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Indigofera suffruticosa has reports of its popular use for analgesy in several cultures. Besides that, all parts of this plant are used for some medicinal outcome. The leaves are used in teas, decoctions, juices and included in baths for treating fever and inflammatory processes.To evaluate the anti-inflammatory activity mechanisms of I. suffruticosa leave aqueous extract (IsAE).Phytochemical screening of IsAE was performed by thin layer chromatography. Total flavonoid content was determined and expressed by milligram of quercetin equivalent per gram of extract (mgEQ/g). 50% of the lethal dose that kills animals (LDPhytochemical analysis revealed the presence of flavonoids and acid gallic in a quantity of 33.9 mg QE/g. Acute toxicity evaluation resulted in a LDIsAE proved to be anti-inflammatory, acting in different parts of the inflammation process, confirming its popular use.

Indospicine, a non-proteinogenic analogue of arginine, occurs only in Indigofera plant species and accumulates in the tissues of animals grazing on Indigofera. Canine deaths have resulted from the consumption of indospicine-contaminated meat but only limited information is available regarding indospicine toxicity in humans. In this study three human cell lines, Caco-2 (colorectal adenocarcinoma), HT29-MTX-E12 (colorectal adenocarcinoma) and HepG2 (hepatocellular carcinoma), were used to investigate the cytotoxicity of indospicine and its metabolite 2-aminopimelic acid in comparison to arginine. Indospicine and 2-aminopimelic acid were more cytotoxic than arginine, displaying the highest toxicity in HepG2 liver cells. Intestinal transport in vitro also revealed a 2-fold higher transport rate of indospicine compared to arginine. The sensitivity of HepG2 cells to indospicine is consistent with observed canine hepatotoxicity, and considering the higher in vitro transport of indospicine across an intestinal barrier, it is possible that similar ill effects could be seen in humans consuming contaminated meat.

Malaria is still a major health problem worldwide. This study aimed to investigate the hepatoprotective role of Indigofera oblongifolia leaf extracts (ILE) against mice hepatic injury induced by Plasmodium chabaudi. Female C57BL/6 mice were treated with 100 mg/kg of ILE after infection with erythrocytes parasitized by P. chabaudi. On day 7 post-infection, the extract improved the histological alteration induced by the parasite. This was evidenced by the decreased histological index induced by ILE. Moreover, ILE was able to increase the hepatic antioxidant capacity and could significantly improve the decrease in erythrocyte count and hemoglobin content in mice blood plasma due to infection. ILE was also able to upregulate the expression of 24 genes related to metabolism and of 3 genes related to the immune response. Furthermore, the extract was able to downregulate the expression of 35 genes related to metabolism and of 82 genes related to immune response. Moreover, the microarray study showed that ILE regulated the change in gene expression induced by the parasite. Among these genes, we quantified the expression of cd209f, cyp7a1, Hsd3b5, Sult2a3, Lcn2, CcI8, Nos2, and saa3-mRNAs. These genes were regulated by ILE. Therefore, our results revealed the protective role of Indigofera oblongifolia against hepatic injury induced by blood stage malaria.

Stachyodin A (1), a rearrangement product of pterocarpan featuring a 6/5/5/6 tetracyclic ring system with an unusual spirotetrahydrofuran ring, and stachyodin B (2), a new dihydrochalcone, along with five known analogues (3–7) were isolated from the roots of Indigofera stachyodes. The structures of 1 and 2 were elucidated on the basis of their HRESIMS and NMR spectroscopic data, and their absolute configurations were determined by X-ray crystallographic analysis and electronic circular dichroism (ECD) data, respectively. Compounds 2–4 showed inhibition of nitric oxide production in lipopolysaccharide-activated BV-2 microglial cells.

