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EDITORIAL BOARD

2009

EVALUATION OF COTTON RELEASED/CANDIDATE VARIETIES UNDER DIFFERENT SOWING DATES AT TANDOJAM CONDITIONS

By Deho, Z.A., S.Laghari, S. Abro, M.A. Arain, S.A. Abro and Fakharuddin

VOL-6 NO-(1-2)

Abstract

A field study was conducted to evaluate the cotton released/candidate varieties under different sowing dates for the qualitative and quantitative characters at Nuclear Institute of Agriculture (NIA) Tando jam. Three sowing dates viz., 15th April, 15th May and 15th June and three varieties and one candidate variety were tested respectively. The result revealed that the highest mean (2983 kg ha-1) seed cotton yield has got sowing date two 15th May where as lowest mean (1996 kgs/ha) seed cotton yield in sowing date 15th June. The mean values of all varieties showed more seed index (8.34g) and highest (90%) germination percentage in sowing date 15th April and 15th May. The highest GOT% (35.42) and germination percent (89%) were recorded in 15th May sown crop. It was found that the delaying sowing date significantly decreased yield and yield contributing parameters.

THE GENETIC VARIABILITY OF THE MICROPROPAGATED SOLENOSTEMA ARGHLE TISSUE CULTURE DERIVED PLANTS USING RAPD TECHNIQUE

By AbdAlla M.M

VOL-6 NO-(1-2)

Abstract

Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic stability of micropropagated plants of ?Solenostema arghle. Eight arbitrary decamers were used to amplify DNA from in vitro plant material to assess the genetic fidelity. All RAPD profiles from micropropagated plants were monomorphic and similar together. No variation was detected within the micropropagated plants. The utilization of RAPD markers both for the assessment of genetic stability of clonal materials and to certify genetic stability throughout the systems of micropropagation is discussed.

THE ROLE OF FERMENTED SOYMILK WITH POTENTIAL PROBIOTIC PROPERTIES IN THE TREATMENT OF DIARRHEA IN YOUNG RATS

By Khiralla1 , Ghada M., Nagwa M.H. Rasmy2 , W.A. El-Malky1 andManar T. Ibrahim2

VOL-6 NO-(1-2)

Abstract

In recent years, novel insights have been made into the role of probiotics in health and inflammatory bowel disease. In the present study, soymilk was fermented with Lactobacillus casei, L. bulgaricus, Bif. longum as single or mixed starters (1:1:1 v/v). The viable counts in probiotic soymilk were ranged from 7 to 8 log CFU/ml. These probiotic soymilk preparations were organolyptically and biologically evaluated. Using these probiotics led to a significant (p>0.05) improvement of the unpleasant taste of raw. The effect of these products against picolax-induced diarrhea in young rats (3 weeks) was also studied. Daily notices, stool frequency, stool fluidity, and death ratio were recorded during 7 days. Biochemical parameters in blood serum of rats including kidney and liver functions, mineral concentrations, glucose and hemoglobin were assayed before and after the onset of diarrhea. Significant reduction (p

CLONING AND SEQUENCING OF CELLULASE-ENCODING GENE (cel 2) FROM LOCALLY ISOLATED BACILLUS SP. STRAIN A4

By IBRAHIM, A. M.1 , Amal A. Abdel-Aziz 2 , Hoda Mahrous3 and U. M. Abdulraouf 4

VOL-6 NO-(1-2)

Abstract

A Bacillus bacterium, Bacillus sp. strain A-4 was isolated from the Egyptian soil and identified according to the methods in Bergey’s Manual of Systematic Bacteriology. The bacterial isolate, Bacillus sp. strain A-4 was a facultative anaerobic, mesophilic, sporeforming, Gram-positive, motile, rod-shaped organism and produced catalase. Bacillus sp. strain A-4 can degrade cellulose, hemicellulose, amylose and other forms of carbon sources. A DNA genomic library constructed from Bacillus sp. Strain A-4 and screened for cellulase (CMCase) activity. Recombinant ?-glycosyl hydrolase activity was detected on the basis of the clearing of hallos around Escherichia coli colons grown on a substrate containing carboxymethylcellulose (CMC). The nucleotide sequence of the cellulase gene (cel 2), corresponded to complete open reading frame 1167 bp that codes for a 389 amino acid and a protein with a molecular mass of 4249Da. From sequence analysis, cel 2 belongs to glycosyl hydrolase family 5 and exhibit high similarity to engD and engO from Clostridium cellulovorans and celB from Ruminococcus albus. The crude enzyme preparation from Bacillus sp. A-4 exhibits activity over a broad range of pH5 to 6 and has good stability across temperatures 50°C and pH6.

