Open Access Others

Regeneration of Shoots from Callus of Ceropegia pusilla Wight and ARN

R. Prabakaran, T. Sasikala, K. Kalimuthu

Biotechnology Journal International, Page 416-423
DOI: 10.9734/BBJ/2013/4048

This study was undertaken to evaluate the most suitable concentration of plant growth regulators and suitable explants for callus induction and subsequent organogenesis of an endangered medicinal plant Ceropegia pusilla. The best performance of callus induction and morphogenesis was found on MS medium supplemented with 6- benzylaminopurine (BA) and naphthalene acetic acid (NAA), from node and internode the maximum account of callus initiation was recorded on MS medium supplemented with BA+NAA (2.22µM + 5.37 µM) and the maximum % of callus induced on subculture is 5.47 ± 0.68. Rooting was best achieved on MS medium augmented with IBA (2.46µM). The maximum number of roots (4.72 ± 0.66 cm) and root length (4.8 ± 0.49 cm) were recorded). Plantlets regenerated in vitro with well-developed shoot and roots were successfully acclimatized in pots containing a mixture of decomposed coir waste, perlite and compost 1: 1:1 ratio and grown in a shade house with 81 ± 3.16 percent survival rates.

Open Access Short communication

Growth Inhibition of Some Phytopathogenic Bacteria by Cell-Free Extracts from Enterococcus sp

Luis C. Mata, Catalina Chavez, Raúl Rodríguez-Herrera, Daniel Hernández-Castillo, Cristóbal N. Aguilar

Biotechnology Journal International, Page 359-366
DOI: 10.9734/BBJ/2013/3024

Aims:  To inhibit of bacterial growth of three important phyto-pathogenic bacteria: Erwinia carotovora, Clavibacter michiganensis sp. michiganensis and Xanthomonas axonopodis by cell-free extracts from submerged cultures of two strains of Enterococcus sp. was tested.

Study Design: A complete randomized experimental design with factorial fix was used to evaluate the efficiency of growth inhibition against the phytopathogenic bacteria.

Place and Duration of Study: Laboratory of Bioprocesses, Department of Food Science and Technology, School of Chemistry, Universidad Autonoma de Coahuila, Mexico, between December 2011 and July 2012.

Methodology: Enterococci strains were isolated from goat milk, buttermilk and whey by typical microbiological procedures and primarily identified based on biochemical tests. Strains were subsequently activated in MRS broth and cells were separated by centrifugation and filtration. Cell-free extracts were tested against plant pathogenic bacteria to determine their growth inhibition potential.

Results: Strains of Enterococcus MII-1 and MIV-2 were able to inhibit the growth of three pathogenic bacteria, demonstrating to be an attractive alternative for biological control assays.

Conclusion: The cell-free extracts of Enterococcus spp. show inhibition potential to inhibit phytopathogenic bacteria that cause diseases in horticultural crops. Further studies are needed to completely evidence the high potential of use of cell-free extracts from Enterococcus MII-1 and MIV-2.

Open Access Original Research Article

Adsorption of Copper (II) Ions from Aqueous Solution onto Synthetic Goethite and Two Naturally Available Red Soils from Yaoundé –Cameroon

Tagne Guy Merlain, NDI Julius Nsami, Ketcha Joseph Mbadcam

Biotechnology Journal International, Page 221-235
DOI: 10.9734/BBJ/2013/3117

The present investigation deals with the utilization of synthetic goethite and two red soils obtained from AHALA-NSAM and NKOLOFANE as adsorbent for the removal of copper from waste water. The synthetic goethite was characterized by IR, XRD and BET where the red soils were characterized by XRD, Chemical Analysis and BET. A series of experiments were conducted in a batch system to evaluate the effect of system variables. The effect of contact time, pH and the initial concentration of copper (II) solution were considered. The results show that the contact time of 180 minutes, 20 minutes and 15 minutes are sufficient to fit the equilibrium for GEO, E1-B1 and NSAM-B2 respectively. The optimal adsorption pH for the adsorption of copper (II) ions is 4.6±1. The maximal quantities adsorbed by every adsorbent were 78 .2 mg/g for GEO, 41.2mg/g for E1-B1 and 32.2mg/g for NSAM-B2. The experimental results obtained have been correlated to four kinetic models including the first order, the second order, Elovich and intra particle diffusion models. The kinetic model that gave the value of the linear correlation coefficient which best fits is that of second order. Three isotherms models amongst which are Langmuir, Freundlich and Tempkin’s have been applied to the experimental data. The Langmuir model best described the adsorption of copper (II) ions in aqueous solution by the three adsorbents.

