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Year 2016 , Volume  48, Issue 1
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1
EFFECT OF EXCESSIVE SOIL MOISTURE STRESS ON SWEET SORGHUM: PHYSIOLOGICAL CHANGES AND PRODUCTIVITY
FEI ZHANGФ, YANQIU WANGФ, HUILIN YU, KAI ZHU*, ZHIPENG ZHANG AND FENG LUAND JIANQIU ZOU*

EFFECT OF EXCESSIVE SOIL MOISTURE STRESS ON SWEET SORGHUM: PHYSIOLOGICAL CHANGES AND PRODUCTIVITY
ABSTRACT:
Sweet sorghum [Sorghum bicolor (L.) Moench] is a potential bioenergy feedstock. Research explaining the response of sweet sorghum to excessive soil moisture (EM) stress at different growth stage is limited. To investigate the effect of EM stress on sweet sorghum antioxidant enzymes, osmotic regulation, biomass, quality, and ethanol production, an experiment was conducted in a glasshouse at the National Sorghum Improvement Center, Shenyang, China. Sweet sorghum (cv. LiaoTian1) was studied in four irrigation treatments with a randomized block design method. The results showed that the protective enzyme, particularly the SOD, CAT and APX in it, was significantly affected by EM stress. EM stress deleteriously affected sweet sorghum growth, resulting in a remarkable reduction of aboveground biomass, stalk juice quality, stalk juice yield, and thus, decreased ethanol yield. EM stress also caused significant reduction in plant relative water content, which further decreased stalk juice extraction rate. Sweet sorghum grown under light, medium, and heavy EM treatments displayed 5, 19, and 30% fresh stalk yield reduction, which showed a significant difference compared to control. The estimated juice ethanol yield significantly declined from 1407 ha-1 (under optimum soil moisture) to 1272, 970, and 734 L ha-1 respectively.

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2
DIVERSITY OF PHYSIOLOGICAL TRAITS IN JERUSALEM ARTICHOKE GENOTYPES UNDER NON-STRESS AND DROUGHT STRESS
RUTTANACHIRA RUTTANAPRASERT1, PORAMATE BANTERNG1*, SANUN JOGLOY1,2, NIMITR VORASOOT1, THAWAN KESMALA1 AND ARAN PATANOTHAI1

DIVERSITY OF PHYSIOLOGICAL TRAITS IN JERUSALEM ARTICHOKE GENOTYPES UNDER NON-STRESS AND DROUGHT STRESS
ABSTRACT:
Physiological traits such as SPAD Chlorophyll Meter Reading (SCMR), specific leaf area (SLA) and harvest index (HI) play an important role in crop yield. The objectives of this work were to study the effect of drought stress on HI, SCMR and SLA and explore genetic variability for these physiological traits in Jerusalem artichoke (JA) (Helianthus tuberosus L.). Field experiments were conducted in the dry period of 2010/11 and 2011/12 in the Northeast of Thailand using a strip plot design with four replications. A horizontal factor was three different water regimes (W1: 100% Crop water requirement (ETcrop), W2: 75% ETcrop and W3: 45% ETcrop) and a vertical factor was 40 JA genotypes. Measurements on HI, relative water content (RWC), SLA and SCMR were conducted at 40, 60 and 70 days after transplantation. Drought stress significantly reduced RWC and SLA but significantly increased SCMR. High variations in SCMR (32-59) and SLA (78-213 cm2 g-1) were found among genotypes. The correlations between HI and SCMR (r = 0.56 to 0.78, p≤ 0.01) were positive and significant, whereas the respective ones between HI and SLA (r = -0.60 to -0.76, p≤ 0.01) were negative and significant as those between SCMR and SLA (r = -0.73 to -0.90, p≤0.01). These findings suggested that SCMR was linked with SLA and HI in JA. SCMR could be used as a physiological trait for indirect selection for HI and productivity under various water regimes in JA.

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3
HALOPHYTIC COMPANION PLANTS IMPROVE GROWTH AND PHYSIOLOGICAL PARAMETERS OF TOMATO PLANTS GROWN UNDER SALINITY
SEMA KARAKAS1, MEHMET ALI CULLU1, CENGIZ KAYA1* AND MURAT DIKILITAS2

HALOPHYTIC COMPANION PLANTS IMPROVE GROWTH AND PHYSIOLOGICAL PARAMETERS OF TOMATO PLANTS GROWN UNDER SALINITY
ABSTRACT:
Salinity becomes a major concern when soil salt concentration becomes excessive in growth medium. Halophytes are capable of accumulating high concentrations of NaCl in their tissues, thus using halophytic plants in crop rotations or even in mixed cropping systems may be a promising management practices to mitigate salt stress related yield loses. Salinity induced yield losses and related physiological parameters on tomato plants (Lycopersicon esculentum Mill. cv. SC2121) grown with or without halophytic companion plants (Salsola soda L. and Portulaca oleracea L.) were investigated in pot experiment. Treatments consist of four soil type (collected from Harran plain-Turkey) with similar physical properties but varying in salinity level: electrical conductivity (EC): 0.9, 4.2, 7.2, and 14.1 dS m-1. The reduction in plant total dry weight was 24, 19, and 48% in soils with slight (4.2 dS m−1), moderate (7.2 dSm−1) and high (14.1 dS m−1) salinity as compared to non-saline soil (0.9 dS m-1), respectively. Leaf content of proline, malondialdehyde (MDA), catalase (CAT) and peroxidase (POX) enzyme activity increased with increasing level of salinity. In tomato plants grown in consociation with Salsola soda, salinity induced DM decrease was only 6, 12 and 28% in soils with slight, moderate and high salinity as compared to non-saline soil, respectively. However, when Portulaca oleracea used as companion plant, no significant change in biomass or fruit yield was observed. This study showed that mixed planting with Salsola soda in high saline soils may be an effective phyto-remediation technique that may secure yield formation and quality of tomato.

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4
METABOLIC IMPLICATIONS OF SALT INDUCED OSMOLYTE ACCUMULATION IN AVICENNIA MARINA
FARZEEN KHAN1, MUHAMMAD YOUSUF ADNAN1,2 AND IRFAN AZIZ1*

METABOLIC IMPLICATIONS OF SALT INDUCED OSMOLYTE ACCUMULATION IN AVICENNIA MARINA
ABSTRACT:
Salt induced solute accumulation is a well known feature for osmotic adjustment in plants under salt stress. Concomitantly, synthesis of compatible osmolytes requires energy which may compromise plant growth. This study highlights the contribution of different solutes to leaf osmolality in Avicennia marina seedlings and saplings and possible metabolic shifts under saline conditions. Salinity was imposed using different seawater strengths (0, 50% & 150% seawater salinity) to evaluate contribution of ions and organic osmolytes and their possible effects on plant growth. Optimum growth was found in 50% seawater salinity (SWS) and decreased under salt stress (150% SWS). Low values for water and osmotic potential could be accounted for an increase in the amount of Na+ which appeared to contribute the most in leaf osmolality in both seedlings (38%) and saplings (43%) under saline conditions. Among tested organic solutes, the contribution of total soluble sugars (TSS) was higher in non-saline control and moderate salinity, followed by glycinebetaine (GB) while proline (PRO) values were lowest. The amount of GB increased to a 4 fold in seedlings and 2 fold in saplings under salt stress. The increased ratio of GB:TSS in seedlings under salt stress indicates their sensitivity over saplings and a likely metabolic shift at the cost of growth.

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5
ARBUSCULAR MYCORRHIZAL FUNGI ENHANCE BASIL TOLERANCE TO SALT STRESS THROUGH IMPROVED PHYSIOLOGICAL AND NUTRITIONAL STATUS
HASHEM ABEER 1,2, ALTERAMI SALWA A.1 , ALQARAWI A.A.3, ABD_ALLAH E.F.3* AND EGAMBERDIEVA D.4

ARBUSCULAR MYCORRHIZAL FUNGI ENHANCE BASIL TOLERANCE TO SALT STRESS THROUGH IMPROVED PHYSIOLOGICAL AND NUTRITIONAL STATUS
ABSTRACT:
Pot experiments were conducted to evaluate the impact of salt stress on some physiological and biochemical attributes in sweet basil (Ocimum basilicum L.) cultivars differing in salt sensitivity and to determine the role of arbuscular mycorrhizal fungi (AMF) in ameliorating the salt stress in plant. Salt stress (250 mM NaCl) reduced the colonization potential of AMF and inhibited photosynthetic pigments, chlorophyll and carotenoids in plant tissue. AMF inoculated plants maintained higher contents of cholorphyll pigments. Salt stressed plants showed increased lipid peroxidation, antioxidant enzyme activities like superoxide dismutase (SOD), ascorbate peroxidase (APX) and peroxidase (POD). Plants inoculated with AMF showed lower lipid peroxidation and enhanced antioxidant enzyme activities. Moreover, the content of lipids, proline, and soluble sugars in basil plants was improved with AMF inoculation. AMF positively affected the uptake of important mineral elements and also reduced the uptake of deleterious ions like Na+. Hence AMF mitigated the impact of salinity on growth by reducing the oxidative damage through strengthening of the antioxidant system. Tolerant cultivar maintained higher activities of antioxidant enzymes and accumulation of osmolytes resulting in better growth than sensitive one.

