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EDITORIAL BOARD

2004

EFFECTS OF DIFFERENT PHYTOHORMONES ON SUGARCANE (SACCHARUM SPP.) REGENERATION

By Imtiaz Ahmed Khan+ , Abdullah. Khatri, Saboohi Raza, Ghulam Shah Nizamani,Muhammad Aquil Siddiqui, Nighat Seema, *Muhammad Umar Dahot and Mazhar H. Naqvi

Vol-1 No-2

Abstract

Three sugarcane clones, AEC82-223, Ghulabi-95 and AEC86-328 were investigated for callus induction and regeneration under different concentrations (2mg/L and 4mg/L) of various auxins i.e. 2,4-dichlorophenoxyacetic acid (2,4-D), indol acetic acid (IAA), 3,6- dichloro 2-methoxy benzoic acid (Dicamba) and picloram. An efficient callus induction was observed on the medium containing picloram followed by 2,4-D but high callus proliferation was achieved with picloram. Non-regenerable callus was produced when calli were transferred for proliferation on Dicamba. The maximum calli induction, its proliferation and plantlets regeneration were recorded in clone AEC82-223, while the minimum in AEC86- 328. The maximum chlorophyll mutation frequency was noted in clone AEC86-328 and minimum in AEC82-223. Among auxin especially 2,4-D induces more genetic variability as compared to other auxins used in this work.

DOUBLED-HAPLOID PRODUCTION TECHNOLOGY IN WHEAT (TRITICUM AESTIVUM L.): ANTHER CULTURE

By M. A. SIAL, MAZHAR H. NAQVI, M. U. DAHOT*, M. A. ARAIN, S. M. MANGRIO** AND A. M. SOOMRO

Vol-1 No-2

Abstract

To achieve homozygosity within the shortest possible period, doubled-haploid technology is a valuable technique to reduce the time and cost of development of new wheat varieties. It involves in vitro development of fixed lines from the parental material; development of each generation of progeny can be initiated before the parents have achieved physiological maturity. The exploitation of recessive genes can be rapidly achieved by microspore culture and anther culture via induction of sporophytic pathway from a gametophytic pollen pathway, resulting in callus or embryoids formation. Anther culture studies were conducted to produce haploid plants in five bread wheat genotypes BW-2, BW-5, BW-8, BW-9, and BW-16. Two media liquid and agar-solidified with different compositions were used to determine their effects on the induction of calli and regeneration of green plants. The potential of callus induction was significantly higher (82.4) in liquid culture medium as compared to agar-solidified medium (55.7) worked out on 100 anther basis. In liquid media, calli induction ranged from 37.8% to 117%, whereas, calli induction ranged from 39.1 to 73.6% on agar-solidified medium. The callus induction and plant regeneration were varied among genotypes on both the media. Genotype BW-8 was more conducive for callus induction to agar medium as compared to liquid medium, however, BW-9 showed less callus differentiation ability on both the media.

DIVERSITY AND SEASONAL OCCURRENCE OF PLANKTONIC ROTIFERS IN KEENJHAR LAKE DISTRICT THATTA SINDH PAKISTAN

By G.A. SAHATO AND K.H. LASHARI

Vol-1 No-2

Abstract

Qualitative and quantitative Zooplankton sampling was carried out every month from November 1998 to October 1999 at three stations in Keenjhar Lake. A total of fifteen rotifer species were identified. Four species belonging to genus Keratella were K. cochlearis, K. volga, K. cochlearis var. tecta, Keratella tropica and four species belonging to genus Brachionus were B. falcatus, B.buidapestinensas, B.quadridentatus and Brubens. The other were Platyias quadriconus, Monostyla sp, Mytilina sp, Lecane sp, Tetramarti opotiensis, Euchlanis sp and Macrochaetis sp, Keratella and Brachionus were present through out the year.

A. W. BALOCH, A.A. RIND AND K. D. JAMALI

By A. W. BALOCH, A.A. RIND AND K. D. JAMALI

Vol-1 No-2

Abstract

Rice is an important cereal crop of Asian countries. It is a major staple food crop Southeast and eastern countries of Asian sub-continent. Knowledge on the genetic constitution of rice plant has immense importance in Plant breeding Programmes. The use of molecular markers has facilitated the selection process. Markers-assisted selection has provided a reliable source for identifying and selecting the desirable genotype in plant breeding programmes. Molecular marker can save time and labour. It is a laborious job to grow a large number of F2 populations and practice selection for morphological markers in conventional plant breeding. Molecular markers will be more useful for selection when (1) the phenotype is difficult or expensive to measure directly, (2) genes of similar phenotype are being pyramided into a single line, or (3) markers are being used to select against the donor genome in a backcrossing programme.

