2010, Volume 4, Issue 1 (Volume 4, Issue 1, January-December 2010)

Development of RAPD markers for authentication of medicinal plant Cuscuta reflexa
Salim Khan, Khanda Jabeen Mirza, Malik Zainul Abdin

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Abstract

Cuscuta reflexa (Convolvulaceae) is a holoparasitic vine that attacks the aerial parts of many shrubs, trees, and is used immensely in the Indian system of medicine. In this study, the randomly amplified polymorphic DNA (RAPD) technique was employed for authentication of Cuscuta reflexa and its adulterant Cuscuta chinensis. Thirty two decamer oligonucleotide primers were used to amplify the genomic DNA isolated from the dried stems as well as seeds of both the species. Out of the thirty two primers used, fourteen did not amplify, eleven gave faint and non-reproducible, while seven gave species-specific reproducible unique bands. The unique bands obtained in PCR amplification clearly discriminated the two species, having similar morphology and thus, RAPD may serve as a complementary tool for quality control.

Keywords: adulterant, Cuscuta reflexa, herbal medicine, molecular marker, RAPD

Citation: Khan S, Mirza KJ, Abdin MZ. Development of RAPD markers for authentication of medicinal plant Cuscuta reflexa. Eurasia J Biosci. 2010;4(1):1-7. https://doi.org/10.5053/ejobios.2009.4.0.1

The antibacterial, antiviral activities and phytochemical screening of some Sudanese medicinal plants
Ietidal El Tahir Mohamed, El Bushra El Sheikh El Nur, Mahasin El Nur Abdelrahman

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Abstract

Methanolic extracts were obtained from 30 parts of 23 different plants belonging to 19 families. Most of the plants are used traditionally to treat different illnesses in Sudan. The extracts were screened for their biological activities against bacterial pathogens including Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae and two animal viruses representing two viral families, Newcastle Disease and Fowlpox Viruses. In addition, the extracts were evaluated for the presence of the major secondary metabolites classes. The highest activity recorded was obtained for Zizyphus spina-christi against S. aureus and K. pneumoniae. Seven of the tested extracts, showed virucidal activity of NDV while eight plant extracts exhibited high activity against replication of PV. Our results support, at least in part, the uses of most plants in ethnopharmacological claims.

Keywords: antibacterial, antiviral, fowlpox, Newcastle disease virus, plant, Sudan

Citation: Mohamed IET, El Nur EBES, Abdelrahman MEN. The antibacterial, antiviral activities and phytochemical screening of some Sudanese medicinal plants. Eurasia J Biosci. 2010;4(1):8-16. https://doi.org/10.5053/ejobios.2010.4.0.2

Coumarins from Zosima absinthifolia seeds, with allelopatic effects
Seyed Mehdi Razavi, Gholamhassan Imanzadeh, Mahdi Davari

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Abstract

Zosima absinthifolia belongs to the Apiaceae family and is found in Iran, Turkey, Iraq and different countries of the Caucasus, Middle East and Central Asia. The fruits are used as food flavoring and as a food spice in Iran. In the present work, an n-hexane extract of the plant seeds was purified by vacuum liquid chromatography and preparative TLC for affording a furanocoumarin named imperatorin and two known coumarins, 7-prenyloxy coumarin and aurapten. The compound structures were elucidated by UV, 1H and 13C NMR data. Our results indicated that all tree compounds, especially imperatorin exhibited fungi toxic activity against Sclerotinia sclerotiorum, a common plant pathogen. The compounds also displayed phytotoxic effects and stunted seed germination, shoot and root growth of lettuce. It could be concluded that the purified compounds play allelopathic roles for the plant and could protect the plant against pathogens and competing herbs.

