RESPONSE OF BALANITES AEGYPTIACA (L.) DEL. VAR. AEGYPTIACA SEEDLINGS FROM THREE DIFFERENT SOURCES TO WATER AND SALINITY STRESSESS
ABSTRACT:
Water and salinity are main co-occurring stresses affecting plant growth and development in arid lands. In this study interactive effects of water and salinity stresses on Balanites aegyptiaca seedlings from three different sources (SD5.1, SD6.2 and KSA) were assessed in potted experiment under greenhouse conditions. The effect was measured on stomatal conductance (Gs), specific leaf area (SLA), seedling quality (Shoot to Root ratio (S/R), Dickson Quality Index (DQI) and Sturdiness Quotient (SQ)), Nutrient uptake (N content, K/Na and Ca/Na ratios) and growth. The seedlings were either watered twice a week (well watered) or every two weeks (water stressed), in addition to four salt concentrations (fresh water as control, 5 dS m-1, 7 dS m-1 and 9 dS m-1 EC). Water and salinity stresses resulted in reduced Gs, SLA, DQ, SQ and S/R, associated with lower height and root collar diameter. However, irrespective of salt concentration, water stressed seedlings displayed substantial reduction in Gs, indicating that Gs is among the most important water conservation strategy for this species. S/R also, remarkably decreased in water stressed seedlings, but, within watering treatment it was increased with increasing salt concentration. SLA and DQI were more affected by salinity stress, due to the increased leaf weight with increasing salinity. N content was more sensitive to water stress than salinity. Both Ca/Na and K/Na ratios were decreased with increasing salt concentration. The three sources exhibited significant variation in their response to water and salinity stresses. SD5.1 displayed higher values in most of studied traits. Gs and S/R may be considered as fitness responses of this species to water stress, while DQI, SLA and K/Na can serve as good indicators to measure response to salt stress.

COMBINED EFFECTS OF DROUGHT STRESS AND NPK FOLIAR SPRAY ON GROWTH, PHYSIOLOGICAL PROCESSES AND NUTRIENT UPTAKE IN WHEAT
ABSTRACT:
The present study investigated the effects of supplemental foliar nitrogen (N), phosphorous (P) and potassium (K) spray, alone or in various combinations, on physiological processes and nutrients uptake in wheat under water deficit conditions. The study comprised of two phases; during the first phase, ten local wheat (Triticum aestivum L.) genotypes were evaluated for their response to PEG-6000 induced osmotic stress. One drought tolerant (Bhakkar-2002) and sensitive (Shafaq-2006) genotype selected from screening experiments were used in the second phase to determine the individual and combined effects of N, P and K foliar spray on physiological mechanisms in wheat under drought stress. The results revealed that limited water supply significantly reduced germination, growth and uptake of N, P and K. Supplemental foliar fertilisation of these macronutrients alone or in different combinations significantly improved the water relations, gas exchange characteristics and nutrient contents in both the genotypes. Bhakkar-2002 maintained higher turgor, net CO2 assimilation rate (Pn), transpiration rate (E), stomatal conductance (gs) and accumulated more N, P and K in shoot than Shafaq-2006. The foliar spray of NPK in combination was effective in improving wheat growth under both well-watered and water-deficit conditions.

CHARACTERIZATION OF ABIOTIC STRESS GENES FROM DIFFERENT SPECIES OF EUCALYPTUS
ABSTRACT:
The stresses causing dehydration damage to the plant cell like cold, drought, and high salinity are the most frequent environmental stresses that influence plant growth, development and restraining productivity in cultivated areas world-wide. Many drought, salinity and cold inducible genes causing tolerance to environmental stresses in many plants include Dehydrin1 (DHN1), Dehydrin2 (DHN2), Dehydrin10 (DHN10), putative phosphate transporter (Ecpt2), choline monooxygenase (CMO) and DREB/CBF1c genes. Gene specific primer pairs were designed for each gene using DNAStar software. These genes were amplified from different species of eucalyptus such as Eucalyptus camaldulensis, E. globulus, E. tereticornis and E. gunii through PCR. Dehydrin2 gene of E. camaldulensis and dehydrin10 gene of E. globulus were cloned using the TA Cloning® Kit with pCR®2.1 vector and sequenced. The Dehydrin genes sequences were submitted to GeneBank: Eucalyptus globulus dehydrin10 gene (Accession No. HG915712) and E. camaldulensis dehydrin 2 gene (Accession No. HG813113). The amino acid sequence of Dehydrin10 from E. globulus showed 97% homology to E. globulus DHN10 (JN052210) and Dehydrin2 from E. camaldulensis presented 94% homology to E. globulus DHN2 (JN052209). These genes can be employed in generating drought resistant crop plants.

DOSE-RESPONSE BEHAVIOUR OF WATER SCARCITY TOWARDS GENETICAL AND MORPHOLOGICAL TRAITS IN SPRING WHEAT (TRICTICUM AESTIVUM L.)
ABSTRACT:
Combining ability was studied in a Line × Tester mating fashion in wheat (Tricticum aestivum L.). Significant differences were observed for all the yield and yield contributing traits. GCA and SCA components of variation were found significant for most of the traits. Under water stressed conditions among lines the genotype Kohistan-97 revealed significant GCA effects for all the traits except spike length. Among testers, the genotype V08172 showed significant effects for the traits spike length, 1000-grain weight and flag leaf area. Based on desirable SCA effects and mean performance the cross combinations Kohistan-97 × V08172, Chakwal-86 × Punjab-81, Fsd-2008 × Punjab-81, Sehar-2006 × V08172 and Chakwal-86 × V08172 behaved best combiner to tolerate the water stress. Results of genetic analysis offered over dominance type of gene action that remained unchanged with the change in water provision for the traits like 1000 grain weight and economic yield. Similarly additive gene action was observed for the trait plant height under both normal irrigation and water stress conditions. However the cumulative genetic effects to control the expression of yield and yield components was shifted due to the changed environments. The study was concluded that due to presence of additive variance, selection could be practiced in early generation whereas in the presence of recessiveness the selection may be delayed up to the later generations. Plant traits associated with water stress tolerance having high heritability and with additive gene action may be used as indirect selection criteria for early selection of water stress tolerant genotypes. The information generated as a result of this study on genetic analysis of important economic traits of wheat under contrasting water availability positions will be of great value to the wheat breeders to design future breeding programmes.

PROTECTION OF PHOTOSYNTHETIC MACHINERY BY UP-REGULATION OF ANTIOXIDANT ENZYMES IN CONTRASTING TOMATO GENOTYPES UNDER DROUGHT
ABSTRACT:
Current study was designed to evaluate the drought effect on some physiological and biochemical properties of tomato plants. Some native and exotic tomato genotypes were subjected to drought stress to investigate the effect on antioxidant enzymes and photosynthetic machinery. The tomato genotypes were exposed to different water regimes viz: 80, 60 and 40% of field capacity. Statistical analysis revealed significant interactions in some physiological parameters including transpiration rate (E), photosynthetic rate (A) and stomatal conductance (gs). Drought stress enhanced the above properties in tolerant varieties like ‘L. pennellii’, ‘L. chilense’, ‘Lyallpur-1’ and ‘CLN1767’ in contrast to rest of the water stress sensitive genotypes. Moreover, same type of significant elevations were also observed when antioxidant enzymes like catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD) were caloremitrcially quantified in drought tolerant tomato varieties. Overall, it was found that some tomato genotypes maintained their degree of water stress tolerance during their growth but with varying mechanism of water stress tolerance. Moreover, the above mentioned physiological and biochemical characteristics can act as valuable markers for selection and breeding programs for development of drought tolerant tomato genotypes.

EFFECT OF MICRONUTRIENTS (Zn, Cu AND B) ON PHOTOSYNTHETIC AND FRUIT YIELD ATTRIBUTES OF CITRUS RETICULATA BLANCO VAR. KINNOW
ABSTRACT:
In this investigation, influence of foliar application of micronutrients (Zn, Cu and B) was studied on the improvement in photosynthetic and fruit yield attributes of citrus (Kinnow) plants. Experiments were conducted in two districts of Punjab (Sargodha and Toba Tek Singh), Pakistan varying in soil properties and agro-climatic conditions. Plants at both sites were subjected to foliar spray of three different levels (i.e. 0.1, 0.2 and 0.3%) of each Zn, Cu and B at three different fruit developmental stages while macronutrients (NPK) were applied at recommended rates as soil amendment. Micronutrients (Zn, Cu and B) application caused a significant improvement in net photosynthetic rate (A), transpiration rate (E), stomatal conductance (gs), Chlorophyll “a”, “b”, “total”, and caroteniods in both the citrus orchards. However, effect of micronutrients i.e. Zn, Cu and B was more pronounced at the levels of 0.3, 0.1 and 0.2%, respectively. These levels of nutrients were also effective in improving fruit yield with better fruit quality.

