Vol. 19, February, 2005.

1/19 EFFECTS OF INFESTATION BY LIRIOMYZA TRIFOLII BURGESS (DIPTERA: AGROMYZIDAE) ON PHYSIOLOGY OF PHASEOLUS VULGARIS PLANTS, WITH SURVEYING THE COMMON BIOLOGICAL CONTROL AGENTS

M.A.M. El-Khawas and S.A.M. El –Khawas*

Plant Protection Research Institute, A.R.C., Dokki, Giza, Egypt.

* Faculty of Science, Ain Shams University, Cairo, Egypt.

       Field studies were carried out to determine the population density of the leafminer Liriomyza trifolii, attacking Phaseolus vulgaris plantsand its common parasitoid species. In addition, the survey included sap sucking insect pests and common predatory species found associated with major insect pests on P. vulgaris plants, during Nili and Summer plantations of the two successive seasons, 2002/2003 and 2003/2004, at Qalubia Governorate. Obtained data revealed that, Nili plantation was more liable to attack by L. trifolii than Summer one. The two parasitoid species Diglyphus isaea and D. crassinervis, were the most common parasitoids parasitizing L. trifoli. among other recorded parasitoids (Neochrysocharis sp., Cirrospilus sp., Hemiptarsenus sp., Chrysocharis sp. and     Halticoptera sp.). As for sap sucking pests, aphids and Empoasca spp. populations in Summer plantation were higher than those of Nili one. While, the population of the whitefly Bemisia tabaci was higher in Nili plantation than in Summer one. The individuals of Thrips tabaci were only recorded in Summer plantation. True spiders were the most common predatory species observed between all other recorded predators on P. vulgaris plants, during both Nili and Summer plantations. Moreover, physiological studies were made to evaluate the effects of feeding by L. trifolii larvae on plant leaflets during early and late infestations by the pest, comparing with non-infested leaflets as control, including their effects on chlorophyll a, chlorophyll b, carotenoids, cytokinins, total soluble sugars, total phenols, percentage of nitrogen, total protein, protein profile and free amino acids. The obtained results indicate that, late infestation by L. trifolii showed significant decrease in most of the previous studied parameters. The most valuable remark is the decrease in amino acids serine and cysteine and the increase of tyrosine and phenylalanine. Furthermore, specific protein bands 48 kDa and 33 kDa were appeared in response to L. trifolii infestation. Finally, it is concluded that, feeding by L. trifolii subjected P. vulgaris   plant to stress. Also, for biological control purpose, the two parasitoids Diglyphus isaea and D. crassinervis   can be laboratory mass reared and released in P. vulgaris fields for controlling L. trifolii. While, the predator Scymnus sp. can be used against aphids with other available control methods. The natural role of biocontrol agents must be encouraged as representing one of the main concepts of recent developed Integrated Pest Management (I.P.M.) programs.

 

 

2/19 THE APPLICATION OF PCR-RFLP-DNA AND PROTEIN PROFILES FOR THE DIFFERENTIATION OF SYNECHOCOCCUS LEOPOLIENSIS AND ITS MUTANTS

N.H. Noaman, M.A. Osman*, A.M. Khaleafa and S.H. Zaki

Botany Department, Faculty of Science, Alexandria University, Alexandria, Egypt.

* Genetic Engineering and Biotechnology Institute, Minoufiya University.

       Using ultraviolet radiation attained four mutants of Synechococeus leopoliensis. The mutants cultures gave no antibiotic activity, which was proved by the parent type. Amplified DNA bands and protein profile of the wild type and its mutants were evaluated. The total number of the attained DNA bands was 21 bands. The number of bands of the parent strain was 5, while this number increased in case of all mutants. Some bands were found to be common between the parent strain and the mutants while, other bands were common between two or three of them. Protein profile of S. leopoliensis and its mutants showed the presence of 35 bands, with different molecular weights. Most of them (24 bands) were found to be of low molecular weight (<50KDa), while the remaining eleven bands had high molecular weight (>50KDa). Mutant number 2 was found to have the highest banding pattern (21 bands), while the lowest number of bands was obtained with mutant number 1. Four bands were found to be common between the parent strain and its mutants. Characteristic bands were also observed for both of the parent strain and each of its studied mutants. The comparison of the obtained similarity indices values between the parent strain and its mutants showed the highest value (40.47%) with the third mutant and the lowest value (31.37%) with the fourth mutant.

 

 

3/19 EFFECT OF GAMMA-RAYS AND MICROWAVE-HEAT TREATMENTS ON THE DETOXIFICATION OF AFLATOXIN B1 IN GROUND BEEF SUPPLEMENTED WITH SOME ANTIMICROBIAL FOOD ADDITIVES.

N.H. Aziz and S.R. Mahrous.

Microbiology Department and Natural Products Department, National Center for Radiation Research and Technology, Nasr City, Cairo, Egypt.

       The influence of gamma-irradiation and/or heat treatments on the detoxification of aflatoxin B1 produced by Aspergillus parasiticus in ground beef supplemented with some antimicrobial food preservatives was investigated. No growth of A.parasiticus and aflatoxin B1 production occurred at 5.0kGy gamma-irradiation dose level. Gamma-irradiation (2.0 kGy), NaCl (4%), potassium sorbate (300 ppm) and sodium benzoate (500 ppm) completely inhibited the growth of A. parasiticus and aflatoxin B1 in ground beef. Microwave heating for 5 min of 2.0 kGy gamma-irradiated ground beef supplemented with NaCl (2%), potassium sorbate (100 ppm) and sodium benzoate (300 ppm), detoxified aflatoxin B1 by 100%. It was concluded that the combined treatment of gamma-rays, heat, and food preservatives at lower levels controlled mould growth and aflatoxin production in ground beef.

 

 

4/19 ISOLATION AND MAPPING OF CHITINASE-ENCODING GENE FROM A LOCAL- ISOLATED CHITINOLYTIC BACTERIUM, SERRATIA SP.

A. M. Ibrahim

Gen. Eng. and Biotech. Res. Inst. (GEBRI),

Microb. Biotechnol. Depart., Minoufiya Univ., Sadat City, Egypt

         A local chitin-degrading bacterium, Serratia sp. was isolated from the Egyptian soils. The isolate is a Gram negative, facultative anaerobe, motile and rod shaped. The chitinase protein was purified from the culture filtrate of Serratia using chitin affinity chromatography, and applied on 10% SDS-PAGE; a protein band was appeared at about 60 kDa. A chitinase gene of Serratia sp. was obtained by PCR technique, and mapped using several restriction enzymes. The obtained PCR fragment is about 900bp, an open reading frame coded for protein of about 294 amino acids, with a predicted molecular mass of 32 KDa.