Open Access Original Research Article

Application of Box-Behnken Design for the Optimization of Citric Acid Production from Corn Starch Using Aspergillus niger

N. A. Amenaghawon, K. I. Nwaru, F. A. Aisien, S. E. Ogbeide, C. O. Okieimen

Biotechnology Journal International, Page 236-245
DOI: 10.9734/BBJ/2013/3379

Citric acid production from hydrolysed corn starch was optimized in this study. Response surface methodology (RSM) was employed for the analysis of the simultaneous effect of substrate concentration, broth pH and fermentation temperature on the concentration of citric acid produced during fermentation of hydrolysed corn starch. A three-variable, three-level Box-Behnken design (BBD) comprising 15 experimental runs was used to develop a second degree statistical model for the optimisation of the fermentation conditions. The optimal fermentation conditions that resulted in the maximum citric acid concentration were substrate concentration; 50 g/L, broth pH; 2.00 and fermentation temperature; 25ºC. Under these conditions, the concentration of citric acid was obtained to be 31.96 g/L. Validation of the model indicated no difference between predicted and observed values.

Open Access Original Research Article

The Application Development of Plant-Based Environmental Protection Plasticizer

Wu Xiaomin, Ge Lan, Lu Yongliang, Pan Weirong, Chen Jianwen, Zou li ming, Wu Jialin

Biotechnology Journal International, Page 246-262
DOI: 10.9734/BBJ/2013/2751

Present research is aimed towards designing and development of environmental protection plasticizers using the fractionation heavy component in the industrial production of corn stalks as raw material. The plant-based rubber oil (PBRO) plasticizers are the reactants of the dihydric alcohols (Polyols) in heavy component with phthalic anhydride (PA) (the molar ratio of Polyols/PA as  2 : 1)  at the temperature range of 180 -190ºC. The main advantage is that this substance does not contain any of the sixteen hazardous substances of PAHs (polycyclic aromatic hydrocarbons) and 38 hazardous substances of PAHs (polycyclic aromatic hydrocarbons) recorded in EU REACH regulation. In plasticization of NBR, the dosage of PBRO can reach 25 phr (parts per hundreds of rubber), but the plasticizing effects of PBRO is inferior to common rubber oil (paraffin oil, naphthenic oil, aromatic hydrocarbon oil and Dioctyl phthalate). In plasticization of SBR, the maximum amount of PBRO is 20 phr. Although the mechanical properties decrease; the aging resistance and thermal stability (the temperature of maximum weight losses of PBRO as 302ºC) are slightly higher than that of common rubber oil.

Open Access Original Research Article

Primary Somatic Embryos from Axillary Meristems and Immature Leaf Lobes of Selected African Cassava Varieties

Jelili T. Opabode, Olufemi O. Oyelakin, Oluyemisi A. Akinyemiju, Ivan L. Ingelbrecht

Biotechnology Journal International, Page 263-273
DOI: 10.9734/BBJ/2013/3088

The study evaluated high-value African cassava varieties for primary somatic embryogenesis using axillary meristems (AM) and immature leaf lobes (LL) on picloram-based medium. The study was conducted at the Central Biotech Lab, International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria between 2006 and 2009. Completely randomized design with four replicates was used for the study. Using LL explants, there were significant (P=.05) differences in percent responding leaf lobes, percent explant with pre-embryogenic structure, PSEF and PSEE among cassava varieties. The PSEF of the only three varieties that produced mature somatic embryo were 93.6, 88.5 and 85.7% for TME 12, Kibaha and Albert, respectively. Similarly, significant (P=.05) differences existed among the varieties in percent enlarged axillary meristem, percent explant with pre-embryogenic structure, PSEF and PSEE when AM was the explant. The PSEFs of the only three varieties that produced mature somatic embryo were 83.6, 77.5 and 72.7% for TME 12, Kibaha and Albert, respectively. The PSEF and PSEE of LL explant were greater than those of AM by an average of 86.1% and 82.7%, respectively. The study concluded that both AM and LL were good explants for production of primary somatic embryo in cassava

Open Access Original Research Article

β 1,4-Xylosidase Production Potential of a Fusarium sp. on Wood Shavings

F. C. Adesina, A. A. Onilude

Biotechnology Journal International, Page 274-285
DOI: 10.9734/BBJ/2013/3078

Aims: This study was aimed at screening fungal isolates from degrading wood samples for β 1,4-xylosidase production, selecting the best isolate based on the screening test to produce the enzyme through solid state fermentation of some wood shavings and determining optimum production conditions for the fungus on best substrate.