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6
EFFECT OF TRANSPIRATION RATE ON SODIUM ACCUMULATION IN RICE (ORYZA SATIVA L.) GROWN UNDER SALINE CONDITIONS
A. SHEREEN1*, R. ANSARI2, S. RAZA3, M.U. SHIRAZI1, M.A. KHAN1 AND S. MUMTAZ1

EFFECT OF TRANSPIRATION RATE ON SODIUM ACCUMULATION IN RICE (ORYZA SATIVA L.) GROWN UNDER SALINE CONDITIONS
ABSTRACT:
Uptake and transporting pattern of toxic ions to the shoot is a crucial response by plant under saline conditions affecting metabolic activities and thereby determines salt sensitivity/tolerance in rice. Experiments were conducted to compare the rate of transpiration and its effect on sodium accumulation in shoot of two inbred rice lines (IR55178 – 163 tolerant and IR55178 -104 sensitive differing in salt tolerance along with a salt tolerant check Shua-92 ) under different levels of salinity (0, 50 and 75 mM). The parameters studied indicated that transpiration rates were sharply decreased in response to salinity in both genotypes. While, sodium uptake continued to increase however, the tolerant genotype maintained transpiration rate quite satisfactorily and also exhibited ability to maintain higher K: Na ratio and low Na concentration in the xylem than the sensitive genotype. It is concluded that transport of sodium ions in the present study was negatively correlated with transpiration (r = -0.806).

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7
APPLICATION OF ACC-DEAMINASE CONTAINING PGPR IMPROVES SUNFLOWER YIELD UNDER NATURAL SALINITY STRESS
MUHAMMAD ZAHID KIANI1, TARIQ SULTAN2, ARSHAD ALI2 AND ZARRIN FATIMA RIZVI3*

APPLICATION OF ACC-DEAMINASE CONTAINING PGPR IMPROVES SUNFLOWER YIELD UNDER NATURAL SALINITY STRESS
ABSTRACT:
Salinity increase ethylene level in root rhizosphere. Plant growth promoting rhizobacteria having ACC deaminase have ability to mitigate the adverse effect of enhanced level of ethylene. Two field experiments were carried out at two different sites during 2013. The site1 had ECe 9.42 dS m-1 and SAR 20.75 (mmolc L-1)1/2, while site2 had ECe 7.51 dS m-1 and SAR 16.25 (mmolc L-1)1/2. Four PGPR strains (KS 44, KS 7, KS 41, KS 42) having ACC deaminase activity were selected for this study. Randomized completes block design (RCBD) was used and hybrid sunflower seeds (SMH-0917) were inoculated with these PGPR strains. The crop was harvested at maturity and data were recorded about plant height, shoot dry weight, head dry weight, number of seed head-1, 1000 seed weight, seed weight head-1 and seed yield kg ha-1. The data were statistically analyzed by using soft ware statistic 8.1. The average increase in grain yield (site 1 + site 2) was 110.07, 108.89, 49.09, 65.77 and 69.70% over control due to inoculation with KS 44, KS 7, KS 41, KS 42 and KS mix treatments respectively. On the basis of statistical analysis, the bacterial strains KS 44 and KS 7, were declared as most promising strains on both sites. Hence these two strains (KS 44 and KS 7) could be used to mitigate negative impact of salinity stress on sunflower.

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8
BIO-INFORMATIC ANALYSIS OF A VACUOLAR Na+/H+ ANTIPORTER (ALaNHX) FROM THE SALT RESISTANT GRASS AELUROPUS LAGOPOIDES
MUHAMMAD ZAHEER AHMED1*, AKIRA KIKUCHI2, KAZUO N. WATANABE2 AND M. AJMAL KHAN1, 3

BIO-INFORMATIC ANALYSIS OF A VACUOLAR Na+/H+ ANTIPORTER (ALaNHX) FROM THE SALT RESISTANT GRASS AELUROPUS LAGOPOIDES
ABSTRACT:
Sodium-hydrogen antiporter (NHX) protein regulates the trans-membrane transport of Na+ in higher plants. Vacuolar-NHX is a type of NHX protein located on tonoplast and minimizes the accumulation of Na+ in cytoplasm by compartmentalizing into vacuole especially in salt tolerant plants. In Aeluropus lagopoides, AlaNHX [NCBI: GU199336, Vacuolar-NHX] plays a vital role for efficient Na+ sequestration into the vacuole and helps in plant survival in saline areas. Therefore, sequence analysis, structural analysis and modeling of AlaNHX were performed through bioinformatics tools. Homology of AlaNHX was 99% similar with the Na+/H+ antiporter of Aeluropus littoralis. Sequence of AlaNHX consisted of 2353 bp including 337 bp of un-translated regions (UTR) at 5΄ and 393 at 3΄ end. In addition, AlaNHX have an “open reading frame” (ORF) of 1623 bp which translated into 59.4 KDa protein containing 540 amino acids (Leucine + Serine contributed in 22% of peptide chain). AlaNHX protein consists of 10 transmembrane domains (TMD; 3 primary and 7 secondary protein structural type) and a long (95 amino acids) carboxyl terminal end in cytoplasmic region. In addition, 3, 5, 7 and 8 TMD regions of AlaNHX were highly conserved. Different glycosylation, phosphorylation and myristoylation sites were also found in AlaNHX protein. Three-dimensional (3D) structure analysis revealed that this protein may be classified as stable and of hydrophobic nature containing a significant proportion of alpha helices. In this study, a three-dimensional structure of AlaNHX protein was predicted by using in-silico3D homology modeling technique. This study provides baseline information for understanding the importance of NHX protein structure in salinity resistance of grasses. This information could help in improving salinity tolerance in salt sensitive grasses through genetic engineering.

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9
CITRIC ACID IN THE NUTRIENT SOLUTION INCREASES THE MINERAL ABSORPTION IN POTTED TOMATO GROWN IN CALCAREOUS SOIL
FABIÁN PÉREZ-LABRADA, ADALBERTO BENAVIDES-MENDOZA*, LUIS ALONSO VALDEZ-AGUILAR AND VALENTÍN ROBLEDO-TORRES

CITRIC ACID IN THE NUTRIENT SOLUTION INCREASES THE MINERAL ABSORPTION IN POTTED TOMATO GROWN IN CALCAREOUS SOIL
ABSTRACT:
Calcareous soils have low availability of mineral nutrients that are essential for crops. The aim of the study was to determine the effect of citric acid (CA) applied in a nutrient solution on the composition of the pore water of a calcareous soil and its correlation with the chemical composition and quality of the tomato plants and fruits. The experimental design was a randomized block; the treatments were different concentrations of CA added to the Steiner nutrient solution. We used CA at 10-2, 10-4 and 10-6 M and a test treatment without CA. A decrease in the electrical conductivity (EC) and oxidation-reduction potential (ORP) was observed in soil pore water (SPW), as well as changes in the concentrations of Ca2+, Mg2+, Na+ and K+ in response to the addition of CA. The concentrations of Zn, Na, Ca, and N increased in leaves, whereas in the fruit, the concentrations of Mn, Na, Mg, and P increased. Significant correlations arise between Na+ and Mg2+ (R=0.60), Na+ and Ca2+ (R=0.68), and Mg2+ and Ca2+ (R=0.76) in the soil pore water, as well as a significant negative correlation (R=-0.73) between the concentration of K+ in the soil pore water and the concentration of Mg in stems and leaves. No relationship arises between the mineral content in the fruits and the mineral composition of the SPW. The addition of CA changed the chemical composition of the soil pore water and plant tissues. CA at 10-6 M increased the fruit production per plant significantly.

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10
PHYSIOLOGICAL AND NUTRITIONAL RESPONSES OF TWO DISTINCTIVE QUINCE (CYDONIA OBLONGA MILL.) ROOTSTOCKS TO BORON TOXICITY
FIGEN ERASLAN1*, MEHMET POLAT2, ADNAN YILDIRIM2 AND ZELIHA KUCUKYUMUK1

PHYSIOLOGICAL AND NUTRITIONAL RESPONSES OF TWO DISTINCTIVE QUINCE (CYDONIA OBLONGA MILL.) ROOTSTOCKS TO BORON TOXICITY
ABSTRACT:
The effects of excess boron (B) on some physiological and nutritional parameters of two distinctive quince (Cydonia oblonga Mill.) rootstocks were investigated. Throughout the world, B toxicity is a widely faced problem of soil in arid and semi-arid environments. In a greenhouse study, boron was applied at the rates of 0 and 40 mg kg-1 soil to quince A and quince C rootstocks. Toxicity of B differentially affected studied parameters and rootstocks. Boron toxicity increased B concentrations of both rootstocks however the increase was more pronounced in quince A rootstock. SPAD readings, (SPAD-meter, Minolta 502 Co Ltd., Japan) as a measure of chlorophyll decreased under B toxicity. Boron toxicity increased membrane permeability and anthocyanin in both rootstocks. Although, there is rootstocks difference, lipid peroxidation (MDA) and proline and TAA (non-enzymatic total antioxidant activity) increased in response to B toxicity. In general, quince C had lower MDA (Malondialdehyde) and TAA but lower level of proline as compared to quince A. Boron toxicity did not affect the concentrations of P, Ca, Zn and Cu however increased B and Mn concentrations. Magnesium (Mg), Mn and Fe concentrations of quince were found higher than that of quince C. Indicating a genotypic effect, quince A and quince C responded to B toxicity differentially.