Indigofera, the third largest genus of Fabaceae, comprises approximately 750 species worldwide with a pantropical distribution. Eight Indigofera species, namely I. calcicola, I. delavayi, I. franchetii, I. hancockii, I. lenticellata, I. pendula, I. rigioclada, and I. szechuensis, are considered a species complex because of their morphological similarities and the phylogenetic analysis based on Internal Transcribed Spacer (ITS). Small populations of these species are allopatrically distributed in Hengduan Mountains in China. Although considerable EST-SSR markers have been developed from the transcriptome of I. szechuensis, no codominant markers have been applied to study population genetic structure of the complex. In this study, we selected 66 EST-SSR markers which were transferable in Indigofera szechuensis complex for estimating polymorphism, of which 44 EST-SSRs (66.67%) were polymorphic. Amplification with selected 23 polymorphic EST-SSRs revealed a moderately high genetic diversity in this complex. The mean value of Observed number of alleles, Expected heterozygosity, Polymorphism information content and Shannon's information index was 10.4783, 0.4335, 0.6228 and 1.4369 respectively. A total of 758 genotypes were detected, with an average of 32.9565 genotypes per locus. The Mantel test showed a significant correlation between genetic and geographic distance (r=0.0748, P=0.0100). High differentiation and limited gene flow were detected among all populations (Fst=0.3589, Nm=0.5168). The PCoA and structure analysis grouped 31 populations of Indigofera szechuensis complex into five main species. I. delavayi was obviously separated from other species, and the result was in accordance with that of morphology and phylogeny. I. pendula was a separate species, and had two distinct phenotypes. Four I. szechuensis populations (Pop23, Pop24, Pop27 and Pop30) were the same species. I. calcicola, I. hancockii, I. rigioclada, I. franchetii, I. lenticellata, and I. szechuensis, distributed in southwest of Jinshajiang, could be considered as the same species; the species of I. franchetii, I. lenticellata, and I. szechuensis distributed in northeast of Jinshajiang could be considered as another separate species. Given the above information, the morphological classification of I. calcicola, I. franchetii, I. hancockii, I. lenticellata, I. rigioclada, and I. szechuensis was incredible.

Indigofera szechuensis Craib, a species endemic to Hengduan Mountains in China, is widely distributed across a range of habitats, including slopes, trailsides, and riverbanks. This species serves as a good model to examine flexible adaptation of plants. In this study, we reported the transcriptome sequence of this species and developed a set of SSR markers for such evolutionary studies. Approximately 19.4 million clean reads from the leaves of I. szechuensis were generated through transcriptome sequencing, and 43,896 unigenes with an average length of 798.54 bp were obtained. A total of 27,320 (62.2%) unigenes were annotated for their functions, and 17,422 and 8017 unigenes could be aligned to the GO and COG database, respectively. By searching against the Kyoto Encyclopedia of Genes and Genomes Pathway database (KEGG), 5682 unigenes were assigned to 118 KEGG pathways that belong to five main categories, i.e., organismal systems, cellular processes, environmental information processing, metabolism, and genetic information processing. We further identified 4052 potential SSR sites. Using 102 randomly selected EST-SSRs, 73 markers were successfully amplified with the expected size in I. szechuensis and 67-71 markers of these 73 SSRs in eight other Indigofera species. We firstly established a public trascriptome platform for the species-rich genus Indigofera. The SSR sites identified here provide additional resources for development of molecular markers for further evolutionary studies for this species and congeners.

Dyeing is one of the stages in tanning process to apply base color on leather to avoid broken finishing coat. One of natural substance used in leather tanning is true indigo (Indigofera tinctoria L.). This substance has blue color and contains tannin to increase the quality of tanned Milkfish skin. The research was aimed to determine the effect of using different concentrations of true indigo application as natural substance to the quality of tanned milkfish skin. This research used completely randomized design (CRD) using three repetitions, the treatment were using the concentration of indigo 20%, 25% and 30%. The result shows that different concentrations of true indigo has significant effect (p < 0.05) to rub resistance on wet and dry coating, fastness to perspiration, tensile strength, elongation, hedonic, and micrograph photos of skin tissue. Tanned milkfish skin using 25% concentration of true indigo has the best quality based on the criteria: rub resistance on wet coating test (3.83%) and dry coating test (4.42%), fastness to perspiration (4.75%), tensile strength (1 843.53 N · cm–2), elongation (47.61%), and skin tissue micrograph photos showing better display of collagen fibers.

The main aim of this work is focused towards possible reuse of both solid and liquid waste generated from the natural indigo dye production process. The solid waste (C/N:15.01) was utilized to produce stable compost with possible re-use in Indigofera cultivation. Among seven compost combinations (C1–C7) using jeevamrutha (JA) and cow-dung (CD) as inoculum, C4 with 8% JA showed higher biomass degradation (51%) and plant growth potential (GI > 125%). Whereas the undiluted liquid waste was treated using algal consortia, bacteria, and indigenous microbial population, achieved a maximum removal of 90% ammonia, 82% nitrate, and 88% phosphorus for its re-use in the dye production process. Hence, incorporation of suitable waste management strategies in natural indigo dye production could help to achieve a zero waste sustainable process.

