf type is uncorrelated to climate as assessed with phylogenetically-informed analyses that related compound vs. simple leaves to the mean values of four Bioclim parameters for 183 species with good occurrence data. Most species occur in open habitats <1000 m alt., and trifoliolate leaves are the ancestral condition, from which unifoliolate and simple leaves each evolved a few times, the former predominantly in humid, the latter mainly in dry climates. Based on chromosome counts for 36% of the 338 sequenced species, most polyploids are tetraploid and belong to a neotropical clade. CONCLUSIONS: An unexpected finding of our study is that in Crotalaria, simple leaves predominate in humid climates and compound leaves in dry climates, which points to a different adaptive value of these morphologies, regardless of whether these two leaf types evolved rarely or frequently in our focal group.

Feeding on Host Plants with Different Concentrations and Structures of Pyrrolizidine Alkaloids Impacts the Chemical-Defense Effectiveness of a Specialist Herbivore.

Sequestration of chemical defenses from host plants is a strategy widely used by herbivorous insects to avoid predation. Larvae of the arctiine moth Utetheisa ornatrix feeding on unripe seeds and leaves of many species of Crotalaria (Leguminosae) sequester N-oxides of pyrrolizidine alkaloids (PAs) from these host plants, and transfer them to adults through the pupal stage. PAs confer protection against predation on all life stages of U. ornatrix. As U. ornatrix also uses other Crotalaria species as host plants, we evaluated whether the PA chemical defense against predation is independent of host plant use. We fed larvae from hatching to pupation with either leaves or seeds of one of eight Crotalaria species (C. incana, C. juncea, C. micans, C. ochroleuca, C. pallida, C. paulina, C. spectabilis, and C. vitellina), and tested if adults were preyed upon or released by the orb-weaving spider Nephila clavipes. We found that the protection against the spider was more effective in adults whose larvae fed on seeds, which had a higher PA concentration than leaves. The exceptions were adults from larvae fed on C. paulina, C. spectabilis and C. vitellina leaves, which showed high PA concentrations. With respect to the PA profile, we describe for the first time insect-PAs in U. ornatrix. These PAs, biosynthesized from the necine base retronecine of plant origin, or monocrotaline- and senecionine-type PAs sequestered from host plants, were equally active in moth chemical defense, in a dose-dependent manner. These results are also partially explained by host plant phylogeny, since PAs of the host plants do have a phylogenetic signal (clades with high and low PA concentrations in leaves) which is reflected in the adult defense.

Pulmonary and hepatic lesions caused by the dehydropyrrolizidine alkaloid-producing plants Crotalaria juncea and Crotalaria retusa in donkeys.

The effects and susceptibility of donkeys to Crotalaria juncea and Crotalaria retusa poisoning were determined at high and low doses. Seeds of C. juncea containing 0.074% of dehydropyrrolizidine alkaloids (DHPAs) (isohemijunceines 0.05%, trichodesmine 0.016%, and junceine 0.008%) were administered to three donkeys at 0.3, 0.6 and 1 g/kg body weight (g/kg) daily for 365 days. No clinical signs were observed and, on liver and lung biopsies, the only lesion was a mild liver megalocytosis in the donkeys ingesting 0.6 and 1 g/kg/day. Two other donkeys that received daily doses of 3 and 5 g seed/kg showed initial respiratory signs 70 and 40 days after the start of the administration, respectively. The donkeys were euthanized following severe respiratory signs and the main lung lesions were proliferation of Clara cells and interstitial fibrosis. Three donkeys ingested seeds of C. retusa containing 5.99% of monocrotaline at daily doses of 0.025, 0.05 and 0.1 g/kg for 365 days. No clinical signs were observed and, on liver and lung biopsies, the only lesion was moderate liver megalocytosis in each of the three donkeys. One donkey that received a single dose of 5 g/kg of C. retusa seeds and another that received 1 g/kg daily for 7 days both showed severe clinical signs and died with diffuse centrilobular liver necrosis. No lung lesions were observed. Another donkey that received a single dose of 2.5 g/kg of C. retusa seeds showed no clinical signs. The hepatic and pneumotoxic effects observed are consistent with an etiology involving DHPAs. Furthermore, the occurrence of lung or liver lesions correlates with the type of DHPAs contained in the seeds. Similarly as has been reported for horses, the data herein suggest that in donkeys some DHPAs are metabolized in the liver causing liver disease, whereas others are metabolized in the lung by Clara cells causing lung disease.