Keywords: allelopathy, auraptene, imperatorin, Sclerotinia sclerotiorum, 7-prenyloxy coumarin

Citation: Razavi SM, Imanzadeh G, Davari M. Coumarins from Zosima absinthifolia seeds, with allelopatic effects. Eurasia J Biosci. 2010;4(1):17-22. https://doi.org/10.5053/ejobios.2010.4.0.3

Indole-3-acetic acid production by Streptomyces sp. isolated from some Thai medicinal plant rhizosphere soils
Sutthinan Khamna, Akira Yokota, John F. Peberdy, Saisamorn Lumyong

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Abstract

A collection of Streptomyces spp. isolated from the rhizosphere soils of 14 Thai medicinal plants were found to produce, the plant growth hormone indole-3-acetic acid (IAA) in a yeast malt extract medium supplemented with 2 mg/mL L-tryptophan. Streptomyces CMU-H009 recovered from soil associated with lemongrass (Cymbopogon citratus) was very effective in producing IAA. Based on the 16S rDNA sequence analysis, this strain was most closely related to Streptomyces viridis NBRC 13373T (99.3% similarity). The IAA production was maximums (300 mg/mL) when the strain CMU-H009 was cultivated in a yeast malt extract broth amended with 2 mg/mL L-tryptophan a pH 7.0, and incubated at 30oC with shaking at 125 rpm for 3d. The culture filtrate from the strain CMU-H009 stimulated a significant increase in the germination and root elongation of maize (Zea mays) and cow pea (Bruguiera parviflora) seeds. The IAA from this strain was extracted, purified and identified by thin layer chromatography.

Keywords: actinomycetes, indole acetic acid, medicinal plant rhizosphere, optimization, Streptomyces viridis CMU-H009

Citation: Khamna S, Yokota A, Peberdy JF, Lumyong S. Indole-3-acetic acid production by Streptomyces sp. isolated from some Thai medicinal plant rhizosphere soils. Eurasia J Biosci. 2010;4(1):23-2. https://doi.org/10.5053/ejobios.2010.4.0.4

Isolation and characterization of salt-tolerant nitrogen-fixing microorganisms from food
Girish Gajanan Jadhav, Dipti Sambhaji Salunkhe, Devidas Punaji Nerkar, Rama Kaustubh Bhadekar

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Abstract

Halophilic microorganisms are already in use for some biotechnological processes, such as commercial production of ß-carotene, polymers (polyhydroxyalkanoates and polysaccharides), enzymes, compatible solutes etc. Considering their commercial importance, food samples (crude salt crystals and raw mango pickle) were used for isolation of halotolerant microorganisms. Two bacterial isolates obtained from food samples were examined for their ability to survive under stressed conditions and their growth response in increasing levels of NaCl (1 to 15% w/v), pH (5.0 to 10.0) and temperature (10 to 70°C). The isolates were rod shaped Gram-positive, salt-tolerant, non-halophilic, nitrogen-fixing strains. Different sugars such as glucose, fructose, maltose, sucrose, xylose and lactose were used to check for acid and gas production. The organisms were studied for their ability to hydrolyse substrates such as casein, starch, gelatin, etc. These organisms (i) grew well in SM basal salt medium and nitrogen-free semi-solid LGI medium (ii) tolerated 10-15% salt concentration (iii) produced acid from D-glucose, D-fructose and sucrose and (iv) utilized glycerol and citrate as carbon source, and v) survived acidic (pH 4-5) and alkaline (pH 9-10) conditions. The results suggested that there is potential to improve their performance as sources of industrially important enzymes. On the basis of morphological attributes and biochemical characteristics the isolates belonged to the genus Bacillus. The results of partial sequencing of 16S rRNA also revealed that the isolates 1 and 2 are closely related to Bacillus subtilis subsp. subtilis NCIB 3610T (97.9% pairwise similarity) and Bacillus sonorensis NRRL B-23154T (99.8% pair-wise similarity) respectively.

Keywords: bacillus, halotolerant, salinity, soil fertility, thermo-tolerant

Citation: Jadhav GG, Salunkhe DS, Nerkar DP, Bhadekar RK. Isolation and characterization of salt-tolerant nitrogen-fixing microorganisms from food. Eurasia J Biosci. 2010;4(1):33-40. https://doi.org/10.5053/ejobios.2010.4.0.5

Screening of elastase and tyrosinase inhibitory ctivity from Jeju Island plants
Ji-Young Moon, Eun-Young Yim, Gwanpil Song, Nam Ho Lee, Chang-Gu Hyun