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11
EVALUATION OF VARIOUS MORPHO-PHYSIOLOGICAL AND GROWTH TRAITS OF DUAL PURPOSE WHEAT UNDER EARLY SOWING DATES
FAZAL MUNSIF1, M.ARIF1, KAWSAR ALI2*, M.J. KHAN3, SAID MUNIR4 AND F. RASUL5

EVALUATION OF VARIOUS MORPHO-PHYSIOLOGICAL AND GROWTH TRAITS OF DUAL PURPOSE WHEAT UNDER EARLY SOWING DATES
ABSTRACT:
The use of wheat as dual purpose crop (for fodder and grain) is considered as promising agronomic management practice fulfilling human and animal need at same time. These expectations are largely found on positive experiences by growing wheat as dual purpose crop throughout the world. However, the validity of these results in Pakistan needs confirmation from field experiments under various sowing date and potential cultivars. The impact of cutting and sowing date on morphological and physiological traits of already existing wheat cultivars was evaluated at Agricultural Research Farm of The University of Agriculture, Peshawar Pakistan in winter 2010-2011 and 2011-2012. Three sowing dates i.e. 15th October, 30th October and 14th November and six wheat cultivars (Ghaznavi-98, Fakhre-Sarhad (FS)-99, Ghaznavi-98, Saleem-2000, Uqab-2000 and Siran-2008) were compared for dual purpose wheat production during the course of experiment. Findings showed that significant reduction were recorded in nodes tiller-1, plant height, spike weight and grains weight spike-1 when sowing of wheat delayed from 15th and 30th October to 15th November. Considerable variations were also noted in nodes tiller-1, internodes length and leaf area index, plant height and lodging score among wheat cultivars but it was mainly due to their genetic characteristic not because of cutting treatment. It was observed that cutting caused 5.5, 12.1, 6.2, 4.1, 25.9, 7.1, 6.6% reduction in wheat leaf area, leaf area index, internode length, plant height, lodging score, spike weight and grains weight spike-1 , respectively over non cut plots in both year of the experiment. Interestingly, no-significant change was noted in spike length and number of nodes tiller-1 under cut and no cut plots. The conclusive findings suggested that optimum sowing date (15th and 30th October) for all wheat cultivars could be utilized as potential source of dual purpose wheat for substantial reduction of lodging without compromising on important morphological and physiological characteristics.

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12
BIOMASS PRODUCTION OF SOME SALT TOLERANT TREE SPECIES GROWN IN DIFFERENT ECOLOGICAL ZONES OF PAKISTAN
KHALID MAHMOOD*, M. ISMAIL CHUGHTAI, ABDUL RASUL AWAN AND RIAZ A. WAHEED

BIOMASS PRODUCTION OF SOME SALT TOLERANT TREE SPECIES GROWN IN DIFFERENT ECOLOGICAL ZONES OF PAKISTAN
ABSTRACT:
A study was carried out to evaluate the biomass production potential of salt tolerant tree species grown in saline environments. For this purpose, 5 sites near Badin, Gawadar, Lahore, Faisalabad and Peshawar in different ecological zones of Pakistan were selected. Plantations of 7 tree species common to all sites including Eucalyptus camaldulensis, Phoenix dactylifera, Acacia nilotica, Acacia ampliceps, Prosopis juliflora, Casurina obesa and Tamarix aphylla were selected for non-destructive biomass measurements. Five trees from each species at each site were assessed for plant height, girth at breast height, canopy area, canopy shape and number of branches. For destructive biomass estimation, six trees of four species (Eucalyptus camaldulensis, Acacia nilotica, Prosopis juliflora and Tamarix aphylla) were harvested at two sites near Lahore and Faisalabad. Biomass of whole tree and its components like stem, branches, twigs, leaves and fruits were determined. Soil and water resources of these sites were also characterized. Results indicated that E. camaldulensis produced maximum average biomass 329 kg in 81/2 years at soil salinity (EC 1:1) 8.5 to 9.4 dS m-1 and T. aphylla produced 188 kg at soil salinity 12.8 dS m-1 in 91/2 years. A. nilotica produced biomass 187 kg at 16.9 dS m-1 in 10 years at Faisalabad; while at Lahore, 369 kg in 18 years under soil salinity level 7.3 dS m-1. P. juliflora produced minimum biomass 123 kg at soil salinity 7.1 dS m-1 in 8 years at Lahore and 278 kg at soil salinity 17.2 dS m-1 in 16 years at Faisalabad. Both soil and water quality was comparatively better at Gawadar and Faisalabad than other sites. Overall, it is concluded that studied tree species are good performer on salt-affected soils and can make saline areas productive.

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13
EFFECTS OF SALICYLIC ACID ON THE GROWTH AND PHYSIOLOGICAL CHARACTERISTICS IN CYCLOCARYA PALIURUS SEEDLINGS
YINFENG XIE*, NAN GUO, YINGYING ZHANG, NANA LIU, TAO WANG, WANXIANG YANG AND SHENGZUO FANG

EFFECTS OF SALICYLIC ACID ON THE GROWTH AND PHYSIOLOGICAL CHARACTERISTICS IN CYCLOCARYA PALIURUS SEEDLINGS
ABSTRACT:
Field studies were conducted to examine the effects of salicylic acid (SA) on the growth and physiological characteristics of Cyclocarya paliurus seedlings by spraying the foliage with 0.0 (control), 0.2, 1.0, and 2.0 mM salicylic acid (SA). Proper concentrations of SA improved the relative growth yield of seedling stems and the soluble protein and sugar content of the leaves. It also increased the content of secondary metabolites including triterpenoids, flavonoids, quercetin and kaempferol, mineral elements K, Ca, Mg, Mn, Fe and Cu. Moreover, it stimulated the activities of superoxide dismutase (SOD), peroxidase (POX) and phenylalanine ammonia-lyase (PAL) in the leaves. The effects of SA on these indices were dose dependent. The relative growth of seedling stem diameter and quercetin content increased gradually with an increase in concentration of SA from 0.0-2.0 mM. A concentration of 0.2 mM was optimal to promote content of soluble protein, sugars, Ca, Mg, Mn, Fe, and Cu, and SOD activity and significanty increased by 38.6%, 22.1%, 17.7%, 8.2%, 20.3%, 23.2%, 15.6%, and 52.4%, respectively, as compared with the control (CK). However, the maximal increase in activities of PAL, POX, and content of triterpenoids, kaempferol, and flavonoids was attained at 1.0 mM treatment, which significantly increased by 76.5%, 78.4%, 76.4%, 96.3%, and 107.4%, respectively, compared with CK. Correlation analysis revealed positive relationships between activities of PAL, POX and content of triterpenoids, quercetin, kaempferol, and flavonoids within a certain concentration range of SA. These results suggested that an appropriate concentration (0.2-1.0 mM) of SA was not only effective in the improvement of physiological function of C. paliurus, but also increased seedling resistance; additionally, it helped to stimulate the synthesis of medicinal components in leaves.

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14
MULTIVARIATE ANALYSIS OF SOME PINE FORESTED AREAS OF AZAD KASHMIR-PAKISTAN
TASVEER ZAHRA BOKHARI1,2, YU LIU1, MOINUDDIN AHMED3, MUHAMMAD FAHEEM SIDDIQUI4, QIANG LI1, ZAHEER UDDIN KHAN5 AND SAEED AHMED MALIK2

MULTIVARIATE ANALYSIS OF SOME PINE FORESTED AREAS OF AZAD KASHMIR-PAKISTAN
ABSTRACT:
Abstracts: Floristic composition and communities in Azad Kashmir area of Pakistan were studied by using multivariate analysis. Quantitative sampling from thirty one sites was carried out in different coniferous forests of Azad Kashmir in order to analyze the effects of past earthquakes and landslides on vegetation of these areas. Though coniferous forests were highly disturbed either naturally or anthropogenic activities, therefore sampling was preferred to those forests which were near fault line. Trees were sampled using Point Centered Quarter (PCQ) method. Results of cluster analysis (using Ward’s method) yielded six groups dominated by different conifer species. Group I and V were dominated by Pinus wallichiana while this species was co-dominant in group III. Other groups showed the dominance of different conifer species i.e. Cedrus deodara, Pinus roxburghii, Picea smithiana and Abies pindrow. Both the cluster analysis and ordination techniques (by two dimensional non-metric multidimensional scaling) classify and ordinate the structure of various groups indicating interrelationship among different species. The groups of trees were readily be superimposed on NMS ordination axes; they were well classified and well separated out in ordination. The present research revealed that these forests had diverse and asymmetric structure due to natural anthropogenic disturbances and overgrazing, which were key factors in addition to natural disturbances. However, some of the forests showed considerably stable structure due to less human interference.