 

 

5/19 EFFECT OF DROUGHT STRESS ON THE VARIATION AND CORRELATION OF VEGETATIVE, CHEMICAL AND YIELD ATTRIBUTES IN DURUM WHEAT.

H.M. Emad El-Din, E.A. Ismail* and M.H. Rashad**

Plant Dept., Fac. Of Science, Cairo University, Giza

* Agronomy Dept., Fac. Of Agric., Cairo University, Giza

** Plant Physiology Section, Botany Dept., Fac. Of Agric., Cairo University, Giza

       Determining the relation between yield components and other morphological and chemical traits under drought stress is important in efficient wheat breeding programs in semi-arid areas. Therefore, twenty two introduced durum wheat genotypes along with an Egyptian check (Beni-suif 1) were evaluated under two contrasting water regimes (non-stressed vs. stressed) in two successive seasons; 1999/2000 and 2000/2001. The experiments were laid out in a split plot design. Vegetation and chemical components were determined in two samples at mid and late season during the first year. Data of yield components were combined over the two years. Phenotypic and genotypic correlations among yield components were estimated from combined data over the two seasons for each water regime separately. Similar correlations were calculated between yield components, vegetative and chemical traits during the first season. Drought stress resulted in a significant reduction in yield components and vegetative attributes. Phenotypic (rp) and genotypic (rg) correlations showed that both spike weight and grain weight per spike were the most related to grain yield per plant under both water regimes. Under non-stressed conditions, grain yield per plant was positively correlated with plant height (rp=0.5 and rg=0.6). Under stress conditions, a different relation was found were grain yield was negatively correlated with both shoot and root dry weight (rp=-0.5 and - 0.6, rg= -0.5 and -0.7, respectively). Variability and accumulation of proline among genotypes increased under drought stress in the mid-season sample. Contrarily, variability and concentrations of chlorophyll b were decreased. Simple correlation coefficients over the two water regimes were highly positive and significant between yield components and chemical traits, particularly nitrogen, phosphorus and potassium (NPK) concentration in the shoot. Results generally revealed the importance and the direct effect of some vegetative, chemical and yield attributes on grain yield of wheat under drought stress.

 

 

6/19 RESPONSES OF IPOMOEA CARNEA TO SOIL SALINITY

E.A. El-Meleigy, M.N. Shehata*, H.Y. Olama* and A.H. Mohammad*

Faculty of Sciences, Botany Department, Suez Canal University,

*Faculty of Sciences, Botany Department, Banha University

       Ipomoea carnea inhabits various habitats ranging from hydric to xeric habitat. Two different habitats namely: canal bank habitat, at Benha and saline habitat at Kafr El-Shiekh were chosen. Soil texture, calcium carbonate content, cations;Na+, K+, Ca2+, Cu2+, Fe,3+ Mg2+, P3+ and zinc were determined. The pH of the soil was found to be 7.35 and 8.00 in the canal bank and saline habitat respectively. The electrical conductivity of samples collected from canal banks habitat was less than 1m mos/cm and more than 1m mos/cm in the latter habitat indicating that there was no development for alkalinity or salinity in the soil supporting Ipomoea carnea. Responses of Ipomoea carnea to saline soil stress was achieved by studying the carbohydrate fractions, nitrogen components, and proteins in leaves and stems as well as photosynthitic pigments of leaves of plants of the two habitats. From the results it is clear that Ipomoea carnea grown in saline habitat is a good salt accumulator from saline soil.

 

 

7/19 MORPHOGENIC AND ULTRASTRUCTURE ABERRATIONS INDUCED BY CYTOCHALASIN B IN SOME MICROORGANISMS

S.M. Ezzat, M.M. Sarhan and A.A. Ismaiel

Botany Department, Faculty of Science, Zagazig University, Zagazig, Egypt.

       Cytochalasin B (CB) had morphogenic effects on cells of Spheronostoc microscopicium decreasing the diameter of both developed hormogonia and heterocysts significantly. Moreover, CB caused a remarkable reduction in the chlorophyll and cartenoid contents of the alga. CB at the concentration of 200 µg /ml caused swelling of the developed conidiophores and conidia of Helminthosporium solani. In addition, this compound induced an irregular increase in the septa of conidiophores with a profuse branching of conidiophores, formed with small vesicles inside conidia and conidiophores accompanied by increase in width of conidia. The response of treated cells of Saccharomyces cerevisiae to CB was detected using light and electron microscopic examination. The diameter of both yeast and bud cells were reduced and the cell wall became thicker than the untreated cells. Furthermore, an irregularity in shape of the treated cells compared with control cells was revealed. Electron - dense round bodies were seen and interpreted as vacuoles. These bodies were absent in the cells treated with CB. However, large number of small sized blisters were clearly seen in the cells treated with CB. These blisters were absent in the control cells. The yeast cell ultrastructure confirmed the hypothesis of integration of cytochalasin particles with the cell membranes and interfering with its function causing disruption of endocytosis and cell deregulation.

 

8/19 sEQUENCE VARIATION OF THE rDNA-ITS REGION WITHIN AND AMONG SOME ANASTOMOSIS GROUPS OF RHIzOCTONIA SOLANI

M.H.M. Abdel-Rahman, S.M.M. Mahmoud*, S.A. Al-Wakeel** andN.M. Gomaa**

Botany Department, Faculty of Science, Cairo University, El Fayoum, Egypt.

*Plant Pathology Institute, Agriculture Research Center, Giza, Egypt.

**Botany Department, Faculty of Science, Cairo University, Giza, Egypt.

       Rhizoctonia solani is the most important pathogen affecting cotton seedling disease severity. The anastomosis-based system of grouping isolates of R. solani, though occasionally imprecise, is beneficial because it enables better understanding of this large and highly heterogeneous species. This work indicated that rDNA-ITS sequencing technique offers an objective supplemental evaluation when used along with hyphal anastomosis reaction, should eliminate erroneous characterization of isolates. ITS-rDNA sequence analyses was not only a useful tool to differentiate among and between different anastomosis groups of R. solani, but also it was useful to differentiate between R. solani from different geographic origin.

 

9/19 SEPARATION OF ERGOLINE ALKALOIDS FROM IPOMOEA VIOLACEA L. (CONVOLVULACEAE) BY CAPILLARY ELECTROPHORESIS

Y. Hussein

Botany Department, Faculty of Science, Zagazig University, Egypt.