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Abstract

To identify new active cosmetics ingredients of natural origin, we screened 299 parts of 263 plant species collected from Jeju Island, the southernmost island of the Korean Peninsula. Plant parts were investigated for their elastase and tyrosinase inhibitory activity for the purpose of identifying anti-aging and skin-whitening ingredients with the potential for use as raw materials in cosmetics. In the anti-elastase inhibition assay, 3 extracts, including Aesculus turbinata, Taxillus yadoriki, and Cornus walteri, showed high inhibitory activity (inhibition concentration ( I C ) 5 0<50 /vg/mL). The I C 5 0 of A. turbinata and T. yadoriki was 43.1 /vg/mL and 36.4 /vg/mL, respectively; C. walteri showed the highest elastase inhibition activity ( I C 5 0 = 26.1 /vg/mL). In the tyrosinase inhibition assay, 4 extracts, including C. walteri(139.2 vg/mL), Maackia fauriei (149.3 /vg/mL), Toxicodendron succedaneum (142.3 /vg/mL), and Sophora flavescens (41.6 v g/mL), showed significantly greater tyrosinase inhibition activity than the positive controls Distylium racemosum (145.9 /vg/mL) and arbutin (180.3 /vg/mL). However, they showed lower activity compared to the positive controls Morus alba (11.9 vg/mL) and Morus bombycis (22 v g/mL). These results suggest that medicinal plants possessing several biological activities may be potent inhibitors of the processes involved in pigmentation increases and aging. Further investigations will focus on in vivo assays and the chemical identification of the major active components responsible for anti-aging and whitening.

Keywords: cosmetics, elastase, Jeju Island, medicinal plants, tyrosinase

Citation: Moon J, Yim E, Song G, Lee NH, Hyun C. Screening of elastase and tyrosinase inhibitory ctivity from Jeju Island plants. Eurasia J Biosci. 2010;4(1):41-53. https://doi.org/10.5053/ejobios.2010.4.0.6

A morphological and anatomical investigation about two rare and endemic Crocus taxa (Iridaceae) from Southern Anatolia
Nezahat Kandemir

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Abstract

The morphological and anatomical characteristics of Crocus reticulatus subsp. hittiticus and Crocus cancellatus subsp. lycius, which are under the danger of extinction, have been investigated. The distributions of the two endemic subspecies are limited in Turkey. The plant samples were collected from the South Anatolia Region. The subsp. hittiticus flowers in the early spring whereas subsp. lycius flowers in the autumn. Their corm tunics are coarsely reticulate-fibrous. The subsp. hittiticus has blackish anthers and purple striped outer segments. The subsp. lycius has flowers with very pale lilac coloured and veined. Also, the style of this subsp. is equal to their anthers. These morphological properties are characteristics for two the subspecies investigated. The transverse-sections of the root, stem and leaf parts of these subspecies have been examined and revealed. Their morphologic and anatomic structures are illustrated. The anatomic properties of the two subspecies have been determined to be similar to the anatomic properties of other species of the genus Crocus. Moreover, the crystal type on the organs of the subsp. lycius was investigated.

Keywords: anatomy, Crocus, endemic, morphology, rare plant

Citation: Kandemir N. A morphological and anatomical investigation about two rare and endemic Crocus taxa (Iridaceae) from Southern Anatolia. Eurasia J Biosci. 2010;4(1):54-62. https://doi.org/10.5053/ejobios.2010.4.0.7

Biological activities of extracts from Andaman Sea sponges, Thailand
Patchara Pedpradab, Wanlapa Molex, Veena Nukoolkarn, Udomsak Darumas

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Abstract

Thirty six organic extracts were prepared from eighteen marine sponges collected from the Andaman Sea, Thailand. The extracts were examined for anti-malaria, anti-Microbacterium tuberculosis, anti-herpes simplex virus, antimicrobial, anti-acetylcholinesterase enzyme and cytotoxic activities. Four extracts showed anti-M. tuberculosis, one anti-malarial, twenty four antimicrobial and one extract exhibited cytotoxic activity. However, anti-acetylcholinesterase enzyme and anti-herpes simplex virus type 1 (HSV-1) activities were not recorded. Dichloromethane extracts prepared from Axinyssa sp., Halichondria sp. and Chondrosia reticulata exhibited potential anti-M. tuberculosis at MIC 50, 100 and 200 mg/mL, respectively. The Hexane part of Phakellia ventilabrum extract showed anti-malarial activity (MIC= 2.8 mg/mL) while the dichloromethane extract showed anti-M. tuberculosis and cytotoxic activity with MIC 200 and IC50 7.1 mg/mL, respectively. Antimicrobial activity was found in both the hexane and dichloromethane parts of extracts.