EFFECTS OF TILLAGE METHODS, CORN RESIDUE MULCH AND N FERTILIZER LEVELS ON THE WHEAT CROP PRODUCTIVITY UNDER THE RAIN FED CONDITION OF LOESS PLATEAU CHINA
ABSTRACT:
A 2 years study was conducted to assess the effects of different tillage methods (Chisel plough tillage, Zero-tillage, Rotary tillage and Mould board plough tillage), two mulch levels (M0 i.e. No corn residue mulch and M1 i.e. Corn residue mulch) and 5 N fertilizer levels (0, 80, 160, 240 and 320 kg N/ha) on the wheat crop productivity under the rain fed condition of Loess Plateau, China. Factorial experiment with three replications, having strip, split-split arrangement, with tillage methods in the main plots, mulch levels in sub- plots and N-fertilizer levels in the sub-sub plots was used for this study. Due to variations in rainfalls, during the year

EFFECTS OF MONO- AND INTERCROPPING ON GROWTH AND BORON UPTAKE OF WHEAT PLANT (TRITICUM AESTIVUM L.) CULTIVATED ON BORON-CONTAMINATED MEDIA
ABSTRACT:
Wheat plant (Triticum aestivum L.) was grown as main crop while clover (Medicago sativa L.) and sunflower (Helianthus annuus L.) were used as intercrops. Boron was applied in the form of Boric Acid (25, 50, & 75 mg L-1). The amount of chlorophyll, carotenoid, proline, B, Ca, K, and P were determined together with % DW and % EC. % DW, total chlorophyll, and carotenoid contents in monocrop and intercrop leaves grown wheat plants decreased in comparison with the control group. However, increases in mentioned parameters were observed in wheat-clover intercrops. Additionally, % EC values and proline contents in the leaves of mono-cultivated wheat plant were higher compared to the control group. Intercropping of wheat plant with clover and sunflower resulted in decreased values. Furthermore, element contents of the mono-cultivated wheat leaves decreased while these values increased when intercropping with clover.

cDNA CLONING AND EXPRESSION ANALYSES OF THE ISOFLAVONE REDUCTASE-LIKE GENE OF DENDROBIUM OFFICINALE
ABSTRACT:
The full length of the isoflavone reductase-like gene (IRL) cDNA of Dendrobium officinale was cloned by using reverse transcription (RT) PCR combined with cDNA library, the IRL function was identified by Bioinformatics and prokaryotic expression analyses, and the IRL expression levels in the organs and tissues of D. officinale plants with different ages were determined by using real-time quantitative PCR (RT-qPCR). The results indicated that the full length of the cDNA of D. officinale IRL, DoIRL, was 1238 bp (accession no. KJ661023). Its open reading frame (ORF) was 930 bp which encoded 309 amino acids with a predicted molecular mass of 34 kDa, the 5′ untranslated region (UTR) was 61 bp and the 3′ UTR containing a poly (A) tail was 247 bp. The deduced amino acid sequence of DoIRL, DoIRL, was forecast to contain a NAD(P)H-binding motif (GGTGYIG) in the N-terminal region, two conserved N-glycosylation sites, a conserved nitrogen metabolite repression regulator (NmrA) domain and a phenylcoumaran benzylic ether reductase (PCBER) domain, to hold the nearest phylogenetic relationship with the PCBER of Striga asiatica, and to share both 73% identity with the isoflavone reductases-like (IRLs) of Cucumis sativus and Striga asiatica. In Escherichia coli ‘BL21’ cells, the DoIRL cDNA expression produced a protein band holding the predicted molecular mass of 34 kDa. DoIRL expressed in all organs and tissues of D. officinale plants with different ages at comparatively low levels, and the expression level in the leaves of the two-year-old plants was the highest.

A COMPARATIVE PROTEOMICS ANALYSIS OF PINUS MASSONIANA INOCULATED WITH BURSAPHELENCHUS XYLOPHILUS
ABSTRACT:
Pine wilt disease caused by Bursaphelenchus xylophilus, also known as pine wood nematode (PWN), is the most devastating disease of pine trees. Test results from different geographical provenance of P. massoniana inoculated with nematodes, indicated that GuangDong5 (GD5) has strong resistance to PWN. This article used GD5 as the experimental material, and sensitive SX1 as comparison. Total proteins were extracted and separated by 2-DE and MALDI-TOF-TOF MS technology from the pine needles of GD5 and SX1, respectively. Differentially expressed proteins in the provenance between inoculated and un-inoculated with PWN for two weeks, were analyzed. At last, 87 differentially expressed proteins were successfully identified by MALDI-TOF-TOF. The results also showed that there were six proteins involved in hydrogen peroxide scavenging capacity and protecting the redox homeostasis system from being damaged. Their up-regulation may be the main cause of the provenance GD5 resistant to PWN.

COMPUTATIONAL IDENTIFICATION OF 18 MICRORNAs AND THEIR TARGETS IN THREE SPECIES OF ROSE
ABSTRACT:
MicroRNAs (miRNAs) are non-protein coding, small endogenous RNAs. Their length ranges from 18-26 nucleotides (nt). The miRNAs convergence property becomes a rational approach for the hunt of novel miRNAs in other organisms by homology search. As presently very little miRNAs are reported for rose species, so this study deals with the identification of miRNAs in different species of rose. Consequently 18 miRNA belonging to 17 miRNA families were identified in 3 species of rose (Rosa hybrid, Rosa chinensis and Rosa virginiana). All of the identified miRNA families (miR156, 160, 164, 166, 398, 482, 831, 837, 838, 841, 847, 3436, 3627, 6135, 6285, 6287 and 6288) are being reported for the first time in rose. Precursors of the identified miRNAs form stable minimum free energy (MFE) stem-loop structures and the mature miRNAs are found in the stem portions of their corresponding precursors. 11 putative targets of the miRNAs have also been identified. The identified targets are various proteins including transcription factors. Identification of 18 miRNAs will be supportive to explore the gene regulation phenomenon in various species of roses and it will be a good contribution for understanding the post transcriptional gene regulation in various stages of the life cycles of roses.

DELETION ANALYSIS OF SUSY-Sl PROMOTER FOR THE IDENTIFICATION OF OPTIMAL PROMOTER SEQUENCE
ABSTRACT:
The promoter region of sucrose synthase (susy-Sl) was identified and isolated from tomato. The 5′ deletion analysis was carried out for the identification of minimum optimal promoter. Transgenic lines of Arabidopsis thaliana were developed by floral dip method incorporating various promoter deletion cassettes controlling GUS reporter gene. GUS assay of transgenic tissues indicated that full length susy-Sl promoter and its deletion mutants were constitutively expressed in vegetative and floral tissues of A. thaliana. The expression was observed in roots, shoots and flowers of A. thaliana. Analysis of 5′ deletion series of susy-Sl promoter showed that a minimum of 679 bp fragment of the promoter was sufficient to drive expression of GUS reporter gene in the major tissues of transgenic A. thaliana.

DETECTION OF THE GENETIC VARIABILITY OF AMARANTHUS BY RAPD AND ISSR MARKERS
ABSTRACT:
RAPD and ISSR markers were used to analyse intra and inter-specific variability of 16 A. caudatus, 18 A. cruentus and 21 A. hypochondriacus accessions. The potential of both approaches was evaluated using three random and three microsatellite primers amplifying in total of 1126 (RAPD), respectively 1013 (ISSR) scorable fragments. Similarity values among accessions of three Amaranthus species ranged from 0.00 to 1.00 in both types of markers. Based on the intra-specific variability the polymorphism percentage varied from 70 to 100% (RAPD) or from 90 to 100% (ISSR) respectively. Resolving power (Rp) of ISSR primers have been higher (5.28 in average) in comparison to RAPD primers (4.84 in average). Percentage of distinguished accessions by ISSR and RAPD primers ranged from 29 to 89% (based on primer’s type) and from 19 to 72%, respectively. Cluster analysis based on RAPD and ISSR data has shown the individual species separation except of three accessions. Two of A. caudatus genotypes originated from India clustered with A. hypochondriacus accessions and one of A. hypochondriacus genotype originated from Nepal clustered with A. cruentus accessions. This study has demonstrated, that a single primer marker systems as RAPD and ISSR are able to generate a sufficient level of informative characters for intra and inter-specific analysis of Amaranthus genus.