       Ergoline alkaloids (lysergic acid amide (LSA) and chanoclavine-I) were detected by thin layer chromatography (TLC) and capillary electrophoresis (CE) in Ipomoea violacea (morning glory) cv. heavenly blue grown at three different growth stages. Ergometrine has not been detected in Ipomoea violacea plant during all growth stages. Ergoline alkaloids (LSA and chanoclavine-I) content of Ipomoea violacea increased sharply from late vegetative stage to fruiting stage and exhibits the highest amount at fruiting stage. Chanoclavine-I and LSA have not been detected in both leaf and stem at fruiting stage, while the fresh fruits accumulate a high percentage of ergoline alkaloids. The content of ergoline alkaloids in Ipomoea violacea stems was found to be higher than in the leaves. Fruits and seeds contain the highest concentration of ergoline alkaloids.

 

 

10/19 RELATIONSHIP BETWEEN LEAF SENESCENCE AND FRUIT DEVELOPMENT AND MATURITY IN FICUS CARICA L.

E.A. El-Meleigy, M.N. Shehata*, H.Y. Olama* and A.N. Mostafa*

Botany Department, Faculty of Science, Suez Canal University

*Botany Department, Faculty of Science, Zagazig University

       During leaf senescence of fig plant, numerous metabolic changes take place. This study investigated the relation between changes in growth regulators, photosynthetic pigments, total carbohydrates, total N and mineral ion   contents accompanying leaf senescence of fig and those occurred in fruit development and maturity. It could be concluded that natural senescence of fig leaves was associated by decrease in growth activators, an increase in ABA, chlorophyll degradation and mobilization of carbohydrates and other nutrients from leaves to other parts of the tree mainly the fruits.

 

 
11/19 ETHANOL PRODUCTION FROM PRETREATED CELLULOSIC WASTES USING DIFFERENT TECHNIQUES.
N.A.Younis 1; N.M. Sidkey 2; A.H. Rady 3 and S.M. Ouda 4

Plant Research Dep,. Nuclear Research Center, Atomic Energy Authority. 1, 3& 4

2 Bot. & Microbiol. Dep., Fac. Of Sci. Al-Azhar Univ., Cairo.
       Saccharomyces cerevisiae, Y10 produced the highest ethanol productivity comparing with other tested S. cerevisiae   strains allowed to grow on peptone yeast   nitrogen (PYN) medium, so it is selected for fermentation process in present study. It is noticed that ethanol production not only   depend on type of fermentation technique but also on the kind of the chemical treatment as well as the cellulosic substrate under investigation. The highest ethanol productivity was recorded from NH4, -pretreated bagasse, HCl-pretreated cellulosic hospital wastes (CHW), while rice straw pretreated with H2O2 recorded reasonable production under Simaltaneous saccharification fermentation (SSF) process which was found to be the preferable tested technique. Hemicellulose resulted from physical & chemical pretreated cellulosic substrates was more easily to be broken than cellulose & lignin, where percentages of change were: 42.4 , 7.41 and 100 % for rice straw, bagasse and CHW, respectively.   Lignin and cellulose contents of treated bagasse and rice straw did not change, while the change resulted in both for treated CHW were 16.7 and 7.2 %, respectively. The ethanol productivity was the maximum at substrate concentration of 4, 8 & 5% (w/v) for rice straw, bagasse and CHW respectively. The maximum ethanol productivity was represented at ratio 1:4 for both (rice straw : H2O2,& bagasse : NH4) and 1:2 for (CHW: HCl). Pretreated cellulosic substrates recorded maximum production at 8 N H2O2, 0.5N NH4 for rice straw &   bagasse respectively while 4 & 6N HCl for CHW. Filtrate 40 % overlime treatment induced the highest ethanol productivity resulted from chemically pretreated rice straw, and   tween 60 at 0.5 % (w/v) induced the highest ethanol productivity resulted from both pretreated bagasse and CHW, percentage of yield increase were 35.7, 9.16 & 25.2 respectively.

 

 

12/19 BIOCONTROL ACTIVITY AND MOLECULAR CHARACTERIZATION OF COLCHICINE-INDUCED AUTOPLOIDY YEAST STRAINS; TILLETIOPSIS PALLESCENS ON POWDERY MILDEW AND RHODOTORULA GLUTINIS ON GREY MOULD IN GREENHOUSE SWEET PEPPER

W.M. Haggag; H.A.A. Mohamed*and M.A. Saker**

Plant Pathology Department, *Microbial Genetics Department, **Plant Biotechnology Department, National Research Center, Dokki, Cairo, Egypt.

       This paper aimed to induce autoploidy strains from the two yeast strains; Tilletiopsis pallescens gokhale and Rhodotorula glutinis Harrison by colchicine treatments and to study the effect of their autoploidy state on the biological control efficiency against Leveillula taurica and Botrytis cinerea, the causal organisms of powdery mildew and grey mould diseases on pepper plants, respectively. Yeast isolates of T. pallescens gokhale is among the more promising biocontrol agents for controlling powdery mildew disease caused by L. taurica on pepper plants. Also, R. glutinis showed promising inhibition effect on grey mould disease caused by B. cinerea. Several types of autoploidy from T. pallescens and R. glutinis isolates have been generated by using 0.1 and 0.2% colchicine treatments and showed variability in reducing the disease severity of powdery mildew and grey mould on plant leaves. Six autoploidy isolates that showed superiority in their growth rates and antagonistic effects against the tested pathogens than their parental strains were selected. The selected isolates and their wild type parents were evaluated for some of biological, morphological, biochemical and molecular criteria.Under the same conditions, cells of autopolyploid strains either from T. pallescens or R. glutinis displayed a large size in comparison to that of their parents. Transmission electron microscope (TEM) revealed that the autoploidy cells increased in diameter, size and density. The ratio of the cellular DNA content of autoploidy strains were increased in amount comparing with their wild types. The selected autoploidy strains showed a significantly higher level of protease, β-1, 3-glucanase and chitinase activity in comparison with their parental strains. DNA fingerprinting of autoploidy yeast strains using randomly amplified polymorphic DNA (RAPD) indicated that some of them have unique DNA banding patterns when compared with standard DNA fingerprints of the wild type. The genetic variations detected at the DNA level between the autoploidy strains, as revealed by the variations in RAPD banding patterns, including the amplification of new DNA bands and disappear of others. These variations were verified using a set of different random primers. Under artificial and natural infection conditions in greenhouses, application of autopolyploid yeasts as foliar sprays, significantly reduced the disease severity and pathogens sporulation on leaves, stems and fruits to minimum level compared with foliar sprays with the parental strains. Furthermore, population counts of autoploidy strains were dramatically increased on plant leaves and improved the plant yield comparing with that of the wild type. These findings led to the conclusion that induction of autoploidy in yeast cells by colchicine is useful for increasing the biocontrol activities.