A new isoflavone 7-hydroxy 3′-methoxyisoflavone ( 1 ) is isolated from the seeds of Indigofera heterantha . The structure of this new compound was established using spectroscopic techniques such as ID, 2D NMR, and mass spectrometry. Density functional theory calculations are carried out for the first time for geometric, electronic and spectroscopic properties of 1 (isoflavone). DFT calculations have been performed at B3LYP/6-311G(d,p) level of theory for obtaining geometric and spectroscopic properties of compound 1 . The simulated vibrational spectrum of compound 1 at B3LYP/6-311G(d,p) shows nice correlation with the experimental IR spectrum after a scaling factor of 0.973. 1 H and 13 C NMR chemical shifts were calculated using Cramer’s re-parameterized function WP04 at 6-311G(d,p) basis set, and show nice correlation with the experimental data. Four conformers were considered for NMR chemical shift calculations. Electronic properties such as band gap, Ionization potential and electron affinities were also simulated for the first time; however, no comparison could be made with the experiment.

Indigofera caerulea Roxb. is a well known shrub among native medical practitioners in folk medicine used for the treatment of jaundice, epilepsy, night blindness and snake bites. It is also reported to have antioxidant and antimicrobial properties. However its actual efficacy and hepatoprotective mechanism in particular is uncertain. Thus the present study investigates the hepatoprotective effect of the methanolic extract of I. caerulea Roxb. leaves (MIL) and elucidation of its mode of action against carbon tetrachloride (CCl4) induced liver injury in rats. HPLC analysis of MIL when carried out showed peaks close to standard ferulic acid and quercetin. Intragastric administration of MIL up to 2000 mg/kg bw, didn't show any toxicity and mortality in acute toxicity studies. During "in-vivo" study, hepatic injury was established by intraperitoneal administration of CCl4 3 ml/kg bw (30% CCl4 in olive oil; v/v) twice a week for 4 weeks in Sprague-Dawley rats. Further, hepatoprotective activity of MIL assessed using two different doses (100 and 200mg/kg bw) showed that intra-gastric administration of MIL (200mg/kg bw) significantly attenuates liver injury. Investigation of the underlying mechanism revealed that MIL treatment was capable of reducing inflammation by an antioxidant defense mechanism that blocks the activation of NF-κB as well as inhibits the release of proinflammatory cytokine TNF-α and IL-1β. The results suggest that MIL has a significant hepatoprotective activity which might be due to the presence of phytochemicals namely analogues of ferulic acid and other phytochemicals which together may suppress the inflammatory signaling pathways and promote hepatoprotective activity against CCl4 intoxicated liver damage.

A new flavan dimer, 2 alpha,3 alpha-epoxyflavan-5,7,3',4'-tetraol-(4 beta -> 8)-flavan-5 '',7 '',4'''-triol (1), and a new flavonol, 3-O-(3-nitropropanoyl)-2,3-cis-5,7,3',4'-tetrahydroxyflavan (2), together with a known compound, 2 alpha,3 alpha-epoxy-5,7,3',4'-tetrahydroxyflavan-(4 beta -> 8)-epicatechin (3), were isolated from the roots of Indigofera stachyodes. Their structures were elucidated by spectroscopic techniques including MS, 1D NMR, and 2D NMR. Compounds 2 and 3 were evaluated to determine their protective effects against carbon tetrachloride (CCl4)-induced hepatotoxicity in the human liver cell line HL-7702. The results showed that 2 and 3 could protect HL-7702 cells from injury induced by CCl4, with cell survival rates of 122.0% and 72.5%, respectively. (C) 2013 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.