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15
ORDINATION STUDY OF VEGETATION ANALYSIS AROUND WETLAND AREA: A CASE STUDY OF MANGLA DAM, AZAD KASHMIR, PAKISTAN
RABAIL UROOJ1, SHEIKH SAEED AHMAD1*, MUHAMMAD NAUMAN AHMAD2, HAROON AHMAD3 AND MUHAMMAD NAWAZ4

ORDINATION STUDY OF VEGETATION ANALYSIS AROUND WETLAND AREA: A CASE STUDY OF MANGLA DAM, AZAD KASHMIR, PAKISTAN
ABSTRACT:
Present study was conducted at Mangla Dam for vegetation ordinal classification by applying multivariate analysis in order to find relationship between vegetation and their edaphic factors. Samples of soil and herbaceous vegetation were randomly collected by using 1*1 square meter quadrats. Total 37 plant species belonging to 17 families were identified. Canonical Correspondence Analysis as direct ordination technique was applied by using CANOCO software. Results of analytical tests revealed that concentration of micro and macro nutrients along electrical conductivity and pH in different soil samples were varying to a greater level in study area while Cynodon dactylon showed higher abundance over broad range of all edaphic factors concentration.

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16
THE BELA FOREST ECOSYSTEM OF DISTRICT JHELUM, A POTENTIAL CARBON SINK
SAJJAD SAEED1, M. IRFAN ASHRAF1, ADNAN AHMAD2* AND ZIAUR RAHMAN2

THE BELA FOREST ECOSYSTEM OF DISTRICT JHELUM, A POTENTIAL CARBON SINK
ABSTRACT:
The present study was carried out in the Bela forest of District Jhelum (Punjab). The study was aimed to estimate the growing stock, biomass and carbon stock of the Bela plantation. Carbon stock in the Bela plantation was assessed in the Upperstory vegetation, understorey vegetation and in soil. The major tree species in the Bela plantation of the study site were Eucalyptus camaldulensis (EC), Dalbergia sissoo (DS), Broussonetia papyrifera (BP), Morus alba (MA) and Acacia modesta (AM). The results of the present study reveled that specie wise stem density ranges from 8±1 to 274±3 trees ha-1 while the mean stem density was 691±13 trees ha-1. The mean height of the trees were in the ranged of 9.51±0.98 m (Morus alba) to 18.97±2.48m (Eucalyptus camaldulensis). The value of basal area ranges from 0.22±0.01 m2 ha-1 to 18.17±0.28 m2 ha-1. The average recorded stem volume was 278.92±7.41 m3 ha-1. The total tree biomass varied between 0.71±0.05 t ha-1 to 176.31±3.19 t ha-1. The total calculated biomass in the shrubs and grasses was 4.93±2.7 t ha-1 while the recorded total carbon stock in the shrubs and grasses was 2.45±1.35 t ha-1. Average soil carbon stock was determined as 30.19 ±12.10 t ha-1 in the study area. Over all the Bela forest of the study site stored about 198.18±18 t ha-1 of carbon. Among the different carbon pools the maximum carbon was stored by the Upper storey vegetation biomass (83.53%) fallowed by soil (15.23%) while the minimum carbon stock was stored in Understory vegetation biomass (1.23%).

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17
MALE SPECIFIC GENE EXPRESSION IN DIOECIOUS PHOENIX DACTYLIFERA (DATE PALM) TREE AT FLOWERING STAGE
ABDULHAFED A. AL-AMERI, FAHAD AL-QURAINY, ABDEL-RHMAN Z. GAAFAR, SALIM KHAN* AND M. NADEEM

MALE SPECIFIC GENE EXPRESSION IN DIOECIOUS PHOENIX DACTYLIFERA (DATE PALM) TREE AT FLOWERING STAGE
ABSTRACT:
Date palm is a long-living and evergreen important tree in the semiarid regions. Its fruit is rich in carbohydrate and fibres. Transcriptional profiling was compared among male and female trees of dioecious date palm at flowering stage. Male specific genes are expressed at flowering stage which was studied using the cDNA–SCoT marker. We developed sequence characterized amplified region (SCAR) markers of size 253 bp from male tree based on cDNA-SCoT fingerprinting. Further, developed SCAR marker was validated on the independently collected samples of both types of trees at flowering stage. The unique and specific band (253 bp) was amplified from male samples only whereas it was absent from female samples.

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18
THE ACTIVITY AND GENE EXPRESSION LEVELS OF P5CS AND δ-OAT IN MEDICAGO SATIVA CV. PIANGUAN UNDER DROUGHT STRESS
HUI-SEN ZHU1, XIU-JU YU1, XIANG ZHAO1, WU-DE YANG2 AND KUAN-HU DONG1*

THE ACTIVITY AND GENE EXPRESSION LEVELS OF P5CS AND δ-OAT IN MEDICAGO SATIVA CV. PIANGUAN UNDER DROUGHT STRESS
ABSTRACT:
Medicago sativa cv. Pianguan, an excellent local germplasm in Shanxi province, China, with the resistance to cold and drought is broadly planted. Many studies have showed that drought resistance of plants is positively correlated with the accumulation of proline, P5CS and δ-OAT play important roles in synthesis of proline. In this study, the proline content, activities and expression of P5CS and δ-OAT mRNA were detected in Medicago sativa cv. Pianguan under drought stress. The results showed that the proline content in Medicago sativa cv. Pianguan increased in the process of drought stress. The expression levels of P5CS and δ-OAT mRNA changed correspondingly at the different stress time point. In conclusion, P5CS and δ-OAT could participate in the drought resistance process by regulating proline content in Medicago sativa cv. Pianguan.

137-142 Download
19
MOLECULAR PHYLOGENY OF TRITICUM AND AEGILOPS GENERA BASED ON ITS AND MATK SEQUENCE DATA
AYTEN DIZKIRICI1, CIGDEM KANSU2 AND SERTAC ONDE2*

MOLECULAR PHYLOGENY OF TRITICUM AND AEGILOPS GENERA BASED ON ITS AND MATK SEQUENCE DATA
ABSTRACT:
Understanding the phylogenetic relationship between Triticum and Aegilops species, which form a vast gene pool of wheat, is very important for breeding new cultivated wheat varieties. In the present study, phylogenetic relationships between Triticum (12 samples from 4 species) and Aegilops (24 samples from 8 species) were investigated using sequences of the nuclear ITS rDNA gene and partial sequences of the matK gene of chloroplast genome. The phylogenetic relationships among species were reconstructed using Maximum Likelihood method. The constructed tree based on the sequences of the nuclear component (ITS) displayed a close relationship between polyploid wheats and Aegilops speltoides species which provided new evidence for the source of the enigmatic B genome donor as Ae. speltoides. Concurrent clustering of Ae. cylindrica and Ae. tauschii and their close positioning to polyploid wheats pointed the source of the D genome as one of these species. As reported before, diploid Triticum species (i.e. T. urartu) were identified as the A genome donors and the positioning of these diploid wheats on the constructed tree are meaningful. The constructed tree based on the chloroplastic matK sequences displayed same relationship between polyploid wheats and Ae. speltoides species providing evidence for the later species being the chloroplast donors for polyploid wheats. Therefore, our results supported the idea of coinheritance of nuclear and chloroplast genomes where Ae. speltoides was the maternal donor. For both trees the remaining Aegilops species produced a distinct cluster whereas with the exception of T. urartu, diploid Triticum species displayed a monophyletic structure.

143-153 Download
20
GENOME-WIDE IDENTIFICATION AND ANALYSIS OF GENES ENCODING PHD-FINGER PROTEIN IN TOMATO
XUEJIN CHEN1,2, SIKANDAR HAYAT1 AND ZHIHUI CHENG1*

GENOME-WIDE IDENTIFICATION AND ANALYSIS OF GENES ENCODING PHD-FINGER PROTEIN IN TOMATO
ABSTRACT:
The PHD-finger proteins are conserved in eukaryotic organisms and are involved in a variety of important functions in different biological processes in plants. However, the function of PHD fingers are poorly known in tomato (Solanum lycopersicum L.). In current study, we identified 45 putative genes coding PHD-finger protein in tomato distributed on 11 chromosomes except for chromosome 8. Some of the genes encode other conserved key domains besides PHD-finger. Phylogenetic analysis of these 45 proteins resulted in seven clusters. Most PHD finger proteins were predicted to PML body location. These PHD-finger genes displayed differential expression either in various organs, at different development stages and under stresses in tomato. Our study provides the first systematic analysis of PHD-finger genes and proteins in tomato. This preliminary study provides a very useful reference information for PHD-finger proteins in tomato. They will be helpful for cloning and functional study of tomato PHD-finger genes.

155-165 Download
21
CHARACTERIZATION AND DIVERSITY OF NOVEL PIF/HARBINGER DNA TRANSPOSONS IN BRASSICA GENOMES
FAISAL NOUROZ1,3*, SHUMAILA NOREEN2 AND J.S. (PAT) HESLOP-HARRISON3

CHARACTERIZATION AND DIVERSITY OF NOVEL PIF/HARBINGER DNA TRANSPOSONS IN BRASSICA GENOMES
ABSTRACT:
Among DNA transposons, PIF/Harbinger is most recently identified superfamily characterized by 3 bp target site duplications (TSDs), flanked by 14-45 bp terminal inverted repeats (TIRs) and displaying DDD or DDE domain displaying transposase. Their autonomous elements contain two open reading frames, ORF1 and ORF2 encoding superfamily specific transposase and DNA-binding domain. Harbinger DNA transposons are recently identified in few plants. In present study, computational and molecular approaches were used for the identification of 8 Harbinger transposons, of which only 2 were complete with putative transposase, while rest 6 lack transposase and are considered as defective or non-autonomous elements. They ranged in size from 0.5-4 kb with 3 bp TSDs, 15-42 bp TIRs and internal AT rich regions. The PCR amplification of Brassica Harbinger transposase revealed diversity and ancient nature of these elements. The amplification polymorphism of some non-autonomous Harbingers showed species specific distribution. Phylogenetic analyses of transposase clustered them into two clades (monocot and dicot) and five sub-clades. The Brassica, Arabidopsis and Malus transposase clustered into genera specific sub-clades; although a lot of homology in transposase was observed. The multiple sequence alignment of Brassica and related transposase showed homology in five conserved blocks. The DD35E triad and sequences showed similarity to already known Pong-like or Arabidopsis ATISI12 Harbinger transposase in contrast to other transposase having DD47E or DD48E motifs. The present study will be helpful in the characterization of Harbingers, their structural diversity in related genera and Harbinger based molecular markers for varietal/lines identifications.

