Vol. 9, January, 2001.

1/9 SYSTEMIC RESISTANCE INDUCING PROTEINS IN

PHASEOLUS VULGARIS LEAVES TREATED WITH

BOTANICAL EXTRACTS OR VIRUS

A.B. Barakat

Microbiology Department, Faculty of Science, Ain-Shams University, Cairo, Egypt

Systemic acquired resistance (SAR) is an inducible plant defense state, the induction of which often requires both the accumulation ofpathogenesis-related (PR) proteins and an enhanced capacity for activating local defense responses upon pathogen attack. The pre inoculation with either botanicals or pathogens induced SAR in distant untreated parts of plant. SR-inducers seem to activate the plant defense genes to encode novel proteins. These proteins were either absent from healthy leaves or present in much smaller amounts. In our study, systemic resistance inducing proteins were detected in Phaseolus vulgaris cv. The prince leaves opposite to those receiving either botanicals from Dianthus sinensis var. heddewigii (DS) and Mirabilis jalapa L (ML) or virus (tobacco necrosis necrovirus TNV). Such proteins have been found to play a key role in inducing strong systemic resistance in susceptible host against viruses. The level of viral resistance observed in untreated parts of plants was correlated well with the amount of novel proteins accumulated and the timing required for accumulation. The maximum inhibitory activity was shown in opposite untreated leaves 3 days after DS, MJ or TNV application. The effect declined after 7 days. Elementary study of DS and MJ revealed that they were potent inhibitors giving 100% inhibition, their dilution-end point was Iff3 and they were heat stable. Polyacrylamide gel electrophoresis analysis revealed the induction of four novel proteins in the homogenates of P.vulgaris leaves opposite to those receiving DS, MJ or TNV 7 days after application. These new proteins had an apparent molecular masses of 14.5, 16.5, 22 and 39.5 KDa, Non of these proteins were detected in water inoculated controls or in leaves treated for 1 or 3 days after application, as well as their corresponding opposite ones. These novel induced proteins resemble ribosome-inactivating proteins extracted from various plant species. In vitro the resistance leaves inhibited local lesion production by TNV.

2/9 SOME MOLECULAR STUDIES ON CALVIN CYCLE GENE

PRODUCTS IN PEA

A. Badr, S.F. Badr, A. Mustafa and W.F. Martin*

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

* Institut fuer Botanik III, Heinrich-Heine Universitaet Duesseldorf, Duesseldorf,

Germany.

Seeds of pea (Pisum sativum L. cv. Rosa Krona) was cultivated and grown under green house conditions. During the seedling and vegetative stages, samples were collected from roots, cotyledons and leaves. After flowering samples were taken from leaves, pericarps and developing seeds during embryogenesis. Each sample was divided into two parts. From the first part, total proteins were extracted and assayed quantitatively, and the enzymatic specific activity of each of the Calvin cycle enzymes was followed in the approprite organ throughout the growth stages. From the second part, total RNA was extracted, denatured and immobilized by dot blotting. The molecular probes of the Calvin cycle enzymes were radioactively labeled, and used to hybridize the immobilized mRNA. In order to study the gene expression of the Calvin cycle enzymes in the approprite organ of the plant. The Calvin cycle enzymes,of ribulose-l,5-bisphosphate carboxylase (RuBisCO), phosphoribulokinase (PRK), NADPH-dependent glyceraldehyde-3-phosphate dehydrogenase and NADP-dependent fructose-1,6-bisphosphatase, which are localized exclusively in the chloroplast, showed remarkable levels of specific activity and gene expression in the root. In addition, high levels of gene expression and specific activity of transketolase (TKL), ribulose-5-phosphate 3-epimerase (RPE) and D-ribose-5-phosphate keto-isomerase (RPI), that are common to both 'he Calvin cycle and the oxidative pentose phosphate pathway (OPPP), were detected in both photosynthetic and non-photosynthetic organs. The relation between gene expression and the specific activity of each enzyme and within all the enzymes was also discussed, taking into consideration previous views about the presence of complexes between some enzymes. Data revealed positive relation between some enzymatic activity and gene expression. However, the activities and expression of other Calvin cycle enzymes show various degrees of correlation, indicating that Calvin cycle enzymes are not randomly distributed in the chloroplast stroma, but rather organized in some fashion.

3/9 EFFECT OF BRASSINOLIDE ON NITROGEN METABOLISM AND NUCLEIC ACIDS LEVEL IN TOMATO PLANT

E.M. Nafie and S.M. El Khallal

Department of Botany, Faculty of Girls, Ain Shams University,

Cairo, Egypt

Modulation of the nitrogen metabolism of tomato plants in response to brassinolide (BR) application was observed as being in concomitance with the activity level of nitrate reductase( NRase) and glutamate dehydrogenase( GDH). Thus, enhanced activity of these enzymes was expressed in a substantial increase in amino-N, total soluble-N, total-N and the protein content. The presence of alliance between the content of NOj and the activity level of NRase refer to the effect ofBR on induction mechanism (de novo synthesis and or / activation) by substrate. Moreover, this result refer to an effect ofBR on NOj uptake. The results attained in the present work refer also to a marked increase in the nucleic acids (DNA & RNA) content of the tomato shoots which was in parellel with each increase in the concentration used of BR. Foliar application was more promising, in this respect, compared with the seed soaking treatment. Thus, while the highest applied concentration (0.2ppm) induced an increase of 142.6 (RNA) (as % of control) and 81.2 (DNA), in case of foliar application, it account to only 54.2 (RNA) and 32.8 (DNA) as a result of seed soaking treatment.