STUDIES ON IN VITRO SURFACE STERILIZATION AND ANTIOXIDANTS ON GUAVA SHOOT TIPS AND NODAL EXPLANTS

By Roshan Zamir+, Syed Tariq Shah, *Nawab Ali, G.S.S. Khattak and T. Muhammad

Vol-1 No-2

Abstract

In vitro studies were carried out to investigate the effect of different surface sterilization agents and antioxidants on the guava (Psidium guajava L.) Cv. Safeda shoots tips and nodal explants. The best sterilant observed for shoot tips was mercuric chloride (HgCl2) at 0.05% for 5 minutes plus 70% ethanol which gave maximum survival percentage of (67%) while minimum were survival (22%) was observed with 2% Sodium hypochlorite for 10 minutes. In case of nodal explants the best sterilant was 4% Calcium Hypochlorite (CaOCl2) for 10 minutes which gave 25% survival. Blackening of media was reduced and gave 55% green shoot tips when 75:50mg/l citric acid and ascorbic acid was supplemented to the medium. While 75:50 mg/l citric acid and ascorbic acid, 100mg/l Polyvinyl pyrolidone (PVP) and 200mg/l Activated charcoal when added to the medium gave 13% green nodal explants.

ISOLATION, CLONING AND PARTIAL CHARACTERIZATION OF THE GENE ENCODING THE POLYGALACTURONASE INHIBITING PROTEIN OF PHASEOLUS VULGARIS CV. NAZ

By A. HOSSAINZADEH COLAGAR*, M. MOTALLEBI and M.R. ZAMANI+

Vol-1 No-2

Abstract

In this research, inhibitory effect of bean (Phaseolus vulgaris cv. Naz and cv. Derakhshan) hypocotyl PGIP on the PG enzymes from highly virulent isolates of Fusarium oxysporum (F15) and Ascochyta rabiei (IK04) was analyzed. It was shown that PGIPs from Naz and Derakhshan cultivars inhibit more effectively the PG activity of F. oxysporum (F15) than PG activity of A. rabiei (IK04). The inhibitory activity of Naz and Derakhshan PGIP on PG activity of these fungi demonstrated that Derakhshan PGIP was able to inhibit the PG activity of F. oxysporum by 55% and A. rabiei by 40%. Naz cultivar PGIP had reduced ability to inhibit PG activity from F. oxyspoum and A. rabiei by 40 and 26%, respectively. The pgip gene was amplified from Naz cultivar genomic DNA, cloned and sequenced. Comparison of DNA sequence and deduced amino acid from Naz cultivar with those of Derakhshan and Saxa cultivars indicates that it differs only in a single nucleotide (G) in position 880 which alters amino acid T to A at position 294. This amino acid is located in LRR9 region of PGIP. Also, the cloned pgip was shown to encode a 37 kDa polypeptide corresponding to the deduced polypeptide molecular weight. The data from this study suggestes possible role for this amino acid change in LRR9 region in specificity of PGIP-PG interaction.

In-Silico ANALYSIS OF THE GENE AND PROTEIN SEQUENCES OF THE ENZYME MUSHROOM TYROSINASE (POLYPHENOL OXIDASE, PPO)

By Mahmud Tareq Hassan Khan*, Atta-ur Rahman and M. Iqbal Choudhary

Vol-1 No-1

Abstract

Tyrosinase (E.C. 1.14.18.1) is a multifunctional copper-containing enzyme which catalyses the biosynthesis of melanin (a huge polyphenolic biomacromolecule) in human, plants and animals. The irregularity in the expressions of this enzyme causes severe clinical problems in humann beings like hyperpigmentation and depigmentation, as well as vitiligo and albinism-type severe dermatological problems. In addition, tyrosinase is known to be involved in the molting process of insect and adhesion of marine organisms. This enzyme and its receptors (TRP1 and TRP2) are involved in the skin melanoma. The gene and protein sequences of this enzyme and their primary, secondary, tertiary and quaternary structures have been analyzed through several bioinformatic approaches.