167-178 Download
22
ANATOMICAL DIFFERENCES OF THE TURKISH STUCKENIA BÖRNER (POTAMOGETONACEAE) AND THEIR TAXONOMIC SIGNIFICANCE
CANDAN AYKURT1* AND ĐSMAĐL GÖKHAN DENĐZ2

ANATOMICAL DIFFERENCES OF THE TURKISH STUCKENIA BÖRNER (POTAMOGETONACEAE) AND THEIR TAXONOMIC SIGNIFICANCE
ABSTRACT:
Anatomical studies of the Stuckenia species occurring in Turkey were conducted. The results showed that the presence or absence of interlacunar bundles and the stelar type were the most important in plant anatomical characters of the stem. The leaves of the all investigated species typically had uniseriate epidermis with a thin cuticle, aerenchyma was composed of arm-shaped chlorophyllous cells, similar to those found in the stem and the peduncle. Two lateral vascular bundles and a central vascular bundle are present in the leaves of all investigated species. However, the number of lateral vascular bundles can vary in S. pectinata according to leaf width and the fiber bundles are also present in the triquetrous leaves of this species.

179-186 Download
23
MORPHOLOGICAL AND ANATOMICAL CHARACTERIZATION AND TRACE ELEMENTS COMPOSITION OF ONOSMA ARGENTATA HUB.-MOR. (BORAGINACEAE) ENDEMIC TO TURKEY
MUSTAFA ÖZKAN1, ERGIN KARIPTAŞ1, CÂNAN ÖZDEMIR2, ALI KANDEMIR3 AND YURDANUR AKYOL4

MORPHOLOGICAL AND ANATOMICAL CHARACTERIZATION AND TRACE ELEMENTS COMPOSITION OF ONOSMA ARGENTATA HUB.-MOR. (BORAGINACEAE) ENDEMIC TO TURKEY
ABSTRACT:
Onosma argentata Hub.-Mor. is an endemic species growing in Turkey. It is under the threat like many other endemic species in Turkey and its IUCN category is evaluated as vulnerable. According to literature, this species was known only from the two collections, in this study the plants were collected from different localities mentioned in literature. Plant is perennial. The stem is 20-35cm, shortly hairy and rigid-setose, clothed at base with remains of leaves.The aim of this study is to provide information on the anatomical and morphological properties of O. argentata. Anatomical sections taken from the root, stem and leaves of the plants. The results have been compared with some species of Boraginaceae. In addition, the concentrations of trace elements O. argentata were determined by using atomic absorption spectrometry. Large amount of sodium (Na), aluminum (Al), potassium (K) and iron (Fe) were present in all flowers, leaf and stem of O. argentata plant.

187-191 Download
24
FACTORS INFLUENCING IN VITRO SEED GERMINATION, MORPHOGENETIC POTENTIAL AND CORRELATION OF SECONDARY METABOLISM WITH TISSUE DEVELOPMENT IN PRUNELLA VULGARIS L.
HINA FAZAL1,2, ZABTA KHAN SHINWARI2, NISAR AHMAD3 AND BILAL HAIDER ABBASI2*

FACTORS INFLUENCING IN VITRO SEED GERMINATION, MORPHOGENETIC POTENTIAL AND CORRELATION OF SECONDARY METABOLISM WITH TISSUE DEVELOPMENT IN PRUNELLA VULGARIS L.
ABSTRACT:
Plant growth regulators (PGRs), polyamines (PAs) and temperature regimes are the key factors that influence morphogenesis and plant architectural development; however, the understanding that how these factors control plant growth and development is still poor and needs further research in Prunella vulgaris. In this study, we monitored the effect of these factors on seed germination, morphogenetic potential and secondary metabolism. Different temperature regimes showed that 25°C is the most suitable temperature for seed germination (88.87±1.76%) on Murashige and Skoog (MS) basal medium. The synergistic combinations of kinetin (Kn), 6-benzyladenine (BA) and putrescine (PUT; 2.0 mg l-1) promoted seed germination (90.22±4.51%) after 24 days of inoculation. A combination of Kn and PUT (1.0 mg l-1) encouraged mean shoot length (11.0±1.95 mm) with the optimum amount of chlorophyll content (23.73±1.8 μg cm-2). However, maximum mean root length (13±0.65 mm) was observed on medium containing Kn and spermidine (SPD, 2.0 mg l-1). Maximum calli (71.56±2.63%) were obtained from root explants on ½ MS-medium containing indole butyric acid (IBA) and α-naphthalene acetic acid (NAA; 0.5 mg l-1). Higher number of shoots (78.5±3.75%) was obtained with Kn and PUT (1.0 mg l-1). IBA concentration of 1.0 mg l-1 was found effective for root formation (74.71±3.3%). Moreover, PGRs and PAs have a significant effect on accumulation of total phenolics, flavonoids and DPPH activity. This protocol is helpful for consistent plantlets and prunellin production in P. vulgaris L.

193-200 Download
25
THE PHYLOGENETIC SIGNIFICANCE OF FRUIT STRUCTURAL VARIATION IN THE TRIBE HETEROMORPHEAE (APIACEAE)
MEI LIU1*, BEN-ERIK VAN WYK2, PATRICIA M. TILNEY2, GREGORY M. PLUNKETT3 AND PORTER P. LOWRY II4,5 AND ANTHONY R. MAGEE6

THE PHYLOGENETIC SIGNIFICANCE OF FRUIT STRUCTURAL VARIATION IN THE TRIBE HETEROMORPHEAE (APIACEAE)
ABSTRACT:
Fruit structure of Apiaceae was studied in 19 species representing the 10 genera of the tribe Heteromorpheae. Our results indicate this group has a woody habit, simple leaves, heteromorphic mericarps with lateral wings. fruits with bottle-shaped or bulging epidermal cells which have thickened and cutinized outer wall, regular vittae (one in furrow and two in commissure) and irregular vittae (short, dwarf, or branching and anatosmosing), and dispersed druse crystals. However, lateral winged mericarps, bottle-shaped epidermal cells, and branching and anatosmosing vittae are peculiar in the tribe Heteromorpheae of Apioideae sub family. Although many features share with other early-diverging groups of Apiaceae, including Annesorhiza clade, Saniculoideae sensu lato, Azorelloideae, Mackinlayoideae, as well as with Araliaceae. Our study shows that fruit anatomy can be used to define the tribe by molecular phylogenetic studies and support that Heteromorpheae are close to Annesorhiza clade and both are placed in the basal position of Apioideae.

201-210 Download
26
ASPECTS OF THE REPRODUCTIVE BIOLOGY OF STENANONA FLAGELLIFLORA (ANNONACEAE)
LILIANA XICOHTÉNCATL-LARA1, DULCE MARÍA FIGUEROA-CASTRO1*, AGUSTINA ROSA ANDRÉS-HERNÁNDEZ1 AND ÁLVARO CAMPOS-VILLANUEVA2

ASPECTS OF THE REPRODUCTIVE BIOLOGY OF STENANONA FLAGELLIFLORA (ANNONACEAE)
ABSTRACT:
Stenanona flagelliflora was described in 2004. There are no studies on its biology. The goal of this study was to document some aspects of its reproductive biology. The particular objectives were to: i) describe the variation on vegetative and floral traits; ii) establish the composition of the community of floral visitors; iii) estimate the mating system and reproductive success; and, iv) establish the relationship between vegetative traits and reproductive success. The study was conducted within the Los Tuxtlas Biosphere Reserve, Veracruz, Mexico. We quantified several vegetative and floral traits; conducted observations and collected floral visitors; and determined the mating system and reproductive success. Stenanona flagelliflora has relatively few stamens and carpels, but a relatively high viability of pollen grains. The most abundant floral visitors were dipterans from the Phoridae Family. Mating system is between xenogamous and facultative xenogamous; thus, pollination depends upon pollen vectors. Fruit-set was relatively high; but seed-set was very low, because most monocarps did not contain seeds. Our results suggest that reproduction of S. flagelliflora is not limited by resource availability, but by pollinator frequency and effectiveness. To our knowledge, this is the first study on the reproductive biology of a species within the Stenanona genus.