Ethnopharmacological relevance Indigofera tinctoria Linn. ( I. tinctoria , Fabaceae) has been widely used for several years in the traditional Indian and Chinese system of Medicine for the treatment of epilepsy, nervous and brain disorders. Aim of the study The effect of SF-6, a compound isolated from I. tinctoria to exhibit neuroprotection in in vitro and in vivo models of Parkinson's disease (PD), was investigated. Materials and methods Using human neuroblastoma SH-SY5Y cells, the effect of SF-6 on α-synuclein- or 6-hydroxydopamine (6-OHDA)-, hydrogen peroxide (H 2 O 2 )-induced cytotoxicity in vitro was investigated. In in vivo studies SF-6 was challenged against 6-OHDA-induced neuronal damage and behavioral deficits in mice. Results SF-6 (1, 5 and 10 μg/mL) significantly inhibited α-synuclein- or 6-OHDA-, H 2 O 2 -induced cytotoxicity and decreased the reactive oxygen species production in SH-SY5Y cells. SF-6 also scavenged hydroxyl free radicals. In in vivo evaluation, SF-6 attenuated the contralateral rotational asymmetry observed by apomorphine challenge in 6-OHDA-lesioned mice. Further, the behavioral deficits evaluated by rotarod test, Y-maze and passive avoidance tasks were reversed by SF-6 and was found more potent compared with standard compound deprenyl. Conclusion Data suggest that SF-6 showed neuroprotection in experimental models of PD due to its potent antioxidant action supporting the traditional claim for its use in nervous and brain disorders.

Poisoning by Indigofera lespedezioides is reported in horses in the state of Roraima, northern Brazil. The main clinical signs are anorexia, sleepiness, unsteady gait, severe ataxia, weakness, stumbling, and progressive weight loss. To induce the disease experimentally, a 7-year-old horse was introduced in a small paddock invaded by the plant. The first nervous signs were observed 44 days from the start of grazing. The animal was euthanized on day 59. No significant gross lesions were observed upon necropsies of the experimental horse as well as one spontaneously affected horse. Upon histologic examination neuronal lipofuscinosis was observed in the brain, cerebellum, and spinal cord. Wallerian-type degeneration was observed on some mesencephalic tracts. Neuronal and axonal degeneration and lipofuscinosis were observed on electron microscopy examination. Indospicine was detected in four samples of I. lespedezioides with concentrations ranging from 63 to 1178 μg/g whereas nitro toxins could be detected in only one of the samples at a concentration of 2.5 mg/g. In conclusion, poisoning by I. lespedezioides is very similar to those poisonings by Indigofera linnaei and Indigofera hendecaphylla. Based on the preponderance of indospince and lack of nitro toxins in the samples it is proposed that indospicine is the toxic compound responsible for the poisoning.

Made available in DSpace on 2013-09-27T14:54:28Z (GMT). No. of bitstreams: 1 WOS000295236700022.pdf: 1054917 bytes, checksum: ade3c93db496ae9bfef43fc129742e42 (MD5) Previous issue date: 2011-09-01 Made available in DSpace on 2013-09-30T18:37:23Z (GMT). No. of bitstreams: 1 WOS000295236700022.pdf: 1054917 bytes, checksum: ade3c93db496ae9bfef43fc129742e42 (MD5) Previous issue date: 2011-09-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:49:42Z No. of bitstreams: 1 WOS000295236700022.pdf: 1054917 bytes, checksum: ade3c93db496ae9bfef43fc129742e42 (MD5) Made available in DSpace on 2014-05-20T13:49:42Z (GMT). No. of bitstreams: 1 WOS000295236700022.pdf: 1054917 bytes, checksum: ade3c93db496ae9bfef43fc129742e42 (MD5) Previous issue date: 2011-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Ethnopharmacological relevance: Indigofera suffruticosa is specie typical of the Cerrado or Brazilian savannah; it is a member of the Fabaceae family - in folkmedicine is used for gastric disorders, infection and inflammation.Aim of the study: Ethyl acetate fraction (AcF) and aqueous fraction (AqF) of the methanolic extract of I. suffruticosa leaves were evaluated against acute gastric ulcer. The AcF fraction was selected to assess its activity in ulcer healing and its gastroprotective effects via mucus and gastric secretion.Materials and methods: The gastroprotective action of AcF and AqF fractions were evaluated in a rodent experimental model. The action mechanisms, involvements of the antisecretory action, mucus and prostaglandin production, toxicological and healing activity of the AcF (100 mg/kg, p.o.) were evaluated. We also used histological analysis (HE and PAS) and immunohistochemical (PCNA and HSP-70) assays to evaluate the effects of I. suffruticosa.Results: AcF significantly inhibited the gastric mucosal damage caused by ethanol. This effect was statistically significant in 100 mg/kg group compared vehicle. AcF did not interfered with gastric secretion, significantly increased the PGE(2) and mucus production (validated in PAS technique). The gastroprotection was attenuated by pretreatment with N-ethylmaleimide, but not L-NAME. In acid-acetic-induced ulcer model AcF accelerated ulcer healing. Immunohistochemistry analysis showed induction of proliferating cell (PCNA) and heat shock protein (HSP 70).Conclusions: These results showed that AcF acted as gastroprotective agent stimulating prostaglandin, mucus and HSP70. (C) 2011 Elsevier B.V. All rights reserved. Univ Estadual Campinas, Dept Anat Biol Celular Fisiol & Biofis, Inst Biol, BR-13083970 Campinas, SP, Brazil Univ Estadual Campinas, Fac Ciencias Med, Dept Farmacol, BR-13083970 Campinas, SP, Brazil Univ Estadual Paulista, Inst Quim, Dept Quim Organ, Araraquara, SP, Brazil Univ Estadual Paulista, Dept Morfol, Inst Biociencias, Botucatu, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Fisiol, Botucaru, SP, Brazil Univ Estadual Paulista, Inst Quim, Dept Quim Organ, Araraquara, SP, Brazil Univ Estadual Paulista, Dept Morfol, Inst Biociencias, Botucatu, SP, Brazil Univ Estadual Paulista, Inst Biociencias, Dept Fisiol, Botucaru, SP, Brazil