Place and Duration of Study: Microbial physiology laboratory, Department of Microbiology, University of Ibadan, Ibadan and Multi Discplinary Central laboratory University of Ibadan, Oyo State Nigeria. Between September 2010 and October 2011.

Methodology: Isolates obtained from degrading wood samples were identified and screened on agar plates of para–nitrophenyl β-xyloside as carbon source. Selected isolates were used to produce β 1,4-xylosidase using shavings of Anogeissus leiocarpus, Gmelina arborea and Terminalia superba moistened with a chemically defined medium as substrates.  Assay was done every 3 days for 15 days. Production dependent parameters such as pH, temperature were varied on the best wood substrate for β 1,4-xylosidase production by selected fungus.

Results: Selected isolate was used for enzyme production on shavings of Anogeissus leiocarpus, Gmelina arborea and Terminalia superba as substrates, each was moistened with chemically defined medium. Assay was done every 3 days for 15 days. Production dependent parameters such as pH, temperature were varied on the best wood substrate for β 1,4-xylosidase production by fungus. Maximum β 1,4-xylosidase activity of fungus on each substrate was; Anogeissus leiocarpus, 31.620Ug-1 on 12th day, G. arborea, 8.935U g-1 on day 9 and T. superba, 6.053Ug-1 on  6th day. Optimum β 1,4-xylosidase production was obtained on A. leiocarpus at 35ºC and pH 6 with 40% and 60% aeration. 50% moisture content of substrate supplemented with soy meal as nitrogen source supported the best β 1,4-xylosidase production by fungus.

Conclusion: Enzyme production was highly enhanced at optimum conditions.

Open Access Original Research Article

Thyroid Hormone Profile in Rwandan Students

Jean Bosco Gahutu

Biotechnology Journal International, Page 286-292
DOI: 10.9734/BBJ/2013/3231

Aim: To illustrate the thyroid hormone profile in young adult students at moderate altitude in Rwanda.

Study Design: A population-based cross-sectional study.

Place and Duration of Study: The study was conducted among healthy university students living at Butare, Rwanda (altitude: 1,768 m, barometric pressure: 629 mm Hg) in December 2011.

Methods: Venous blood was collected in the morning, after overnight fasting. TSH, total T3 and total T4 hormonal assays were performed in the laboratory of Butare University Teaching Hospital by classical sandwich ELISA technique for TSH and competitive ELISA technique for total T4 and total T3.

Results: The results of 111 subjects (65 males and 46 females; mean age: 22 years, age range: 18-28 years) are as follows (mean ± SD): total T3: males: 0.9 ± 0.3 ng/mL, females: 1.3 ± 0.3 ng/mL; total T4: males: 6.4 ± 1.1 µg/dL, females: 9.3 ± 2.2 µg/dL; TSH: males: 1.8 ± 1.2 mIU/L, females: 1.9 ± 1.4 mIU/L.

Conclusion: The results of our study compare well with findings of other studies, with a slight increase in serum TSH concentration as compared to sea level values. A comprehensive study needs to be done to establish reference intervals.

Open Access Original Research Article

Effect of Gas Flaring on the Phytochemical and Nutritional Composition of Treculia africana and Vigna subterranean

C. O. Ujowundu, L. A. Nwaogu, F. N. Ujowundu, D. C. Belonwu

Biotechnology Journal International, Page 293-304
DOI: 10.9734/BBJ/2013/3033

Aims: The effect of petroleum exploration and exploitation activities on the phytochemical and nutritional composition of African breadfruit (Treculia africana) and Bambara groundnuts (Vigna subterranean) harvested from polluted and unpolluted environment were studied.