MOLECULAR MARKERS IN WHEAT

By Karim Dino Jamali, A.M. Soomro and Syed Ashraf Ali

Vol-1 No-1

Abstract

The progress made in DNA markers technology has been tremendous and exciting. DNA markers have provided valuable tools in various analysis ranging from polygenetic analysis to the positional cloning of genes. The development of high density molecular maps which has been facilitated by the PCR-based markers have made the mapping of almost any trait possible. The size and structure of the wheat genome makes it one of the most complex crop species for genetic analysis. The development of molecular technique for genetic analysis, in particular the use of molecular markers to monitor DNA sequence variation between varieties, and races, and wild relatives of wheat and related grass species, has led to a dramatic expansion in our understanding the structure and behavior of wheat genome.

UP TAKE OF GLYCOLIPIDS BY MYCOPLASMA CAPRICOLUM MEBRANES

By A. Sattar Khan

Vol-1 No-1

Abstract

Mycoplasma capricolum was grown with and without DGDG containing cholesterol on Edwards modified media. Total lipids were extracted with chloroform: methanol (2:1). Membrane protein and phospholipids were determined by Lowery and Chen methods respectively. Total lipids were separated on TLC and glycolipids were located with iodine while diphenylamine and orcinol test was performed for lipid sugar. Three positive sugar spots for microorganisms grown on DGDG and cholesterol were found. Either DGDG is being metabolized to new components or new glycolipids are being synthesized from DGDG.

PRODUCTION OF AMYLASE BY FUNGI THROUGH SUBMERGED FERMENTATION

By A. Sattar Qureshi, M. Umar Dahot and A.U.Rehman*

Vol-1 No-1

Abstract

Amylase has versatile application in food and industries, which inspired to produce this enzyme by less expensive culture medium using agricultural or forest waste. In the initial stage, amylase production was carried out by different fungal species such as Mucor geophillus, Aspergillus niger, Aspergillus fumigatus, Penicillium lilacinum and mixed culture (A. niger + A. fumigatus) using mineral medium with and without glucose. Highest amount of amylase (34.4 & 1.36 units/ml both) was produced by A. fumigatus when grown at 31 + 2 °C with in 48 hours when mineral medium was supplemented with and without glucose respectively in comparison to other fungal species.

BIOSYNTHESIS OF -GLOCOSIDASE PENICILLIUM EXPANSUM

By Khalid Bhatti, M. Hanif Noomrio, *M. Umar Dahot, K.D. Ujjan and M. Uris Siyal

Vol-1 No-1

Abstract

Sugarcane bagasse and leaves were utilized after treatment with 0.6N NaOH and KOH as a substrate for the growth of Penicillium expansum and the production of -glucosidase. It was observed from the results that maximum yield (4750 units/ml) of -glucosidase was achieved at 240 hrs when 0.6N NaOH treated sugarcane bagasse was used as carbon source.

KINETRIC ANALYSIS F NUTRITIONAL STRATEGIES FOR NVERTASE PRODUCTION BY SACCHAROMYCES CEREVISIAE KR18

By Kiran Shafiq, Sikander Ali and Ikram-ul-Haq

Vol-1 No-1

Abstract

Fermentation conditions for invertase secretion by Saccharomyces cerevisiae strain were optimized. Submerged fermentation technique was employed for present investigation. 6.76±0.5 U of invertase ml -1 of fermented broth were obtained by Saccharomyces cerevisiae KR18 when fermentation medium was supplemented by 15.0 mg sucrose ml-1 and incubated at 25oC at initial pH 6.0. Sugar consumption and cell mass production (mg ml-1 ) at this pH level were 11.21±0.5 and 4.67±0.5, respectively. One ml of vegetative inoculum (1.2 × 103 cells) developed for 12 h was used. Kinetic analysis of fermentation results was made to check out the feasibility of fermentation process. All the kinetic parameters i.e. product (Yp/x, Yp/s) and growth rate (Yx/s) coefficients as well as specific rates (µ h-1 ) were favorable for optimized conditions.

SELECTION OF ACID AND BASE TREATMENT OF SUGARCANE WASTE AS CARBOON SOURCE FOR THE PRODUCTION OF AMYLASE BY PENICILLIUM EXPANSUM

By K.D.Ujjan, M. Hanif Noomrio, *M. Umar Dahot and M. Uris Siyal

Vol-1 No-1

Abstract

Sugarcane wastes were (bagasse and Leaves) pretreated with 0.6N acid and base and the hydrolysates were utilized as carbon source for the growth of Penicillium expansum and the production of amylase. It was found that maximum yield of amylase was achieved (2.6208 units/ml) at 168 hours when Penicillium expansum was grown on pretreated KOH sugarcane leaves mineral medium.