Ethnopharmacological relevance: Although traditional antimalarial plant remedies in herbal markets are a very important component of the health care system in Ghana this has not been previously studied to allow for the formulation of effective strategy for malaria control in Ghana. Aim of study: The main objective of the present study was to collect and analyse data on the antimalarial plant remedies in herbal markets in southern Ghana. Materials and methods: Herborists were interviewed using a validated questionnaire and species of plants were identified using a combination of field photo guides, local names and voucher specimens. Results: A total of 71 herborists (95.8% female) were interviewed. There were potential correlations between different parameters and variables such as ethnic groups, type of vendor and age-groups. The study revealed 29 species of plants belonging to 22 families being sold for the treatment of malaria. The detailed use of these plants is documented. The most frequently mentioned species of plants were Morinda lucida Benth., Indigofera sp. and Nauclea latifolia Sm. The majority (82.8%) of the plant materials were sold in the dried state and 6.9% were sold in fresh state. About 76.2% of the herbal remedies were sold throughout the year while 23.8% were scarce in the dry season. The cost of treatment of malaria using the herbal remedies ranged from 1 to 2 United States Dollars (USD). Conclusion: Standardization of names and authentication of plant materials using organoleptic, phytochemical and DNA barcoding techniques as well as further research on efficacy, safety and dosage prescriptions for both fresh and dried plant materials being sold for the treatment of malaria in southern Ghana are needed.

By considering controversial discussions in the literature with regard to gland denomination in Indigofera species, as well as the taxonomic value of secretory structures in Leguminosae, we aim to morphologically detail glands that had been previously observed in I. microcarpa and I. sabulicola, and to investigate the occurrence of glands in vegetative and reproductive organs of other six Neotropical species that belong to the genus. Glands analyzed through scanning electronic microscopy (SEM) in combination with anatomic analyses correspond to secretory trichomes that are classified into seven types. Main variations in relation to types occurred with regard to head shape and peduncle size. Trichome heads were multicellular, with a thin cuticle. Hollow heads with conspicuous inner space characterized only one type (type 1); the other trichome types had massive heads. Peduncles, which varied from biseriate to multiseriate, had thick, pecto-cellulosic cell walls. Trichomes were found on stems, stipules, petioles, rachis, petiolules, leaflets, bracteoles, sepals, standards and fruits, more commonly along the margins. Each of the eight Indigofera species analyzed had at least two different trichome types out of the seven types that occurred in reproductive and vegetative organs of these taxa. Various types of secretory trichomes were found in I. campestris, I. lespedezioides, I. microcarpa, I. spicata, I. suffruticosa and I. truxillensis. Stems and rachis were the vegetative organs in which a greater variety of trichomes occurred, and sepals were parts of reproductive organs with the same status. Five out of the seven secretory trichome types occurred on both vegetative and reproductive organs. Distribution and gland types differed between species and these gland distribution patterns can be used as diagnostic characters. Reports of glands in Indigofera campestris, I. hirsuta, I. lepedezioides, I. suffruticosa, I. spicata and I. truxillensis, their recognition as secretory trichomes, and the morphological variety of types found for such trichomes are novel data for Indigofera.