Study Design:  Plant samples were collected from Izombe, an environment highly polluted by gas flaring and Eziobodo, an environment not known for gas flare exposures. Both locations are within the (Niger-Delta) rain forest region of Imo State, Nigeria.

Methodology: The processed samples were subjected to proximate and phytochemical screening. The vitamins, micro and macro minerals contents were determined using standard biochemical methods and equipment.

Results: Some of the phytochemicals and nutritional contents showed varying levels of significant difference (P<0.05) in samples from the two localities. Bambara groundnuts from polluted area was significantly lower (P<0.05) in moisture, ash, protein and carbohydrate when compared to samples from unpolluted environment. The concentrations of antinutrients such as oxalate, phytate, tannin and cyanogenic glycoside were significantly higher (P<0.05) in samples from polluted areas when compared to samples from unpolluted areas. The results showed increased accumulation of microminerals and reduced macrominerals in samples from polluted environment. Riboflavin, vitamins A, C and E were also significantly higher (P<0.05) in samples obtained from unpolluted environment.

Conclusion: Data obtained revealed that T. africana and V. subterranean are highly nutritious and their nutrient composition are responsive to pollution due to gas flaring.

Open Access Original Research Article

Antibacterial and Antiviral Activities of Essential Oils of Northern Moroccan Plants

Naima El Moussaoui, Gloria Sanchez, El Ouardy Khay, Mohammed Idaomar, Ahmed Ibn Mansour, Jamal Abrini, Rosa Aznar

Biotechnology Journal International, Page 318-331
DOI: 10.9734/BBJ/2013/3596

Aims: This study was designed to evaluate the antimicrobial activity of five essential oils (EOs) extracted from the aerial parts (leaves and flowering tops) of three species growing in the north of Morocco: Origanum elongatum, Thymus capitatus and Mentha suaveolens.

Study Design: Chemical analysis of EOs, antibacterial and antiviral activities of EOs.

Place and Duration of Study: Department of Biology (Faculty of Sciences), Institute of Agrochemistry and Food Technology (IATA), between September 2009 and December 2009.

Methodology: The EO constituents were extracted by hydrodistillation and analysed by GC-MS. The antibacterial activity of EOs was tested against three reference strains, Salmonella enterica subsp. enterica CECT 915T, Listeria monocytogenes CECT 4031T, and Escherichia coli O157:H7 CECT 4267, and two food isolated strains Salmonella sp. S64 and Listeria monocytogenes L23, using the diffusion method and the microtitration assays. The antiviral effect of EOs was evaluated for the inactivation of murine norovirus (MNV-1), a human norovirus surrogate.

Results: GC and GC/MS analyses revealed that thyme EO predominantly contains carvacrol (58.77-68.63%), p-cymene (4.84-5.63%), γ-terpinene (2.78-3.75%) and β-caryophyllene (2.62-2.91%). Oregano EO was mainly constituted by carvacrol (19.21-40.12%), thymol (3.57-14.24%), p-cymene (16.08-16.19%) and γ-terpinene (7.27-13.48%). While, mint EO was characterized by piperitenone oxide (41.84%), (-)-isopulegol (11.95%) and limonene (7.35%). All extracts exhibited an antibacterial activity at different levels against strains reported as the causal agents of foodborne diseases, but a low antiviral activity (0.87-0.50 log10 TCID50/ml reduction) was observed.

Conclusion: Results suggest the potential use of tested EOs as bio-preservatives in the food industry. However, their antiviral activity needs to be further investigated

Open Access Original Research Article

Bacteriological Study of Urinary Tract Infections with Antibiotics Susceptibility to Bacterial Isolates among Honeymoon Women in Al Qassim Hospital, Babylon Province, Iraq

