USE OF MOLECULAR MARKER TECHNOLOGY: WHEAT (AN OVERVIEW)
By M. A. Sial, S. M. Mangrio* , M. U. Dahot**, Mazhar H. Naqvi, B. Nisa Mangan M. A. Arain, and Shabana Memon***
Molecular markers technology has significant impact in identification; isolation of genes or genomic regions associated with any discrete trait and manipulated genomic regions through marker associated selection. Selection of elite material through marker-assisted selection (MAS) provides an efficient complementary breeding tool. Genotypes selected on the basis of desired traits can be differentiated through molecular markers. Molecular markers allow cultivars identification in early stages of plant development as being neutral to environmental effects. A large number of molecular markers associated with different traits have been identified in various plants. This paper will focus a brief overview on molecular markers their useful in plant breeding particularly for wheat
BIOLOGICAL CONTROL OF SIX SOIL-BORNE FUNGI OF COTTON USING ANTAGONISTIC STREPTOMYCES ISOLATES
By Mansour M.T.M.1 and Sonya H. M. Hussein
In this study, antagonistic activity of fifteen isolates of Streptomyces, in vitro or in vivo, was determined against six fungi causing cotton seedling diseases. Data showed that all Streptomyces isolates were highly antagonistic in-vitro studies, isolate Sc-2 was the most effective against Fusarium solani (antagonism distance (AD) was 1.72 mm), on the other hand, isolates Qa-53, Ps-12 showed the lowest antagonism against F. solani (AD were 0.80, 0.83 mm, respectively). Rhizoctonia solani was the most sensitive fungus in vitro to the antagonism of most Streptomyces isolates (Qa-84, Qa-51, Is-10, Ps-12, Si-1, Si-4, Si-6, Si-8 and Si-9) with their AD ranged from 2.27 to 2.70 mm. However, Sc-2 was the least effective with AD as low as 0.93 mm. F. moniliforme was the least sensitive fungus to the antagonism of Streptomyces isolates. Antagonisms of Streptomyces isolates against F. solani; Sclerotium rolfsii and R. solani were positively correlated with their antagonisms against F. oxysporum (P0.01), R. solani (P0.05) and Macrophomina phasulina (P.0.01), respectively, but negatively correlated against F. moniforme. Treated fuzzy seeds with Streptomyces significantly reduced seedling disease in Sc-11, Ma-13, Ps-12, Si-1 and Si-6. All Streptomyces isolates were ineffective in controlling the disease when the seeds were acid delinted. Some Streptomyces isolates showed no efficiency on reducing the seedling disease wheather the seeds were fuzzy or acid delinted such as Sc-2, Qa-44, Qa-51, Qa-53, Qa-84, Da-3, Is-10, Si-4, Si-8 and Si-9.
A NOVEL POINT MUTATION IN rpsL GENE OF STREPTOMYCES COELICOLOR WITH ENHANCED BLUE PIGMENT PRODUCTION
By Hechun Zhang, Haizhen Wu, Huizhan Zhang, and Yuanxing Zhang*
Three strains showing enhanced blue pigment production were screened among thirty streptomycin resistant mutants of Streptomyces coelicolor 100. Their yields of blue pigment were all improved, especially for mutant S-3. Analysis of the mutants revealed that a point mutation occurred in mutant S-3. In rpsL gene, A to G transition at the position 128 resulted in Lys-43 to Arg substitution in the ribosomal protein S12 encoded by rpsL gene. There was no mutation in rpsL gene of other two streptomycin resistant mutants. Compared with the wild type strain, the mutant S-3 showed increased blue pigment yield and slightly decreased growth rate. This strategy of streptomycin resistance induction could be especially effective for improving the blue pigment production from the wild type strain of Streptomyces coelicolor.