MASS PRODUCTION OF BANANA (MUSA SP.) THROUGH BIOTECHNOLOGICAL TECHNIQUES

By A. Khatri, I.A. Khan, G.S. Nizamani, M.A. Siddiqui, M.H. Khanzada, N.A.Dahar, N. Seema and M.H. Naqvi

Vol-1 No-1

Abstract

Meristematic tip with leaf primordial from 13 clones of desert banana (Musa spp.) were evaluated for in-vitro propagation techniques accelerates the production of disease-free banana (Musa spp.) clones through shoot tips. It was observed that apices cultured on solid MS media supplemented with 20μM BAP/1produced 3-8 shoots. Longitudinal splitting of shoots induced shoot clusters. Shoot elongation and rooting were obtained on media containing 1μM IBA/1 and 40gm/1 sugar (commercial). The regenerated plants were transferred to jiffy pots and kept in growth chamber for two weeks for acclimatization. Plants were then transplanted in field.

EFFECT OF GAMMA RAYS ON THE ROOTING OF GUAVA SHOOT TIPS

By R. Zamir, G. S. S. Khattak, T. Mohammad, S. A. Shah, I.Ali and N. Ali.

Vol-1 No-1

Abstract

In-vitro mutagenesis followed by micro propagation via axillary bud proliferation in shoot tips of guava (Psidium guajava L.) cultivar Safeda was carried out. Shoot tips were irradiated with 15 to 90 Gy gamma rays using 60Co cell source and cultured on MS medium containing 3.0% sucrose, 6-benzyleamino purine (BAP), and Lglutamine. The Shoot proliferation was observed after 7 weeks of culturing. Best shoot proliferation was recorded on MS medium supplemented with 1.0 mg /l (BAP) and 250 mg/l L-glutamine. Rooting of the cultured shoots were observed on half strength MS medium supplemented with Indole-3-acetic acid (IAA) and Indole-3-butyric acid (IBA). Radio sensitivity was assessed by determining the percentage shoot tips survival and shoot proliferation. The LD50 (50% of the population kill does) was observed on 45 Gy. The doses above 75 Gy were found lethal to explants

IN-VITRO CULTURE STUDIES IN SUGARCANE

By I.A.Khan, A. Khatri, G.S.Nizamani, M.A. Siddiqui, M.H. Khanzada, N.A. Dahar, N. Seema and M.H. Naqvi

Vol-1 No-1

Abstract

Eleven sugarcane clones were compared for callusing and regeneration potential. All these clones showed varied response to the traits under study. However, the highest callus formation and plantlets regeneration were recorded in clone NIA-98 while the lowest in CP67-412 followed by SPSG-26. The maximum chlorophyll mutation frequency was noted in clone AEC82-1026 and minimum in AEC81-0819.

In vitro MUTAGEGENESIS IN OIL SEED BRASSICA

By Iftikhar Ali and Mumtaz Ahmad

Vol-1 No-1

Abstract

Two Concentrations of EMS mutagens were used to induce the genetic variations in microspore culture of two genotypes of oilseed rape (Brassica napus). Embryos derived from mutated microspores were placed in liquid medium NLN. Within a few days only surviving embryos were transferred to solid medium B5 for further development. The effects of various core of EMS concentrations on the production and survival of embryos and plants in both genotypes has been discussed and it was concluded that the lower concentration had the desirable mutagenic effects for a practical mutation breeding programme of genetic improvement of oilseed brassicas.

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, 2004 to December 31, 2004 for Vol. 1 No. 1 and 2, 2004.
Prof. Dr. Richard M. Cooper Bath, U.K.
Prof. Dr. Asif AliFaisalabad, Pakistan
Prof. Dr. J.J.Zhong Shanghai, China
Prof. Dr. Jian He XuShanghai, China
Prof. Dr. Muhammed Umar Dahot Uni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Rafi ArainUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Atta-u-RehmanUni. Of Sindh, Jamshoro, Pakistan
Prof. Dr. Rehmatullah Choudhary Uni. Of Sindh, Jamshoro, Pakistan
Dr. Tayyab Hussain Lahore, Pakistan