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EDITORIAL BOARD

2007

EFFECT OF SEMI-DWARFISM ON YIELD AND YIELD COMPONENTS IN WHEAT (TRITICUM AESTIVUM L.)

By Jamali K.D, Saima Arain, M.H. Naqvi, A.M. Soomro, M.A. Arain, and Syed Ashraf Ali

Vol-4 No-(1-2)

Abstract

Yield and yield component studies were conducted for semi-dwarf wheat genotypes. The yield comparison showed that line 03 (525 g) had the highest grain yield per plot compared with other lines and tested varieties. This could be due to early heading date and increased number of grains per spikelet. The subsequent lines of higher grain yields were 15 (512.5 g), 01 (462.5 g), 14 (462.5 g), 5 (437.5 g), 9 (412.5 g) and 13 (425 g). The better number of grains per spikelet (fertility) was the second reason beside the early heading date for high grain yield of line 15. The higher grain yield in line 1 could be due to medium heading date, higher number of spikelets per spike and increased number of grains per spike. The combined correlation studies suggested that plant height had positive and highly significant correlation with spike length, number of spikelets per spike and main spike grain yield. Spike length had positive and highly significant correlation with number of spikelets, number of grains per spike and main spike grain yield. Number of spikelets per spike had positive and highly significant correlation with number of grains per spike and main spike grain yield. Number of grains per spikelet had highly significant negative correlation with spike length and number of spikelets per spike. Grain yield of main spike is a very important character which had highly significant positive correlation with characters plant height, spike length, number of spikelets per spike and number of grains per spike.

CITRIC ACID FERMENTATION OF HYDROLYSED SWEET POTATO STARCH BY ASPERGILLUS NIGER

By Andleeb Anwar Sardar, Sikander Ali*, Samina Anwar and Ikram-ul-Haq

Vol-4 No-(1-2)

Abstract

The present study deals with the screening of Aspergillus niger from soil for citric acid production by submerged fermentation. A. niger IIB-A6 produced 23.32±1.44 g/l citric acid. Sweet potato starch was found as a better substrate compared to maize starch. The kinetic parameters Yp/x (1.96 g/g cells), Qp (0.13 g/l/h) and qp (0.014 g/g/h) were 8-10 fold higher with sweet potato starch than the maize starch. Sugar conc. (150 g/l), incubation period (168 h) and initial pH (6.0) were optimized. The maximum citric acid production obtained during the course of study was 28.40±1.07 g/l. Sugar consumption and dry cell mass were 121.33±0.91 and 14.29±0.88 g/l, respectively. The mycelia were small pellets.

EFFECTIVE DETECTION OF BANANA BUNCHY TOP NANOVIRUS USING DNA PROBES

By Khalil S.M.1 , M.I.1 Salama, I.A. Abdel-Hamid1 , Hanan A. Nour El-Din1 and A.S. Sadik1,2

Vol-4 No-(1-2)

Abstract

According to DNA sequences of components one and four of BBTV isolate, that were isolated from AGERI cultivated area, Egypt, two pairs of primers were designed to amplify BBTV replicase (BBTV-rep) and movement protein (BBTV-mv) genes. The amplified fragments were cloned into pGEM-T easy vector and two new plasmids were created, i.e., pBBTV-Rep and BBTV-MV and transformed into Escherichia coli. Results showed that PCR products with a size of about 1.1 Kbp, which represented BBTV-rep and BBTV-mv, were successfully amplified. In case of BBTV-mv, a number of 17 clones were subjected to minipreparation followed by restriction endonuclease analysis. Results showed that 9 (1-5, 12-14 and 16) out of the 17 clones were found to be recombinant and 1.1 Kbp fragments were released. For Southern blotting analysis, the previous gel was southern blotted onto a membrane and the same 9 clones gave signals when the membrane was hybridized with 32P-labeled-PCR probe of BBTV-mv amplified from DNA of BBTV-infected plants. On the other hand, similar probe was prepared from pBBTV-MV plasmid and used for virus detection in BBTV-infected plants. On the other hand, a PCR product amplified from the pBBTVRep plasmid was labeled with 32P and used as a probe for detection of BBTV in virusinfected plants as well as the healthy as control. Findings of this study could be concluded in developing a rapid, effective and sensitive detection of BBTV in nucleic acids of either PCR amplified from BBTV-infected plants or 32P-DNA-PCR labeled probe via Southern blot hybridization