A COMPARATIVE STUDY BETWEEN A NUMERICAL AND RAPD-PCR METHODS FOR THE IDENTIFICATION OF SOME STREPTOMYCETE STRAINS
By Sonya H.M. Hussein1 , Hassan I. Abdel-Fattah2 and Zubeda Chaudhry3
Streptomyces is a gram-positive bacterium that undergoes morphological differentiation. To taxonomy or classify the streptomycetes according to their properties some systems have been proposed. In this study, a comparative study between a numerical method and the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) technology for identification of 19 streptomycetes belonging to different series color (gray, 4 strains; red, 8 strains divided into three groups; white, 3 strains; violet, 2 strains and yellow, 2 strains) was carried out. Results showed that units of utilization of carbon sources (UCS) were effective for some Streptomyces strains. As expected the violet Streptomyces strains (Si-9 and Si-1) of group 5 showed the lowest total units. Similarity differences based on the numerical and RAPD-PCR analyses of the four gray Streptomyces strains (group 1: ST10, ST11, ST12 and ST13) were ranged from 1.7 to 9.5%. The phylogenetic tree reflected the relationship between the identified Streptomyces strains (group 3: ST1, ST2 and ST3), as the strains were arranged as follows: RF/C-/SM; RF/C+/SM and RA/C+/SM. Interestingly, the similarity between the phylophenetic and phylogenetic trees of the Streptomyces strains (ST08, ST14 and ST15) was 100% and strains were organized in a compatible situation with their characters. Data also showed reasonable differences (9.5% and 6.8%) between the applied analysis methods in case of Streptomyces strains of groups 5 and 7, respectively. Finally, the phylophentic data of this study provided a basis for reduction of the number of Streptomyces species now known to occur in nature.
GENETICAL STUDIES AND ANTAGONISTIC EFFECTS OF A NEWLY BACTERIAL FUSANT AGAINST MELOIDOGYNE INCOGNITA, ROOT-KNOT NEMATODE, AND A PLANT PATHOGEN FUSARIUM OXYSPORUM INFECTING SUNFLOWER
By El-Hamshary, O.I.M1 ; El-Nagdi, W.M.A. 2 and Yousuf M.N.A2
All tested bacterial concentrations of P.aeruginosa and fusant strain (Psa::Psf), between Pseudomonas fluorescens and Pseudomonas aeruginosa, affected Meloidogyne incognita juveniles J2 survival in invitro. The mortality percentages of nematode were dependent on the bacterial concentration and exposure time. The fusant strain (Psa::Psf), was used as soil drench or as seed soaking for controlling root-knot nematode Meloidogyne incognita, which was associated with sunflower plant (Helianthus annus) under greenhouse conditions. The fusant strain proved to be more effective than its parental strain, P. aeruginosa, in reducing different nematode parameters(give names) as well as enhancement of plant growth. Soil drench treatment was more effective in controlling root-knot nematode than the seed soaking treatment. The genetic stability of antagonist against Fusarium oxysporum of the fusant and its parental strain were 83% and 72%, respectively. RAPD-PCR studies indicated the same DNA-patterns among the fusant and its reisolated offspring. Three fusant isolates were selected, from which two isolates showed the same band when using two primers, while one isolate did not produce any band. Plasmid profile studies revealed the occurrence of one plasmid, of about 23 kb, in the fusant and its offspring while no plasmid has been found in the parental strain. The corresponding genes of antagonist against Fusarium oxysporum were located on the bacterial chromosome. From the present results, it could be advised to use modified genetically bacteria for management of the root-knot nematode.
INTRACELLULAR INVERTASE AND SUCROSE HYDROLYSIS BY CALCIUM ALGINATE ENTRAPPED MUTANT CELLS OF SACCHAROMYCES CEREVISIAE NA-47
By Aafia Aslam, Sikander Ali and Ikram-ul-Haq
Biocatalyst by entrapping the whole mutant yeast cells of Saccharomyces cerevisiae NA-47 into calcium-alginate for the production of inverted syrup (glucose and sucrose) from sucrose as substrate. Out of M1-M8, the medium M2 containing glucose (2.0 %) as carbon source was selected for cell growth and intracellular invertase after the time interval of 48 h. The optimum conditions for sucrose hydrolysis were as sucrose (50 %), alginate beads were repeatedly used of 26 days after every 18 h of incubation time. The beads were also stored at 4°C for 6 months without appreciable loss of the invertase activity
DETECTION AND IDENTIFICATION OF BEET MOSAIC POTY VIRUS (BTMV) INFECTING SUGAR BEET PLANTS IN EGYPT
By R.A. Omar*, S.A. Sidaros*, S.A. El-Kewey*, Hayam S. Abd El-Kader**, A.A. Deif** and Jehan M. Abass**
The virus was isolated from naturally infected sugar beet plants and identified as BtMV according to symptomatology, host range, and modes of transmission, electron microscopy and immunological and molecular detection. Host range studies revealed that, BtMV infected 28 plant species belonging to 6 families. BtMV was not easily sap transmitted, not transmitted through seeds but was readily transmitted by Myzus persicae Sulz. and Aphis fabae Scorp. The electron microscope preparations revealed flexuous filamentous virus particles 715-775 nm in length. The purified virion protein of BtMV and the total protein extracted from infected beet tissues showed a strong cross reaction with Bean Yellow Mosaic virus (BYMV) polyclonal antibodies in dot blot immunoassay indicating a higher degree of relatedness. First strand cDNA synthesis of BtMV was synthesized by priming the 3 terminal poly (A) tail of the viral RNA with Oligo (dT). For cDNA amplification, Potyvirus specific primers corresponding to the non-coding region upstream of the coat protein gene was used. A PCR fragment of the expected size approximately 220 bp was amplified and its size was estimated by agarose gel electrophoresis.