211-221 Download
27
SEED MORPHOLOGY OF ZANTHOXYLUM (RUTACEAE) IN CHINA AND ITS SYSTEMATIC SIGNIFICANCE
MING CAO1*, FENG-JUAN MOU2, AMIN SHAH3**, SARVAT RAHIM3 AND JIAN-SHE CHEN1

SEED MORPHOLOGY OF ZANTHOXYLUM (RUTACEAE) IN CHINA AND ITS SYSTEMATIC SIGNIFICANCE
ABSTRACT:
Seed morphology of 31 species belonging to the genus Zanthoxylum in China were examined using light microscopy and scanning electron microscopy. The macro- and micro-morphological characters of seed, including seed surface, colour, shape, size and coat sculpture were examined. Under light microscopy, the results indicate that seed surface is more or less smooth; seed colour is black or brown to red; seed shape is spheroidal or elliptical and size is large. Under scanning electron microscopy, the seed epidermal cellular arrangement is netted and the epidermal cell shape ranges from tetragonal to hexagonal in all taxa. The fine relief of the outer cell wall is rough, smooth and finely folded sculpturing. The systematic significance of these seed characters was evaluated. It was proved that the seed coat sculpture was a valuable character which offers evidence to solve some taxonomical problems of Zanthoxylum. Based on the seed morphological data, the present study proved that subgen. Fagara and subgen. Zanthoxylum shared many common seed characters, suggesting a close relationship between the two subgenera. The value of some characters of seed in identifying species of Zanthoxylum is also discussed here.

223-230 Download
28
MUTATIONAL, PHYLOGENY AND EVOLUTION ANALYSES OF SALVIA COPALYL DIPHOSPHATE SYNTHASE
DA CHENG HAO1*, RAMESHA B. THIMMAPPA2 AND PEI GEN XIAO3

MUTATIONAL, PHYLOGENY AND EVOLUTION ANALYSES OF SALVIA COPALYL DIPHOSPHATE SYNTHASE
ABSTRACT:
The cyclization of geranylgeranyl diphosphate (GGPP) is catalyzed by copalyl diphosphate synthase (CPS), a class II diterpene synthase (diTPS), to form copalyl diphosphate (CPP), which is an essential substrate of a variety of diterpenes in secondary metabolism of angiosperm including Salvia medicinal plants. The protein environment of the N-terminal class II active site stabilizes the carbocation intermediates and maintains the catalytic activity of angiosperm class II diTPS. The virtual modeling and mutagenesis of the class II diTPS of Salvia miltiorrhiza (SmCPS) were accomplished to illuminate the catalytic activity of SmCPS. Terminal truncations and point mutations established the role of the βγ domain and α domain, i.e., they facilitate the flexible conformational change of the class II active site after substrate binding. E203 and K238 in the N-terγ domain of SmCPS1are functional in the substrate binding and conformational transition and might be essential in catalysis. Similar to other CPSs, the ensuing protonation of the GGPP substrate and coordination of the diphosphate group are governed by highly conserved residues in the DxDD motif of SmCPS, e.g., D372 of CPS1. Moreover, F256 andY505stabilize the carbocation and control the enzymatic activity during CPP formation. The amino acids of the predicted active sites, despite under purifying selection, vary greatly, corresponding to the functional flexibility of angiosperm CPSs. Molecular phylogeny and evolution analyses suggest early and ongoing evolution of labdane-related diterpenoid metabolism in angiosperm.

231-239 Download
29
A MULTI-GENE PHYLOGENY OF CERATOCYSTIS MANGINECANS INFECTING MANGO IN PAKISTAN
ASMA RASHID1*, SHAZIA IRAM2 AND IFTIKHAR AHMAD1

A MULTI-GENE PHYLOGENY OF CERATOCYSTIS MANGINECANS INFECTING MANGO IN PAKISTAN
ABSTRACT:
Mango trees (Mangifera indica L.) are affected by a serious wilt disease, recognized as mango sudden death first time reported in Muzafargargh Punjab, Pakistan in 1995. Its prevalent is in almost all mango growing areas with severity ranged from 2-5% in Punjab and 5-10% in Sindh. Survey and sampling was conducted during the year 2011-12, on mango orchids in different distracts of Punjab and Sindh and no location was found free from this Disease. For molecular identification, DNA was successfully extracted and was then amplified by using ITS, BT, TEF (600-800)primers through Polymerase Chain Reaction (PCR) assay andnucleotide evidence of Pakistani isolates (45 for each gene) exhibiting the maximum genetic homology with Ceratocystis manginecans (99-100%) followed by C. fimbriata (97%) and C. omanensis (80%) respectively. On the basics of morphological tools and comparison of nucleotide evidence of multi-genes, C. manginecans is different from C. fimbriata and C. omanensis which infect mango in Pakistan. The availability of disease-free planting material and management in combination with fertilization and proper irrigation system would help in improving orchard management system.

241-247 Download
30
ANDIROXYLON THANOBOLENSIS SP.NOV. A NEW SPECIES OF FOSSIL WOOD OF FAMILY FABACEAE FROM MANCHAR FORMATION EXPOSED NEAR THANOBOLA KHAN, DISTRICT JAMSHORO, SINDH, PAKISTAN
SHABAB ALI KHAN1, BASIR AHMED1*, MUHAMMAD TAHIR RAJPUT2 AND SYEDA SALEHA TAHIR1

ANDIROXYLON THANOBOLENSIS SP.NOV. A NEW SPECIES OF FOSSIL WOOD OF FAMILY FABACEAE FROM MANCHAR FORMATION EXPOSED NEAR THANOBOLA KHAN, DISTRICT JAMSHORO, SINDH, PAKISTAN
ABSTRACT:
Andiroxylon thanobolensis sp. nov., a new silicified wood has been described and identified from Tertiary Manchar formation exposed near Thanobola Khan, district Jamshoro, Sindh, Pakistan. The xylotomical features of present fossil shows close resemblance with the genus Andira of the Family Fabaceae. The absence of growth ring indicated that plants were growing in tropical type of climate.

249-253 Download
31
SYSTEMATICS OF CHROOCOCCUS FROM PAKISTAN
MUBASHRAH MUNIR1, RAHMATULLAH QURESHI1, *, MUHAMMAD ILYAS1, MAHMOODA MUNAZIR1 AND M.K. LEGHARI2

SYSTEMATICS OF CHROOCOCCUS FROM PAKISTAN
ABSTRACT:
The present paper reports taxonomic enumeration of genus Chroococcus belonging to the Chroococcaceae family from the Kallar Kahar Lake, Chakwal, Pakistan. In all, 18 species have been recorded from the study area. Of them, 5 species viz., C. microscopicus, C. planctonicus, C. polyedriformis, C. subtilissimus and C. vacuolatus are new records for Pakistan and South Asia. Their taxonomic description and geographic distribution is provided and discussed in this paper.

255-262 Download
32
INTRA AND INTER SPECIFIC PROFILING OF PAKISTANI QUERCUS SPECIES GROWING IN THE HILLY AREAS OF DISTRICT DIR KHYBER PAKHTUNKHWA
MOHAMMAD NISAR1*, ABDUL GHAFOOR2, SYED FAZAL WADOOD1, AJMAL IQBAL AND NAUSHEEN1*

INTRA AND INTER SPECIFIC PROFILING OF PAKISTANI QUERCUS SPECIES GROWING IN THE HILLY AREAS OF DISTRICT DIR KHYBER PAKHTUNKHWA
ABSTRACT:
The intra and inter genetic diversity among 20 genotypes of Pakistani Quercus species viz., ten genotypes of Q. incana, eight Q. baloot and two Q. dilatata were analyzed using morphological characterization and proteomic profiling. A total of 14 morphological traits were scored for estimation of genetic diversity through descriptive statistics, traits similarity index and cluster plotting. Similarly, seven loci (bands) were detected in the collected germplasm of Quercus sp. Intra species locus contribution to genetic diversity (LCGD) was 42.9% in Q. baloot and 14.2% in Q. incana. Similarly, inter species LCGD was 71.43% in the collected germplasm. Out of seven loci, locus-1, 5 and 6 showed polymorphic in Q. baloot and locus 6 in Q. incana. Importantly, locus 3 and 4 was monomorphic in all collected lines and marked as generic specific locus for Quercus (sp.). SDS-PAGE profiling based on one-way cluster plotting successfully resolved the three species into separate clusters. The present data reflect that though the Quercus sp. showing intra and Inter species genetic diversity, but maintained species specific identity in the area regardless of environmental fluctuation.