4/9 THE INTERACTION EFFECTS OF SALINITY AND

CERTAIN BIOREGULATORS ON THE PROTEIN METABOLISM OF TWO DIFFERENT RICE CULTIVARS

H. Khattab

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

Two differently salt sensitive rice (Oryza saliva L.) cultivars. Giza 178 and Sakha 102 were imbibed in H^O, GAs, ascorbic acid or nicotinic acid for 6 hours then, the grains of each treatment were exposed to 0, 100 or 200 mMNaCl to induce salt stress. Seed germination, total soluble protein content, protein patterns, total free amino acids and the proteolytic activity of proteases enzymes were studied. Protein breakdown and turnover was delayed by the NaCl as compared with control. The specific activity of proteases decreased due to the delayed breakdown of proteins under salt stress. Salinity modulates the production of selected groups of proteins in rice, especially cv. Giza 178 (relative sensitive cultivar). Some specific radicle emergence polypeptides bands disappeared (107 KD in cv. Giza 178 and 90 KD in cv. Sakha 102) under salt stress. Salt stress induced the synthesis of mid and high molecular weights new polypeptides (salt induced polypeptides) with molecular weight 61KD in cv. Giza 178 and 116 KD in cv. Sakha 102. These results suggest that the expression of salt stress proteins is related to the adaptation process of grains to salinity as well as the genetic constitution of selected salt tolerant genotype.

5/9  PATHOGENESIS-RELATED PROTEIN AS AN INDEX FOR DETECTION OF POTATO PLANT PATHOGENS

M.A. Tag El-Din; Kh.A. El-Dougdoug* and R.M. Taha**

Biochem. Dep. Fac Agric., Ain Shams Univ., Cairo, Egypt.

*Microbiol. Dep. Virology Lab., Fac. Agric., Ain Shams Univ., Cairo, Egypt.

**Botany Dep. Fac. Science, Cairo Univ., El-Fayoum Branch, El-Fayoum, Egypt.

Alteration in soluble protein constituents of infected potato leaves with Alternaia solani (A. solani); Pseudomonas syringae (Ps. Syringae); Potato virus X (PVX) and mild and severe potato spindle tuber viroid (m-PSTV& s-PSTV) could be detected using SDS-PAGE. It was found that peroxidase and polyphenol oxidase activities were considerably higher in infected potato leaves than that of healthy ones. PSTV infection exhibited higher activity of peroxidase followed by PVX, Ps. syringae and A. solani infection. While the polyphenol oxidase activity was higher in case of A. solani infection in comparison with Ps. syringae, PVX and PSTV infections. The alteration in protein fractions from healthy and infected potato leaves exhibited six polypeptides relative to s-PSTV, four relative to m-PSTV, seven relative to PVX, six relative to Ps. syringae and six to A. solani. It was found that, the 55.6; 37.8 and 14.3 kDa proteins were the most prominent related to these pathogens. These related proteins that bound to pathogens were present only in detectable levels in the soluble protein fractions of infected potato leaves. The alteration that was detected in the protein pattern of infected potato leaves as judged by the intensity of their staining resulted in the enhancement of 55.6; 37.8 and 14.3 kDa for all pathogens. In addition, 50.3 for PVX andPSTVd; 28.9 for A. solani and Ps..syringae; 31.2 for PSTVd; 26.6 for PVX, 104.8 for A. solani and 28.9 for Ps. syringae.

6/9 GENOMIC FINGERPRINTING USING RANDOM AMPLIFIED POLYMORPHIC DNA FOR DISCRIMINATION BETWEEN PROTEUS MIRABILIS STRAINS

M.S.M. Mansy

Department of Microbiology, Faculty of Pharmacy, University of Al-Azhar, Nasr City,

Cairo, Egypt.

The suitability of random amplified polymorphic DNA (RAPD) for the detection
of differences between Proteus mirabilis strains was evaluated. Genomic DNAsfrom
35 well-defined isolates of Proteus mirabilis from human, animal and sewage
sources and 4 reference strains of Proteus species, were examined by PCR-based
RAPD fingerprinting analysis using the oligonucleotide primer (GCCAGGCTCCAG). Strain-specific arrays of DNA fragments were generated, which allowed us to distinguish closely related strains of Proteus mirabilis. Strains of Proteus vulgaris, Proteus penneri and Proteus myxofaciens yielded arrays of DNA fragments that differed markedly from those generated from the Proteus mirabilis strains using the same arbitrary primer. This approach was compared with the phenotypic and genotypic typing methods such as biotyping, susceptibility resistance pattern, proticin typing, PCR-based RFLP profile, protein pattern and ribotyping. Thirty-two RAPD profiles were obtained by PCR-based RAPD genomic fingerprinting from the 39 strains of Proteus species. The results showed that the PCR-based RAPD fingerprinting method distinguishes genetically different strains of Proteus mirabilis and may be valuable for monitoring transmission of this pathogen.