 

 

13/19 CHARACTERIZATION AND ACTION OF ANTIPHYTOVIRAL AGENT PRODUCED BY CERTAIN STREPTOMYCETES SPECIES AGAINST ZUCCHINI YELLOW MOSAIC VIRUS

M.F. Ghaly, N.M. Awny, A.M. Galal and A. Askora

Botany Department, Faculty of Science, Zagazig University

       Two streptomycete isolates were isolated, identified as Streptomyces albovinaceus and Streptomyces sparsogenes and tested for their capacity to protect Cucumis sativus from zucchini yellow mosaic virus (ZYMV). Foliage treatment with culture filterate of Streptomyces albovinaceus and Streptomyces sparsogenes reduced mosaic symptoms by 95 and 100% respectively. The filterates of the two tested strepomyces species showed more virus inhibitory activity when applied before virus inoculation than when applied after inoculation. The first 12h after inoculation was the most sensitive period of ZYMV reproduction cycle. The most favourable conditions for higher antiphytoviral production by Streptomyces albovinaceus and Streptomyces sparsogenes were attained after incubation for 6 and 7 days respectively at 30oC andpH7. Morphological and biochemical changes of pre-treated Cucumis sativus leaves infected by ZYMV were detected. The antiphytoviral agent produced by S. sparsogenes have molecular formula C18H33ClN2O5S with molecular weight 424K.Da.It is designated as clindamycin which is one of licosamide class of antibiotics.

 

 

14/19 FLORAL VASCULARIZATION AND THE STRUCTURE OF THE CORONA IN PASSIFLORA CAERULEA L. AND PASSIFLORA SUBEROSA L.(PASSIFLORACEAE)

S.M. Sharawy and E.A.K. Karakish

Botany Department, Faculty of Science, Ain Shams University, Cairo, Egypt.

       The floral vascularization of Passiflora caerulea L. and Passiflora suberosa L. was investigated for an understanding of the structure and nature of the corona. The vascular architecture in both species did not manifest serious deviations from that of a standard angiospermic flower. Fusion of the carpellary ventral bundles in P. suberosa was considered to be an advance over P. caerulea in which the similar bundles were found to be distinct. The outer four whorls in P. caerulea viz. radii (two whorls), pali and operculum were considered to be belonging to the perianth, or of petaline nature if the latter is described to be of sepals and petals. The annulus and the limen (fifth and sixth whorls) were considered to be receptacular outgrowths. The coronary whorls in P. suberosa were considered to be of tepaloid nature.

15/19 LONG-TERM EXPOSURE TO MICROWAVES OF CELL-PHONE RELAY STATIONS: EFFECT ON THE REDOX STATUS IN VITAL ORGANS AND SERUM MONOAMINES

R.A. Taha, O.S.M. Ali*, G.K. Helal, W.A. Ahmed**, K.M. Tohamy***

and F.M.A. Hamada

Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University; *Department of Biochemistry, Faculty of Pharmacy, Al-Azhar University;**Department of Cancer Biology, NCI, Cairo University;***Department of Physics, Faculty of Science, Al-Azhar University, Cairo, Egypt.

       The hazards of exposure to microwaves are observed on different tissues, the mechanistic pathways by which microwaves can produce such effects still need to be delineated. The present work aims to monitor the possibility of modulation in the oxidation-reduction balance in the examined tissues after exposure to microwaves produced from mobile base station at frequency of 900 MHz, power density of 0.05 mW/Cm2 and at a distance of 15 meters. Male albino rats were classified into control and exposed groups (8/group) for periods of one, two and three months. This study revealed that, reduced glutathione (GSH) was markedly decreased in liver, heart and spleen of exposed animals in a time dependent manner. The effect was observed in the liver and heart after the first month of exposure but delayed to the second month in the spleen when compared to their respective controls. The activity of super-oxide dismutase (SOD) in the three organs showed a different pattern; it was initially significantly increased after one month of exposure in all tissues but then markedly reduced after two and three months when compared to their respective controls. Serum alanine amino transferase (ALT) and creatine phosphokinase-MB (CPK-MB) activities were significantly increased. The level of lipid peroxides in the liver showed marked increase allover the study period, while in the heart and spleen the peroxides shows significant rise only after the first month. Moreover, exposure to microwaves induced ECG changes and an increased heart rate. Serum monoamines, namely serotonin, norepinephrine and dopamine, showed decrement in their concentration in a time dependent manner. In conclusion, exposure to microwaves resulted in disturbance in oxidation-reduction balance in the tested organs and increased activity of ALT and CPK-MB in addition to decreased serum monoamines level. These observations may be considered as some of the mechanisms that may be implicated in what is so called microwaves disorders of various organs and tissues.  

 

 

16/19 ADAPTATION OF SACCHAROMYCES CEREVISAE TO WASTE WATER AND SIMULTANEOUS PRODUCTION OF INVERTASE

O.M. Gomaa

Radiation Microbiology Department, National Center for Radiation Research and Technology, P.O. Box 29- Nasr City, Cairo, Egypt

       The waste water from   meat & poultry cold cut factories represent a major environmental problem in the industrial sector in Cairo due to the elevated levels of the Biological Oxygen Demand (BOD), the Chemical Oxygen Demand (COD) and offensive odor. Saccharomuces cerevisiae was considered for invertase production using only the waste water supplemented with 2% molasses. The maximum invertase obtained was 246.99 U / gm of cells (with 1.86 fold increase to the initial production) when a pre-culture media was used (to decrease the lag phase and increase the exponential phase) at 150 rpm, 40oC after 12 hours incubation. Cell wall distortion was evident after 24 hours incubation and this affected the internal integrity of the cell membrane, thus, the intracellular enzyme production. The nitrogen content of the waste water was enough to act as an invertase inducer and the molasses increased the production through carbon catabolite repression. The use of waste water as fermentation media for invertase production is considered a suitable treatment for decreasing COD, BOD and offensive odor in an economical way.
 
