REGENERATION OF PLANTS IN EMS TREATED LOCAL MUNG BEAN (VIGNA RADIATE L. WILCZEK) UNDER SALT STRESS
By Muhammad Rafiq, Mahadev Mali, S. H. Ahmad Naqvi, M. Umar Dahot, Hafiza Faiza and Arsala Khatari
The regeneration of EMS treated seeds of two local varieties of mung bean was investigated under salt stress conditions. Three explants e.i. cotyledon, leaf and shoot apical meristem of mung bean obtained from in vitro grown EMS treated seeds of varieties NM-92 and Khalood were inoculated on MS medium containing various concentrations of 2, 4-D and BAP and IBA. The highest callus proliferation (98.3%) was obtained in EMS treated leaf of NM-92 followed by 96.5% in shoots of NM-92 control on MS medium supplemented with 1.8ÁM 2, 4-D and 3.56ÁM BAP. The increase in callus fresh weight was higher in leaf explants of both treated varieties. Similarly 57.3▒1.3 callus cultures of treated leaf explants of NM-92 were survived on media containing 50mM NaCl. Similarly the highest regeneration (65.5▒2.1%) was observed in NaCl selected callus cultures raised from shoots of EMS treated seeds.
DETECTION OF BANANA BUNCHY TOP VIRUS IN VIRUS-INFECTED PLANTS USING POLYMERASE CHAIN REACTION
By Sadik, A.S.1,2,3 , El-Afifi Sohair I.1 , Harding, R.M.2 , Dale, J.L.2 and Allam, E.K.1
Polymerase chain reaction (PCR) was developed for the detection of Banana bunchy top virus (BBTV) at maximum after 210 min and at minimum after 90 min using Pc-1 and Pc-2, respectively. PCR detection of BBTV in crude sap indicated that the freezing of banana tissue in liquid nitrogen (LN2) before extraction was more effective than using sand as the extraction technique. BBTV was also detected using PCR assay in 69 healthy and diseased plants using Na-PO4 buffer containing 1 % SDS. PCR detection of BBTV in nucleic acid extracts using seven different extraction buffers to adapt the use of PCR in routine detection in the field was studied. Results proved that BBTV was detected with high sensitivity in nucleic acid extracts more than in infectious sap. The results also suggested the common aetiology for the BBTV by the PCR reactions of BBTV in nucleic acid extracts from Australia, Burundi, Egypt, France, Gabon, Philippines and Taiwan. Results also proved a positive relation between the Egyptian-BBTV isolate and abaca bunchy top isolate from the Philippines, but there no relation was found with the Cucumber mosaic cucumovirus (CMV) isolates from Egypt and Philippines and Banana bract mosaic virus (BBMV) were found.
STREPTOMYCES SPECIES ABLE TO UTILIZE SOME HERBICIDES AS NITROGEN AND CARBON SOURCES
By Zaki M.M.1 , E.A. Saleh1 , A. Rahal2 and Sonya H. Mohamed2,3
The present work was designed to isolate and identify some actinomycetes able to degrade Basta (glufosinate) and Sencor (metribuzin) herbicides, which are widely used for weed control in Egypt. Results showed that 100 isolates of actinomycetes were isolated and purified from the rhizosphere soils of 11 different crops (barley, broad bean, clover, cotton, corn, grape, cantaloupe, pepper, sesame, tomato and wheat) treated with pesticides. The tolerance of the actinomycete isolates for Basta and Sencor herbicides were determined. Results showed that, 70 out of the 100 actinomycete isolates were able to grow on the recommended dose of Sencor (0.75 g/L) but 24 out of them were showed a good growth on the ten folds of the recommended dose of Sencor (7.5 g/L). At the same time, 38 actinomycete isolates grew on the recommended dose of Basta (2 g/L) and 18 of them appeared a moderate growth on 20 g/L of Basta herbicides. In addition, the ability of the 70 isolates to utilize the Sencor as carbon and/or nitrogen source was studied. Results showed that, 9 out of 70 actinomycete isolates gave a good growth on the starch nitrate agar medium containing the Sencor as a sole nitrogen source, while no isolates were found to be able to grow on the same medium with the Sencor as a sole carbon source.In this study, 5 isolates were biologically identified and found to be strains of Streptomyces rectiviolaceus, S.roseolus, S.albosporeus subsp abilomycaticus, S. herbaricolor and S. aureomonopodiales.
BIODEGRADATION OF SENCOR HERBICIDE BY SOME TREPTOMYCETES IN LIQUID CULTURE
By Sonya H. Mohamed1 , A. Rahal1 ; Zaki M.M.2 ; E.A.Saleh2 and A.S. Sadik2,3
This work was designed to study the role of streptomycetes in the biodegradation of Sencor herbicide and its persistence in liquid culture. Testing the biodegradation abilities of nine isolates of Streptomyces in vitro revealed that they differed greatly in the number of degradation-products produced from Sencor. Gas liquid chromatographic analysis revealed that all tested isolates began to degrade Sencor after 15 days from incubation. Number of compounds varied from one to five depending upon the isolate used, as the streptomycete isolates varied greatly in their abilities to degrade Sencor as indexed by the number of compounds produced from Sencor degradation. The products of Sencor degradation were more demonstrated after 30 and 45 days from incubation. Half life period of the herbicide was reached after 15 days in S. aureomonopodiales and mixture treatments, while was reached in the other treatments after 30 days. The present results could be considered as an additional prove for the in-vitro abilities of these streptomycete isolates to degrade and utilize Sencor as a sole nitrogen source in culture medium.