7/9 APPLICATION OF SOME NATURAL SUBSTANCES TO CONTROL YELLOW LEAF CURL DISEASE IN TOMATO PLANTS

Kh.A. El-Dougdoug; R.M. Taha* and A.M. Kheder**

Micrbiol. Dep., Fac. Agric., Ain Shams Univ., Cairo, Egypt.

*Botany Dep. Fac. Science, Cairo Univ., El-Fayoum Brach, El-Fayoum, Egypt.

**Plant Pathology Dep.Fac. of Agric., Zagazig Univ.

The inhibition effect of spraying mixture of khella (Ammi visnaga) extract and fermented yeast (Saccharomyces cervisae) solution against tomato yellow leaf curl gemnivirus (TYLCV) was assayed TYLCV samples were collected from naturally infected tomato (Lycopersicon esculentum cv. Castle rock). The virus (TYLCV-D) was identified according fo symptomatology, reaction on some differential hosts, mode of transmission, polymerase chain reaction (PCR) using specific primers for old world whitfely-transmitted gemnivirus and SDS-PAGEpathogenesis-relatedprotein (RP). Spraying tomato plants with khella extract and fermented yeast solution showed a notable reduction in the percentage of viral infection and revealed an improvement in the rate of growth and yield compared with non sprayed ones. There was a reducton in virus concentration in sprayed infected plants compared with non-sprayed ones as detemined by infectivity assay and indirect-ELISA.

8/9 INFLUENCE OF BRASSINOLIDE ON THE CHANGES OF ENDOGENOUS GROWTH HORMONES AND THE ACTIVITIES OF CERTAIN OXIDATIVE ENZYMES IN TOMATO PLANTS.

S.M. El-Khallal and E.M. Nafie

Botany Department, Faculty of Girls, Ain Shams University,

Cairo, Egypt

Brassinolide (BR) showed an important role in the growth and development of tomato plants by overlapping or controlling the activities of endogenous growth hormones. BR caused variable changes in the levels of the endogenous growth regulating substances of tomato plants. Data showed that both seed soaking and spraying tomato plant with BR increased the contents of auxins, gibberellins and cytokinins, while ABA content was markedly reduced. The obtained increase of auxin content was related to the inactivation of IAA oxidase (IAA-O) andpolyphenol oxidase (PP-O) and the simultanous increases in peroxidase (P-O) and catalase (CAT) activities. It is worthy to mention that spraying tomato plants with BR was more effective on the appropriate endogenous growth hormones than seed soaking at similar concentrations ofBR.

9/9 EFFECT OF GAMMA RADIATION ON THE GROWTH OF

ASPERGILLUS FLAWS AND AFLATOXIN Bj IN WHEAT

FLOUR AND SOME BAKARIES

B.M. Youssef and S.R. Mahrous

National Center for Radiation Research and Technology

P.O.Box 29 Nasr City, Cairo, Egypt.

The occurrence of aflatoxin B/ and aflatoxins-producing moulds in wheat flour and the effect of'/-radiation on the prevalence of aflatoxin bi in wheat flour and some bakery products were investigated. Aspergillus flavus isolates were recorded at high frequency in wheat flour and of 48 isolates 50% produced aflatoxin B]. The growth of Asp. flavus in wheat flour was completely inhibited by 5.0 kGy dose level. Aflatoxin bi was detected in 11 and 7 out of 20 samples, eachofwheatflourof82and72% extraction respectively. Irradiation at dose level of 12.5 kGy markedly decreased the concentration of aflatoxin bi in wheat flour. The concentrations of aflatoxin bi showed a visual decrease in all the different bakery products prepared from 2.5, 5.0 and 7.5 kGy gamma irradiated flour. Higher destruction of aflatoxin bi (84-91%) was obtained after baking the products made of 7.5 kGy gamma irradiated wheat flour.

10/9 SEED PROTEIN DIVERSITY, ISOZYME VARIATION AND CHROMOSOME NUMBER OF SOME EUCALYPTUS SPECIES

IN EGYPT

A.A. El-Ghamery, S.F. Khalifa* and W.T. Kassem

Botany & Microbiology Department, Faculty of Science, Al-Azhar University,

Madient Nasr, Cairo, Egypt.

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

Seed protein diversity as reveal by variation in SDS-PAGE, chromosome number and morphological characters of five species of genus Eucalyptus belonging to two section: Macroantherate and For anther iodae are discussed to reassess the taxonomic relationship of taxa of these species of the Eucalyptus L. The relationship of the studied taxa as reveal by the producedphenogram, based on numerical analysis of criteria are evaluated in relation to previous classification of Eucalyptus. According to the numerical analysis method, the studied taxa are splitted into three groups: Group I (E. camaldulensis accessions), Group II (E. globulus, E. hemipholia and E. nicholii) and Group III (E. citriodora accessions). According to esterase isozyme reseults, E. hemipholia is splitted from others in a separate group.The results obtained in this work reinforce the previous taxonomic treatment of the genus Eucalyptus.

11/9 EFFECT OF AGEING ON PROTEIN METABOLISM

OF BARLEY GRAIN

M.E.H. Osman, W.A. Kasim, S.S. Elfiky, RJ. Fido* and P.R. Shewry*

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

*IACR, Long Ashton Research Station, Department of Agricultural

Sciences, University of Bristol, Long Ashton, Bristol, BS18 9AF, UK.