17/19 STUDIES OF SOME PHYSIOLOGICAL RELATIONSHIPS BETWEEN THE PARASITE OROBANCHE CRENATA FORSSK AND SOME OF ITS HOSTS (VICIA FABA L., TROPAEOLUM MAJUS L. AND ANAGALIS ARVENSIS L.)

S.A. El-Khawas,

Botany Department, Faculty of Science, Ain Shams University, Abbassia,

Cairo, Egypt.

       Clearly, understanding the biochemical and molecular interactions occurring at the penetration stage of parasitism is an important key to manipulating host resistance. Applications of modern tools in biotechnology enrich this standing. The influence of Orobanche crenata Forssk on host metabolites and DNA fingerprinting is being studied in the present work using three hosts (Vicia faba L., Tropaeolum majus L. and Anagalis arvensis L.). In general parasitism decreases the water content, the total chlorophylls and consequently the soluble and insoluble sugars of the hosts. The total nitrogen and protein showed the same pattern .Conversely the carotenoids and total phenols were increased. The RAPD patterns showed that, the total number of fragments produced using five arbitrary primers is 14 with molecular sizes ranging between 800 and 100bp. The highest number of bands (10) in all samples was recorded using primer OPB-07. The lowest number of bands (2) in all samples was scored using primers OPA-11 and OPB-02. These are two monomorphic bands with molecular size   100 and 200 bp. the identification of complete sequence of these bands may indicate the relationships between the parasite Orobanche and the three hosts. The highest number of bands (7 and 6) of separate samples was recorded in Orobanche and Tropaeolum, respectively using primer OPB-07. Qualitative changes include the scoring of four unique bands in Tropaeolum using primers OPA-01 (molecular size: 450, 330, 330 and 100 bp), OPA-02 (molecular size: 450 bp) and OPB-07 (molecular size: 800 bp). One common band with molecular size 320 bp was scored for conjunction between Orobanche and the three used hosts using primer OPB-07.There is also another band at molecular size 350 bp scored for conjunction between Orobanche and Anagalis using primer OPB-07. This work should lead to the development of strategies for minimizing the devastation caused by parasitic weeds in the Egyptian and International agriculture.

 

18/19 LANTIBIOTIC BACTERIOCIN FROM STREPTOMYCES XANTHOCIDICUS Z22; ELUCIDATION OF PURIFICATION, MOLECULAR SIZE AND AMINO ACID COMPOSITION OF XANTHOCIN Z22

G. Enan and I. El-Azouni

Botany Department, Faculty of Science, Zagazig University, Zagazig, Egypt.

       Xanthocin Z22, a bacteriocin produced by Streptomyces xanthocidicus Z22, was purified sequentially by ammonium sulphate precipitation, gel filtration and reverse-phase chromatography. This purification protocol led to a 49-fold increase in xanthocin Z22 bioactivity. On the basis of electrophoretic pattern in SDS-PAGE, xanthocin Z22 appeared as a single peptide band of about 32 kilo Daltons. Amino acid composition analysis of purified xanthocin Z22 revealed the presence of lanthionine residues, suggesting that xanthocin Z22 is a member of the lantibiotic family of antimicrobial peptides which termed as class I bacteriocins

 

 

19/19 ENHANCING THE HERBICIDAL ACTIVITY OF BENSULFURON METHYL AGAINST RADISH BY ANTIOXIDANTS

M.B. Mahmoud and H.M.S. Khalifa

Plant Protection Department, Faculty of Agriculture, Al-Azhar University

       Effect of some antioxidants, i.e., selenium (Se) at 5 and 10 µg/ml pipronyl butoxid (PBO) at 20 and 40 µg/ml and butylated hydroxy anisole (BHA) at 5 and 10 µg/ml on the herbicidal activity of bensulfuron-methyl was studied. The tests were carried out either in agar or spraying 7-day-old radish seedlings. The results indicated that the bensulfuron-methyl at high concentrations decreases root growth rather effectively than shoot either in agar or as spray treatment. The same result was obtained by low concentrations of the herbicide when the antioxidants were added. It was found that 10µg/ml of the herbicide reduced shoot and root growth in agar medium by 53.45 and 90.11% respectively. Interestingly, 1.0 µg/ml of the herbicide reduced shoot and root by 62.06 and 100% respectively when 10µg of selenium was added. The same results were also recorded in the case of the other two antioxidants, either in agar medium or as spray treatment. BHA seemed to be slightly less effective than selenium or PBO. To confirm these results some metabolic products, i.e. total carbohydrates, proteins and lipids were determined in treated radish seedlings. The results demonstrated that the herbicide reduced these products, 10 days after spraying. Interestingly this reduction effect was promoted by adding the antioxidants. Thus it could be mention that the addition of the tested antioxidants reduce the usage of the recommended rate of the herbicide (240 ug/ml) without decreasing the herbicide activity.

 

 

20/19 EFFECT OF AZOSPIRILLUM INOCULATION AND NITROGEN FERTILIZATION ON THE GROWTH, YIELD COMPONENTS AND GRAIN QUALITY OF SORGHUM

M.M. Hanna, N.A. Naguib* and S.J. Latif*

Soil, Water and Environment Res. Inst. Agricultural Research Center. (ARC),

Giza Egypt.

* Field Crops Res. Inst., Agricultural Research Center. (ARC), Giza Egypt.

       The goal of this study was to recognize the benefits of Azospirillum lipoferum inoculation with or without different levels of N fertilizer (0, 50, 75, 100 kg Nfed-1) on growth, Rihzospher microbial activities, yield components, quality and chemical composition of grain sorghum (Shandaweel -6 hybrid (SH-6 h) and Dorado cultivar). Therefore, two field experiments were conducted at Agricultural Research Center (ARC) farm, Giza, Egypt during 2002 and 2003 growing seasons. Grain sorghum were incorporated into the soil alone or coated with A. lipoferum inoculums. Results showed that morphological characters (root volume and fresh and dry weight of root) were significantly higher in their values in treatments inoculated with either Azospirillum alone or combined with different N- levels than the control ones. Shadaweel-6 hybrid surpassed Dorado cultivar in plant height, grain yield, weight and number of the grain /panicle. No significant differences were found between applying 100 kg N fed-1 or 75 kg N fed-1 +Azospirrillum inoculation or Azosperrillum alone applied to Shandaweel -6 hybrid (SH-6 h) in plant height, leaf area index (LAI), 1000 kernel weight, grain yield, fodder yield, days to 50 % flowering, panicle grain yield and number of grains panicle-1. Insignificant differences were also found between applying Azospirrillum only or with 75 kg N fed-1 on Dorado cv. in LAI, 1000 kernel weight, fodder yield, days to 50 % flowering and panicle grain yield. Shandaweel -6 hybrid significantly surpassed Dorado cv. in shoot length and seedling dry weight. Whiles no significant differences were found between both SH-6 h and Dorado cv. in germination percent, root length and electrical conductivity (EC). Adding 100 kg N fed-1 produced higher values than the other treatments without significant differences in germination percent, root and shoot length, seedling dry weight and (EC). Shandaweel -6 hybrid significantly surpassed Dorado cv. in crude protein, total carbohydrates and crude oil. Adding 100 kg N fed-1 significantly exceeded other treatments in crude protein and crude oil. No significant differences in crude oil were detected between inoculation only or inoculation + 50 kg N fed-1.   In conclusion, the biofertilizer application is considered to be excellent as the effective tools to reduce chemical fertilizer and consequently the cost and environment pollution.