Keywords: Andaman Sea, anti-malaria, antimicrobial, biological activity, marine sponges

Citation: Pedpradab P, Molex W, Nukoolkarn V, Darumas U. Biological activities of extracts from Andaman Sea sponges, Thailand. Eurasia J Biosci. 2010;4(1):63-9. https://doi.org/10.5053/ejobios.2010.4.0.8

Effects of hydrogen peroxide on the germination and early seedling growth of barley under NaCl and high temperature stresses
Kürsat Çavusoglu, Kudret Kabar

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Abstract

In this work, the effects of a hydrogen peroxide (H2O2) pretreatment on the germination and early seedling growth of barley under salt (NaCl) and temperature (20º, 25º, 30º and 35ºC) conditions was studied. H2O2 enormously removed the germination-delaying and inhibiting effects of temperature increases. In addition, it was rather effective on alleviation of the reductive effects of temperature increases on the parameters of growth such as the percentages of coleoptile emergence, elongations of radicle, and coleoptile and fresh weights (FWs) of the seedlings. H2O2 pretreatment became very successful in the overcoming of the germination-delaying and preventing effects of the increases in both salt and temperature levels. It markedly alleviated the inhibitions of salt on seedling growth at all temperatures as well. H2O2 both removed the blockage of abscisic acid (ABA) on the germination and shortened the time required for germination at 20ºC, an optimal temperature, by its influsion into the seeds with pretreatment or its presence in the medium of germination. On the other hand, H2O2 relatively alleviated ABA inhibitions on the radicle elongations and FWs while it had no effect on ABA suppression on the emergence percentage and elongation of the coleoptile.

Keywords: barley, hydrogen peroxide, salt stress, seed germination, seedling growth, temperature stress

Citation: Çavusoglu K, Kabar K. Effects of hydrogen peroxide on the germination and early seedling growth of barley under NaCl and high temperature stresses. Eurasia J Biosci. 2010;4(1):70-9. https://doi.org/10.5053/ejobios.2010.4.0.9

Molecular variability among the isolates of Sclerotium rolfsii causing stem rot of groundnut by RAPD, ITS-PCR and RFLP
Saddala Durga Prasad, Shaik Thahir Basha, Narreddy Peddanarappa, Gari Eswara Reddy

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Abstract

Genetic variability among the virulent isolates of Sclerotium rolfsii was studied using molecular techniques like RAPD, ITS-PCR and RFLP. The RAPD banding pattern reflected the genetic diversity among the isolates by formation of two clusters. A total of 221 reproducible and scorable polymorphic bands ranging approximately as low as 100 bp to as high as 2500 bp were generated with five RAPD primers. ITS region of rDNA amplification with specific ITS1 and ITS4 universal primers produced approximately 650 to 700 bp in all the isolates confirmed that all the isolates obtained are Sclerotium rolfsii. Studies by ITS-RFLP indicated that there was no polymorphism in restriction banding pattern among the isolates with the restriction endonucleases used.

Keywords: groundnut, molecular variability, Sclerotium rolfsii, stem rot

Citation: Prasad SD, Basha ST, Peddanarappa N, Reddy GE. Molecular variability among the isolates of Sclerotium rolfsii causing stem rot of groundnut by RAPD, ITS-PCR and RFLP. Eurasia J Biosci. 2010;4(1):80-7. https://doi.org/10.5053/ejobios.2010.4.0.10

Plant growth promoting activities of phosphatesolubilizing Enterobacter asburiae as influenced by fungicides
Munees Ahemad, Mohammad Saghir Khan