MOLECULAR CHARACTERIZATION OF FOUR POTATO CULTIVARS (SOLANUM TUBEROSUM L.) COMMERCIALLY GROWN IN EGYPT
By Badr1 , E. A.; S.A. Riad1 ; Hadia A. Heikal2 and F.A. Aly1
The present study aims to identify four different cultivars (i.e. Cara; Lady Rossita; Nicola and Spunta) of potatos (Solanum tuberosum L.) that are commonly planted in Egypt the tissue culture of each. In this respect, the tissue culture of each was established, while Spunta explants were considered best than others for invtro responses, when cultured on. Than different tissues (green leaves; calli and regenerated shoots) each cultiver used for biochemical and molecular analysis to measure genetic relationship among them when MS medium supplemented with 0.5mg/L NAA; 2.24mg/L 6-BAP and 8mg/L GA3. Each cultivar differed for isoesterase and isoperoxidase electrophoretic patterns, as the highest differential expression of esterases and peroxidases were screened for regenerated shoots and callus tissues, respectively which may be due to a differential protein activation during with developing tissues. Furthermore, RAPD analysis fingerprints provided a number of useful DNA markers to distinguish and identify the four cultivars. According to the data of the six primers for all cultivars, the highest percentage of DNA polymorphism was screened for regenerated shoots. Such feature may be due to the somaclonal variation for in vitro culture, which is producing sequence dissimilarity among the original genotypes. Probably, due to this phenomenon, RAPD based dendrograms for green leaf; callus and regenerated shoot genomes revealed different genetic relationships among the four cultivars. Therefore, it may be concluded that plant leaves are preferred to design dendrograms of similarities and relationships among different cultivars. However, generally the somaclones may be a way of generating useful genetic variation and selection of desired traits for in vitro Plantlets.
ROLE OF BIOFERTILIZERS IN ACCUMULATION OF NICKEL IN SUNFLOWER (HELIANTHUS ANNUUS L.) AND THE EFFECT OF NICKEL ACCUMULATION ON ENDOGENOUS LEVEL OF HORMONES IN SUNFLOWER
By * S.Tatheer Naqvi, *Asghari Bano, *Safia Ahmed, **Nisar Ahmed, *Irfan Ali and *Jahangir Asad
The objective of the study was to determine the effect of Biofertilizers i.e. Phosphate solublizing bacteria (PSB), Effective Microorganism (EM) and Rhizobium in the uptake and accumulation of nickel by different parts of sunflower (Helianthus annuus L.). The results revealed that all biofertilizers has increased accumulation of nickel significantly as compare to control during both stages. At different concentrations of nickel that were applied externally to the plants, different fertilizers show different behavior. At lowest external concentration of nickel Rhizobium treated plants has maximum accumulation while at highest external concentration of PSB and EM treated plants accumulate maximum nickel. Translocation of nickel in treated plants was also increased as compare to control plants during vegetative stage of development while insignificant during 50% flowering stage. Accumulation of nickel was also increased as the external concentration increased. In addition, effect of nickel accumulation on Gibberellins (GA) and Indole-acetic acid (IAA) contents were also observed in leaves of sunflower at vegetative stage. Both GA and IAA contents were affected by nickel accumulation.