 

 

21/19 GLUCONIC ACID PRODUCTION BY SOME FUNGI

A.A. Shindia, G.A. El-Sherbeny, A.A. El-Esaway and Y.M.M.M. Sherif*

Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig, Egypt.

*Sugar and Integrated Industries Co. (SIIC) Hawamedia, Giza, Egypt.

       The capacity of forty-one fungal species belonging to15fungal genera, isolated and identified from Egyptian soil and sugar cane waste samples, to produce total acid and gluconic acid in their culture filtrates, when grown on selected substrates was tested. For, the first screening samples of culture filtrates were tested by paper chromatography analysis and the positive results were further estimated by quantitative means. Despite the most tested fungi have potentiality for total acid production in their filtrates; it was found that nearly one-half (51%)of them showed positive results as to production of gluconic acid. Aspergillus niger was distinguished by its capacity to produce substantial amounts of gluconic acid when cultivated on selected medium. Its growth and gluconic acid yield were affected by the composition of the culture medium. Under controlled conditions, the optimal gluconic acid yields resulted from submerged culture at 30oC and initial pH6 following 7days of incubation. Among the various substrates used, glucose (14%) was found to be suitable carbon source for maximal gluconic acid during fermentation. Further, both gluconic acid and fungal biomass were at maximum in the presence of 1% peptone as the sole nitrogen source. Moreover, influence of the concentration of some inorganic salts as well as the rate of aeration on the gluconic acid and biomass production is also discussed.

 


22/19 GENETIC VARIATIONS AND THE RELATIONSHIPS AMONG SOME SPECIES OF THE GENUS BRASSICA L. BASED ON DNA (RAPD) ANALYSIS

E.A. Kamel, H.Z. Hassan*, A.I. El-Nahas and S.M. Ahmed

Department of Biological Sciences and Geology, Faculty of Education, Ain Shams University, Roxy - Cairo, Egypt.

*Department of Botany, Faculty of Science, Ain Shams University, Abassia,

Cairo, Egypt.

       Molecular genetic fingerprints for 18 Brassica L. samples belonging to six species (Brassica alba, B. nigra, B. oleracea, B. rapa, B. napus and B. tournefortii) were conducted to elucidate relationships among these samples. Extracted DNA from fresh leaf samples was used to identify the molecular fingerprints of Brassica L. genotypes. Fifteen 10-mer arbitrary oligonucleotide primers were initially used to establish their randomly amplified polymorphic DNA based on polymerase chain reaction (RAPD-PCR) fingerprints and to discriminate among them. Only six primers were successfully generated reproducible polymorphic products. The primers are; UBC 43, 51, 66, 71, 77 and 91. The fingerprints generated by these primers revealed characteristic profiles for each Brassica L. genotypes, in terms of number and position of RAPD bands. The results revealed that both the number and size of the amplified products varied considerably with the different primers. A sum of 110 polymorphic bands was generated by these primers. A total of 19 unique bands were identified out of the polymorphic ones and these unique bands were used to discriminate among the studied Brassica L. samples. Most of the studied samples of the genus Brassica L. were discriminated by one or more unique bands.

 

23/19 FACTORIAL DESIGN FOR OPTIMIZATION OF

11Α-HYDROXYPROGESTERONE PRODUCTION BY

MUCOR RACEMOSUS NRRL 3639

H.A. El- Refaie

Chemistry of Microbial and Natural Products Dept. National Research Center,

Cairo, Egypt

         Optimization of biotransformation of progesterone to 11α- hydroxy-progesterone by the filamentous fungus; Mucor racemosus NRRL 3639 was investigated adopting statistically based experimental designs. The fungus was examined for its potentiality of the bioconversion process under different culture conditions planed according to the Plakett-Burman factorial design. Peptone and L-aspargine were the most significant factors affecting progesterone bioconvertibility. A verification experiment was carried out, which confirmed the validity of the predicted near optimum conditions. In the second optimization step, the steepest ascent method was applied for determination of optimal levels of the two significant factors that led to maximum 11α-hydroxyprogesterone output. Based on the results of this experiment, maximum 11α-hydroxyprogesterone (11α-HP) yield (65.20%) could be achieved by using the following composition of cultivation medium (g/l): peptone 5, yeast extract 0.5, glucose 30, MgSO4.7H2O 0.1, KH2PO4 1.5, and L-aspargine 1.14; pH 6.0.


24/19 BIOLOGICAL CONTROL OF BACTERIAL SPOT DISEASE OF TOMATO CAUSED BY XANTHOMONAS CAMPESTRIS PV. VESICATORIA BY USING RAHNELLA AQUATILIS

H.H. El-Hendawy, M.E. Osman, N.M. Sorour

Botany and Microbiology Department, Faculty of Science, Helwan University, Ain Helwan, Cairo, Egypt.

       Xanthomonas campestris pv. vesicatoria strain 2 was isolated from infected tomato seedlings grown in open field in Egypt. This strain was able to produce irregular yellow-necrotic areas on tomato leaves as well as spotting of the stem.   In an attempt to control this disease biologically, four experiments were conducted and tomato seedlings were pretreated, before the pathogen, with either of two antagonistic strains of Rahnella aquatilis through leaves, roots, soil or seeds. In all experiments, seedlings pretreated with R. aquatilis showed reduced susceptibility toward X. campestris. pv. vesicatoria. They also contained reduced protein concentration and showed reduced number of protein bands on SDS-PAGE as well as increased fresh and dry weight relative to control seedlings inoculated with the pathogen only. This indicates that R. aquatilis reduced the deleterious effect and the stress exerted by X. campestris pv. vesicatoria on tomato seedlings. Foliar application of R. aquatilis was the most effective method in disease reduction which could be attributed to the direct effect of the antagonistic bacteria on the pathogen. On the other hand, the highest amount of fresh and dry weight was obtained from root treatment which might suggest that root bacterization could provide more protection than could be provided with foliar, soil or seed treatment.