TOBACCO RANDOMLY INSERTED TT8 DIFFERENLY ENHANCE LIGHT SIGNALS AND FLAVONOID ACCUMULATION
ABSTRACT:
The individual lines of tobacco over-expressing TT8, a bHLH gene, were constructed and cultured under tissue culture condition radiating with photosynthetically activation radiation (PAR) or PAR+UVA. They were compared to wild type (WT). Leaf of treated plants was extracted and analyzed for flavonoid accumulations using a spectrophotometer. The extract of TT8 plants significantly contained flavone, flavonol and anthocyanin level, higher than the WT extract did. The petal extracts of mature transgenic under PAR had a similar absorbance profile of each substance, but these extracts had higher flavonoid contents than the leaf extracts did. All flavonoid subgroups and p-coumaric acid biosynthesis were significantly enhanced after the additional UVA radiation to plant. This UVA condition slightly stimulated an accumulation of these substances in normal plant. Some transgenic greatly increased flavonoid accumulation in responding to PAR+UVA, but the others were slightly different compared to WT. The distinct insertion site is directly affected TT8 gene expression. Transgenic seeds had a dark brown color more than WT seed, which indicated high content of polymer flavonoids (proanthocyanins). This over-expressing TT8 in transgenic tobacco may directly or indirectly enhance the signal transductions of PAR and UVA and raise up flavonoid accumulation.

STUDIES ON THE CHANGES OF PROTEIN SECONDARY STRUCTURE AND CARBOHYDRATE CONTENTS IN SEEDLING-STAGE OF ABRUS CANTONIENSIS HANCE IN DROUGHT STRESS BASED ON FTIR AND CHEMOMETRICS
ABSTRACT:
To study the response mechanism of seedling-stage Abrus cantoniensis to drought stress from the level of infiltration conditions, this research uses Fourier infrared spectroscopy based on such mathematical approaches as Fourier self-deconvolution and Gaussian curve-fitting to analyze the broad characteristic absorption bands of the protein secondary structure and carbohydrates in the root, stem and leaf of seedling-stage under four drought stress treatments. The results show that the relative absorption of every type of secondary structure (namely, α-helix, β-sheet and β-turn), the α-helix/β-sheet ratio and the relative contents of starch, soluble sugars and cellulose show significant differences in the different parts of seedling-stage A. cantoniensis under different drought treatments. These differences provide a theoretical basis for investigating the response mechanisms of seedling-stage A. cantoniensis to drought stress and for advancing rational cultivation measures for this species. A new method that is fast, efficient and sensitive is presented for the study of the response mechanism of seedling-stage A. cantoniensis to drought stress.

EARLY FIELD PERFORMANCE OF DROUGHT-STRESSED SCOTS PINE (PİNUS SYLVESTRİS L.) SEEDLINGS
ABSTRACT:
Scots pine (Pinus sylvestris) has a large natural distribution throughout the world, including semi-arid areas of Turkey, where it is being used for afforestation. Determining the drought resistance of Scots pine provenances can increase the success of afforestation efforts in semi-arid regions. In the first stage of this study, water-stress treatments were applied to ten provenances of one-year-old Scots pine seedlings in their second vegetation period (between April and November). The diameter and height of the seedlings were evaluated in the nursery in order to determine their morphology. The four drought-stress treatments consisted of once-weekly irrigation (IR1), twice-weekly irrigation (IR2-Control), biweekly irrigation (IR3) and open field conditions (IR4). Later, the water-stressed seedlings were planted in a semi-arid district in Bayburt, Turkey, and their survival and growth performances were evaluated over a five-year period. The nursery study showed that drought stress and provenance as well as the interaction of the two significantly affected the morphological characteristics of the seedlings. Under water-stress conditions, the best growth performance was found in the Dokurcun, Degirmendere and Dirgine provenance seedlings. Water-stress and provenance factors and their interaction also affected the open field performance of the seedlings, where the Degirmendere, Dirgine and Dokurcun provenances again exhibited the best performance. Consequently, these Scots pine provenances can be recommended for afforestation sites having conditions similar to those of the study site.

DNA CONTENTS OF EMBRYO STRUCTURES OF BUTIA CAPITATA GERMINATING IN VITRO
ABSTRACT:
We determined the DNA contents and assessed the quality of the flow cytometry analyses of different structures of Butia capitata arising at various stages during the In vitro germination and growth of zygotene embryos, characterizing them both morphologically and anatomically. The experiments evaluated 16 organs and stages, with the embryo, cotyledon petiole and haustorium being evaluated at 8, 16 and 27 days, the primary root and leaf sheath at 16, 27 and 37 days, and the leaves after 37 days; the mature leaf was used as a control. DNA contents were found to vary according to the structures analyzed. After 27 days of In vitro culture, the cotyledon petiole, leaf sheath and radicle showed the lowest coefficients of variation and had the same amounts of DNA. The haustorium and cotyledon petiole had higher DNA values at eight days, but with coefficients of variation, indicating less reliability of these analyses. Anatomical sections of both structures after eight days showed idioblasts with large quantities of phenolic compounds that probably interfered with the analyses.

ESTABLISHMENT OF DNA FINGERPRINTING IN CLONAL TEA IMPROVED CULTIVARS FROM YUNNAN OF CHINA USING ISSR MARKERS
ABSTRACT:
In this study, DNA fingerprints were constructed by using ISSR markers for 20 clonal improved varieties developed by two breeding institutes in Yunnan province. Seven core ISSR primers were selected from 15 primers. A total of 110 bands were generated by PAGE with seven core primers, 93 of which were polymorphic bands, the percentage of polymorphic band (PPB) was 84.54%, and the mean value of polymorphism information content (PIC) reached 0.417; the genetic similarity coefficient of the cultivars was 0.574-0.854. The two primers, UBC835 and ISSR2, had high PIC values, and could be used to distinguish all cultivars, presenting the most efficient single primers. Among the all of primer combinations from the seven core primers, the three combinations, UBC835/UBC811, UBC835/ISSR2, and UBC835/ISSR3 showed lower similar coefficients, and more efficient in identifying the 20 improved varieties than the other primer combinations. Then these three primer combinations were further scored in 15 traditional cultivars. The results showed that UBC835/ISSR2 was the optimal primer combination, which could be used to distinguish each material among the 20 clonal improved varieties and 15 traditional cultivals. Finally, the DNA fingerprints of the 20 clonal improved varieties were constructed based on country and region code, breeding institute, core primer name and ISSR marker data. The established fingerprints could provide reliable scientific base for the protection of intellectual property right for these clonal improved varieties, and the important molecular information contained in these fingerprints would be useful for the authenticity identification and genetic relationship analysis of tea varieties.

DIFFERENTIATION OF RICIN USING RAPD MARKERS
ABSTRACT:
The aim of this study was to assess genetic diversity within the set of 111 ricin genotypes using 13 RAPD primers. For differentiation of 111 ricin genotypes 13 RAPD primers were used. Amplification of genomic DNA of 111 genotypes using RAPD analysis yielded 102 fragments, with an average of 7.85 polymorphic fragments per primer. Number of amplified fragments with RAPD primers ranged from 3 to 13, with the size of amplicons ranging from 100 to 1500 bp. The polymorphism information content (PIC) value ranged from 0.491 to 0.898 with an average of 0.764 and diversity index (DI) value ranged from 0.576 to 0.900 with an average of 0.776. The dendrogram based on hierarchical cluster analysis using UPGMA algorithm was prepared. In dendrogram separated unique genotype RM-32 from other 110 genotypes which were further grouped into 3 subclusters (1, 2, 3). Only four genotypes were not distinguished. Using more polymorphic RAPD markers genetically close genotypes can be distinguished. Knowledge on the genetic diversity of castor can be used for future breeding programs for increased oil production to meet the ever increasing demand of castor oil for industrial uses as well as for biodiesel production.