Nada Khazal Kadhim Hindi, Shaimaa O. Hasson, Shatha Khazal Kadhim Hindi

Biotechnology Journal International, Page 332-340
DOI: 10.9734/BBJ/2013/3573

Urinary Tract Infections (UTIs) is an infection caused by the presence and growth of microorganism anywhere in the urinary tract. This study aims at determining the most common microorganism that causes UTI in honeymoon women, detection the most effective antimicrobial agents that causative agents of UTI and select the best antibiotics for treated UTI. One hundred and thirty specimens were collected from women suspected with UTI {honeymoon (95) and other women (35)} in al-Qassim hospital; during the period (January 2012 to 30th September 2012).The bacterial isolates were identified tested for antibiotics sensitivity test6. Honeymoon women were showing higher rate of UTI than other women in (87 from 95) (91.5 %) and (19 from 35) (54.3%) respectively. E. coli (46%) and Staphylococcus aureus (42%) were predominant isolate in honeymoon. Regarding Antibiotics sensitivity test, E. coli and Staphylococcus aureus showed highly sensitive to Nitrofurantoin (100%), (94.5%) followed by Amikacin (90%), (85%), Gentamicin (88.2%), (75%) and ciprofloxacin (80.7%) (88%) respectively, while Oxacillin were not effective against tested isolates. In summary, Honeymoon women were showing higher rate of UTI than other women. E. coli was isolated more than other bacteria that causing UTI in honeymoon women. Most bacterial isolates was showed highly sensitive to Nitrofurantoin and resistance to Oxacillin.

Open Access Original Research Article

Light Microscopic and Immunohistochemical Study Revealed Sex-Dependent Kidney Morphology in DBA/2crSlc Mice

M. A. Awal, A. Yabuki, O. Ichii, M. Matsumoto, S. Suzuki, J. Alam

Biotechnology Journal International, Page 341-349
DOI: 10.9734/BBJ/2013/3100

Aim: Sex-dependent differences in kidney histology have been observed in different species of the laboratory animals. The present study was conducted to evaluate the sex and strain-dependent changes in DBA/2CrSlc mouse kidney morphology by using light microscopy and immunohistochemistry.

Methods: A total of 12 DBA/2CrSlc male and female mice of 2 months of age were used in this study. Mice were sacrificed by exsanguination under anesthesia using a mixture of Ketamine and Medetomidine. Both right and left kidneys were removed aseptically and central slices including hilum were cut perpendicular to the long axis of the organ and preserved in Zamboni solution. Paraffin blocks were made and tissue sections were stained with hematoxylin and eosin, and PAS stains to observe the general morphology of the kidney glomerulus. Immunohistochemistry was performed to detect renin positive sites, expression of Cyclooxygenase-2 (COX-2) and Nitric Oxide Synthase (nNOS). Number of renin, COX-2 and nNOS positive sites were counted and tabulated. The data were statistically analyzed for any significant differences between male and female mice.

Results: Our results reveal that the glomerular capsule of male mouse kidney was consisted of a single layer of simple cuboidal epithelium whereas it was a single layer of simple squamous epithelium in the female kidney. PAS-positive granules (small and giant granules) were observed in PST epithelium and collecting ducts in female kidney, but this feature was absent in male kidneys. Strong nNOS positive reaction for PST epithelium and collecting ducts was observed in female, but this character was absent in male kidney. The total number of glomeruli, renin, COX-2, and nNOS positive sites was comparatively higher in female kidneys then that in male. However, statistical analysis revealed no significant differences of the areas of renin, nNOS and COX-2-positive sites between the male and female kidneys (P<0.05).

Conclusion: Light microscopic and immunohistochemical study revealed sex-dependent histological morphology of the DBA/2CrSlc mouse kidney. DBA/2CrSlc female mouse kidney revealed renin, COX-2 and nNOS -positive reactions in the present study but male mice showed nNOS-negative reaction. The reason for nNOS-negative reaction in male is not clearly understood. It is suggested that this species can be experimentally used in the laboratory for investigating kidney function and related pathological studies.

Open Access Original Research Article

Screening of Locally Isolated Aspergillus species for Their Cellulolytic Potential and Their Optimization on Vigna mungo in Solid State Fermentation

Umbrin Ilyas, Faryal Gohar, Sumaria Saeed, Zanib Bukhari, Hira Ilyas

Biotechnology Journal International, Page 350-358
DOI: 10.9734/BBJ/2013/2849

Aims: To evaluate cellulolytic potential of locally isolated Aspergillus species and emphasis the importance of screening on qualitative bases. Also, to enhance cellulase production by optimizing the process parameters of the solid state fermentation of Vigna mungo with the isolated Aspergillus species.

Place and Duration of Study: Institute of Biological Sciences, Department of Botany, University of Gujrat, Gujrat, Pakistan, from February, 2012 to November, 2012.