263-270 Download
33
MACRO AND MICRO-NUTRIENT CONTENTS OF 18 MEDICINAL PLANTS USED TRADITIONALLY TO ALLEVIATE DIABETES IN NUEVO LEON, NORTHEAST OF MEXICO
RATIKANTA MAITI1, HUMBERTO GONZALEZ RODRIGUEZ2, CH ARUNA KUMARI3 AND N C SARKAR4

MACRO AND MICRO-NUTRIENT CONTENTS OF 18 MEDICINAL PLANTS USED TRADITIONALLY TO ALLEVIATE DIABETES IN NUEVO LEON, NORTHEAST OF MEXICO
ABSTRACT:
Although several medicinal plants has been documented to alleviate diabetes in Nuevo Leon, Northeast of Mexico, no systematic study has been undertaken to determine the efficacy of these plant species. The present study was undertaken to determine micronutrients (Cu, Fe and Zn) and macro-nutrients (K, Mg and P), C, N and C/N and to select plants with high macro and micronutrient contents for high efficacy in 18 medicinal plants collected from botanical gardens of Forest Science Faculty, UANL, Mexico used in Nuevo Leon in Northeast of Mexico, at the experimental station of Facultad de Ciencias Forestales, Universidad Autonoma de Nuevo Leon. Following standard protocols, carbon and nitrogen were determined using a CHN analyzer (Perkin Elmer, model 2400). Mineral contents were using the wet digestion technique (Cherney, 2000). The present study indicated the presence of large variation in the contents of several macro and micronutrients among these 18 species of medicinal plants utilized traditionally to control diabetes and other diseases in Nuevo Leon, Mexico. Among these species containing high nitrogen content (%) are Moringa oleifera (6.25), Melia azadirachta (5.85), Marrubium vulgare (4.56) and Phoradendron villosum (4.9). The C/N values ranged from 8 to 30. The species having high C/N were Agave macroculmis (30), Arbutus xalapensis (26) and Rhus virens (22). The species Melia azadirachta, Marrubium vulgare, Buddleja cordata, Tecoma stans, Hedeoma palmeri, Phoradendron villosum, Opuntia ficus-indica, Arbutus xalapensis exhibited large variations in the contents of macro and micronutrients which, could be considered to be used effectively for the control of diabetes. Few species viz. Marrubium vulgare, Buddleja cordata, Tecoma stans, Hedeoma palmeri, Phoradendron villosum, Opuntia ficus-indica and Arbutus xalapensis on the basis of high nutrient content with respect to C, N, C/N, Cu, Fe, Zn, K, P and Mg are selected and recommended to control diabetes.

271-276 Download
34
EVALUATION OF ANTIBACTERIAL AND ANTIOXIDANT ACTIVITY OF LEAVES, FRUIT AND BARK OF KIGELIA AFRICANA
TANVEER HUSSAIN2 , IRAM FATIMA1*, MUHAMMAD RAFAY2*, SAMMIA SHABIR1, MUHAMMAD AKRAM3 AND SABAHAT BANO1

EVALUATION OF ANTIBACTERIAL AND ANTIOXIDANT ACTIVITY OF LEAVES, FRUIT AND BARK OF KIGELIA AFRICANA
ABSTRACT:
In vitro antibacterial activity of extracts was tested against six bacterial strains viz. Staphylococcus aureus, Proteus vulgaris, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Citrobacter amalonaticus by agar-disc diffusion method. Ethanol and n-hexane were used as negative control and oxytetracycline was used as a positive control. Ethanolic and aqueous extracts of bark and leaves of Kigelia africana showed remarkable activity against various bacterial strains as compared to n-hexane. S. aureus and E. coli were proved as highly sensitive strains while K. pneumonia was the resistant strain as the extracts formed no inhibition zone against it. The percentage of antioxidant activity of different parts of Kigelia was assessed by DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical assay. Quercetin was used as a standard antioxidant which showed 93.6% inhibition. Kigelia bark extract showed good antioxidant activity i.e., 67.33% inhibition, fruit extract possess moderate antioxidant activity i.e., 62.66% inhibition while leaves showed the poor antioxidant activity i.e., 59.66% DPPH inhibition respectively. Overall, the comparative analysis revealed that bark extract exhibited the most remarkable antibacterial as well as antioxidant activity as compared to leaves and fruit extracts.

277-283 Download
35
ESTABLISHMENT OF AN EFFICIENT AND REPRODUCIBLE REGENERATION SYSTEM FOR POTATO CULTIVARS GROWN IN PAKISTAN
AAMIR IQBAL12*, ALI RIZWAN12, ZAHID MUKHTAR12, SHAHID MANSOOR12, ZAFAR MEHMOOD KHALID3 AND SHAHEEN ASAD12

ESTABLISHMENT OF AN EFFICIENT AND REPRODUCIBLE REGENERATION SYSTEM FOR POTATO CULTIVARS GROWN IN PAKISTAN
ABSTRACT:
The present study was carried out to assess the effect of growth regulators in three different combinations on mass propagation of currently grown three potato cultivars cv. Desiree, Kuroda and Cardinal. Varietal response on In vitro regeneration under different hormonal combinations. For callus induction, internodes of potato cultivars were cultured on modified Murashige & Skoog (MS) medium, added with different growth hormonal combinations. Callusing frequency for all these treatments and cultivars were recorded and in callus induction medium (CIM1) explants showed significantly higher callus formation as compared to two other combinations. For shoot induction, calli were cultured on Murashige & Skoog (MS) modified medium, supplemented with different hormonal combinations. Shoot induction medium (SIM3) gave best shoot induction frequency as compared to other media combinations. On the same media, an average number of shoots per explant were obtained for cultivar Desiree which is significantly different from the other two media combinations. Overall, the In vitro regeneration and multiplication potential was highest in the variety Cardinal followed by Kuroda and Desiree. The interaction between different hormonal combinations and varietal response for all the parameters showed significant differences.

285-290 Download
36
MICROPROPAGATION TO RESCUE ENDANGERED PLANT MORINGA CONCANENSIS NIMMO (MORINGACEAE)
HUMA FATIMA1, ANJUM PERVEEN2 AND MUHAMMAD QAISER3

MICROPROPAGATION TO RESCUE ENDANGERED PLANT MORINGA CONCANENSIS NIMMO (MORINGACEAE)
ABSTRACT:
Efficient micropropagation was developed for an endangered plant; Moringa concanensis Nimmo. The plant has many medicinal properties. It is an antioxidant, anti-diabetic and a powerful tonic alternative. A high frequency and maximum number of shoots were produced in MS supplemented with the combination of Benzylaminopurine (BAP) and α-Naphthalene Acetic Acid (NAA) (0.10mg/l +0.05mg/l). Rooting was achieved by the inoculation of regenerated shoots on half strength MS without addition of hormone.

291-294 Download
37
DEVELOPING A PLANT CULTURE MEDIUM COMPOSED OF VINASSE ORIGINATING FROM HAEMATOCOCCUS PLUVIALIS CULTURE
ANDRÉ LUIZ GOLLO1, ANDRÉ LUÍS LOPES DA SILVA1, KHIOMARA KHÉMELI DELLANI DE LIMA1, JEFFERSON DA LUZ COSTA2, MARCELA CÂNDIDO CAMARA1, LUIZ ANTONIO BIASI3, CRISTINE RODRIGUES1, LUCIANA PORTO DE SOUZA VANDENBERGHE1, VANETE THOMAZ SOCCOL1 AND CARLOS RICARDO SO

DEVELOPING A PLANT CULTURE MEDIUM COMPOSED OF VINASSE ORIGINATING FROM HAEMATOCOCCUS PLUVIALIS CULTURE
ABSTRACT:
The mineral nutrients in vinasse provide support for algal and plant growth. Algal culture releases organic compounds into its liquid culture medium. These organic and inorganic substances can be useful for formulating a plant tissue culture medium, because tissue culture medium is composed of organic and inorganic components. Therefore, the aims of this study were to develop a plant culture medium by using the vinasse that is employed for Haematococcus pluvialis culture (algal filtrate); to investigate the possible beneficial effects of the biocompounds in the micropropagation of Nidularium procerum (Bromeliaceae), to evaluate quercetin content, total phenolics content in vinasse and to evaluate the cytotoxicity of the media by performing a bioassay with Artemia salina. The vinasse that originated from H. pluvialis culture can be used to formulate plant tissue culture at a 3% dilution, and its mineral nutrients can support In vitro plant growth, but some nutrients must be supplemented to enhance its efficiency. An efficient micropropagation protocol was developed for N. procerum. The micropropagated plants were suitable for transfer to the field (they were acclimatized). This culture medium provides a way to reuse wastewater, gives a rational alternative to vinasse disposal and adds value to what is currently considered to be an undesirable residue. Moreover, this process can reduce the production costs of clonal seedlings and/or bioactive compounds in biofactories. There was no apparent biostimulatory effect by the algal filtrate on morphogenesis; however, it did increase quercetin production. The H. pluvialis culture that was grown in the vinasse decreased the cytotoxicity and phenolic compound contents, which prevented explant tissue necrosis and represented a treatment for this residue for safer disposal in the environment.

295-303 Download
38
OPTIMIZING THE RAPID TECHNIQUE FOR PROPAGATION OF CERASUS CAMPANULATA BY TISSUE CULTURE
BIHUA CHEN1,2*, JIANMIN LI1,2, JUAN ZHANG1,2, ZHUOXI WU1,2, HUIHUA FAN1,2 AND QIANZHEN LI1,2

OPTIMIZING THE RAPID TECHNIQUE FOR PROPAGATION OF CERASUS CAMPANULATA BY TISSUE CULTURE
ABSTRACT:
The initiation medium for Cerasus campanulata was 1/2MS+1.0 mg L-1 BA+0.1 mg L-1 NAA +30 g L-1 sugar, and the average initiation rate was 94.4%. The proliferation medium composed of MS+0.3 mg L-1 BA+0.2 mg L-1 NAA+30 g L-1 sugar provided proliferation rate 3.6 and average shoot length 2.3 cm. The rooting medium composed of 1/2MS+0.6 mg L-1 IBA+20 g L-1 sugar provided rooting rate 85.6%, average root number per individual 5.9 roots, and average root length 2.7 cm. The dishwashing detergent instead of Tween-20 as the spreader reduced the contamination rate for the explant disinfection. The red-core soil adopted to be the plantlet transfer medium provided 91.3% survival rate. The light media LZ and SK were not eligible to be the transfer medium for the species.