THE FIRST INSIGHT INTO THE SALVIA (LAMIACEAE) GENOME VIA BAC LIBRARY CONSTRUCTION AND HIGH-THROUGHPUT SEQUENCING OF TARGET BAC CLONES
ABSTRACT:
Salvia is a representative genus of Lamiaceae, a eudicot family with significant species diversity and population adaptibility. One of the key goals of Salvia genomics research is to identify genes of adaptive significance. This information may help to improve the conservation of adaptive genetic variation and the management of medicinal plants to increase their health and productivity. Large-insert genomic libraries are a fundamental tool for achieving this purpose. We report herein the construction, characterization and screening of a gridded BAC library for Salvia officinalis (sage). The S. officinalis BAC library consists of 17,764 clones and the average insert size is 107 Kb, corresponding to ~3 haploid genome equivalents. Seventeen positive clones (average insert size 115 Kb) containing five terpene synthase (TPS) genes were screened out by PCR and 12 of them were subject to Illumina HiSeq 2000 sequencing, which yielded 28,097,480 90-bp raw reads (2.53 Gb). Scaffolds containing sabinene synthase (Sab), a Sab homolog, TPS3 (kaurene synthase-like 2), copalyl diphosphate synthase 2 and one cytochrome P450 gene were retrieved via de novo assembly and annotation, which also have flanking noncoding sequences, including predicted promoters and repeat sequences. Among 2,638 repeat sequences, there are 330 amplifiable microsatellites. This BAC library provides a new resource for Lamiaceae genomic studies, including microsatellite marker development, physical mapping, comparative genomics and genome sequencing. Characterization of positive clones provided insights into the structure of the Salvia genome. These sequences will be used in the assembly of a future genome sequence for S. officinalis.

IDENTIFICATION OF A NOVEL GA-RELATED BUSH MUTANT IN PUMPKIN (CUCURBITA MOSCHATA DUCHESNE)
ABSTRACT:
Pumpkin (Cucurbita moschata Duchesne) bush mutant plants were characterized by short stems. The sensitivity of pumpkin bush mutant plants to exogenous hormones was identified in this study. Results revealed that internode elongation of bush mutant plants could respond to gibberellins (GA4+7 and GA3), but not to indole acetic acid (IAA) and brassinosteroids (BR); by contrast, the mutant phenotype of bush mutant plants could not be fully rescued by GA4+7 and GA3. The internode of bush mutant plants yielded a lower KS expression level than that of vine plants. Therefore, pumpkin bush mutant plants were designated as GA-related mutant plants eliciting a partial response to GAs; the action of IAA and BR might not be involved in the internode growth of pumpkin bush mutant plants, specifically Cucurbita moschata Duch.

MOLECULAR CHARACTERIZATION AND DIVERSITY OF A NOVEL NON-AUTONOMOUS MUTATOR-LIKE TRANSPOSON FAMILY IN BRASSICA
ABSTRACT:
Transposable elements (TEs) are capable of mobilizing from one genomic location to other, with changes in their copy numbers. Mutator-like elements (MULEs) are DNA transposons characterized by 9 bp target site duplications (TSDs), with high variability in sequence and length, and include non-conserved terminal inverted repeats (TIRs). We identified and characterized a family of Mutator-like elements designated as Shahroz. The structural and molecular analyses revealed that family had a small number of mostly defective non-autonomous MULEs and has shown limited activity in the evolutionary history of the Brassica A-genome. The Shahroz elements range in size from 2734 to 3160 bp including 76 bp imperfect TIRs and 9 bp variable TSDs. The individual copies have shown high homology (52–99%) in their entire lengths. The study revealed that the elements are less in numbers but active in Brassica rapa genomes and PCR amplification revealed their specificity and amplification in A-genome containing diploid and polyploids Brassica. The phylogenetic analysis of Brassica MULEs with other plant Mutator elements revealed that no correlation exists between Brassica MULEs and other elements suggesting a separate line of evolution. Analyzing the regions flanking the insertions revealed that the insertions have showed a preference for AT rich regions. The detailed study of these insertions revealed that although less in number and small sizes, they have played a role in Brassica genome evolution by their mobilization.

VEGETATION PATTERNS AND PLANT COMMUNITIES DISTRIBUTION ALONG AN ALTITUDINAL GRADIENT AT ASIR MOUNTAIN, SOUTHWEST SAUDI ARABIA
ABSTRACT:
Vegetation composition, main plant communities and altitudinal vegetation zones at Asir Mountain southwest Saudi Arabia were studied, and in relation to various environmental factors including climatic, geological, ecological and topographic. A total of 74 plots were sampled according to Braun-Blanquet method along an altitudinal gradient. Floristic and environmental (topographic, geological and climatic) data were collected and analyzed using numerical classification and ordination. Two main climatic regions, four altitudinal vegetation zones and fifteen vegetation groups were identified: (1) two groups in an arid coastal plains mainly on an alluvial substratum between 0 and 200m (2) seven groups in the interspersed hills and plateaus of Tihamah arid region from 200 to 1100m; (3) three groups in the escarpments from 1100 to 2000m in the semi-arid region; and (4) two groups in a semi-arid high mountains zone from 2000m to the summit. Geo-altitudinal gradient was affected by the climatic factors (rainfall and temperature) and vegetation structure (species richness and coverage) on a regional scale, while edaphic factors played the leading role in the distribution of different vegetation groups on a local scale. These findings coincide with those reported about the distribution of vegetation at other tropical mountains. Human activates have a significant effect on modifying the distribution and abundance of plant species.

UNDERSTORY PLANT DIVERSITY IN MIXED AND PURE PLANTATIONS OF JATROPHA CURCAS VS. NATIVE VEGETATION IN THE LOWER-MIDDLE REACHES OF THE LANCANG-MEIKONG RIVER WATERSHED, CHINA
ABSTRACT:
22 plots at the Xiaoheijiang base, located in the lower-middle reaches of the Lancang-Meikong River in China, were investigated to analyze the understory biodiversity of Jatropha curcas plantations. Two kinds of mixed modes of J. curcas (mixed plantation with Macadamia integrifolia and mixed plantation with shrub species) and a pure plantation of J. curcas were planted, while the native vegetation served as a control. The plots were distributed along the gradients of forest management, succession and elevation by CCA analysis. Species richness was not significantly different for the different types of plantation, but the evenness of species could be affected, especially for the total community and the understory by planting J. curcas. The diversity and evenness indices of species were affected for the mixed plantation with different proportions of M. integrifolia, especially for the shrub layer, the Shannon diversity index and Pilou evenness index showed significant differences. And for the different mixed shrub species, only the Shannon diversity index and Pilou evenness index were significantly different. Finally, from the perspective of biological diversity, J.curcas plantation with shrub species would be a recommended planting model for ecological restoration in a dry-hot valley area, while J. curcas plantation with M. integrifolia would be an effective planting model to balance crop yield and food security.

VEGETATION DESCRIPTION AND PHYTOCLIMATIC GRADIENTS OF SUBTROPICAL FORESTS OF NANDIAR KHUWAR CATCHMENT DISTRICT BATTAGRAM
ABSTRACT:
This paper communicates an analytical exploration of the vegetational profile in the subtropical zone of Nandiar Khuwar catchment area, District Battagrtam, Pakistan. On the basis of physiognomy of vegetation the study area was divided into 16 stands. Six plant communities were recognized by TWINSPAN classification. Among biological spectrum nanophanerophytes was dominated with 36 species and leaf size spectra were dominated by microphyll contributing 63 species. Similarity index was maximum (33.61) between Pinus, Micromeria, Rubus community and Pinus, Rubus, Cynodon community. In Bray-Curtis ordination the maximum ordination scores were recorded for axis 2 (0.921). The gradient length was maximum (3.35) for axis 1 with eigenvalue 0.50. Total variance (inertia) in the species data were 2.92, explanatory variables account for 100%. Among environmental variables the maximum positive strength were recorded for altitude (0.818) and Phosphorous (0.801) while maximum negative strength were recorded for wind speed (-0.864), barometric pressure (-0.825) and temperature (-0.820).