EXPRESSION OF SOME APOPTOTIC INHIBITORS IN CASES OF ADULT ACUTE MYELOID LEUKEMIA

By Abir A. Saad1 §, Shaden M. Hanafi2 , Samir A. El-Masry2 and Magdy M. Hassan3

VOL-6 NO-(1-2)

Abstract

The expression of antiapoptotic genes; bcl-2, Survivin and Aven was examined in Egyptian cases of acute myeloid leukemia (AML). Flow cytometry (FCM) was used to detect the expression of bcl-2 and reverse transcriptase-PCR was used to examine the expression of Survivin and Aven mRNA. Bcl-2 was highly expressed in all cases of AML than controls (p< 0.001). The percentage of bcl-2+ cells was significantly higher in M1 (60%), M2 (54%) and M3 (56%) than in M4 (43%) and M5 (40%) subtypes (p=0.01). The AML patients showed expression of Survivin mRNA but no expression of Aven mRNA was observed. Out of 30 patients, 23 (76.6%) showed detectable levels of Survivin expression. The ratio between Survivin mRNA and beta-Actin mRNA was analyzed quantitatively in AML samples showing expression bands. Different expression levels of Survivin mRNA could be observed between different FAB subgroups. M1, M4 and M5 AML had a relatively higher expression level of Survivin mRNA (range: 0.56-0.81) than M2 and M3 patients (range:0.04-0.1). There was also a significant correlation between Survivin expression and PB blast % in cases of M5 subtype of patients (P=0.02). The Survivin-positive AML cases showed a higher percentage of bcl-2 expression and a significantly higher Bcl-2 MFI (mean fluorescence index) in comparison with Survivinnegative ones (p= 0.01). Our findings suggest that over-expression of both apoptotic inhibitors Bcl-2 and Survivin might be involved in AML pathogenesis.

PRODUCTION OF PROTEASES BY STAPHYLOCOCCUS EPIDERMIDIS EFRL 12 USING COST EFFECTIVE SUBSTRATE (MOLASSES) AS A CARBON SOURCE

By Qureshi, A. Sattar and Dahot, M. Umar

VOL-6 NO-(1-2)

Abstract

Staphylococcus epidermidis EFRL 12 capable of producing extracellular protease isolated from soil. The protease production was investigated in batch wise submerged fermentation. The protease production was studied with varying carbon sources with (0.5 and 1.0%) concentrations, peptone (0.25-1.5%) and different nitrogen sources (1.0%).The effect of sugarcane bagasse, rice husk, molasses and date syrup were checked along with pure carbon sources. The best production was obtained when microorganism was grown on mineral medium containing 1.0% molasses as carbon source and 1.0% ammonium nitrate as nitrogen source after 6 hours of incubation at 37°C and initial pH was adjusted to 7.5.

APPLICATION OF LASER MICROBEAM CELL SURGERY AND AGROBACTERIUM–MEDIATED GENE TRANSFORMATION SYSTEMS IN MELON (CUCUMIS MELO L)

By Khames 1 G., 1 A.A. Mona, 2 M.H. Gihan, 3,4A.S.Sadik and 1 Y.Badr

VOL-6 NO-(1-2)

Abstract

In the current investigation, the application of laser microbeam cell surgery as a transformation system in melon has been studied and compared with transformation via Agrobacterium tumefaciens as well as a combine of the two systems. All these transformation treatments have been carried out on the hypocotyl proximal zone of melon cv. Shahd El-Dokki using the plasmid pISV2456 that harboring gus-intron and bar genes. The treated explants were regenerated on shoot regeneration medium supplemented with 250 µg/l bialaphos. Results showed that there was no significant difference of shoot regeneration between the laser and Agrobacterium treatments, however, the regeneration percentage varied among the treatments, the laser microbeam treatment showed highest percentage than the Agrobacterium and combined methods, (76, 72 and 68 % respectively). The transformation systems were evaluated by detecting the expression of the gus gene using histochemical assay, while the integration of the both genes (gus and bar) were confirmed by the PCR assays, indicating that the two genes were successfully transferred to the plants in the three methods. Seven shoots out of 30 tested shoots were PCR-positive, representing 23.33%, in both Agrobacterium and laser treatments, while, only 6 out of 30 tested shoots were PCR-positive, representing, 20%. This study demonstrates using laser tool as a new transformation method in melon cv Shahd El-DOkk, indicating that laser technique could be considered a competitive transformation method with the Agrobacterium-system.

ESTABLISHMENT OF REGENERATION AND TRANSFORMATION SYSTEMS OF THE CULTIVAR 'M9681' OF SUGAR BEET (BETA VULGARIS L.)