The present study aims to demonstrate the effect of accelerated ageing with different temperature treatments on germination-mediated biochemical processes of barley. The results show a non-significant reduction in the content of albumin and globulin of grains incubated at 10 °C and 20 °C for 6 weeks, but a significant reduction was recorded with grains incubated at 40 °C, starting from the first week. Hordein I and Hordein II show a significant reduction in case of grains incubated at 30 "Cfor 5 and 6 weeks, while grains incubated at 40 °C showed a sharp reduction in Hordein content. There was no obvious change either in the pattern or intensity of protein bands in grain stocks aged at any temperature. The polypeptides in the C-Hordein region started to disappear from most stocks except those incubated at 40 °C for 2,3,4,5, and 6 weeks which were characterized by more intense B-Hordein.

12/9  PURIFICATION AND CHARACTERIZATION OF

XYLANASE AND MANNANASE FROM GREEN BEAN (VICIA

FABA) POD AND ENZYMATIC DEGRADATION OF SOME

AGRICULTURAL WASTES AND SAWDUST

S.T. El-Sayed

Biochemistry Department, National Research Center, Tahrir St, Dokki, Cairo, Egypt.

Xylanase andmannanase enzymes were extracted and purified from green bean (Vicia faba) pod with specific activities 16 and 17.3 U/mg protein and 57.5 and 40% overall recovery respectively. Purification process included ammonium sulfate precipitation (30-80% saturation) of the crude extract and gel filtration on two successive Sephadex G-120 columns with 34.04 and 91.1 purification fold for xylanase and mannanase, respectively. Each enzyme was shown to be homogenous chromatographically. Xylanase and mannanase enzymes showed apparent molecular weights of 68 and 48 KD respectively as estimated by gel filtration on Sephadex G-120 column. The pH's optima for their activity were 7.0 and5.0 for xylanase and mannanase, respectively at 50°C. Both enzymes were fairly stable at temperatures up to 50°C. Eighty and sixty five percent of the activity remained after xylanase and mannanase, respectively had been heated at 60°Cfor 180 min. Xylanase, which is active on xylans and mannanase, which is active on galactomannans are not associated with any detectable cellulose activity. Both enzymes showed no activity
toward glycogen, starch, lichenins, laminarin, chitin, pectin, yeast glucan, avicel,
carboxymethyl cellulose, cellobiose, maltose, lactose and sucrose. By thin layer
chromatography of the xylan and galactomannan hydrolysed products, the enzymes
were classified as an endo-xylanase and endo-mannanase. Data show that, advancing
time of enzymatic hydrolysis of 'unsoaked agricultural wastes and sawdust from 2 to
24 h, the yield of sugars increased from 1.9-5.2% to 5.7-12.9%. The soaked
agricultural wastes and sawdust showed higher rates of digestibility, besides, the
yield of sugars exhibited 7.4-15.4% and was higher than those of unsoaked samples.
The processes of agitation during the enzymatic hydrolysis of the soaked agricultural
wastes and sawdust yielded increase of the resultant sugars (14.2-25.0 %). 

13/9 A NEW RECORD OF THIOPHENE DEGRADING ENZYME SYSTEM [THIOPHENASE(S)] OF NOVEL MICROBIAL ISOLATES CAPABLE OF NORMAL GROWTH ON THIOPHENE AS A SOLE CARBON AND ENERGY SOURCE

M.S. Ammar, H.G. Solimnn and O.A. Hassabalah

Botany and Microbiology Dept., Faculty of Science, Al -Azhar

University ,Cairo, Egypt

An investigation concerning the microbial degradation of thiophene as a sole carbon source and a representative of sulfur- containing heterocyclic compounds as well as the isolation, purification and experiment of the responsible enzyme (thiophenase) on the degradation of such complex substrates has been undertaken.optimal parameters controlling the biodegradation of thiophene by the intact cells are: pH 7, thiophene, 1%, v/v,incubation temperature 32°c and incubation period 2 days. Also, all isolates succeeded to grow normally on dibenzothiophene (0.5%, v/v) which indicates that these isolates have an enzymatic system specific to sulfur- bond with the thiophene heteroring independently of the hydrocarbon portion of the sulfur - heterocyclic compounds. In order to confirm the degradation power by the intact cells, the most potent two isolates, Streptomyces albaduncus, A7 and Pseudomonas pseudomallei, Bl were used for thiophenase (s) production, isolation, purification and experimentation in the reaction mixture of thiophene (specific substrate). Both crude form (cell free filtrate) and purified one (after sephadex G- 200 column) degraded thiophene. The cell free filtrate (c. f.f) of A 7 exhausted thiophene (1%) mixed in water or even gasoline within 3 days whereas c.f.f. ofBl did so within 3 days with water and 5 days with gasoline. Purified preparations of thiophenases from A7 and Bl on the same column of sephadex G 200 were located infractions 28-29 for A 7 and fraction 23 for Bl which indicates that there are two forms of thiophenases obtained from two different microorganisms under the same specified production conditions. The remarkable degradation power of the present crude and pure preparations of thiophenases refers to their possible application in the safe and clean decontamination of sulfur impurities of heterocyclic compounds which represents a new trend for the possibility of having an enzyme process route to clean up sulfur in oil.