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Abstract

Four fungicides, tebuconazole, hexaconazole, metalaxyl and kitazin were applied at recommended and higher rates to examine their effects on the plant growth promoting activities of the fungicide tolerant Enterobacter asburiae strain PS2 under in vitro conditions. Enterobacter asburiae strain PS2 was isolated from the mustard rhizosphere and was assessed for the fungicide-tolerance and production of plant growth promoting traits (phosphate solubilization, siderophores, indole acetic acid, exo-polysaccharides, hydrogen cyanide and ammonia production) both in the presence and absence of fungicides. Enterobacter asburiae strain PS2 showed plant growth promoting activities even in the presence of fungicides which however, decreased progressively with the increase in fungicide concentration. Fungicides at recommended dose had little effect while the dose higher than the recommended one adversely affected the physiological traits, like, phosphate solubilization, siderophore, and indole acetic acid synthesis. Among all fungicides, tebuconazole in general, showed maximum toxicity to plant growth promoting activities. Fungicide-tolerant E. asburiae strain PS2 with the inherent ability to produce growth regulators even in the presence of fungicides can be exploited as a bio-inoculant to increase the productivity of crops grown in fungicide stressed soils.

Keywords: enterobacter, fungicide, phosphate solubilization, plant growth promoting rhizobacteria (PGPR)

Citation: Ahemad M, Khan MS. Plant growth promoting activities of phosphatesolubilizing Enterobacter asburiae as influenced by fungicides. Eurasia J Biosci. 2010;4(1):88-95. https://doi.org/10.5053/ejobios.2010.4.0.11

Effects of 24-epibrassinolide on growth, chlorophyll, electrolyte leakage and proline by pepper plants under NaCl-stress
Samira Ibn Maaouia Houimli, Mounir Denden, Bouthaina Dridi Mouhandes

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Abstract

Brassinosteroids are steroidal phytohormones that have the ability to overcome plant environmental stress. This study was carried out to investigate the role of 24-epibrassinolide in inducing pepper plant salt tolerance as measured by a range of physiological parameters: growth, chlorophyll, electrolyte leakage and proline. Capsicum annuum cv. Beldi seedlings were sprayed with 24-epibrassinolide both in the presence or the absence of NaCl and were sampled, 28 days after treatments. As a result of analysing the cultures under salinity stress, it was determined that the biomass and the chlorophyll decreased significantly, while the electrolyte leakage and the proline concentration increased considerably under salinity stress. However, the application of 24-epibrassinolide significantly ameliorated the adverse effects of salinity on the examined parameters, confirming the suppositions of previous authors who have claimed that exogenously applied 24-epibrassinolide can increase growth and protect the integrity of the cellular membrane in stressed plants.

Keywords: Brassinosteroids, chlorophyll, electrolyte leakage, proline, salt tolerance

Citation: Houimli SIM, Denden M, Mouhandes BD. Effects of 24-epibrassinolide on growth, chlorophyll, electrolyte leakage and proline by pepper plants under NaCl-stress. Eurasia J Biosci. 2010;4(1):96-104. https://doi.org/10.5053/ejobios.2010.4.0.12

Cytological effects of the root extracts of Boerhaavia diffusa on root tips of Crinum jagus
Ndubuisi Moses Chikere Nwakanma, Bosa Ebenezer Okoli

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Abstract

The mitotic effects of the root extracts of Boerhaavia diffusa on the root tips of Crinum jagus were investigated. The results of this study showed several chromosomal abnormalities including stickiness of chromosomes (both at metaphase and anaphase), C-metaphase, lagging chromosomes, and sticky bridges. The trend of the results showed that the higher the concentration of the extracts for treatment, the more inhibitory the effect on mitosis with more pronounced chromosomal aberrations. The root extract of B. diffusa was found active at concentrations of 0.1%, 0.2%, 0.4%, and 0.8%. The extract was found to accumulate metaphase and could thus be of immense help in cytological work. These results are discussed in the light of their reported use in cancer therapy, the treatment of other diseases in traditional medicinal practice and its' possible use as an alternative to colchicine in cytological work.