By Attia, A.O.1 ; Roba M. Ismail1 ; Eman M. Fahmy2 and A.S. Sadik1,3,4

VOL-6 NO-(1-2)

Abstract

Sugar beet is one of the most important arable crops. In Egypt, sugar beet plant is a second main source of sugar after sugarcane plant. In this study, the regeneration and Agrobacterium-transformation systems of the M9681 sugar beet cultivar were established. The effect of the combinations of 6-bezylaminopurine (BAP) as cytokinin and indole-3-acetic acid (IAA) as an auxin on adventitious shoot regeneration from petiole explants excised from the seedlings, and the effect of NAA and IAA on root formation with the aim of improving the procedures for regeneration. gus-intron and bar as a reporter and selectable marker genes, respectively, were used for the establishment of Agrobactirum-transformation system. The integration of transgenes into sugar beet genome was detected by PCR. Two fragments of about 2070 and 540 bp were amplified regarding the two genes, respectively. Furthermore, the expression of gus-intron gene was also tested by GUS histochemical assay

EVALUATION OF TRITICUM DURUM FOR SALT TOLERANT

By AbdAlla, M.M.

VOL-6 NO-(1-2)

Abstract

This work was conducted to evaluate salt tolerance in Triticum durum wheat varieties by selecting the high tolerant genotypes from the total genotypes under salt stress by using mature embryos. Mature embryos of four Triticum durum were grown on MS medium for germination under 4 concentrations of salt solutions (zero, 4000, 6000 and 8000ppm). The germination percent at the high salt concentration (8000ppm) was 77.7% S1, followed by77.1%D2, 66.2% ID12 and finally 64.5% D1 respectively. Plantlets on these media were transferred to pots with sand and irrigated with the same salt solutions. The produced grains were recultured to obtain some grains for propagation. Among the saline tolerance cultures proline accumulation was increased. SDS-page profile revealed an increasing in band intensity in 8000ppm than in the control and presented two bands with molecular weight 32and 28kda related to proline accumulation in the cells. The four Triticum durum (wheat) varieties had the ability to resist against salt stress.

IN VITRO PROPAGATION OF CARNATION (DIANTHUS CARYOPHYLLUS L.) UNDER SALT STRESS

By Faouzi Haouala* and Faiza Jaziri

VOL-6 NO-(1-2)

Abstract

Herbaceous micro-cuttings of Carnation (Dianthus caryophyllus L., cv Yellow Liberty and Crimson Tempo) were rooted on half-strength Murashige and Skoog medium (MS/2) supplemented with different concentrations of NaCl (50, 100 and 150mM). Cultures were maintained for eight weeks with monthly subculture. On NaCl deprived medium (Control), the rooting rate was better for the cultivar Crimson Tempo (94.1%) than for Yellow Liberty (86.3%). Salinity affected significantly the rooting of cuttings. In fact, the rooting rate and the length of the main root were clearly reduced in presence of NaCl. Crimson Tempo was more tolerant to NaCl than Yellow Liberty. Rooting rate of its cuttings at 150mM NaCl was 36.2%. No roots were obtained with the cv Yellow Liberty. Vegetative growth of cuttings was also reduced by adding 100 and 50mM NaCl to the culture medium for Crimson Tempo and Yellow Liberty, respectively.

COMPARISON ON THE EFFECT OF TREATMENT AND SUBCULTURING ON SHOOT REGENERATION FROM SHOOT TIP SEEDLINGS OF Psidium guajava L. var. Beaumont

By Tajul A. Abdullah1, Maheran A. Aziz *2 , Azmi A. Rashid2 , Ghizan Saleh3 , S.M.A. Elhory2 , Syaiful B Panjaitan2 , Hailmi M.S. 1 and Noor Muzamil M.

VOL-6 NO-(1-2)

Abstract

An experiment was carried out in order to study the effect of treatment containing 0.5 and 1.0 mg/L 6-benzylaminopurine (BAP) and effect of subculture on percentage of shoot formation, number of shoot per explant and mean shoot height of shoot tip explant of Psidium guajava L. var. Beaumont. MS medium (Murashige and Skoog, 1962) was used with 30 g/L of sucrose and 7g/L agar type 900. Shoot tips of elongated shoots were decapitated to abolish its apical dominance prior to sub-culturing was carried out to a fresh medium every 4 weeks. The increase in mean number of shoots per explant and the shoot height was observed for every subculture. Treatment 1.0 mg/L BAP in comparison to treatment 0.5 mg/L BAP showed higher mean number of shoot per explant per subculture whereas treatment 0.5 mg/L BAP showed higher mean shoot height per subculture. Data were analyzed using T-test until the fifth subculture.