14/9 EFFECT OF DIMETHYL DIPHENYL BICARBOXYLATE (DDE) ON HEPATIC ANTIOXIDANT STATUS AND ON CARBON TETRACHLORIDE INDUCED HEPATOTOXICITY IN RATS

H.A. El-Bahrawy, S. El-Sawy* and A. El-Shafay**

Department of Biochemistry, Faculty of Pharmacy, Tanta University

*Department of Physiology, Faculty of Medicine, Tanta University

**Department of Pathology, Faculty of Medicine, Tanta University

Dimethyl diphenyl bicarboxylate (DDE) is a synthetic analogue ofSchizandrin C, one of the components isolated from Fructus Schizandrae, a traditional Chinese tonic. DDE has been widely prescribed for improvement of liver functions and as hepatoprotective agent inpatients with hepatitis or under cancer therapy. The purpose of this study was to evaluate the effect ofpretreating male albino rats with DDE at increasing oral daily doses (50, 150, 300 mg/kg) for 21 days on hepatic glutathione redox status in control and carbon tetrachloride (CCl^-intoxicated rats. Although, treating ruts with DDE did not produce any significant alteration in serum alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and sorbitol dehydrogenase (SDH) activities, it produced a gradual decrease in serum malondialdehyde (MDA) content. Pretreating rats with DDE at the same dosage regimen caused dose-dependent increase in hepatic glutathione reductase (GRD) activity. However the activities of glucose-6-phosphate dehydrogenase (G6PDH) and glutathione peroxidase (GPX) were down regulated to varying degrees with biphasic changes in hepatic reduced glutathione (GSH) level. CC\4 treatment caused drastic increase in serum ALT, LDH and SDH activities with concomitant increase in MDA level. Moreover, CCl4-intoxication was associated with marked alteration of the hepatic glutathione redox status. It was found that, the same DDE pretreatment regimen caused amelioration to the CCl4-induced hepatotoxicity. This hepatoprotection was assessed as marked drop in the activities of both ALT and LDH but not SDH, and it was associated with significant enhancement in the hepatic GSH redox status, as indicated by the substantial increase to the depleted GSH levels. Where the activity of GRD was increased over that of CC^ intoxicated rat; there was also a minimal elevation in G6PDH activity upon pretreatment with increasing doses of DDE. In contrast DDE has no remarkable beneficial effects on the appearance of CC14-induced histopathological changes. The ensembles of results suggests that the hepatoprotection afforded by DDE pretreatment may be mainly attributed to the improvement of the liver functions and to the improvement of hepatic GSH redox status.

15/9 MOLECULAR CHARACTERIZATION OF A SALMONELLA PARATYPHIB GENE (ROXA) WHICH CAUSES MULTIPLE-DRUG RESISTANCE IN ESCHERICHIA COLI

M. A. M. Yassien, H. Ewis*, C. Lu* and A.T. Abdelal*

Department of Microbiology and Immunology, Faculty of Pharmacy, Ain-Shams

University, Cairo, Egypt.

*Department of Biology, Faculty of Arts and Science, Georgia State University, Atlanta, Georgia, USA.

A multiple-drug resistant (MDR) clone E. coli DH5a (pNFXR42) was obtained by transformation of E. coli DH5a with the recombinants .obtainedfrom ligation of DNA fragments of MDR Salmonella, paratyphi B. mutant with pUC19. The sequence insert in pNFXR42 fragment was found to be 5.6 Kbp in length and contained an open reading frame which showed by computer analysis 100% identity in the nucleotide sequence to a putative S. typhimurium roxA gene (suggested from GenBank databases). When roxA gene was PCR amplified, ligated to pGEMT and expressed in E. coli DHSa strain (pNFXroxA), the same MDR phenotype was observed. Comparative analysis of the outer membrane proteins profiles of E. coli DHSa pNFXR42, E. coli DHSa pNFXroxA, and E. coli DHSa parent strain revealed that expression of roxA in E. coli DHSa caused disappearance of OmpFprotein and overproduction of TolC protein . Alignment of the amino acid sequence ofroxA proteins with other related proteins showed high degree of similarity to Klebsiella pneumoniae ramA, E. coli marA, and E. coli SoxSproteins (89%, 34%, and 38% identity, respectively). The potential helix-turn-helix DNA-binding domain found near the N-ter minus of ramA, marA, and SoxS, was also found in roxA. The results of the mobility shift assay showed that rox-A protein can recognize marA box and bind to it, indicating that development of multiple-drug resistance phenotype in pNFXR42 may be carried out by a mechanism similar to that which occurs in marA mutants. Accordingly, roxA can be considered as a newly identified member related to XylS-AraC family of transcriptional activators. The sequences ofroxA gene that was PCR amplified from DNA genomes of the S. paratyphi B parent and its mutant showed no difference in the sequence indicating that roxA gene was not responsible for the development of MDR phenotype in MDR S. paratyphi B mutant although its expression caused MDR phenotype in E. coli DHSa.

16/9  FLAVIPIN AND RED PIGMENTS FROM ASPERGILLUS SPECIES AND THEIR APPLICATIONS

S.M. Abou El-Souod, M. Bedaiwy, Y.A.-G. Mahmoud and E.H. Abd El-Zaher

Botany Department, Faculty of Science, Tanta University, Tanta, Egypt

Flavipin, red 1 and red 2 pigments were extracted from Aspergillus carneus, A. Flavuas var columnar is and A. ochraceus, respectively. The characters of pigments were studied by TLC, UV, visible spectra, solubility in solvents and IR spectrum. The effect of time course, temperature, pH, carbon source and nitrogen source onflavipin were studied. Crude extracted pigments were tested as dyeing material for dyeing the raw textile fabrics and their fastness to light, washing and rubbing were studied. The extracted pigments were also screened for their antimicrobial activity.