SIMULTANEOUS DETERMINATION OF MONOPHENYLTIN, DIPHENYLTIN AND TRIPHENYLTIN BY HPLC WITH A UV DETECTOR AND ROLE OF THIOL COMPOUNDS
By Wen-Qiang Zhou1 ; Guo-Xin Sun1 ; Yue-Jiang2 and Jian-Jiang Zhong3*
Simultaneous determination of monophenyltin (MPT), diphenyltin (DPT) and triphenyltin (TPT) by a UV detector was developed using high performance liquid chromatography (HPLC) with methanol–acetic acid–water mixtures containing thiol compounds as mobile phase. Peaks of TPT, DPT and MPT were completely separated and they were detected using UV detector at 257 nm. The effects of thiol compounds, i.e. dithiothreitol (DTT), dithioerythritol (DTE) and 2-mercapoethanol (ME) on the peak heights and retention times of phenyltin compounds were investigated, and the optimum performance was achieved with methanol–acetic acid–water (60:10:30 [v/v/v]) containing 5 mM DTE. The different affinity of thiol group with different phenyltins was found helpful to the separation of phenyltins. The developed method was successfully applied to the analysis of phenyltins in organotin biodegradation by Pseudomonas aeruginosa.
BREEDING FOR HIGH GRAIN YIELD THROUGH MUTAGENESIS IN RICE (ORYZA SATIVA L.)
By A.W. Baloch, A.M. Soomro, H.R. Bughio, M.S. Bughio, I.A. Odhano, M.A. Asad and S.J. Abbasi
A high yielding mutant IR6-25A was selected from gamma rays irradiated material of coarse (non-aromatic) rice variety IR6 and was evaluated at the experimental farm of Nuclear Institute of Agriculture (NIA) Tando Jam. The mutant strain IR6-25A was significantly (P 0.05) better than its mother variety IR6 and check variety Shadab in the entire yield contributing parameters. It gave better paddy yield as compared to its parent, check other promising mutant strains in Micro, Preliminary and Station Varietal Trials at NIA, Tando Jam during 1996, 1997 and 1998. On the basis of three years average, it has shown 16% increase in paddy yield over its parent-IR6 as well as check variety Shadab. In Zonal Varietal Trials, conducted for three subsequent years at 9 different locations with diverse agro-climate conditions, it maintained superiority over all the entries by yielding 6816, 6208 and 6530 kg of paddy per hectare during 1999, 2000 and 2001 respectively. It gave 20% and 23% higher paddy yield than IR6 and check variety Shadab.
DETECTION AND IDENTIFICATION OF AN EGYPTIAN ISOLATE OF CUCUMBER MOSAIC VIRUS (CMV) AFFECTING SUGARBEET USING NON-RADIOACTIVE RNA PROBE
By 1Abdelkader, H. S.; 1Khatab, E. H.; and 2Abd El Fattah A.I.
Detection and identification of cucumber mosaic virus (CMV) RNA was carried out from sugar beet leaves by using reverse transcription and polymerase chain reaction (RT-PCR). The coat protein gene (RNA 3) was amplified selectively by using cucumovirus-specific coat protein gene primers. No DNA product of any length was produced when healthy leaves or tobacco mosaic virus RNA was used as templates. Dot-blot hybridization using CMV-CP/RNA probe also confirmed the presence of CMV genome in sugarbeet-infected tissues. The double-stranded PCR product (650 bp) was cloned into pGEM-T-Easy vector and the insert was partially sequenced. The multiple nucleotide sequence alignment of the CMV/CP-EG displayed 75 % homology with CMV subgroup II published nucleotide sequences of Fny-CMV, NT9-CMV, Q-CMV and Trk7-CMV strains.
Acknowledgment of Reviewers
|Prof. Dr. Alexandre Semenov||Moscow, Russia|
|Prof. Dr. A.A.Saboury||Tehran, I.R. Iran|
|Prof. Dr. Yousefi||Tehran, I.R. Iran|
|Prof. Dr. Jian He Xu||Shanghai, China|
|Prof. Dr. M.R.Zamani||Tehran, I.R. Iran|
|Prof. Dr. Hossain Massumi||Kerman, Iran|
|Prof. Dr. Muhammed Umar Dahot||Uni. Of Sindh, Jamshoro, Pakistan|
|Prof. Dr. Aijaz Rasool Sheikh||Karachi, Pakistan|
|Dr. M. Motallebi||Tehran, Iran|
|Prof. Dr. Rafi Arain||Uni. Of Sindh, Jamshoro, Pakistan|
|Prof. Dr. Atta-u-Rehman||Uni. Of Sindh, Jamshoro, Pakistan|
|Prof. Dr. Asif Ali||Faisalabad, Pakistan|
|Prof. Dr. Imtiaz Ahmed Khan||Faisalabad, Pakistan|
|Dr. Muhammed Ibrahim Rajoka||Faisalabad, Pakistan|
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