IDENTIFICATION AND QUANTIFICATION OF AMINO ACIDS FROM PSORIATIC AND NORMAL EPIDERMIS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

By Shah Muhammad Mahesar*, M. Umar Dahot**, Muhammad Yar Khuhawar*** and Javeed Anwar*****

VOL-6 NO-(1-2)

Abstract

In this study, a modified fluorescence technique high performance liquid chromatography (HPLC) was adapted to separate the amino acids from the hydrolyzed keratin samples. These samples obtained from the epidermal layer of the normal and psoriatic human subjects. The keratin extracts were quantified in gram percentage of the dried skin and the amino acids concentrations were measured in µg/g, mean retention time (tR), slope value and the coefficient of determination (r2) of each eluted amino acid was calculated. The coefficients of variation for amino acid standards ranged from 0.12% to 0.28%, mean, standard deviation of peak area and coefficients of variation of peak area were calculated. From the normal hydrolyzed keratin protein fraction, 12 amino acids were determined and identified as aspartic acid, glutamic acid, asparagines, serine, glutamine, glycine, histidine, citrulline, arginine, ?-alanine, tyrosine and valine. These amino acids were also determined in psoriatic samples while standard deviations (SD), standard error mean (SEM) and coefficient variation (CV%) of normal and psoriatic samples were also calculated. The higher concentration of amino acids in normal samples against psoriatic samples determined as glutamic acid 92.76±16.83/50.87±9.88, glutamine 198.05±18.74/19.74±3.74 while higher concentrations of amino acids was determined in psoriatic samples against normal samples such s asparagines 81.06±10.62/29.98±3.64/, arginine 164.42±35/46.14±8.46, tyrosine 214.38±29.61/59.64±8.82 and valine 169.7± 19.35/128.06±15.14. It is concluded that the absolute concentration of amino acids in psoriatic skin indicated a number of variations as compared to normal skin samples.

GENETIC CHARACTERIZATION OF INDIAN MUSTARD (Brassica juncea) GENOTYPES TOLERANT TO HEAVY METALS

By Ibrahim1 , I. A., A.A. Guirgis2 , E. A. El-Absawy3 , Amal Abd El-Aziz2 and Azza2 M. Mhmod

VOL-6 NO-(1-2)

Abstract

In the present research work, ethyl methanesulfonate (EMS) agent was applied to select highly tolerant Indian mustard (Brassica juncea) plants to some heavy metals (zinc, cadmium, nickel, and molybdenum). Four hyper-accumulator lines were selected, that proved to be more tolerant to the corresponding heavy metal when compared to the wildtype control, for genetic analysis in an attempt to elucidate the genetic bases of heavy metal tolerance in higher plants. Total soluble protein electrophoretic profiles indicated that metallothioneins (5-8 kDa) were expressed in Cd, Mo, Ni, and Zn hyperaccumulator lines when compared with their corresponding wild-type plants. Peroxidase and super oxide dismutase isozyme patterns were not conclusive. The expression of glutathione synthetase (GSH) gene(s) was monitored in the four genotypes lines and the wild-type control, using RT-PCR technologies. Increases ranging between 5 to 2 folds were encountered in the genotypes lines relative to the controls. According to the present findings, it seems that B. juncea hyperaccumulator genotypes have evolved a number of mechanisms to cope with heavy metal stress such as the expression of several metallothioneins and the over expression of GSH gene(s). However, the results of oxygen radical scavengers were not conclusive, and need further justifications.

Acknowledgment of Reviewers

The Editor, Pak. J. Biotechnol. is very grateful to the following scientists who dedicated their considerable time and expertise to the journal by serving as reviewers from January 1, 2009 to December 31, 2009 for Vol. 6 No. 1 and 2, 2009.
Prof. Dr. Alexandre SemenovMoscow, Russia
Prof. Dr. Atef S. SadikTaif, Saudi Arabia
Prof. Dr. Sher Muhammed MangrioUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Jian He XuShanghai, China
Prof. Dr. Ma WeiShanghai, China
Prof. Dr. Wei WangShanghai, China
Prof. Dr. Muhammed Umar Dahot Uni. Of Sindh, Jamshoro, Pakistan
Dr. S. A. Anitha ChristyHouston, USA
Dr. Mostafa RahimnejadUniv. Babool, Iran
Prof. Dr. Nisa RachmaniaBogor, Indonesia
Prof. Dr. Azara KhanumRawalpindi, Pakistan
Prof. Dr. Asif AliFaisalabad, Pakistan
Dr. S. Habib Ahmed NaqviUni. Of Sindh, Jamshoro, Pakistan
Dr. Imtiaz Ahmed KhanNIA Tando Jam, Pakistan
Dr. Mehboob Ali sialiNIA Tando Jam, Pakistan