ANALYSIS OF GENETIC STABILITY OF MICROPROPAGATED OCHRADENUS BACCATUS

By Ghareb, Heba E. 1; Gaber, Mahdia F. 1; Abdallah, Naglaa A. 2,3

Vol-4 No-(1-2)

Abstract

Ochradenus baccatus (Del.), an important medicinal plant, belongs to the family Resedacea. Presently this is an endangered species therefore nonconventional methods for their propagation are requested to prevent its eradication. We developed a micropropagation method for mass multiplication of O. baccatus. Explants collected from South Sinai were used to establish cultures of O. baccatus. Surface sterilized nodal shoot segments and shoot tips were used for micropropagation by culturing on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of BA and 2ip. Stem node sections were superior in enhancing formation of sub axillary shoots on MS medium supplemented with 1.0 mg/l BA + 1.0 mg/l 2ip. The multiplied shoots were then rooted on WPM supplemented with 1.0 mg/l IBA. The plantlets were acclimatized ex vitro. Genetic stability of plants produced in vitro was evaluated using random amplified polymorphic DNA (RAPD) analysis. Plants produced under different concentrations of hormones, mother plant and the plants grown ex vitro were analyzed by RAPD using 8 random 10-mer DNA primers. Results showed that 73 RAPD bands were obtained which gave 39.72 % polymorphism. The production of variant plants is not good for commercial propagation. However, it could be minimized by reducing the subculture cycle or by reducing hormone level in media. Additionally, this variability could be used for further improvement and selection of new cultivars and is an important source of variability to be exploited.

ISOLATION AND CHARACHTERIZATION OF CRUDE PROTEASE INHIBITORS FROM RICINUS COMMUNIS (Castor Beans)

By Fahmida A. Soomro, M. Umar Dahot and *A.U. Rehman

Vol-4 No-(1-2)

Abstract

Protease inhibitors are frequently found in different sources such as animals, plants and microorganisms and they inhibit the activity of proteases. In our studies protease inhibitors (PIs) were isolated and characterized from plants tissues such as seeds. Ricinus communis has shown highest inhibition activity as compared to other seeds (Glycin max, Triticum astevium, Zey mays, Helianthus annus, Pennisetium typheides, Avena sativa, Brassica compestris etc.) at optimized conditions. Crude protease inhibitor sample (extracted from castor beans) incubated for 60 minutes at 35 ºC, using sodium phosphate buffer pH =7.5, soluble casein as a substrate and trypsin enzyme were used for maximum inhibition activity. The crude protease inhibitors were characterized at different parameters such as effect of time period, substrate concentration, enzyme and sample concentration, variable temperature and pH control.

STUDIES ON STABILITY OF SUGAR BEET GENOTYPES UNDER OLD AND NEW RECLAIMED LANDS

By 1 Shalaby N.M.S., 2 I.E El-Beially, A.H.S. 3Al-Labbody and 4 S .H.M Abd- El- Haleem

Vol-4 No-(1-2)

Abstract

The stability measures are useful in characterizing genotypes by showing their relative performance in various environments. Ten sugar beet genotypes were evaluated across eight different environments in old lands and four environments in new land reclaimed with two sowing dates and two locations. Two years to study their yield potential and stability. Four statistical parameters (Wricke’s Ecovalence, Shukla’s (two measures), kang and Coefficient of variation) were used for determining yield and other traits stability of the genotypes. Highly significant differences due to genotypes (G) in old land and all environments for all studied traits except sucrose % in combined data were found. Environments (E) and GxE interaction had the same trend in all lands and combined data except sucrose % in old land, which is suggesting differential responses of the genotypes. Results showed that the statistics parameters were differed in their efficiency for determining the true stable genotypes. But all proposed parameters were in line for determining stability of G4 in old land for root yield and sucrose yield and over all the environments with considering mean yield, so it could be recommended for growing under conditions of old land and G6 for root yield and G3 for sugar yield in new reclaimed land. Concerning sucrose (%), the genotype G9 in old land and the genotypes (G4, G5, G6, G9 and G10) concerning yields in new reclaimed land were recorded by all proposed parameters (C.V.,  2 i, S2 i, Eco. and Ysi) for stability. So they could be recommended for growing under these conditions. The relative small value for the heritability in all traits studied means that the large effect by environmental on the studied traits.