ANALYSIS OF FUNGAL DIVERSITY IMPACTS ON PINUS ROXBURGHII SEEDS FROM PINE FOREST AND PLANT NURSERIES OF AZAD KASHMIR, PAKISTAN
ABSTRACT:
Pinus tree plays a pivotal role in commercial revenue generation, domestic lives of rural communities and sustaining of climate of Azad Kashmir. Pinus grows in forest as wild species but due to harsh environmental parameters it is also cultivated in nurseries for propagation and plantation. In this research, injurious impacts of mycofloral diversity on seed germination of Pinus roxburghii Sarg. in nature (forest) and nurseries were explored from different localities of Azad Kashmir, Pakistan. In the analysis two protocols viz., blotter method (BM) and agar plate method (APM) were employed and 11 fungal species of nine genera were isolated. APM was found better (66a±0.32) than BM (60a±0.09). The prevalence of different isolated taxa was as: Aspergillus niger (42.75%), Aspergillus flavus (24.0%), Botrytis sp (14.25%), Botryosphaeria sp. (17.75%), Cladosporium cladosporioides (32.75%), Drechslera sp. (5.75%), Fusarium sp. (47.50%), Penecillium sp. (7.25%), Rhizopus stolonifer (11.50%), Rhizopus oryzae (13.0%) and Mucor sp.(7.0%). Pathogenicity analysis depicted that Fusarium was the most harmful (15.75e±0.54), followed by Aspergillus flavus (20.50d±0.32), Aspergillus niger (25.75c±0.42) and Rhizopus sp. (35.75b±0.12). Different pathogenicity results of analyzed fungal species were found in different areas and it was highest in Muzaffarabad (52.0%), Kotli (45.6%), Samahni (42.4 %) and least in Bhimber (36.0 %). Radical length (mm) of Pinus roxburghii was severely affected by Aspergillus flavus (46.6a±0.44) in Muzaffarabad, Rhizopus sp. (44.1a±0.72) in Samahni, Fusarium sp.( 42.5a±0.28) in Kotali, Aspergillus niger (37.8a±0.44) in Samahni, respectively. The tested species showed that plumule length (mm) of samples was most retarded in Muzaffarbad (37.98%) and least affected in Mirpur (24.58%). The results depict that fungi do cause damage to seed germination and growth of seedlings in nature and nurseries and these findings will be useful for forest and nurseries researchers to produce good quality seeds for growth of pinus trees.

DISTRIBUTION PATTERNS OF SEGETAL WEEDS OF CEREAL CROPS IN TAJIKISTAN
ABSTRACT:
Using the literature data and field research conducted in 2009-2013 the distribution patterns, habitat conditions, phytogeographical characterisation and endangerment of weeds occurring in cereal crops in Tajikistan were analysed. We found out that Tajik weed flora of cereal crops counts 686 taxa. The most species rich families include Asteraceae, Poaceae and Fabaceae. The highest number of cereal weeds were noted in large river valleys of Syr-Daria, Amu-Daria and their tributaries in south-western and northern Tajikistan. This subregions have the warmest climate conditions and extensive arable lands. The greatest weed species richness was observed in submontane and montane elevations between approx. 700 and 1,900 m a.s.l. Cereal weeds occur frequently outside segetal communities in Tajikistan. They were noted usually in screes, wastelands, xerothermophilous grasslands, river gravel beds and in steppes habitats. The assessment of threat status reveals that ca. 33% of total cereal weed flora in Tajikistan are disappearing or occur very rarely. According to the chorological data we find that in the cereals of Tajikistan, 35 endemic and 14 subendemic species occur. The most numerous chorological elements of threatened weed flora of Tajikistan are Irano-Turanian (55%), pluriregional (16%), cosmopolitan (14,5%), Mediterranean (9%) and Eurosiberian (5%) species. Further research is suggested to explore the distribution patterns of all weed species in Tajikistan as it should be useful for economy and effectiveness of crop production as well as conservation of most valuable species.

ORDINAL CLASSIFICATION OF VEGETATION ALONG MANGLA DAM, MIRPUR, AJK
ABSTRACT:
Vegetation plays an important role in ecosystem maintenance. But the construction of dams transform the riparian vegetation into impoundment region. The present study was conducted to identify and quantify herbaceous flora around the vicinity of Mangla dam. Study area was divided into two zones on the basis of distance from the dam boundary. Pattern of vegetation distribution and their association in area was grouped in to different communities by using ordination techniques. Two ordination techniques TWINSPAN and DECORANA were used. A total of 37 species belonging to 17 families were identified from fifty quadrats. Random sampling was done by using 1 ×1 m sized quadrat. Percentage of vegetation was assessed by using Domin cover scale. TWINSPAN classified two groups and four communities in Zone-I, while in Zone-II two groups and six communities were formed. Dominance curve showed that Cynodon dactylon, Desmostachya bipinnata and Rhynchosia minima were dominant species in Zone-I and Croton bonplandianus, C. dactylon, D. bipinnata and Brachiaria decumbens were frequent species in Zone-II. DCA as indirect multivariate technique based on reciprocal averaging determined the environmental gradients that affect the species richness and also verified the groups of species and indicated four communities in both Zones. Monte Carlo test of significance was used to analyze stress in relation to number of axis/dimensionality under Non-metric Multidimensional Scaling (NMS) through p-value. This study provided the significant results of least abundant and most abundant herbaceous species around the dam which will be helpful for biodiversity conservation and in decision making for further land planning.

FIELD SCREENING OF GUAR [CYAMOPSIS TETRAGONOLOBA (L.) TAUB.] ACCESSIONS FOR ENHANCED FORAGE PRODUCTION ON HOT DRYLANDS
ABSTRACT:
To explore the high biomass producing accessions of guar [Cyamopsis tetragonoloba (L.) Taub.], under hot and dry conditions, an experiment was conducted under natural field conditions located at Research Station, University College of Agriculture, Sargodha, Pakistan using 36 different accessions. Twenty-day old plants grown under normal irrigation were subjected to control and drought (withholding of irrigation for 20 and 40 days) conditions. It was observed that plant fresh and dry biomass, plant height, net CO2 assimilation rate (Pn), transpiration rate (E) and inter CO2 conc. (Ci) of all guar accessions under study decreased considerably on withholding of irrigation for both 20 and 40 days. Of all guar accessions, Acc. BR99, 5597, BWP 5595, BWP 5596, BWP 5609, BWP 5611, 24323, Chiniot Black, Khushab Black were relatively higher in plant height and plant biomass (fresh and dry) under water stress conditions. However, a significant increase particularly in BR99, BWP 5595, BWP 5599, BWP 5609, 41671 followed by Sialkot White was observed in the relative membrane permeability of all guar accessions under water limited conditions. All accessions varied significantly in terms of Pn, E and Ci. Accessions BR90, BR99, BWP 5596, 24323, 24332, Chiniot Black, Khushab Black, 41671, 24323, BWP 5611, Khushab White, Mardan, Khanewal Local 2 and Sialkot White were better in Pn, Ci and E. No significant change was observed in the water-use efficiency of guar accessions under water-limited conditions. Overall, BR99, 5597, BWP 5595, BWP 5596, BWP 5609, BWP 5611, 24323, Chiniot Black and Khushab Black were higher, whereas accessions BWP 5595, 24321, Khushab White, Silanwali White, Sialkot Black and Khanewal Local lower in plant biomass and different gas exchange characteristics under water limited conditions, so the earlier mentioned accessions can be recommended as high biomass producing guar accessions for their cultivation on hot drought-prone areas.

THE EPIPHYTIC BRYOPHYTE FLORA AND VEGETATION OF ZONGULDAK-GÖBÜ VILLAGE (NORTHWEST TURKEY)
ABSTRACT:
The epiphytic bryophyte flora and vegetation of the Zonguldak-Göbü village was investigated in this study. Between the years 2012-2013, as a result of identification of the 236 epiphytic bryophyte samples collected from the trunks of trees; total of 29 taxa (8 liverworts, 21 mosses) belonging to 23 genera and 17 families were determined. As a result of evaluation of the 33 relevés taken from the trunk of trees by Braun-Blanquet method, the Plagiothecietum neglecti association, Pylaisietum polyanthae typicum and Pylaisietum polyanthae -lophocoleetosum heterophyllae subassociation are identified. From these syntaxa, Plagiothecietum neglecti has been reported for the first time in Turkey, in addition Pylaisietum polyanthae -lophocoleetosum heterophyllae subassociation is new for knowledge. The life forms and life strategies of syntaxa examined in terms of ecological and floristic have been analyzed. Mat and weft are dominant of the life forms while Ag is dominant of the life strategies. In addition, the most taxa of floristic list are determimed mesophytic, sciophyt and acidic as a character.

THE PHYLOGENETIC RELATIONSHIPS AMONG GERMPLASM RESOURCES OF WILD RAMIE (BOEHMERIA NIVEA L. GAUD) IN CHINA BASED ON trnL-F AND ITS SEQUENCES
ABSTRACT:
Ramie (Boehmeria nivea L. Gaud) is an important fiber crop in China, which also possesses many wild species in genus Boehmeria Jacq. However, the taxonomic position of these species has not been settled. To determine the evolutionary relationships among the members of the genus Boehmeria, the combination of ITS and trnL-F sequences were used for molecular phylogenetic analyses of 31 ramie accessions (28 species and three varieties) including multiple materials collected in high-altitude regions that have not been previously reported (B. clidemioides var. diffusa, B. bicuspis and B. longispica). The ITS and trnL-F trees produced showed that Boehmeria was classified into four separate clusters. The Sect. Duretia, which has a high evolutionary level, clustered with Sects Zoilingeriana and Phyllostachys. The grouping pattern of clustering differed from traditional taxonomy and indicated possible interspecific hybridization among Boehmeria. We found that B. malabarica Wedd. var. leioclada of Sect. Boehmeria clustered into a clade with Sect. Tilocnide, providing solid support for the expansion of wild ramie core germplasm resources. The molecular results did not support the intraspecific geographic migration of Boehmeria. This study, therefore, established relationships among wild species which will help in ramie crop improvement programs. The results will be important for the collection and conservation of germplasm resources of Chinese wild ramie.