Methodology: Aspergillus species were isolated from different deteriorated plant materials and maintained on potato dextrose agar. The purified isolated species were qualitatively screened on caboxymethylcellulose agar plates. The cellulolytic ability was further tested by culturing Aspergillus species on V. mungo in solid state fermentation. Various nutritional and cultivation parameters were optimized for the production of cellulases by Aspergillus species on V. mungo.

Results: Most of the isolated Aspergillus species showed potential for cellulase production. However, A. terreus gave qualitatively, the highest cellulase activity by fermenting V. mungo of 0.566 IU/g while A. niger gave quantitatively, the highest cellulase activity of 0.435 IU/g. Cellulase production by A. terreus reached at its maximum with 0.1% of urea as nitrogen source and 80% of initial moisture level at 35°C after 144 hours of fermentation.

Conclusion: A. terreus was identified as highest cellulase producer among the isolated Aspergillus species. The cellulolytic ability of Aspergillus specie greatly depends upon the nutritional and cultivation parameters of fermentation.

Open Access Original Research Article

Characteristics of Penicillin G Acylase Immobilized onto Iron Oxide Nanoparticles

Momen Seyed Bahman, Atyabi Seyed Mohammad, Azim Akbarzadeh, Salimi Mona, Sedigeh Hatami Gigloo, Farhangi Ali, Norouzian Dariush

Biotechnology Journal International, Page 367-376
DOI: 10.9734/BBJ/2013/2987

Penicillin G acylase was immobilized onto iron oxide nanoparticles coated with polyethyleneimine and then cross linked with glutaraldehyde solution. The FTIR spectrum of immobilized enzyme showed peak at 1648cm-1 which can be attributed to the C=N bonds of Schiff’s base linkage formed between glutaraldehyde and amino group of penicillin G acylase. By considering the FTIR spectrum of nano particle coated with polyethyleneimine, adsorption of penicillin G acylase has taken place and then glutaraldehyde cross linked enzyme onto activated support. Catalytic properties of nano penicillin G acylase were improved upon immobilization as compared to its free counterpart. The optimal pH and temperature were determined to be 7.0, 10.0, 50 and 75ºC for free and immobilized penicillin G acylase, respectively. Thermal stabilities of both nano and free penicillin G acylase were studied .The Km value of immobilized nanozyme was calculated from Lineweaver Burck plot to be 0.23 μM while that of free penicillin G acylase was 0.28μM. In this way nano penicillin G acylase with improved catalytic properties was developed as compared to its soluble counterpart.

Open Access Original Research Article

In vitro and In vivo Effects of the Leaf Extracts of Cassia tora and Cassia sophera in Reducing the Cytotoxicity and Angiogenesis

Sumangala Rao, C. Suresh

Biotechnology Journal International, Page 377-389
DOI: 10.9734/BBJ/2013/4244

Aim: Consumption of C. tora and C. sophera of family caesalpinaceae as leafy vegetables by local folklore has initiated this study to investigate the cytotoxic effect and antiangiogenic effect of their leaf extracts on human cancer cell lines and chick embryo respectively.

Study Design: For in vitro studies colon and hepatocarcinoma cell lines were used. For in vivo studies chicken egg chorioallantoic membrane (CAM) assay is used.

Methodology: Methanol, ethyl acetate and petroleum ether extracts of leaves were tested for antiproliferative activity using MTT assay, on HCT15 and Hep G2 cell lines. In vivo CAM model was used to study anti-angiogenesis activity of extracts using filter paper discs.  

Results: All extracts of both plants were toxic to tested cell lines with increasing concentrations and time. Ethyl extract of Cassia tora was found to be most toxic on Hep G2 cell line, though ethyl extract and methanol extracts of both plants were found to give strong antiproliferative activity. Hep G2 was found to be most sensitive to all extracts than HCT15 as inhibited by low IC50 values. Ethyl acetate extracts of both plants exhibited greater anti-angiogenesis activity than other extracts.