 

 

25/19 CYTOLOGICAL, PHYSIOLOGICAL AND BIOPHYSICAL STUDIES ON THE EFFECT OF LOW FREQUENCY ELECTRIC FIELD ON WHEAT PLANT

M.S. Hanafy, H.A. Mohamed* and E.A. Abd El-Hady**

Dept. Phys., Fac. Sci. Zagazig University, Zagazig, Egypt.

*Dept. Botany, Fac. Sci. Zagazig University, Zagazig, Egypt.

**Dept. Genetic and Cytolgy, National Research Center, Dokki, Giza, Egypt.

       Two exposure systems of an extremely low frequency electric field were used in this work, the first was an experimental model (50 Hz-6 Kv/m strength) and the second was the high voltage transmission lines passing through an open agricultural field (50 Hz - 66 Kv/11m). The effect of the two exposure systems were investigated on mitosis, meiosis and pollen grains viability in the wheat plant to reflect the different abnormalities in the chromosome behaviour. Also, the effect of that two fields on some morphological characters such as the stem length, the spike number, spike weight per plant and the number of grains per spike together with some physiological parameters such as the chlorophyll content, the elements level, carbohydrate content, and total crude protein were estimated for exposed and unexposed grains. Classification of the water soluble protein (WSP) extracted from the exposed and unexposed grains as well as their molecular weight distribution were also investigated by using SDS polyacrelimide gel electrophoresis technique. The absorption spectra of the WSP also measured at wavelength range 200-600nm. The results indicated that the electric field caused a marked effect on the frequency of the different mitotic phases. Both of the two exposure systems showed a decrease in the mitotic index of wheat root meristematic cells with induction of high frequency of chromosomal abnormalities in one hand, and in the other hand the treated wheat flower buds showed an increase in the percentage of abnormal pollen mother cells (PMCs) and a marked increase in the frequency of the nonviable pollen grains. The results also indicated remarkable changes in the morphological characters of the exposed grains where the stem length increased but the spike weight and the number of grains in the spike decreased. Further, the data showed an increase in the total chlorophyll content and the total carbohydrates in the grains. Furthermore, the amount of total protein in the exposed grains decreased, also, pronounced changes were occurred in the concentration of the elements due to the exposure processes. On the other hand, the data indicated that the molecular structure of the extracted WSP changed, the amount of protein in the bands of exposed grains decreased and their molecular weight changed and also fluctuation in the electrophoretic mobility of the protein bands.

 

 

26/19 EFFECT OF PEPTIC HYDROLYSIS ON THE ELECTROPHORETIC PATTERNS AND ANTIBACTERIAL ACTIVITY OF HUMAN, CAMEL MILK AND EWES MILK

F.A. Fathey, K.M. El-Deib* and S. El-Behery**

Dairy Science Department, National Research Centre, Dokki, Egypt

*Molecular Biology Department ,**Food Evaluation & Food Science Dept.

National Organization for Drug Control & Research (NODCAR)

       The effect of peptic hydrolysis on the electrophoresis patterns and antibacterial activity of human, camel and ewe’s milk were studied. The Human, camel and ewe's milk samples were obtained from healthy lactating animals and women. Milk samples were defatted by centrifugation and peptic hydrolysis were carried out by the direct addition of pepsin after pH adjustment to pH 2 and were incubated at 37ºC for 45 min followed by heating at 80 ºC for15 min, then pH were adjusted to pH 7 and centrifuged at 5000 rpm for 5 min. Supernatant was filtrated through 0.45 µm and used for experimental study. The electrophoresis analysis data identified the major proteins in as lactoferrin (77, 84 & 79 kDa), serum albumin (66, 74, 68 kDa) , sIgA heavy chain (55, 54 & 53 kDa), β-casein (30, 35 & 40 kDa), sIgA light chain (25, 35 & 24 kDa) and α-lactalbumin (14 , 16 & 15 kDa) in   human, camel and ewes milk proteins respectively. While, the obtained data of digested human, camel and ewes milk with pepsin identified the different bands as 75, 72, 44, 15 & 8 kDa, 81, 66, 50, 35, 23, 20 & 13 kDa and 78, 60, 53 48, 40, 24 & 15 in human, camel and ewe’s milk proteins digest respectively. The obtained data from the antibacterial activity study showed that human, camel, ewe's milk proteins and its peptic digest had little antibacterial action against gram positive and gram negative bacteria and no antibacterial action against all bacterial strains grown in fat-free milk medium at the used concentrations (30-100 µg/ml for peptic milk protein digest and 1 to 2 mg/ml for whole milk protein), while this activity can be easily demonstrated in 1% peptone or other simple growth medium and the inhibitory activity against these bacteria at the same concentration were increased and mixed with 0.25 mM EDTA.


27/19 MORPHOLOGICAL AND PHYTOCHEMICAL STUDIES ON CESTRUM PARQUI L' HERIT. (SOLANACEAE)

M. E. Tantawy; S. F. Khalifa; A. K. Youssef* and I. M. Damlakhi**

Department of Botany., Faculty of Science, Ain Shams University, Cairo, Egypt

*Desert Research Center, Cairo, Egypt

**Department of Botany, Faculty of Science, Aleppo University, Syria

       Macro, and micromorphological characters of the vegetative organs of Cestrum parqui L' Herit. (Solanaceae) were investigated for morphological, anatomical purposes and accurate identification of the species. The macromorphological characters of both vegetative and floral organs and the anatomical characters of the vegetative organs only are on the same ground plan of Solanaceae. Phytochemical investigation of the dried powder of the aerial parts indicated the presence of fatty acids, hydrocarbons, sterols, carbohydrates and amino acids which were identified by Gas-Liquid Chromatography (GLC) and microanalysis techniques. Two flavonoid compounds (quercetin-3-O-α-L-rhamnoside (1-6) β-D-glycoside and kaempferol-3-rutinoside) were isolated   and identified by UV spectral data and 1H-NMR analysis for the first time.