CYTOLOGICAL OBSERVATION OF SOLANUM PIMPINELLIFOLIUM L. MICROSPORE DEVELOPMENT
ABSTRACT:
Tomato (Solanum lycopersicum L.) is one of the most important horticultural crops worldwide, and has also been adapted as a model plant in a wide range of research disciplines. However, stamen development in tomato is less known than other model species, such as Arabidopsis thaliana and Oryza sativa L. (rice). In order to understand tomato flower development in more detail, the key goal of this study was to establish a precise correlation between visual morphological features and cytological events. To this end, we characterized a wild tomato species, S. pimpinellifolium (accession LA1585), to define stamen developmental stages using semi-thin sectioning. Based on distinct stages of flower development, S. pimpinellifolium anther/stamen development was divided into ten stages, and characteristic morphological and cytological features in stamens at each developmental stage were identified. In this way, flower characteristics and microspore development were correlated. In addition, specific cytological key events in S. pimpinellifolium were compared with those in A. thaliana and rice at corresponding stages, as well as with those in domesticated tomato (S. lycopersicum L.), and the differences are discussed.

THE MORPHOLOGICAL STUDY OF THE FRUIT, SEED AND SEEDLING OF HYDROCHARIS DUBIA (HYDROCHARITACEAE)
ABSTRACT:
The structure of the fruit, seed and seedling of Hydrocharis dubia in Hydrocharitaceae were studied with light microscope. The results show that the fruit has incompletely six locules with superficial placentation. The seed has anchor-like trichomes occur on the surface and with no endosperm. The seed embryo develops on the water. The hypocotyl hairs appear earlier than the primary root which develops weakly. The adventitious roots are formed with the leaf development. The paracytic stomata are present in the upper epidermis of the young leaf and the air canals are present in the mesophyll. The petiole has a sheath-like base. The seedlings usually die and only a few could develop into mature plants and the development of the species is similar to those of the other taxa in Alismatales. The study provides the morphological evidence for the placement of the Hydrocharitaceae in the order Alismatales.

MORPHOLOGICAL AND ANATOMICAL FEATURES OF CYPSELA OF SOME CREPIS TAXA (ASTERACEAE) FROM TURKEY AND THEİR TAXONOMIC IMPORTANCE
ABSTRACT:
Fruit morphology and anatomy have taxonomic importance in Asteraceae. The fruits structures of Crepis from Turkey, which include five species (C. alpina, C. smyrnaea, C. pulchra, C. zacintha, C. sancta) and two subspecies (C. foetida subsp. foetida and C. foetida subsp. rhoeadifolia) were studied for fruit morphological and anatomical characters with one-way analysis of variance, cluster analysis and principal component analysis. Fruit size, shape, color, and the presence of beak were observed with stereomicroscopy. Whereas the surface patterns of fruit and pappus were examined using scanning electron microscopy (SEM). Furthermore, pericarp structure, thicknesses of testa and endosperm, and number of rib, cotyledon width in cypsela were studied anatomically. Results indicated that cypsela sizes, the presence or absence of beak on the cypsela, fruit and pappus surfaces, pericarp. Thickness of testa and endosperm, and number of ribs are of major importance to illustrate interspecific relations among the examined taxa. Also, this investigation is a preliminary study, which was performed to use fruit morphological and anatomical characters for their practicality on the classification of taxa within the genus.

IN VITRO MICROPROPAGATION OF SOLANUM VILLOSUM─A POTENTIAL ALTERNATIVE FOOD PLANT
ABSTRACT:
Solanum villosum Miller is annual to biennial herb which is used as potherb as well as fodder/forage that limits its distribution in Pakistan. The aim of this study was to develop a suitable protocol for S. villosum through direct organogenesis. Leaf, stem node and shoot tip explants from the tested plant were inoculated in three different hormonal combinations of BAP (6-benzyl amino purine) alone along NAA (α-naphthalene acetic acid) and Kin (Kinetin). Maximum shoot induction was recorded for stem node and leaf (91% each) in MS medium comprising of BAP (1.9 mg/l) and NAA (0.1 mg/l), while shoot tip showed somewhat moderate (81%) response. The highest mean number of shoot (9.1±0.12) was also obtained for the same medium using leaf explants. Plantlets were successfully rooted in auxin free medium and shifted to green house for multiplication after acclimatizing them. This study may contribute in providing quick and disease free propagation of this neutraceutically and economically potential plant.

FLOWER CHARACTERISTICS AND PHENOLOGY OF ANDROMONOECIOUS JATROPHA CURCAS
ABSTRACT:
Andromonoecious Jatropha curcas (Jatropha) has male and hermaphrodite flowers in one inflorescence. Jatropha is a potential source for biofuel raw material. The development of this plant has various constraints, such as lower fruit set that causes low seed productivity. The existence of andromonoecious Jatropha with high fruit set provides a great opportunity to overcome the obstacles. Understanding floral biology is a requirement for developing a high fruit set andromonoecious Jatropha. This study was aimed at understanding the flower’s characteristics and phenology. The andromonoecious Jatropha flower’s characteristics, number and viability of pollen, and phenology were observed and compared to those of monoecious Jatropha. Andromonoecious Jatropha had inflorescence with a dichasial cyme pattern. An inflorescence produced two main branches and several secondary and tertiary branches, where a hermaphrodite flower was found at whose ends. The number of inflorescence branches in andromonoecious Jatropha was higher than that of monoecious Jatropha. The diameter and size of the sepal and stalk in hermaphrodite flowers were found to be larger than those in male and female flowers. The development of andromonoecious Jatropha flowers was divided into 4 phases, namely initiation (0-3 dai = days after initiation), small bud (3-7 dai), large bud (7-20 dai), and flower blooming (20-39 dai) phases. The anthesis and anther dehiscence of hermaphrodite flowers occurred at the same time between 06:00 to 06:59 am, facilitating self-pollination of the hermaphrodite flowers. The average pollen viability during the anthesis phase was 96.26%, and 88.79% of them survived until the next day.

A STUDY OF SOME HEAVY METALS FOUND IN MEDICINAL PLANTS (EUPHORBIA CORNIGERA, RHAZYA STRICTA AND CITRULLUS COLOCYNTHIS) IN TURBAT REGION OF BALOCHISTAN WITH REFERENCE TO PREVENTION OF ENVIRONMENTAL POLLUTION
ABSTRACT:
The aim of this paper is to present the accumulation content of toxic heavy metals such as Cd, Zn, Cu, Ni and Pb in the shoots of three plants species (Euphorbia cornigera, Rhazya stricta and Citrullus colocynthis) collected from Turbat (Balochistan). These therapeutic plants play an important role in the traditional medication system that has been existed for several years and continuously provide new and novel remedies to the human beings. Moreover, these medicinal plants are used throughout the world for the purpose of treatment of many diseases and also a significant source of raw material for pharmaceutical and agricultural industries. Further, the various concentrations of heavy metals in medicinal plants was recorded as Pb > Zn > Cd > Cu > Ni. In the present research the results were related with the safety standards recommended by the World Health Organization (WHO).

PROTEIN ESTIMATION AND PALYNLOGICAL STUDIES OF CANNABIS SATIVA L. POLLEN IN RELATION TO RESPIRATORY ALLERGIES
ABSTRACT:
Airborne pollen allergies and asthma are on a rise in the metropolitan city of Islamabad. Knowledge of allergenic pollen is limited in the area. Cannabis sativa L. or commonly known as Hemp is widely spread weed in the city. Morphological studies performed via light microscopy and SEM have shown that the pollen of Cannabis sativa are 21µm long having triporate aperture, spheroidal in shape and scaberate exine. Quantitative and qualitative analysis of pollen proteins has also be done in to recognize allergenic protein bands. Bradford’s analysis for proteins quantification has shown that the hemp pollen has 30.69 mg/g protein in fresh weight of pollen. While SDS-PAGE analysis showed 11 bands of various protein size ranging from 17kDa to 150kDa. The research findings indicate that Cannabis sativa, could be a potent allergenic pollen-producing weed that might cause serious health problems in the population of Islamabad.