Two experiments were completed to determine the influence of season/year and species on forage quality of Indigofera accessions. In Experiment 1, leaf material was collected from five Indigofera species (i.e., I. amorphoides, I. arrecta, I. brevicalyx, I. costata and I. cryptantha) by harvesting six middle plants of a plot in the autumn of 2003 and the autumn of 2004. In Experiment 2, edible forage (leaves +

An investigation of aliphatic nitro-compounds in Indigofera carlesii resulted in the identification of two blended novel compounds: 2- O -acryl-3,6-di- O -(3-nitropropanoyl)-α- d -glucopyranose and 6 -O -acryl-2,3-di- O -(3-nitropropanoyl)-α- d -glucopyranose. The structures of the new compounds were elucidated on basis of spectral analysis.

A review is given of the legume tribe Indigofereae for the region of southern Africa including Namibia. The overall distribution, taxon statistics, phylogenetic relationships, uses, bibliography and a checklist of taxa in the tribe are provided covering the c. 340 taxa of Indigofereae within the region.

Bacteria were isolated from root-nodules collected from indigenous legumes at 38 separate locations in the Gascoyne and Pilbara regions of Western Australia. Authentication of cultures resulted in 31 being ascribed status as root-nodule bacteria based upon their nodulation of at least one of eight indigenous legume species. The authenticated isolates originated from eight legume genera from 19 sites. Isolates were characterised on the basis of their growth and physiology; 20 isolates were fast-growing and 11 were slow-growing (visible growth within 3 and 7d, respectively). Fast-growers were isolated from Acacia, Isotropis, Lotus and Swainsona, whilst slow-growers were from Muelleranthus, Rhynchosia and Tephrosia. Indigofera produced one fast-growing isolate and seven slow-growing isolates. Three indigenous legumes (Swainsona formosa, Swainsona maccullochiana and Swainsona pterostylis) nodulated with fast-growing isolates and four species (Acacia saligna, Indigofera brevidens, Kennedia coccinea and Kennedia prorepens) nodulated with both fast- and slow-growing isolates. Swainsona kingii did not form nodules with any isolates. Fast-growing isolates were predominantly acid-sensitive, alkaline- and salt-tolerant. All slow-growing isolates grew well at pH 9.0 whilst more than half grew at pH 5.0, but all were salt-sensitive. All isolates were able to grow at 37deg;C. The fast-growing isolates utilised disaccharides, whereas the slow-growing isolates did not. Symbiotic interactions of the isolates were assessed on three annual, one biennial and nine perennial exotic legume species that have agricultural use, or potential use, in southern Australia. Argyrolobium uniflorum, Chamaecytisus proliferus, Macroptilium atropurpureum, Ononis natrix, Phaseolus vulgaris and Sutherlandia microphylla nodulated with one or more of the authenticated isolates. Hedysarum coronarium, Medicago sativa, Ornithopus sativus, Ornithopus compressus, Trifolium burchellianum, Trifolium polymorphum and Trifolium uniflorum did not form nodules. Investigation of the 31 authenticated isolates by polymerase chain reaction with three primers resulted in the RPO1 primer distinguishing 20 separate banding patterns, while ERIC and PucFor primers distinguished 26 separate banding patterns. Sequencing the 16S rRNA gene for four fast- and two slow-growing isolates produced the following phylogenetic associations; WSM1701 and WSM1715 (isolated from Lotus cruentus and S. pterostylis, respectively) displayed 99% homology with Sinorhizobium meliloti, WSM1707 and WSM1721 (isolated from Sinorhizobium leeana and Indigofera sp., respectively) displayed 99% homology with Sinorhizobium terangae, WSM1704 (isolated from Tephrosia gardneri) shared 99% sequence homology with Bradyrhizobium elkanii, and WSM1743 (isolated from Indigofera sp.) displayed 99% homology with Bradyrhizobium japonicum.