Glucose, stem dry weight variation, principal component and cluster analysis for some agronomic traits among 16 regenerated Crotalaria juncea accessions for potential cellulosic ethanol.

The objectives of this research were to identify candidate sunn hemp accessions having high concentrations of cellulose for use as parents in breeding for cellulose and to determine variability for glucose content and some important agronomic traits among sunn hemp accessions. Since sunn hemp is an under-utilized species, glucose content and agronomic trait variation is essential for the identification of superior sunn hemp accessions for use as potential ethanol for biofuel. Sixteen sunn hemp accessions including the following plant introductions (expressed as glucose concentration) and stem dry weights were studied. "Sixteen sunn hemp accessions including the following plant introductions (expressed as glucose concentration) and stem dry weights were studied." In addition, to verify variability, these traits plus morphological, phenological, and seed reproductive traits were analyzed using multivariate and cluster analysis. The accessions, PI 250487, PI 337080, and PI 219717 produced the highest glucose concentrations (859, 809, and 770 mg g(-1) stem dry weight, respectively), however PI 468956 produced the highest stem dry weight (258 g). Branching significantly correlated with foliage (r(2) = 0.67**) and relative maturity (r(2) = 0.60*), while maturity had a significantly negative correlation with seed number (r(2) = -0.67**) and plant width (r(2) = -0.53*) as well. Seed number significantly correlated with plant width (r(2) = 0.57*). Average linkage cluster analysis grouped the 16 sunn hemp accessions into well-defined phenotypes with four distinct seed-producing groups and one outlier. Based on multivariate and cluster analysis, sufficient variation among these16 sunn hemp accessions exists to support the development of cellulosic ethanol producing cultivars with improved architecture, early maturity, seed yield, glucose concentrations, and stem dry weights.

Influência da adubação verde e diferentes adubos orgânicos na produção de fitomassa aérea de atroveran (Ocimum selloi Benth. ) / Influence of green manure and different organic manure on production of aereal phytomass of Ocimum selloi

Este trabalho teve por objetivo verificar o efeito de diferentes adubos orgânicos em associação ou não com adubo verde na produção de folhas de Ocimum selloi Benth., planta nativa do Brasil. O experimento foi conduzido na área experimental da Embrapa Meio Ambiente (CNPMA), localizado no município de Jaguariúna, em duas áreas distintas, sendo uma delas submetidas anteriormente ao plantio e incorporação de adubo verde (Crotalaria juncea). Os tratamentos utilizados foram T1 - testemunha (solo sem adubação), T2 - cama de aviário (5 kg m-2), T3 - hidrolisado de peixe (produto comercial Fishfértil - 5 mL m-2) e T4 - composto orgânico (4 kg m-2). A colheita foi realizada 180 dias após o plantio, em janeiro de 2011, sendo colhidas as plantas úteis (quatro plantas por parcela). Avaliou-se o rendimento de fitomassa fresca e seca. O delineamento experimental adotado foi o esquema fatorial 2x4, com quatro repetições (blocos). As médias obtidas foram submetidas à análise de variância seguida de teste de médias (Tukey). Com base nos resultados obtidos, pode-se concluir que a cama de aviário apresentou resultados mais satisfatórios quanto à produção de folhas de Ocimum selloi quando comparados aos demais tratamentos orgânicos, não se observando incremento nos resultados pela associação com o adubo verde Crotalaria juncea.