BIOSYNTHESIS, PURIFICATION AND CHARACTERIZATION OF COMMERCIAL ENZYME BY PENICILLIUM EXPANSUM LINK
ABSTRACT:
Ever growing biotechnological industry has motivated the research towards the comprehensive survey of microorganisms, which could be used in extreme conditions of industry. In the present work optimization parameters in submerged fermentation, purification and characterization of invertase from Penicillium expansum Link using agricultural wastes (sunflower waste, cotton stalk and rice husk) as well as agro-industrial wastes (date syrup and molasses) as sources of carbon. Maximum production of invertase (7.03 U/mL) was observed when the strain was grown on culture medium (CM1) containing yeast extract as a source of nitrogen, date syrup as a source of carbon after 48 h of incubation at initial pH 5.0, temperature 35ºC, inoculum size of 6x106 conidia in 50 mL of culture medium and agitation rate of 150 rev/min. After optimization the enzyme was also purified partially and then characterized. Kinetic constants (Km 2.57 mM and Vmax 178.6 U/mL/min) were determined by Lineweaver-Burk Plot and molecular mass (110 kDa) by 10% SDS-PAGE. Invertase showed maximum activity at pH 5.5 (128.7 U/mL) and at the temperature of 60ºC (114.6 U/mL). BaCl2 (21.9%), MgSO4 (42.6%), MnCl2 (46.8%) and EDTA (8.3%) enhanced the relative activity of enzyme while HgCl2 (-90.9%), CuSO4 (-82.3%) and CuCl2 (-78.7%) were proved inhibitors.

COMPARATIVE STUDIES ON CONVENTIONAL (WATER-HOT ACID) AND NON-CONVENTIONAL (ULTRASONICATION) PROCEDURES FOR EXTRACTION AND CHEMICAL CHARACTERIZATION OF PECTIN FROM PEEL WASTE OF MANGO CULTIVAR CHAUNSA
ABSTRACT:
Pectin, a naturally occurring heteropolysaccharide, is widely used as a functional ingredient in food and pharmaceutical industries due to its gelling and stabilizing properties. During the present study pectin was extracted from peel of mango (cultivar Chaunsa) using conventional (water-hot acid) and non-conventional (ultrasonication) methods. In conventional method, HNO3, H2SO4, or HCl was used under variable conditions of pH (2.0, 2.5, 3.0), temperature (70, 80, 90, 100oC), duration of extraction (30, 60, 90, 120 min), and solvents (ethanol, methanol, acetone, isopropyl alcohol). Maximum yield of 16.6 g pectin 100 g-1 peel was obtained with HNO3 at pH 2.5, 90oC, 90 min extraction, and ethanol. Whereas in non-conventional method, ultrasonication was used for different time intervals (10, 20, 40 min) using HNO3 at pH 2.5 and 90oC. Maximum yield of 15.8 g pectin 100 g-1 peel was obtained by this method in 20 min. Pectin extracted by the above two methods was found to be of high quality as was determined in respect of methoxyl and galacturonic acid contents, degree of esterification, equivalent weight, and FTIR spectra. Extraction of pectin from mango peel by employing non-conventional method (ultrasonication) was observed to be an energy efficient method due to its less extraction time (20 min as compared to 90 min in conventional method) suggesting its suitability on commercial scale for the extraction of pectin from mango and other available fruit peel wastes.

RHIZOSPHERE MICROBIAL COMMUNITIES FROM RESISTANT AND SUSCEPTIBLE WATERMELON CULTIVARS SHOWED DIFFERENT RESPONSE TO FUSARIUM OXYSPORUM F. SP. NIVEUM INOCULATION
ABSTRACT:
Fusarium oxysporum f. sp. niveum (FON), a soil-borne pathogen of watermelon (Citrullus lanatus), can cause substantial production losses worldwide. In this study, plate culture and PCR-denaturing gradient gel electrophoresis (DGGE) methods were used to evaluate the effects of inoculation of Fusarium oxysporum f.sp. niveum on rhizosphere microbial communities of different watermelon cultivars to FON. Two methods indicated that the effects of watermelon rhizosphere microbial community of different resistance cultivars to FON were much different. Populations of culturable bacteria and actinomycetes in the rhizosphere of susceptible watermelon cultivar were significantly lower than in the resistant cultivar after inoculation (P<0.05), but the opposite result was observed for fungi. Principal component analysis of bacterial and fungal community structure also showed that the cultivar of FON-inoculated soil treatment were separated from the non-inoculated controls after inoculation, and there was clear discrimination between the susceptible cultivars and the resistant cultivars. Sequence analysis of specific bands from DGGE profiles showed that specific rhizosphere bacterial and fungal groups differed between watermelon cultivars after inoculation . Both methods demonstrated that different resistant watermelon cultivars occupied different rhizosphere microbial communities, and and disease suppression might be correlated with high microbial diversity. F. oxysporum f. sp. Niveum alters the structure and functional diversity of microbial communities associated with watermelon rhizosphere.

EFFICACY OF WILD PLANT IN COMBINATION WITH MICROBIAL ANTAGONISTS FOR THE CONTROL OF ROOT ROT FUNGI ON MUNGBEAN AND COWPEA
ABSTRACT:
Present work was carried out to investigate the efficacy of Aerva javanica in combination with different microbial antagonists namely Rhizobium meliloti, Pseudomonas aeruginosa, Trichoderma harzianum and Aspergillus niger. Soil amended with A. javanica stem, leaves, flower powder @1% w/w and seeds of cowpea (Vigna unguiculata L.) and mungbean (Vigna radiata L.) were coated with microbial antagonists for the control of root infecting fungi like Macrophomina phaseolina (Tassi) Goid, Fusarium spp. and Rhizoctonia solani Kühn. Infection of M. phaseolina and R. solani were completely suppressed when seeds were coated with P. aeruginosa, T. harzianum, A. niger, R. meliloti and A. javanica leaves powder mixed in soil @1% w/w. All antagonists showed reduction in combination with A. javanica leaves powder @1% but T. harzianum and P. aeruginosa in combination with A. javanica leaves showed promising results in complete reduction of R. solani and M. phaseolina on both crops. All growth parameters were maximum when soil was amended with A. javanica leaves powder @1% w/w and seeds were coated with T. harzianum and P. aeruginosa.

ISOLATION, IDENTIFICATION OF ANTAGONISTIC RHIZOBACTERIAL STRAINS OBTAINED FROM CHICKPEA (CICER ARIETINUM L.) FIELD AND THEIR IN-VITRO EVALUATION AGAINST FUNGAL ROOT PATHOGENS
ABSTRACT:
Plant growth promoting rhizobacteria (PGPR), are associated with roots, found in the rhizosphere and can directly or indirectly enhance the plant growth. In this study soil was collected from rhizosphere of chickpea fields of different areas of Rawalpindi division of Pakistan. PGPR were isolated, screened and characterized. Eight isolates of rhizobacteria (RHA, RPG, RFJ, RC, RTR, RT and RK) were isolated from Rawalpindi division and were characterized. The antagonistic activity of these PGPR isolates against root infecting fungi (Fusarium oxysporum and Verticillium spp.,) was done and production of indole acetic acid (IAA), siderophore and P-solubilization was evaluated. The isolates RHA, RPG, RFJ, RC, RRD and RT were found to be positive in producing siderophore, IAA and P-solubilization. Furthermore, most of the isolates showed antifungal activity against Fusarium oxysporum, and Verticillium spp. The rhizobacterial isolates RHA, RPG, RFJ, RC, RRD, RTR, RT and RK were used as bio-inoculants that might be beneficial for chickpea cultivation as the rhizobacterial isolates possessed the plant growth promoting characters i.e. siderophore, IAA production, phosphate solubilization. In in vitro tests, Pseudomonas sp. and Bacillus spp. inhibited the mycelial growth of the fungal root pathogens. The isolates (RHA and RPG) also significantly increased (60-70%) seed germination, shoot length, root length of the chickpea. The incidence of fungi was reduced by the colonization of RHA and RPG which enhanced the seedling vigor index and seed germination. The observations revealed that isolates RHA and RPG is quite effective to reduce the fungal root infection in greenhouse, and also increases seed yields significantly. These rhizobacterial isolates appear to be efficient yield increasing as well as effective biocontrol agent against fungal root pathogen.