Aqueous extract of leaves of Indigofera suffruticosa (AELIs) were studied for adverse effects in preimplantation mouse embryos. Two-cell mouse embryos were cultured for 94 h in human tubal fluid medium (HTF), and AELIs at a concentration of 5 or 10 mg/ml. On Day 4 of culture, morulae and blastocysts were collected for morphological analysis of blastomeres. We found that embryos exposed to the higher concentration of AELIs (10 mg/ml) did not develop and all embryos persisted at the two-cell stage. Those embryos exposed to the lower concentration (5 mg/ml) showed development until morula, blastocyst and hatched blastocyst stages that were similar to the controls. These results suggest that use of AELIs may be hazardous to humans who make use of it in folk medicine.

The effect of intraperitoneal injection of 400 mg/kg of ethanolic extract of Indigofera aspalathoides (EEIA) against Dalton's ascitic lymphoma has been evaluated in Swiss albino mice. A significant increase in the life span, a decrease in the cancer cell count and body weight were noted in the tumour-induced mice after treatment with EEIA. These observations indicate that the plant is having protective effect in Dalton's ascitic lymphoma.

Summary On a defined medium, containing indoleacetic acid and benzylaminopurine, numerous shoot-buds differentiate from excised leaves, green cotyledons, and hypocotyl of immature seeds of Indigofera . The shoot-buds differentiated from leaves, however, failed to grow further whereas shoot-buds from other explants developed into plants which flowered in vitro. These observations are suggestive of differential morphogenic potential of different parts of the same plant. For differentiation the auxin, IAA, and the cytokinin, benzylaminopurine, were required. Substitution of benzylaminopurine either by furfurylaminopurine or zeatin failed to induce differentiation of shoot-buds. Replacement of indoleacetic acid by 2,4-dichlorophenoxyacetic acid or naphthaleneacetic acid induced callusing. The callus induced on 2,4-dichlorophenoxyacetic acid medium on transfer to differentiation medium, containing IAA and BAP, formed shoot-buds.

Summary Callused leaf explants of Indigofera potaninii , but not of I. amblyantha or I. delsiana , were induced to form buds and shoots on NAA/BAP and IAA/BAP combinations in Murashige and Skoog (MS), Uchimiya and Murashige (UM) and Gamborg (B5) media. Medium UM containing 0.2 mg 1 −1 NAA and 5.0 mg 1 −1 BAP was used routinely for bud multiplication and shoot production. Prolific and prolonged production of shoots occurred on this medium. Shoots were rooted under light on growth regulator free B5 medium after a 3 day period on half strength B5 medium containing 1.0 mg 1 −1 IBA. The effect of light and contact time on root induction was investigated. Callus derived from suspension cultures of I. potaninii also produced shoots on UM medium containing 0.2 mg 1 −1 NAA and 5.0 mg 1 −1 BAR These shoots could not be induced to develop into mature plants. The response of I. potaninii leaf explants to differing combinations of NAA and BAP was assessed and compared with Nicotiana tabacum . cv. Xanthi n.c. explants.

Further proof is offered that the toxic constituent of creeping indigo is β-nitropropionic acid. This compound was synthesized and compared with the isolated material. Both compounds were toxic in the chick test, both had the same melting point, and both gave the same infrared absorption spectra. A sensitive chemical test for the quantitative analysis of this compound in plant material is discussed. This test is sufficiently sensitive so that individual plants may be tested for the presence of this toxic compound. A survey of the nitrogen compounds in creeping indigo revealed a high nitrite concentration. Nitrites were shown to be relatively toxic in the chick test and so may add to the toxicity of creeping indigo.

Chromatography on silica gel of a toxic acetone extract of leaves and stems of Indigofera endecaphylla Jacq. has provided, in addition to 3-nitropropanoic acid, ethyl 3-nitropropanoate, succinic acid, and methyl β- D -glucopyranoside, a series of nine 3-nitropropanoate esters of glucose. Six of these, endecaphyllins A, A 1 , A 2 , B, B 1 , and C are isomeric triesters while three, C 1 , D, and E, are diesters. A fourth nitro-containing glucose diester, endecaphyllin I, was also obtained by chromatography and is suggested to contain as an esterifying acid a nitro-acid other than 3-nitropropanoic. The eleventh member of this group, endecaphyllin X, was isolated directly from the crude extract and found to be a glucose tetra-(3-nitropropanoate) ester. Some chemical and physical properties of these compounds are described.