ANTAGONISTIC AND INSECTICIDAL ACTIVITIES OF SOME STREPTOMYCES ISOLATES

By Osman G.1 ; Mostafa S.1 and Sonya H. Mohamed2

Vol-4 No-(1-2)

Abstract

Fifteen local isolates of Streptomyces named, S01, S02, S03, S04, S05, S06, S07, S08, S09, S10, S11, S12, S13, S14, and S15, isolated from different soils and geographical areas in Egypt were used in this investigation. These isolates were propagated for screening studies and to evaluate their efficiency as antagonistic agents against some phytopathogenic fungi (such as, Rhizopus nigricans, Aspergillus niger, Fusarium oxysporum and Helminthisporum gramenium) and insect pest, (cotton leaf worm, Spodopetra littoralis). The Streptomyces isolates were grown on starch nitrate broth medium under shaking condition at 28oC for 6 days. Culture filtrates were then tested against the five phytopathogenic fungi. Results revealed that most of the isolates were varied in their antagonistic activities. Isolate S08 was active against R. nigricans, A. niger and F. oxysporum while isolates S01, S05, S11 and S14 were active against A. niger and F. oxysporum and isolates S04, S09 and S13 were active against R. nigricans and A. niger. On the other hand, no antifungal activity was found against Helminthisporum sp. The insecticidal activity of both culture filtrates and cell pellets were tested against cotton leaf worm. The experimental results showed that the pellets of some Streptomyces isolates were more active against cotton leaf worm than culture filtrates. Generally, isolates S05, S08, S10 and S15 showed 80, 100, 70 and 80% mortality against cotton leaf worm, respectively. The protein(s) of isolate S08 cells was purified through ammonium sulfate saturation 40, 60 and 80%. Results of SDS-PAGE analysis showed that a 40 KDa protein was purified and showed high activity against four instars of the cotton leaf worm. This result demonstrated the ability to use such Streptomyces isolates as effective biopesticide agents.

EFFECT OF SOME PLANT GROWTH REGULATORS ON BIOCHEMICAL GENE EXPRESSION, GROWTH AND YIELD OF FABA BEAN

By Ibrahim1 , I.S.; A.B. Abdel-Razik and M. Ebeed Naglaa

Vol-4 No-(1-2)

Abstract

In the greenhouse experiment Vicia faba L. cv. Giza 716 were treated with 25 and 50 ppm of paclobutrazol (PP333), kinetin (Kin), naphthalene acetic acid (NAA) and 50 and 100 ppm of ethephon (Et) as foliar spray at the onset of flowering and fruiting. Plant height, branches and flowers number were recorded after 104 days from sowing. Two samples were taken after 90 days from planting and at harvest. At the 1 st sample date, biochemical analyses were conducted in leaves for determination of total soluble sugars, phenols, indoles, free amino acids, water soluble protein and isozymes. At harvesting time (2nd sample), number of pods, pods and seeds dry weights/plant were taken. Total seed proteins and soluble carbohydrates were evaluated in dry faba bean seeds. PP333 at 50 ppm had significant and negative effect on plant height, flowers number, total phenols and indoles. The high concentration of Kin, Et and NAA treatments increased the number of pods, the dry weight of pods and seeds per plant whereas the opposite occurred in the same concentration of PP333 treatment. Seed yield was correlated with the number of pods per plant and the dry weight of pods per plant. In general, NAA at 25 ppm has strongly stimulating effect on total soluble sugars, phenols, indoles and free amino acids (in leaves), soluble carbohydrates (in seeds) and seed index (100-seed weight). It seems that growth promoter NAA enhance the mobilization of photoassimilates to filling seeds. Protein banding patterns and isozymes of superoxide dismutase, peroxidase, catalase, α, β- esterases, acid phosphatase separation by electrophoresis were successfully used to faba bean subjected to PP333, Kin, Et and NAA. The applied plant growth regulators affected the rate of gene expression in leaves more than in seeds. Their effects on growth, fruit set, yield, correlated components, proteins SDS-PAGE and isozymes were discussed below.