17/9  VITAMIN C MEDIATED MINIMISATION OF ROGOR AND MALATHION INDUCED MITOINHITION AND CLASTOGENY IN VICIA FABA

A.I. Shehata

Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia

Administration of vitamin C, either concurrently or aspre- and post-treatment to the pesticide exposure, was very much helpful in minimising mitoinhibition and clastogeny induced by two organophosphorus pesticides, Malathion and Rogor on Vicia faba-root tip cells. The concurrent treatment with vitamin C was more effective than the two other modes of its supplementation.

18/9  ISOLATION, SCREENING AND INDUCED MUTANTS BY

GAMMA IRRADIATION OF BACTERIAL ISOLATES

PRODUCING P-GALACTOSIDASE

M.R. Abo Shady*, M.Z. El-Fouly, T.M. El-Mongy and H.M. EL-Shafey. *Microbiology Department, Faculty oi Science. - Ain Shams University.

National Center for Radiation Research and Technology, P. O. Box 29, Nasr city,

Cairo, Egypt.

Escherichia coll W3, isolated from salt whey, was selected as the most active isolate for ft-galactosidase production, among twenty tested bacterial isolates, isolated from different local environmental sources. The best medium supported the production of the enzyme by this isolate was found to be Volbrecht broth medium. Utilization of sweet whey fortified with 2% yeast extract as a fermentation medium yielded an enzyme production which is nearly two-third that obtained using Volbrecht broth medium. The d10 value of E. coli W3 was calculated and found to be 0.395 kGy. Two mutants among eighteen tested isolates, after exposure to the sublethal irradiation doses, produced fi-galactosidase more than the wild type, however, they lost the advantage of increasing the production of enzyme on storage for 30 days at 4oC+1.


19/9  ULTRASTRUCTURAL CHANGES OF NICOTIANA

TABACUM LEAVES SYSTEMICALLY INFECTED WITH

YELLOW SEVERE STRAIN OF TOBACCO MOSAIC VIRUS

MM. El-Shamy and E.T.A. Sayed*

Botany Department, Faculty of Science, Menoufia University * Botany Department, Faculty of Science, Cairo University

Electron micrographs ofmesophyll cells ofNicotiana tabacum leaf systemically infected with Yellow Severe Strain (YSS) of Tobacco Mosaic Virus (TMV) revealed several ultrastructural changes. Virus particles occurred in the cytoplasm of both palisade and spongy cells, either singly or aggregated and sometimes in longitudinal. X-profile body containing viral protein in the form of bands were found either singly or in groups. The infected nucleus does not seem to contain virus particles but it sometimes developed an irregular boundary. The number of mitochondria increased. Deformed mitochondria were disrupted, their cristae and bounding membrane were disintegrated. Infected chloroplasts did not contain virus particles, showed breakdown of bounding membrane, degeneration of grana and disorganization of intergrana lamellae. Virus infection induced an increase in the number of osmiophilic globules and a presence of one or more very large starch grains. Virus particles distributed in the lumen of mature sieve elements and inside xylem vessel cavity. Sometimes very thick cell wall appeared.

20/9  EFFECT OF BENZYLADENINE OR COUMARIN ON SOME METABOLIC ACTIVITIES ASSOCIATED WITH THE GROWTH

OF RADISH SEEDLINGS

E.A.A. Foda and A.A. Mohsen

Botany department Faculty of Science, Tanta University, Tanta, Egypt

Study was conducted on radish seedling (Raphanus sativus L. var. egyptiaca) to investigate the impact of benzyladenine or coumarin on the growth parameters, chlorophyll content, transferase enzymes, nitrogen and nucleic acid content. The results indicated that percentage germination, radicle length and seedling dry weight of the high does of either benzyladenine or coumarin was equally suppressive. The results of the chlorophyll content indicated that the higher level of benzyladenine was less effective than its lower one, whereas, the both levels of coumarin induced the same increase in chlorophyll content. Glutamic - oxalacetic acid transaminase (GOT) activity was attenuated with benzyladenine and coumarin applications, whereas coumarin was more effective than benzyladenine. benzyladenine application resulted in glutamic-pyruvic acid transaminase (GPT) activity reached maximum activity whereas insignificantly affected by coumarin. The low doseofbenzyladeninewas slightly suppressive whereas the higher dose was slightly stimulatory to nitrogen accumulation a response that was reversed with coumarin application. Again the lowest does of benzyladenine remarkably increased RNA and DNA accumulation whereas the reverse the case with coumarin and the higher does ofBA.


21/9 EFFECT OF ENVIRONMENTAL FACTORS AND GAMMA

IRRADIATION ON THE PRODUCTION OF p-GALACTOSIDASE

ENZYME FROM ESCHERICHIA COLI

M.Z. El-Fouly*, T.M. El-Mongy*, M.R. Abo-Shady**and H. M. EL-Shafey*.

* National Center for Radiation Research and Technology, P. O. Box 29, Nasr city,

Cairo, Egypt.