DNA FINGERPRINTING OF SOME HERBICIDE-TOLERANT STREPTOMYCES SPECIES AND PCR-ISOLATION OF BAR GENE
By Saleh, E.A.1 ; Zaki M.M.1 ; Rahal, A.2 ; Mohamed Sonya H.2,4; Bahieldin A.M.3 and A.S. Sadik1,3,4
In this study, differentiation between five identified Streptomyces species was carried out on the basis of the nucleotide sequence of DNA using random amplified polymorphisms of DNApolymerase chain reaction (RAPD-PCR) technique. Results revealed that these species differed in their DNA sequence as indexed by differences in numbers, sizes and unique amplified fragments. Such dissimilarities could be considered as an additional proof that identified isolates were different species of Streptomyces. In addition, the herbicide resistance (bar) gene was isolated by PCR technique in the DNA extracts of the five identified Streptomyces isolates (Streptomyces albosporeus subsp abilomycaticus, S. herbaricolor and S. aureomonopodiales) to be available for using as a selectable marker in the transformation of the transgenic plants using the Basta herbicide. It is the first evidence to detect this gene in local Streptomyces species in Egypt.
CHARACTERIZATION OF STREPTOMYCETES HAVING ANTIBIOSIS ACTIVITIES ISOLATED FROM SOIL IN WESTERN REGION OF KSA
By Shori Ghadeer B.O.1 , Mohamed Sonya H.1,2, Abdel-Salam Shimaa M.3 and Sadik A.S.1,4
In this study we are focused on the color groups of actinomycetes, in particularly streptomyctes, in soil of western region of KSA. Therefore, we collected soil samples from different climate locations in KSA (Taif, Makah and Jeddah).The color groups of the isolated actinomycete isolates were determined. The antagonistic activities of the isolated actinomycetes were also tested against seven microorganisms including, bacteria and fungi. The highest active isolates were identified as strains of S. polychromogenes (isolate 08), S. chattanoogensis (isolate 14), S. lucensis (isolate 20), S. violaceus (isolate 21), S. violans (isolate 32), S. griseorubiginosus (isolate 34), and S. antibioticus (isolate 35). It was show that the 7 selected streptomycete isolates were able to grow in the presence of 7% NaCl in the starch nitrate agar medium. At concentration of 10.5% NaCl, four isolates grew with weak growth (+) and three isolates showed in-doubt growth (▒). Keywords: Actinomycetes, Streptomyces, Identification, Taif, KSA
EFFECT OF SODIUM AZIDE ON THE GROWTH OF CAPSICUM ANNUUM (CHILI)
By M. Umar Dahot, M. Rafiq, Ambreen M. Arif and S. H. Ahmed Naqvi
The present study was carried out to check the mutagenic effects of 0.5, 1.0 and 2.0% NaN3 on the germination and growth of Capsicum annum (Chili). The results showed that 0.5% NaN3 was most appropriate for the creation of mutagenesis and was not highly lethal. The higher germination rate was observed in control and 0.5% NaN3 treated seeds but germination rate was decreased in 1.0% and 2.0% NaN3 treatment. The variation was observed in plant fresh and dry weight, total and reducing sugars and total protein contents of plants in different concentrations of NaN3 treatments.
Isolation and Identification of some halotolerant Actinomycetes having antagonistic activities against some plant pathogens (i.e., Tobacco mosaic virus, Aspergillus Sp., Fusarium Sp.) from soil of Tai
By Mohamed Sonya H.1 , W.M.Omran 1 , M.Abdel-Salam Shimaa2 , A.S.Al-Shehri1 and A.S.Sadik
Actinomycetes are considered one of the important bacteria due to their ability to produce several substances as secondary metabolites, found to be effective in the control of some plant pathogens. In this study, a number of soil samples were collected from different locations of Taif as well as Jeddah and Makah. This was followed by determination of the microbial total counts, i.e., bacteria, fungi and actinomycetes. Actinomycetes were isolated and purified and their color groups were determined. The actinomycete isolates were identified based on their cultural and morphological properties as Streptomycetes. The salt tolerance range of the purified Streptomyces isolates were determined by growing them on starch nitrate agar medium supplemented with different NaCl concentrations ranged from 3.5 to 10.5%. The highly tolerant streptomycete isolates were grown on a starch nitrate broth medium for 6 days under shaking conditions at 28-30oC. Then, their antiviral and antifungal activities against TMV, Aspergillus sp. and Fusarium sp. were determined. The halotolerant Streptomycete isolates having antifungal and/or antiviral activities were completely identified based on their cultural, morphological and physiological properties. The DNA of the identified isolates was extracted and used for determination of DNA fingerprinting of these isolates using the RAPD-PCR molecular tool.
Acknowledgment of Reviewers
|Prof. Dr. Alexandre Semenov||Moscow, Russia|
|Prof. Dr. Atef S. Sadik||Taif, Saudi Arabia|
|Prof. Dr. Sher Muhammed Mangrio||Uni. Of Sindh, Jamshoro, Pakistan|
|Prof. Dr. Jian He Xu||Shanghai, China|
|Prof. Dr. Muhammed Umar Dahot||Uni. Of Sindh, Jamshoro, Pakistan|
|Dr. S. A. Anitha Christy||Houston, USA|
|Dr. Mostafa Rahimnejad||Univ. Babool, Iran|
|Dr. Muhammed Rafiq||Uni. Of Sindh, Jamshoro, Pakistan|
|Dr. S. Habib Ahmed Naqvi||Uni. Of Sindh, Jamshoro, Pakistan|
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