MUTATION OF CEPHALOSPORIUM ACREMONIUM NRRL-2443 THROUGH UVIRRADIATION FOR CEPHALOSPORIN-C PRODUCTION

By Shumaila Butt, Sikander Ali* and Ikram-ul-Haq

Vol-4 No-(1-2)

Abstract

The present study is concerned with the selection of a mutant variant of Cephalosporium acremonium for the optimal production of cephalosporin-C. The parental strain C. acremonium NRRL-2443 was subjected to UV-radiations for 10-80 min. Among all the mutants tested, C. acremonium GCUV-6 gave maximum antibiotic i.e. 454 mg/ml, which was 3.91 fold improved compared to the wild (116 mg/ml). The cultural conditions and nutritional requirements for cephalosporin-C production by selected mutant were optimized by submerged culture technique. The incubation period (72 h), initial pH (6.5) and soybean meal (6.0 %, w/v) supported maximum production of cephalosporin-C (493 mg/ml).

USE OF INTER SIMPLE SEQUENCE REPEAT-POLYMERASE CHAIN REACTION (ISSR-PCR) TECHNIQUE FOR DETERMINATION OF THE EFFECT OF FAST NEUTRON IRRADIATION ON VICIA FABA SEEDS

By Hussein, T.S.1 and A. El-Shourbagy Gehan2

Vol-4 No-(1-2)

Abstract

The effect of low doses of fast neutrons on the DNA genome of broad bean (Vicia faba) mature seeds was determined. In this study, inter simple sequence repeat (ISSR)-polymerase chain reaction (PCR) tool was used to analyze the genetic alteration of treated seeds of V. faba using six ISSR primers. Results showed that each primer gave rich and clear patterns with bands ranged from 179 to 1256 bp. Among the 61 ISSR fragments, 21 were representing the effect of the treatment of V. faba seed samples. ISSR markers clearly and easily distinguish all the tested samples and also discriminate between fast neutron-treated and untreated seeds. The dissimilarity values ranged 42 to 47% between the fast neutron-treated seeds and the control seed (untreated) and from 27 to 38% between the fast neutron-treated seeds. The dendrogram resulting from a UPGMA cluster analysis comprised two main distinct clusters, the first one including the control V. faba seed (untreated) and the second, includes all samples from the same kind of treatment divided into three groups: the S4 (group A), S2 and S3 (group B) and S1 (group C), these groups being separated at a similarity level.

IN-VITRO MULTIPLICATION OF BANANA (MUSA SPP.) UNDER DIFFERENT NaCl STRESSES

By Ikram-ul-Haq*, Faheeda Soomro, M. U. Dahot, Shahrrukh and Um-e-Aiman

Vol-4 No-(1-2)

Abstract

Salinity is a major abiotic factor. Its effects were assessed on the micropropagation efficiency in Pakistani banana (Musa spp.) variety Basrai. Micropropagation rate was decreased with an increase in NaCl level. Due to it, numbers of plantlets per explant, pseudostem diameter was decreased. Bio-chemical contents were also affected due to salinity such as K+ was decreased while Na+ and Clincreased significantly. So salinity is reducing not only micro-propagation efficiency in banana as well as decreasing its yield.

REGENERATION AND AGROBACTERIUM-MEDIATED TRANSFORMATION OF WATERMELON

By Zakaria, H.M. 1, Hussein, Gihan M. 1, Sadik, A.S. 1,3 and Abdallah, Naglaa A. 1,2

Vol-4 No-(1-2)