Keywords: Boerhaavia diffusa, chromosom, Crinum jagus, cytological effect, mitosis

Citation: Nwakanma NMC, Okoli BE. Cytological effects of the root extracts of Boerhaavia diffusa on root tips of Crinum jagus. Eurasia J Biosci. 2010;4(1):105-11. https://doi.org/10.5053/ejobios.2010.4.0.13

Colonization of tomato roots by some potentially human-pathogenic bacteria and their plant-beneficial properties
Dilfuza Egamberdieva

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Abstract

Many studies provide evidence for the occurrence of opportunistic pathogens in the rhizosphere of plants. The presence of a competitive microflora in soil may restrict root colonization by potentially pathogenic bacteria. The aim of the present work was to study the competitive tomato root tip colonization of potentially human pathogenic bacteria such as Bacillus cereus, Staphylacoccus saprophyticus, Pseudomonas aeruginosa, Acinetobacter sp. against best known root tip colonizer P. fluorescens WCS365 and to characterize for their ability to stimulate plant growth and to suppress tomato foot and root rot disease caused by Fusarium oxysporum f.sp. radicis-lycopersici. In competitive tomato root tip colonization assays all tested bacterial strains showed poor colonization of the tomato rhizosphere except P. aeruginosa which was better competitive colonizer than reference strains. All five bacterial strains stimulated shoot length (up to 35%) and dry matter (up to 58%) of tomato and only one strain B. cereus TSAU80 showed statistically significant disease control (27%) of tomato foot and root rot in comparison to the Fusarium-infected control plants (47%). These results show that the opportunistic pathogens possess beneficial effect on plant growth and are able to colonize the rhizosphere of tomato, potentially resulting risk of contamination for tomato greenhouses. However, their competitive colonization abilities in the rhizosphere are poor, whereas only P. aeruginosa appeared to be better competitive colonizer than reference strain.

Keywords: human pathogenic bacteria, plant growth promotion, root colonization, tomato

Citation: Egamberdieva D. Colonization of tomato roots by some potentially human-pathogenic bacteria and their plant-beneficial properties. Eurasia J Biosci. 2010;4(1):112-8. https://doi.org/10.5053/ejobios.2010.4.0.14

A novel set of highly polymorphic chloroplast microsatellite and ISSR markers for the biofuel crop Jatropha curcas
Neha Mittal, Alok Kumar Dubey

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Abstract

A novel set of polymorphic ISSR and chloroplast microsatellite markers were identified to mark the genetic relationship among different accessions of Jatropha curcas. A total of 12 polymorphic ISSR primers used, amplified a total of 26 fragments among ten different accessions of J. curcas. The cross-species amplification of microsatellite loci is timesaving as well as a cost-effective approach for developing locus specific markers for new species. Further, in an attempt to reveal a high degree of polymorphism and genetic variation among different accessions of J. curcas chloroplast microsatellite primer pairs developed for Acorus calamus (Acoraceace) were examined for cross-species amplification and validation among them. Out of 18 cpSSRs, six (33.33%) were identified as resulting in good amplifications and polymorphism. The number of amplified alleles per cpSSR primer pair ranged from 2 to 4 with the total number of 18 size variants as whole. The 18 size variants at 6 loci among 10 accessions were combined in 4 different haplotypes. The total chloroplast DNA diversity (HT) and the differentiation (GST) were found as 0.700 and 0.560 respectively. Whereas, Shannon's information index (I), Nei's genetic diversity among different accessions of J. curcas using ISSR and cpSSR markers was estimated at 0.33±0.28; 0.22±0.19 and 0.94±0.35; 0 . 5 5 ± 0 . 1 8 respectively. The genetic relationship among the accessions was established using UPGMA cluster analysis, which were grouped into two major clusters denoted as cluster-I and cluster-II. The implication of the results in devising the importance of a identified novel set of markers for the genetic relationship and genetic diversity analysis in J. curcas is discussed.

Keywords: accessions, Acorus calamus, cross-species amplification, genetic diversity, Jatropha curcas

Citation: Mittal N, Dubey AK. A novel set of highly polymorphic chloroplast microsatellite and ISSR markers for the biofuel crop Jatropha curcas. Eurasia J Biosci. 2010;4(1):119-31. https://doi.org/10.5053/ejobios.2010.4.0.15