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

Studying growth conditions for producing high levels of fl-galactosidase by Escherichia coll W$ isolate showed that the optimum period of incubation, incubation temperature, and initial pH of the medium were 24 hours, 30 °C, and 7.5, respectively. Lactose (3%) was found to be the best carbon source supported the enzyme production, while yeast extract (0.6%) was found to be the best nitrogen source supported the enzyme production. Adding ofl% sodium chloride to the fermentation medium stimulated the enzyme production. Using the optimum growth conditions together resulted in almost duplication of the enzyme production by the isolate Escherichia coll Wj. Low doses of gamma irradiation, up to 0.25 kGy resulted in slight increases of infra-cellular enzyme production, it reached its maximum production at 0.15 kGy with increasing of activity 34.26% than the control, while the production of extra-cellular enzyme decreases with all dose levels used.

22/9 PURIFICATION AND CHARACTERIZATION OF P-GALACTOSIDASE FROM ESCHERICHIA COLI

T. M. El-Mongy*, M.R. Abo-Shady**, M.Z. El-Fouly* and H.M. EL-Shafey*.

* National Center for Radiation Research and Technology, P. O. Box 29, Nasr city,

Cairo, Egypt.

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

Partial purification offt-galactosidase, was achieved by fractional precipitation with ammonium sulphate, acetone and ethanol. Ammonium sulphate method yielded the highest ft-galactosidase activity. Partially purified enzyme was purified 31.21 fold by fractionation on Sephadex G-100 as compared with the crude enzyme. Optimum temperature, pH and assay time for both crude and purified enzyme were found to be 40 °C, 7.2 and 20 minutes, respectively. The crude enzyme was found to be stable at the 4°C ± I for at least 60 days, while the purified enzyme lost 46.89% of its activity after 9 days. The enzyme activity (intracellulary) increased slightly with increasing doses of gamma irradiation till reached its maximum activity at 0.12 kGy, then the activity decreased with increasing irradiation doses. Meanwhile the enzyme activity (extracellulary) decreased with increasing doses of gamma irradiation and it lost 58.48% of its activity at 0.25 kGy.

23/9  EFFECT OF COPPER AND CADMIUM ON SOME GROWTH CRITERIA AND PHYSIOLOGICAL ASPECTS OF SORGHUM BICOLOR

W.A. Kasim

Botany Department, Faculty of Science, Tanta University, Tanta, Egypt

The application of Cu (Iff5 M), Cd (10~6 M) and a mixture of both (1:1 v/v) as sulphates to Sorghum bicolor (L.) Moench. Plants (broom corns) for two months from the beginning of germination in sandy soil irrigated with halfHoagland solution lead to. significant reductions in some growth criteria (fresh and dry weights and the water content of roots and shoots). There is a clear correlation between the extent of reduction caused by all 3 heavy metal treatments in leaf area andphotosynthetic activity. Myristic and heptadecanoic acids predominate the fatty acid composition, with 9 others in trace amounts. The former disappeared by Cufrom roots, by Cdfrom shoots to be replaced by the unsaturated linolenic acid, and from both organs by the combined treatment. The latter disappeared from roots by Cu, decreased in shoots and roots by Cd, and reaches about 86% of the total fatty acid content by Cu+Cd. Palmitic and arachidic acids replaced myristic and heptadecanoic acids when they disappeared from roots by Cu. Four minor fatty acids (caprilic, capric, stearic, oleic) disappear from roots and shoots by any metal treatment. Catalase and ascorbate peroxidase activities diminished with all metal treatments, with Cu having the most severe effect. All 3 treatments significantly increased the Ca2+ and 1C content of the roots, and decreased their Na+ content. The results are discussed in the light of previous studies.

24/9 UTILIZATION POSSIBILITY OF ECHINOCHLOA CRUS-GALLI FOR DECREASING SALINITY AND WASHING PURPOSE IN SALINE SOILS UNDER AGRICULTURAL RECLAMATION

E.A.A. Foda*, M.M. Migahid** and M.A. Elhaak* *Botany department Faculty of Science, Tanta University, Tanta, Egypt.

**Biology department Faculty of Education, Alexandria University,

Alexandria, Egypt.

Dry -matter production, leaf area, and some relevant growth parameters in addition to the nutrient elements accumulation were studied in Echinochloa crus-galli (Poaceac) growing as a common weed in rice fields. The plant grows with healthy vigor showing a remarkable tolerance to salinity up to 3 mmhos/cm. But salinity linearly reduced the plant shoot height, root length, dry weight of root, shoot and plant, leaf area, number of leaves as well as number of branches per individual. These inhibitory effects were concomitant with reductions in plant relative growth rate (RGR), net assimilation rate (NAR), leaf area ratio (LAR), leaf weight ratio (LWR) and specific leaf area (SLA) and enhancement in specific leaf weight (SLW). Most outcome of metabolism was directed toward root growth and accumulation of osmoregulatory compounds. The photosynthetic apparatus (number ofstomata and content of pigments) of the plant was not disturbed significantly by the imposed salinity stress. The plant accumulated considerable amount of salts, greater in shoot than in root. The study concluded the importance of using E. crus-galli plant in recovering the soil salinity which increases by extensive use of inorganic fertilizers and bad drainage in the soil of the Delta region.