 

 

28/19 POTENTIAL APPLICATION OF THE EXTRACELLULAR CHITINASE PRODUCED BY BACILLUS ALVEI IN BIOCONTROL

SH.M. Abdel-Aziz and F.E. Moafi *

Microbial Chemistry Dept. and *Microbial Biotechnology Dept.,

National Research Center, Dokki, Cairo, Egypt.

         Bacillus alvei, a local bacterial isolate, was used for production of chitinase usingoriginal flaked chitin in comparison with chitin recovered from shrimp shells as a sole carbon source. Maximum chitinase activity in the culture supernatant obtained after 48h, with no obvious differences in the enzyme production between original or recovered chitin underoptimal growth conditions in which 30°C, 150 rpm, pH7.0 and 3d time were applied. Chitinase produced by B. alvei was found to be active at 50°C and a wide range of pH values from 7-10 with a maximum activity at pH 9.5. The enzyme was able to hydrolyze colloidal chitin, soluble chitosan with different degrees of acetylation, crystalline cellulose, CM-cellulose, or xylan as substrates.The enzyme was highly activated by N-acetylglucosamine, Cu2+, Ca2+, or ethanol; slightly inhibited by Zn2+, Hg2+, or acetone. Antifungal activity of the enzyme on some pathogenic fungi was investigated in growing cultures. Fungal lysis was found to be 95, 97, and 94% for Rhizopus sp., Mucor rouxii and Fuserium solani, respectively. Both Aspergillus flavus and Pythium sp. were the least affected i.e., fungal lyses were 20 and 25%, respectively.Chitinase produced by B. alvei may contribute to the biological control, might play a role against some larve (larvicidal activity), or it might has potential on opportunistic fungal infections in burn wound caused by Phycomyces or Aspergillus.

 


29/19 UTILIZATION OF SHRIMP AND CRAB SHELLS FOR PRODUCTION OF AN ALKALINE PROTEASE BY

BACILLUS SUBTILIS

S.M. Abdel-Aziz and F.E. Moafi*

Microbial Chemistry Dept. and *Microbial Biotechnology Dept.,

National Research Centre, Dokki, Cairo, Egypt

       The application of protease produced by Bacillus subtilis as a laundry detergent additive is frequently reported. The strain was adapted to produce an alkaline protease using shrimp and crab shells (SCS) as a sole source of carbon and nitrogen, without any nutrition additives. Optimal culture conditions for the enzyme production were 32°C, 150 rpm, pH 8, and 48h growth. The optimal pH and temperature for protease activity were found to be 10.5 and 55oC, respectively. The enzyme exhibited a unique property; it did not require Ca2+ for activity and thermal stability. The enzyme was relatively stable in presence of detergents and oxidants. It attained more than 95% of activity in presence of X-TRA or Biocleana and 80% of its activity in a concentration of 10% chlorine. The enzyme activity was, however, inhibited by EDTA, suggesting that this protease is a metallo-enzyme. Thus, the protease produced by B. subtilis could be ideal for detergent applications.

30/19 PRODUCTION OF PHB-co-PHV FROM COMPOST Biodegradation BY PURPLE NON-SULFUR BACTERIA

M.S. Shabeb, M. Younis, A. Amara* and A. Shoreit**

Botany Department, Faculty of science, South Valley University, Aswan, Egypt.

*Protein Department,Institute for Genetic engineering and Biotechnology, Mubarak City for Scientific Research and Technology Applications, Alexandria, Egypt.

**Botany Department, Faculty of science, Assiut University, Assiut, Egypt.

      For the first time we reported the production of PHAs using compost water extract, where eight strains of purple non-sulfur bacteria (Rhodospirillaceae) were isolated from mud of different parts of Eichornia crassipes area at different localities from River Nile (Aswan- Egypt) have been investigated for their ability to accumulate PHAs when grown on compost water extract. CWE: compost water extract (C:N 13:1) has been used only at different concentrations as a sole carbon source. All strains showed the best production of PHAs when CWE used at concentration of 75%. Rhodospirillum rubrum ASEC 39 produced the highest content of PHAs (46.8%), which represented 5.5% of the 3-hydroxybutyrate (3-HB) and 40.2% of 3-hydroxyvalerate (3-HV). Rhodopseudomonas palustris ASEC 12 represented the second one, which produced 43.5% of 3-PHAs (28.5% 3HB and 12.5% 3HV). Rhodopseudomonas palustris ASEC 47 and Rhodospirillum rubrum ASEC 36 produced a content of 43.1% and 39.7% 3-PHAs these represented 31.5% and 3.35% of 3HB; 11.3% and 34.5% of 3HV respectively. The production of PHAs by photosynthetic bacteria has become interesting commercial prospect due to lower carbon feedstock coasts. Moreover, photosynthetic bacteria work as scavengers of the pollution by organic compounds bioremediation. Compost extract is considered a new strategy for PHAs production.

31/19 PURIFICATION AND CHARACTERIZATION OF THERMOSTABLE CHITINASE FROM MALBRANCHEA CINNAMOMEA ML2-STRAIN

A.A. Shindia, S.A. Khalaf, G.A. Helal and A.S. Abdel-Fatah

Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig, Egypt

       Malbranchea cinnamomea ML2, a thermophilic fungal strain capable of utilizing chitin as a carbon source, was isolated from compost samples. Three types of exochitinases (Chi I, Chi II and Chi III) produced from the culture supernatant of M. cinnamomea ML2 were purified to an apparent homogeneity identified by SDS-PAGE through ammonium sulphate as well as organic solvent precipitation, membrane technology and sephadex G200 gel filtration. Molecular weights of the enzymes were 64.5 KDa (Chi I), 41.7 KDa (Chi II) and 26.3 KDa (Chi III). The chitinase (II) showed high specific activity and protein peak. The quantitative composition of the amino acid of chitinase (II) was analyzed in relation to it’s thermostability. The optimum temperature and pH of the 41.7KDa chitinase (Chi II) were found at 60°C and pH 6.0. The enzyme showed thermal (50-70°C) and pH (5-7) stabilities. Hydrolysis process increase with increasing the enzyme concentration, reaction time as well as colloidal chitin concentration up to 15 mg in the enzymatic reaction mixture. The impact of shaking of reaction mixture on enzyme activity and the antifungal activity of purified enzyme were also discussed. In addition, the chitinase activity was activated by Fe2+ and Zn2+ ions, while K+, Na+ and Ca2+ ions markedly inhibited the enzyme activity. It was also found that, the purified chitinase is stable in frozen state for two months.
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