Abstract

In this study, a rapid regeneration system for two Egyptian watermelon cultivars (Giza 1 and Giza 21) was developed using the proximal zone of the hypocotyls. An Agrobacterium-mediated transformation system was also established for both cultivars. Results showed that the highest regeneration frequency of both cultivars (Giza 1 and Giza 21) was detected on explants that cultured on medium 5MSBAABA, as it revealed a percentage of 75 (Giza 1) and 72 (Giza 21) with 4-6 shoots/explants. It was also clearly observed that carbon source has an effect on the shoot regeneration frequency. Data showed that the hypocotyls explant revealed a regeneration percentage of 96.6 instead of 73.3 % when the sucrose was only used in 5MSAABA medium. Also, when 1.5g/L phytagel and 3g/L agar were used in 5MSBAABA medium the vitrification phenomenon of the developed shoots was decreased. A percentage of ~80 % of the shoots formed roots on the medium contained 40 µg/L NAA. At the level of transformation experiment, results showed that a number of 20 (16.7%) explants out of 120 explants of both watermelon cultivars were survived on the bialaphos- selection medium, and produced 42 (for Giza 1) and 33 (for Giza 21) bialaphos-resistant shoots. The presence of gus-intron (2070 bp) and bar (540 bp) genes was confirmed via polymerase chain reaction while their expressions were detected by histochemical GUS assay and leaf painting with 1 mg/L Basta herbicide.

BATCH CULTURE THERMO TOLERANCE OF SACCHAROMYCES CEREVISIAE DURING INVERTASE PRODUCTION

By Sikander Ali*, Aafia Aslam and Ikram-ul-Haq

Vol-4 No-(1-2)

Abstract

Saccharomyces cerevisiae strains IIB-12 (wild) and NG-46 (mutant) were compared on the basis of kinetics and thermo tolerance for invertase production. NG-46 showed significantly higher (p≤0.05) values of Qp and Yp/s compared to other yeast strains. Thermodynamics revealed that cell system exerted protection against thermal inactivation during product formation. The enthalpy (∆H, KJ/mol) and entropy of activation (∆S, J/mol/K) for Invertase production substantially improved specificity constant activation energy, free energies for transition state formation and substrate binding for sucrose fermenting. The results highlighted an improvement in thermo stability (>40°C) of microbial endogenous metabolism for enzyme secretion.

PRODUCTION OF POLYCLONAL ANTIBODIES SPECIFIC TO CITRUS TRISTEZA VIRUS (CTV) USING THE 6X HIS-TAGGED FUSION COAT PROTEIN

By Salama, M.I.1 ; El-Domyati, F.M. 2 ; Salem, R.E.1 ; Nour El-Din, Hanan1 and Sadik, A.S.1,3

Vol-4 No-(1-2)

Abstract

Citrus tristeza virus (CTV) is one of the most destructive and economically important diseases of commercial citrus worldwide. In this study, the coat protein (cp) gene of CTV was subcloned into PQE-30 vector at Bam HI and Hind-III restriction sites. The constructed plasmid that called pRMAF2 was transformed into Escherichia coli M15 strain for its expression and production of fusion protein for CTV-cp gene. The suitable condition for inductions of CTV-cp was in the presence of 1 mM IPTG and overnight incubation time. The bacterial culture was harvested and the expressed fusion protein(s) was purified by QIA express kit. Polyclonal antibodies (PAbs) were raised by injection of the purified fusion protein(s) into two white mice followed by western blotting analysis using the expressed fusion protein(s) and the CTV-infected citrus extracts as controls. It can be concluded that the immuno-enzymatic detection based on the produced antibodies against CTV-cpORF fusion proteins were safe, speed and sensitive for the detection of the virus.

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, 2007 to December 31, 2007 for Vol. 4 No. 1 and 2, 2007.
Prof. Dr. Alexandre SemenovMoscow, Russia
Prof. Dr. Atef S. SadikTaif, Saudi Arabia
Prof. Dr. Jian He XuShanghai, China
Prof. Dr. Muhammed Umar Dahot Uni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Aijaz Rasool SheikhKarachi, Pakistan
Dr. M. MotallebiTehran, Iran
Prof. Dr. M.R. ZamaniTehran, I. R. Iran
Prof. Dr. Atta-u-RehmanUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. A.A. SabouryTehran, I.R. Iran
Prof. Dr. Iftikhar Ahmed KhanFaisalabad, Pakistan
Dr. Muhammed Ibrahim RajokaFaisalabad, Pakistan