A PERSPECTIVE OF LEAF RUST RACE FHPRN AND ITS IMPACT ON LEAF RUST RESISTANCE IN PAKISTANI WHEAT VARIETIES
ABSTRACT:
Leaf rust infected leaves of a widely growing variety Seher-06 were collected in wheat season of 2011-12. The leaf rust isolates were assessed on Thatcher derived Lr isogenic lines and a race FHPRN was identified. Seventy six wheat varieties/lines besides Lr isogenic lines were screened against this race for seedling in glass house and for adult plant resistance at Bahawalpur and Faisalabad during 2012-13. Lr1, Lr2a, Lr9, Lr19, Lr24, Lr10+27+31 (Gatcher) and Lr28 were found completely resistant at both stages against FHPRN. Molecular screening of the wheat varieties/lines indicated the presence of leaf rust resistance genes Lr9 (0%), Lr13 (43%), Lr19 (1%), Lr20 (0%), Lr24 (4%), Lr26 (23%), Lr28 (0%), Lr34 (38%), Lr37 (1%) and Lr47 (1%) in them. Field data suggested that As-02 (Lr10+26+34), Bhakar-02 (Lr13) and Shafaq-06 (Lr10+13+27) were resistant; Pasban-90 (Lr10+13+26+27), Chenab-2000 (Lr10+13+26+27+31+34), Fbd-08 (Lr10), Millat-11 (unknown) and Punjab-11 (unknown) were found moderately resistant; Blue silver (Lr13+14a), Pak-81 (Lr10+23+26+31), Bahawalpur-97 (Lr13+26) and Lasani-08 (Lr13+27+31) were susceptible while Sh-88 (unknown), Auqab-2000 (Lr10+23+26+27+31), Iqbal-2000 (Lr3+10+13+26+27+31), Bahawalpur-2000 (Lr34) and Seher-06 (Lr10+27+31) were found highly susceptible against FHPRN. Present and previous studies revealed the presence of Lr3, 10, 13, 14a, 23, 26, 27, 31 and 34 in the Pakistani wheat varieties yet lacking Lr9, 19, 24 and 28. Therefore, the latter genes and their effective combinations should be incorporated in Pakistani varieties to combat leaf rust effectively.

WHITE BLISTER RUSTS AND DOWNY MILDEWS FROM BAJAUR AGENCY FATA, WITH SOME NEW RECORDS FROM PAKISTAN
ABSTRACT:
In a species diversity study of Oomycyctes of Bajaur Agency FATA, Pakistan, infection of white blister rusts and downy mildews recorded on three cultivated and four wild plants. Capsella bursa-pastoris showed mixed infection of Albugo candida and Hyaloperonospora parasitica (syn: Peronospora parasitica). Similarly, A. candida and H. brassicae (syn: P. brassicae) parasitized Brassica campestris. Wilsoniana portulacae (syn: Albugo portulacae) and W. occidentalis com. nov. (syn: Albugo occidentalis) recovered from Portulaca oleracea and Spinacia oleracea, respectively. Bremia taraxaci, B. sonchicola and B. saussureae recorded on Taraxicum officinale, Sonchus sp., and Saussurea sp., respectively. All these obligate parasites are new records from Bajaur Agency, while H. parasitica, W. occidentalis, B. taraxaci, and B. saussureae on the mentioned hosts are new records from Pakistan.

ANTIFUNGAL ACTIVITY OF EVERNIA PRUNASTRI, PARMELIA SULCATA, PSEUDEVERNIA FURFURACEA VAR. FURFURACEA
ABSTRACT:
The aim of this study was to investigate the In vitro efficacy of 96% alcohol extracts of Evernia prunastri and Pseudevernia furfuracea var. furfuracea that were in foliose-fruticose form and Parmelia sulcata in foliose form against important plant pathogens. The growth of fungal colonies in Petri plates amended with lichen extracts at 25°C was measured a day before covering all surface of Petri plate in control treatment. Data were analysed according to statistic analysis test LSD at p≤0.05. The in vitro efficacy of extracts of E. prunastri, P. sulcata and P. furfuracea var. furfuracea showed a significant inhibition against mycelia and spor growth of Aspergillus niger, Botrytis cinerea, Fusarium culmorum, F. solani, Macrophomina phaseolina, Penicillium expansum and Rhizoctonia solani. The level of inhibition among extracts showed variation. It was concluded that secondary metabolites of lichens may be used as biological chemicals against some plant pathogens.

ARBUSCULAR MYCORRHIZAL FUNGI AND THEIR INFLUENCING FACTORS FOR AEGICERAS CORNICULATUM AND ACANTHUS ILICIFOLIUS IN SOUTHERN CHINA
ABSTRACT:
Our study aimed to explore Arbuscular mycorrhizal fungi (AMF) colonization and spore density for Aegiceras corniculatum and Acanthus ilicifolius across five mangrove ecosystems in southern China, focusing mainly on the relationships between AMF and biotic/abiotic factors. Soil physicochemical properties and seawater salinity, as well as the numbers of culturable soil microbes (bacteria, fungi and actinmycetes) were measured to analyze their potential effects on AMF colonization. The results showed that AMF were very common for both plant species in the investigated mangrove ecosystems, and hyphae were the dominant structures for both species. Total AMF colonization rates (TC%) ranged from 0.33% to 36.50%, while the average TC% for A. ilicifolius (13.47%) was slightly higher than for A. corniculatum (9.47%). The average spore density for A. corniculatum was 49.0 spores per 25g air dried soil, and 51.7 for A. ilicifolius. Soil physicochemical analysis showed that soil in mangroves was with high moisture and organic matter content, slightly acidic pH, low levels of total and available P and high levels of N content. Microbial counting experiment recorded high microorganism numbers in mangroves. Data analysis revealed that soil available P content and seawater salinity may be important factors influencing AMF in mangroves. The two mangrove species showed different correlations with microbial numbers, which may illustrate that host plant is a key factor influencing AMF and other microbes.

ANTIMICROBIAL ACTIVITY OF NERIUM OLEANDER L. AND NICOTIANA TABACUM L.: A COMPARATIVE STUDY
ABSTRACT:
The antimicrobial activity of aqueous ethanolic extract of two plant species i.e. Nerium oleander L. and Nicotiana tabacum L. from Multan district were checked, against three pathogenic bacteria viz: Staphylococcus aureus (gram positive), Escherichia coli and Pseudomonas aeruginosa; (gram negative). Disc diffusion technique was used to check the antimicrobial activity. The bacterial strains under question were found susceptible to plant extracts. Nerium oleander and Nicotiana tabacum exhibited strong antibacterial action against tested bacteria. The ethanolic extract of Nerium oleander leaves showed highest antibacterial action against Pseudomonas aeruginosa at 900mg/ml concentration, whereas, Nicotiana tabacum showed maximum zones of inhibition against Staphylococcus aureus at 900mg/ml concentration. The results showed that the leaves extracts of these plants have great power as anti-microbial agents against these common bacterial isolates. However, for future pharmacological implication of these plants more research is needed for the isolation of active ingredient of these plants. The results obtained could be attributed to the differential environmental conditions on the ecology of these plants.

ANTIMICROBIAL ACTIVITY OF ELAEIS GUINEENSIS LEAF EXTRACT AGAINST GANODERMA BONINENSE OF OIL PALM BASAL STEM ROT
ABSTRACT:
Basal stem rot (BSR) which is caused by Ganoderma boninense is the most serious disease faced by oil palm Elaeis guineensis in Malaysia. Hitherto, many control measures such as cultural practices, fungicides, and biocontrol agents were attempted, yet, the result were unsatisfactory. This study was conducted to investigate potential antimicrobial properties in oil palm leaf extract against G. boninense. Oil palm leaf were extracted with different solvents namely methanol, acetone, chloroform and petroleum ether. The antimicrobial activities and fungitoxicity of leaf extracts against G. boninense were expressed in inhibition of mycelia growth of G. boninense on Potato Dextrose Agar (PDA) incorporated with the four different solvent leaf extracts with range concentration of 0-40 mg mL-1. Acetone leaf extract was found to be very fungitoxic to G. boninense at concentration of 40 mg mL-1, the highest concentration tested in this study. Methanol and petroleum ether leaf extracts were having inhibitory effect with 40 mg mL-1in comparison to the absolute control. Further separation of active bioactive compounds was conducted using thin layer chromatography (TLC) bioassay for methanol and acetone leaf extracts. Acetone extract was found to possess good antifungal activity with properties of Rf 0.09, 0.21, 0.24, and 0.60 compared to methanol extract, which only showed inhibition at Rf0.74 against Aspergillus niger. The Rf values suggested that potential antimicrobial compound might be mixtures of polar and non-polar properties. The results suggested oil palm leaf extract might potentially explore as an important allelopathic agent against Ganoderma boninense.