The aim of this work was to avail the effect of different manures in association or nor with green manure on yield of leaves of Ocimum selloi Benth. The assay was accomplished on experimental area of Embrapa Environmental (Jaguariúna district), at two different spaces (with or without green manure Crotalaria juncea). The treatments used were T1 - witness (no manure), T2 - chicken manure (5 kg m-2), T3 - commercial product Fishfértil - 5 mL m-2) and T4 - composting (4 kg m-2). The cut was realized on 180 days after the planting (january - 2011), and were collected four plants/ plot. The yield of dried and fresh Ocimum selloi phytomass was availed. The experimental design was factorial scheme (2x4), with four repetitions. The treatment with chicken manure showed best results on Ocimum selloi leaves yield than the others treatments, but did not was influence by using Crotalaria juncea.

Crotalaria medicaginea associated with horse deaths in northern Australia: new pyrrolizidine alkaloids.

Crotalaria medicaginea has been implicated in horse poisoning in grazing regions of central-west Queensland, which resulted in the deaths of more than 35 horses from hepatotoxicosis in 2010. Liver pathology was suggestive of pyrrolizidine alkaloidosis, and we report here the isolation of two previously uncharacterized pyrrolizidine alkaloids from C. medicaginea plant specimens collected from pastures where the horses died. The first alkaloid was shown by mass spectometric and NMR analyses to be 1ß,2ß-epoxy-7ß-hydroxy-1α-methoxymethyl-8α-pyrrolizidine, which, like other alkaloids previously isolated from C. medicaginea, lacks the requisite functionality for hepatotoxcity. The second alkaloid isolated in this investigation was a new macrocyclic diester of otonecine, which we have named cromedine. The (1)H and (13)C NMR spectra of cromedine were fully assigned by 2D NMR techniques and allowed the constitution of the macrocyclic diester to be assigned unambiguously. C. medicaginea specimens implicated in this investigation do not belong to any of the three recognized Australian varieties (C. medicaginea var. neglecta, C. medicaginea var. medicaginea, and C. medicaginea var. linearis) and appear to be a local variant or form, referred to here as C. medicaginea (chemotype cromedine).

Pyrrolizidine alkaloid-containing toxic plants (Senecio, Crotalaria, Cynoglossum, Amsinckia, Heliotropium, and Echium spp.).

Pyrrolizidine alkaloid (PA)-containing plants are found throughout the world and are probably the most common plant cause of poisoning of livestock, wildlife, and humans. PAs are potent liver toxins that under some conditions can be carcinogenic. This article briefly introduces high-risk North American PA-containing plants, summarizing their toxicity and subsequent pathology. Current diagnostic techniques, treatments, and strategies to avoid losses to PA poisoning are also reviewed.

[ Nodulation of certain legumes of the genus Crotalaria by the new species Methylobacterium]. / Certaines légumineuses du genre Crotalaria sont spécifiquement nodulées par une nouvelle espèce de Methylobacterium.

We studied a collection of 126 rhizobial isolates from eight species of Crotalaria (C. comosa, C. glaucoides, C. goreensis, C. hyssopifolia, C. lathyroides, C. perrottetii, C. podocarpa, and C. retusa) growing in Senegal. Nodulation and nitrogen-fixation tests on nine Crotalaria species revealed two specificity groups within the genus Crotalaria. Group I consists of plants solely nodulated by very specific fast-growing strains. Group II plants are nodulated by slow-growing strains similar to promiscuous Bradyrhizobium spp. strains already reported to nodulate many tropical legumes. SDS-PAGE studies showed that slow-growing strains grouped with Bradyrhizobium while fast-growing strains constituted a homogeneous group distinct from all known rhizobia. Amplified ribosomal DNA restriction analysis (ARDRA) of 10 representative strains of this group using four restriction enzymes showed a single pattern for each enzyme confirming the high homogeneity of group I. The 16S rDNA sequence analysis revealed that this specific group belonged to the genus Methylobacterium, thus constituting a new branch of nodulating bacteria.