Open Access Original Research Article

SSR- Based Genetic Diversity Assessment in Tetraploid and Hexaploid Wheat Populations

Hanaa Mahdy Abouzied, Samah M. M. Eldemery, Kamal Fouad Abdellatif

Biotechnology Journal International, Page 390-404
DOI: 10.9734/BBJ/2013/4340

Molecular analysis for a set of hexaploid (Triticum aestvium) and tetraploid (Triticum durum) wheat cultivars was investigated by applying 11 SSR primers set. The plant materials consisted of 45 genotypes 15 of which were Triticum aestivum and 30 of T. durum obtained from four different regions Egypt, Greece, Cyprus and Italy. PCR products were separated on a 6% denaturing polyacrylamide gel electrophoresis and produced a total of 3840 DNA fragments which were used for the molecular analysis. The estimated parameters computed by POPGENE (Version 1.32) within the two population indicated that the Nei’s genetic diversity (H) was 0.2827, and the Shannon's Information index (I) was 0.4533 with standard deviation ± 0.0699 and ± 0.0852 respectively. The analysis of population structure revealed that genetic diversity within populations (Hs=0.2761) represented 97.7% of the total genetic diversity (HT=0.2827). The proportion of the total genetic diversity that was attributed to the population differentiation was low (Gst=0.0233) within population. ANOSIM (ANalysis Of Similarities), results showed that R was equal to 0.9048 (P<0.0001) indicated that all the most similar samples of genotypes are within the same population. The wheat varieties from the four distinct regions were clustered according to SSR data into two main clusters, durum wheat varieties and bread wheat varieties, the principal coordinate analysis (PCOORDA) validated the results of the dendrogram. This study showed that the two populations still had moderate considerable level of genetic diversity and show little genetic differentiation among them. Understanding genetic variation within and between populations is essential for the establishment of an effective breeding program concerning the intraspecific and interspecific hybridization.


Open Access Original Research Article

Efficient Micropropagation of Spilanthes acmella (L.) Murr.: A Threatened Medicinal Herb

Shiwali Sharma, Anwar Shahzad

Biotechnology Journal International, Page 405-415
DOI: 10.9734/BBJ/2013/3556

The present study describes an efficient and reproducible protocol for micropropagation of S. acmella. Shoot tips taken from 3 week-old aseptic seedlings were cultured on Murashige and Skoog (MS) semi-solid medium supplemented with different concentrations of TDZ. Among various concentrations, 0.25 µM TDZ was found to be optimum for shoot regeneration as it induced a maximum of 30.0 shoots per explant however with retarded growth (1.0 cm). Among different volumes of culture media, 15 ml liquid culture medium favored best response wherein a maximum of 80.2 shoots per explant with an average shoot length of 7.0 cm were induced after 6 week of subculturing. Successful in vitro rooting was induced on 2.5 µM NAA containing half-strength MS medium. Almost 96% rooted plants successfully transferred and acclimatized ex vitro under green house conditions. Morphological and physiological parameters compared with the in vivo-grown seedlings of the same age appeared to be ‘normal’ in respect to the fundamental characteristics examined.

Open Access Review Article

African Cassava: Biotechnology and Molecular Breeding to the Rescue

Michael A. Gbadegesin, Charles O. Olaiya, John R. Beeching

Biotechnology Journal International, Page 305-317
DOI: 10.9734/BBJ/2013/3449

Cassava is an important African food crop, where it is a staple to about 250 Million people. It is a household name in Nigeria, the world largest producer of the root crop. It is propagated from stem cuttings and well known for its adaptation to wide range of adapho-climatic conditions and including those unfavourable for other crops. However cassava production, exploitation, utilization and acceptance are limited by diseases and pests, cyanogenesis, low protein content and quality, and post-harvest physiological deterioration. The breeding research activities of IITA (International Institute of Tropical Agriculture) Ibadan, Nigeria, CIAT (International Centre of Tropical Agriculture) located in Cali, Colombia and National Root Crop Research Institute (NRCRI), Umudike, Nigeria have transformed cassava to double as a food security crop as well as a cash and industrial crop. Of recent, Bio Cassava Plus, an initiative sponsored by Bill and Melinda Gates, has been using experimental biotechnology approaches to address several of the main constraints to African cassava. This review presents the many advantages of cassava to the small-scale farmer and its potentials for industrial applications. It also describes the roles of biotic and abiotic factors hampering the production yield, root quality, nutritional adequacy, marketability and acceptance, and commercial processes. The use of conventional breeding and biotechnology in unravelling the milieu of these constraints is discussed as well.