Analysis of Genetic Diversity and Structure Pattern of Indigofera Pseudotinctoria in Karst Habitats of the Wushan Mountains Using AFLP Markers.

Indigofera pseudotinctoria Mats is an agronomically and economically important perennial legume shrub with a high forage yield, protein content and strong adaptability, which is subject to natural habitat fragmentation and serious human disturbance. Until now, our knowledge of the genetic relationships and intraspecific genetic diversity for its wild collections is still poor, especially at small spatial scales. Here amplified fragment length polymorphism (AFLP) technology was employed for analysis of genetic diversity, differentiation, and structure of 364 genotypes of I. pseudotinctoria from 15 natural locations in Wushan Montain, a highly structured mountain with typical karst landforms in Southwest China. We also tested whether eco-climate factors has affected genetic structure by correlating genetic diversity with habitat features. A total of 515 distinctly scoreable bands were generated, and 324 of them were polymorphic. The polymorphic information content (PIC) ranged from 0.694 to 0.890 with an average of 0.789 per primer pair. On species level, Nei's gene diversity (Hj), the Bayesian genetic diversity index (HB) and the Shannon information index (I) were 0.2465, 0.2363 and 0.3772, respectively. The high differentiation among all sampling sites was detected (FST = 0.2217, GST = 0.1746, G'ST = 0.2060, θB = 0.1844), and instead, gene flow among accessions (Nm = 1.1819) was restricted. The population genetic structure resolved by the UPGMA tree, principal coordinate analysis, and Bayesian-based cluster analyses irrefutably grouped all accessions into two distinct clusters, i.e., lowland and highland groups. The population genetic structure resolved by the UPGMA tree, principal coordinate analysis, and Bayesian-based cluster analyses irrefutably grouped all accessions into two distinct clusters, i.e., lowland and highland groups. This structure pattern may indicate joint effects by the neutral evolution and natural selection. Restricted Nm was observed across all accessions, and genetic barriers were detected between adjacent accessions due to specifically geographical landform.

Genetic diversity and population structure of Indigofera szechuensis complex (Fabaceae) based on EST-SSR markers.

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.

The demographic response of a deciduous shrub (the Indigofera bungeana complex, Fabaceae) to the Pleistocene climate changes in East Asia.

East Asia harbors the highest level of floristic diversity among the world's temperate regions. Despite the increase in phylogeographic studies of temperate plants in East Asia, far less attention has been paid to widely distributed deciduous shrubs that widespread across several floral regions. We sequenced two chloroplast DNA (cpDNA) fragments (ndhJ-trnF and trnD-trnT) and one nuclear DNA (Pgk1) of 472 individuals from 51 populations of such a group, the Indigofera bungeana complex. We used population genetic data as well as ecological niche modelling to examine the evolutionary history and glacial refugia during the Last Glacial Maximum (LGM) of this group. We recovered 133 cpDNA and 68 nuclear haplotypes. The star-phylogeny of the recovered cpDNA and nuclear haplotypes and demographic analyses suggested distinct range expansion of I. bungeana complex have occurred during the early and middle Pleistocene. The climate change of the LGM might have affected little on the distribution of this complex based on the niche modelling. However, these climate changes and geographic isolation probably resulted in fixtures of the private haplotypes and genetic differentiations between regions. Our results suggested that this arid-tolerant species complex may have different responses to the Quaternary climate changes with those climate-sensitive species.