25/9  THE ROLE OF POLYAMINE PRECURSORS; ARGININE, ORNITHINE OR METHIONINE IN AMELIORATING THE INHIBITORY EFFECT OF NaCl ON WHEAT PLANT

FA. El-Shintinawy and *R.A. Hassanein

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

*Botany Department, Faculty of Science, Ain-Shams University, Cairo, Egypt

Wheat plants subjected to different levels of NaCl showed a pronounced growth inhibition. Roots treated with 100 or 150 mMNaCl showed 40 and 68% reduction in their fresh weights relative to their respective control roots. Moreover, the fresh weight of shoots exposed to the above two salinity levels decreased to 28 and 12%, respectively compared with the control. Addition of 4 mM arginine, ornithine, methionine or phenylenediamine to the 100 mM NaCl-treatedplants increased their shoot fresh weights to 84, 74, 68 and 102%, respectively relative to the control. Also, they increased the salinized root fresh weights to 92, 136, 112 and 92%, respectively relative to control roots. Moreover, only 4 mM arginine, among the all tested chemicals, retarded the senescence of wheat leaves via preventing the loss of total chlorophylls induced by 100 mM NaCl. Meanwhile, salinity treatment with 100 mM NaCl induced 13 and 4.5 fold increases in both spermine (Spm) and spermidine (Spd) contents coupled with 4.5 fold decrease inputrescine (Put) level in the plant roots. In addition, 15 and 1.4 fold increases in Spm and Spd contents associated with 2.6 fold decrease in Put content were observed in the plant shoots. Addition of 4 mM methionine to the 100 mM NaCl-treated roots increased spermine content from 1391.75 to 589.5 jug g'1 fresh weight with NaCl only. Moreover, ratio of total polyamine/Put increased 16 and 36 times in salinized roots and shoots, respectively compared to their non-salinized organs. Addition of 4 mM arginine or ornithine to the shoots or roots exposed to 100 mM NaCl decreased the ratio of total polyamine/Put almost to that of control of both organs. Moreover, 4 mMphenylenediamine in the presence of 100 mM NaCl decreased the ratio of total polyamine/Put of the two organs compared to those recorded in salinized plants, since the greater ratio is an indication of the more deleterious injury induced by salinity stress.

26/9 GERMINATION POTENTIAL, EARLY GROWTH, AND MODULATION OF PROTEIN PATTERNS OF JOJOBA UNDER SALINITY STRESS AND POSSIBLE AMELIORATION BY GIBBERELLIC ACID

MM. El-Araby,   A.H. Nassar, and   H.F.M. Shaaban

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

Jojoba (Simmondsia chinensis (Link) Schneider) is a newly introduced industrial crop in Egypt. Seed germination and subsequent growth of seedlings were found to be sensitive to salinity within the first 6 days, but adaptation seemed to be taking place afterwards where the adverse effect of salinity, even at 250 mM, was markedly attenuated after 12 days. Moreover, amelioration could be achieved by soaking the dry seeds in a SOppm gibberellic acid (GAj) solution for 6 hours prior to sowing. Concomitant changes in protein banding patterns mainly manifested induction and repression of certain treatment-specific proteins (Mwts : 97,78,42 and 27 KDa). The two with higher molecular weights (97 and 78 KDa) were assumed to be NaCl-repressible and could be reverted by GAs (SOppm). The latters (42 and 27 KDa) were induced by NaCl treatments (50, 150 and 250 mM) where densitometer tracing showed a progressive enhancement with increase of salinization level. Reversibly, the occurrence of these two proteins could be blocked by GA$ treatment, inspite of subsequent supply with NaCl solutions.


27/9  MODULATION OF PROTEIN PATTERNS, GENOMIC DNA AND PHYTOHORMONES BY POLYAMINES DURING ROOTING OF COW PEA HYPOCOTYL CUTTINGS

M.M. El-Araby

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

Spermidine (Spd) at 0.15 mM induced maximum enhancement of adventitious rooting than putreseine (Put) at 0.5 mM and spermine (Spm) at 0.15 mM, in hypocotyl cuttings of Vigna sinensis. Endogenous changes in polyamine (PA) concentrations were recorded irrespective of exogenously applied types. Thus, exogenous supply of Spd was associated with a substantial accumulation of endogenous Spm and this pattern underlyed maximum rooting potential after two days of enhancement. More or less similar trends were observed with endogenous PAs in association to exogenous supply with Put or Spm. The results concluded diminish of Put and elevation of higher PAs, especially Spm during the earlier stage of rooting and Spd, at the subsequent later stage. Modulation of protein patterns was recorded during enhancement of adventitious rooting of cow pea hypocotyl cuttings, in response to treatments with 0.5 mM putrescine (Put), and 0.15 mM of each ofspermidine (Spd) and spermine (Spm). It could be revealed from the results obtained that a modification of the gene expression was assumed to be achieved. This could be confirmed by the concomitant changes in genomic DNA fragments, in response to Put, Spd and Spm treatments. PCR-RAPD analysis using jive arbitrary decamer primers indicated the induction of three DNA fragments by either Put or Spd, whereas five fragments were induced in response to treatment with Spm. The modulation of endogenous PA concentrations at different stages of rooting was generally concomitant with enhancement of endogenous levels oflAA, ABA, cytokinins and coumarins, particularly at the earlier stage of rooting induction. The changes in GA$ concentration indicated a more pronounced elevation at the later stage. The different results are discussed from the view of possible role of PAs in adventitious root formation.

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