Beneficial effects of methanolic extract of Indigofera linnaei Ali. on the inflammatory and nociceptive responses in rodent models

ABSTRACT Indigofera linnaei Ali. (Tamil Name: Cheppu Nerinjil) belongs to the family Fabaceae, used for the treatment of various ailments in the traditional system of medicine. In the present study, the beneficial effects of methanol extract of whole plant of I. linnaei (MEIL) were evaluated on inflammation and nociception responses in rodent models. In vitro nitric oxide (NO), lipoxygenase (LOX) and cyclooxygense (COX) inhibitory activities were also performed to understand the mode of action. MEIL at the dose of 200 & 400 mg/kg, p.o. significantly inhibited carrageenan induced rat paw volume and reduced the weight of granuloma in cotton pellet granuloma model. The results obtained were comparable with the standard drug aceclofenac. The anti-nociceptive effect of MEIL in mice was evaluated in hot plate and acetic acid induced writhing model. The plant extract significantly reduced the number of writhes and the analgesic effect was higher than that of the standard drug aspirin. However, the extract fails to increase the latency period in hot plate method suggesting that the extract produce nociception by peripheral activity. The extract produced inhibitory effect on NO, LOX and COX in concentration dependent manner. The extract exhibited pronounced and selective COX-2 inhibition. Altogether, these results suggested that the methanol extract of Indigofera linnaei could be considered as a potential anti-inflammatory and analgesic agent.

RESUMO Indigofera linnaei Ali pertence à família Leguminosae e é utilizada para o tratamento de várias doenças na medicina tradicional. No presente estudo, os efeitos benéficos do extrato metanólico da planta inteira de I. linnaei (MEIL) foram avaliados em respostas inflamatórias e nocicepção em modelos de roedores. Testes in vitro de atividade inibitória do óxido nítrico (NO), lipoxigenase (LOX) e ciclooxigenase (COX) também foram realizados para compreender o modo de ação. MEIL nas doses de 200 e 400 mg/kg, p.o. inibiu significativamente o volume da pata de rato induzido por carragenana e reduziu o peso do granuloma no modelo de pélete de algodão. Os resultados obtidos foram comparáveis ao do fármaco padrão, aceclofenaco. O efeito anti-nociceptivo de MEIL foi avaliado em camundongos no modelo de placa quente e de contorção induzida por ácido acético. O extrato da planta reduziu significativamente o número de contorções e o efeito analgésico foi maior do que o do fármaco padrão, ácido acetilsalicílico. Porém, o extrato não conseguiu aumentar o período de latência no método da placa quente, sugerindo que este produz nocicepção por atividade periférica. O extrato produziu efeito inibitório sobre o NO, LOX e COX dependente da concentração. O extrato exibiu inibição acentuada e seletiva da COX-2. No seu conjunto, estes resultados sugerem que o extrato metanólico de Indigofera linnaei poderia ser considerado como agente anti-inflamatório e analgésico potencial.

Polyphenol contents and antioxidant activities of five Indigofera species (Fabaceae) from Burkina Faso.

Aqueous acetone extracts prepared from five Indigofera species of Burkina Faso, namely Indigofera colutea (Burm.) Murril., I. macrocalyx Guilld et Perr., I. nigritana Hook f., I. pulchra willd. and I. tinctoria L., were investigated for their phytochemical composition and their antioxidant activities. Standard methods and TLC were used to screen the phytochemical composition. The total phenolic and flavonoid content of extracts were assessed by Folin-Ciocalteu and AlCl3 methods, respectively. These extracts were also evaluated for their antioxidant potentials using ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) assays. Flavonoids, saponins, quinones, sterols/triterpenes and tannins were present in all these species except for I. pulchra where quinones were not found. Gallic acid, caffeic acid, rutin and myricetin in I. colutea; gallic acid, quercitrin, myricetin in I. tinctoria; galangin and myricetin in I. macrocalyx were identified by thin layer chromatography. Among these, I. colutea, I. tinctoria, I. nigritana and I. macrocalyx, which had the highest phenolic content, were also found to possess the best antioxidant activities. The results indicated a good correlation between antioxidant activities and total phenolic content (p<0.05 for FRAP/DPPH and DPPH/ABTS and p<0.01 for FRAP/ABTS). These plants represent promising sources of natural antioxidants and these findings give scientific bases to their ethnopharmacological uses.