305-309 Download
39
PERFORMANCE AND ECONOMICS OF GROWING MAIZE UNDER ORGANIC AND INORGANIC FERTILIZATION AND WEED MANAGEMENT
ABBAS ALI1, MUHAMMAD AZIM KHAN1*, ASHIQ SALEEM2, K.B. MARWAT3, ABBAS ULLAH JAN1, DAWOOD JAN1 AND SHAHID SATTAR1

PERFORMANCE AND ECONOMICS OF GROWING MAIZE UNDER ORGANIC AND INORGANIC FERTILIZATION AND WEED MANAGEMENT
ABSTRACT:
Weed competition and imbalanced fertilizers are important yield reducing factors in maize. To investigate the impact of weed management and combinations of fertilizers on yield and net income of maize, a field trial was conducted at National Agricultural Research Centre, Islamabad, Pakistan during summer 2014. Randomized complete block design with split-plot arrangement was used with three replications. Four weeds pressures viz. (1) hand weeding at 25 cm on both sides of each row of maize plants, (2) No hand weeding at 25 cm on both sides of maize rows, (3) application of Primextra gold (atrazine plus S-metolachlor) @ 1.44 kg a.i. ha-1 as pre-emergence and (4) weedy check (control) were assigned to main plots. While different combinations of NPK were assigned to sub plots. Data revealed that dry weed biomass close to maize plants were significantly lower (140.4 kg ha-1) as compared to weeds 25 cm away from maize plants (153.2 kg ha-1). However, the application of atrazine plus S-metolachlor showed promising results by decreasing the weed biomass (53.6 kg ha-1) as compared to 155.6 kg ha-1 in control. Combination of fertilizers, also significantly affected the weed biomass. As compared to control, all the fertilizers (N, P and K) significantly increased weed biomass. Presence of weeds close to the crop rows, proved more harmful for grain yield of maize. Overall, application of herbicide in combination with NPK showed promising results in term of weed control and grain yield. Net income was higher when herbicide in combination with N, P or NP was used.

311-318 Download
40
OPTIMIZATION OF AN EFFICIENT NON-TISSUE CULTURE TRANSFORMATION METHOD FOR BRASSICA JUNCEA
IJAZ NAEEM1, IQBAL MUNIR1*, FARHATULLAH2 AND AQIB IQBAL1

OPTIMIZATION OF AN EFFICIENT NON-TISSUE CULTURE TRANSFORMATION METHOD FOR BRASSICA JUNCEA
ABSTRACT:
the major hurdles in successful in vitro transformation of Brassica juncea through standard tissue culture (STC) method are: culture contamination, somaclonal variations, and lack of expertise. Moreover, the current STC method is time consuming and needs continuous electricity. In the present study, the in planta transformation method through floral dip with or without vacuum infiltration was optimized for successful transformation of B. juncea. The B. juncea CV “RAYA Anmol” was used for transformation through Agrobacterium tumefaciens strain GV3101 harboring the binary vector plasmid pBinGlyBar4-EADcT. Based on the resistance reaction to the herbicide Basta, 20 and 40 resistant seedlings were obtained from ~2000 seed germinated from the plants transformed through floral dip and vacuum infiltration methods, respectively. The PCR analyses further confirmed the presence of transgene in 3 floral dipped plants without vacuum infiltration and 17 floral dipped plants with vacuum infiltration, giving the transformation frequencies of 1.5x10-3 and 8.5x10-3, respectively. This method, which avoids tissue culture, will reduce the somaclonal variation accompanying prolonged culture of cells in a dedifferentiated state, will facilitate functional genomics and improvement of Brassica juncea with novel desirable traits while reducing time and expense.

319-324 Download
41
NO EVIDENCE FOR DIFFERENTIAL BIOMASS AND MINERAL CONTENT IN ADULT PLANTS GROWN FROM DIMORPHIC SUAEDA ARALOCASPICA SEEDS
LEI WANG1, HONG-LING WANG1, CHANG-YAN TIAN1* AND ZHEN-YING HUANG2

NO EVIDENCE FOR DIFFERENTIAL BIOMASS AND MINERAL CONTENT IN ADULT PLANTS GROWN FROM DIMORPHIC SUAEDA ARALOCASPICA SEEDS
ABSTRACT:
The production of two or more seed types by a single plant is known as seed heteromorphism. There have been many comparisons of seed traits or growth between plants grown from heteromorphic seeds. However, information is scarce regarding the mineral contents of adult plants from heteromorphic seeds. We herein present biomass and mineral profiles of adult plants grown from dimorphic seeds (non-dormant brown seeds and black seeds with non-deep physiological dormancy) of the annual desert halophyte Suaeda aralocaspica at different nutrient and salinity levels. Although nutrient and salinity treatments affected dry weight and mineral content, there were no significant differences among S. aralocaspica seed-dimorphic plants under the same experimental conditions. This study is one of the few to compare the physiological responses between seed-heteromorphic plants, and reveals that mineral status corresponds with growth performance in these plants.

325-330 Download
42
ECONOMIC EVALUATION FOR INTEGRATED USE OF GLYPHOSATE HERBICIDE AND TILLAGE COMBINATIONS APPLIED BEFORE SOWING OF RAIN-FED WHEAT (TRITICUM AESTIVUM L.)
SAFDAR ALI1*, M. AZIM MALIK1, M. ANSAR1 AND RAHMATULLAH QURESHI2

ECONOMIC EVALUATION FOR INTEGRATED USE OF GLYPHOSATE HERBICIDE AND TILLAGE COMBINATIONS APPLIED BEFORE SOWING OF RAIN-FED WHEAT (TRITICUM AESTIVUM L.)
ABSTRACT:
Low average yield, scarce soil moisture and less soil fertility are major problems of rain-fed wheat. Economic feasibility of different tillage systems integrated with glyphosate herbicide and wheat crop productivity was determined through field experiments conducted at the University Research Farm of Pir Mehr Ali Shah Arid Agriculture University Rawalpindi, Pakistan during summer and winter seasons of 2012-13 and 2013-14. Different combinations of tillage and glyphosate herbicide were used in the fallow period (summer season) that were consisted of following treatments viz. T1 = 1 Mould board Plowing + 8 Cultivations, T2 = No-Till + Glyphosate, T3 = 1 Mould board Plowing + Glyphosate, T4 = 1 MB Plowing + 4 Cultivations, T5 = 1 Disc Harrowing + Glyphosate, T6 = 1 Disc Harrowing + 4 Cultivations and T7 = 1 Chiseling + Glyphosate. Results showed that the highest yield viz. 3.5132, 3.1242 t ha-1 were obtained in the case of conventional tillage (T1) and reduced tillage (T4), respectively with a net profit of 888.92 and 839.35 $ ha-1. The yield was positively affected by tillage intensity. In conclusion, T1 is recommended for getting maximum net return from wheat grown in rain-fed areas of Pakistan.

331-337 Download
43
AGRO-ECONOMIC OPTIMIZATION OF FERTILIZER LEVEL AND RHIZOBIUM STRAINS FOR ENHANCED GRAIN YIELD IN MUNGBEAN (VIGNA RADIATA L.)
MUHAMMAD MANSOOR1, ZAFAR ISLAM2, SHER MUHAMMAD3, ASGHAR ALI4, AND MUHAMMAD EHSAN ELAHI1

AGRO-ECONOMIC OPTIMIZATION OF FERTILIZER LEVEL AND RHIZOBIUM STRAINS FOR ENHANCED GRAIN YIELD IN MUNGBEAN (VIGNA RADIATA L.)
ABSTRACT:
To explore the effect of PGPR for improvement in grain yield of mungbean, different levels of fertilizer (00:00, 20:50 and 20:70 kg N:P2O5 kg ha-1) were tested on four strains (CB 1015 Australia, Vm-L1, Vm-M1 and Vm-M2) at Agricultural Research Institute, Dera Ismail Khan. A Randomized Complete Block Design with split plot arrangement was used to conduct the experiment by keeping three fertilizer levels in the main plot and Rhizobium strains in sub-plot. The data were recorded for different parameters like number of branches plant-1, number of clusters plant -1, number of pods plant-1., number of grain pod-1, pod length (cm) and grain yield kg ha-1. The response of fertilizer levels and inoculation with Rhizobium strains were found significant in most of the parameters under study. The strain Vm-M1 produced the highest number of branches plant-1 (5.42), number of clusters plant-1 (22.92), number of pods plant-1 (77.64), pod length (8.37 cm) when applied in combination of N:P2O5 @ 20:70 and 20:50 kg ha-1. Although the grain yield (1421 kg ha-1) recorded in treatment (20:70 kg N:P2O5 ha-1 + Inoculation with strain Vm M1) was highest but economically the treatment (20:50 kg N:P2O5 ha-1+Inoculation with strain Vm M1) with net return of Rs. 13618 ha-1and BCR of 2.52 was on top.

339-343 Download
44
SYMBIOTIC NITROGEN FIXATION IN ALFALFA (MEDICAGO SATIVA L.) BY SINORHIZOBIUM MELILOTI AT AL-QASSIM REGIONS, SAUDI ARABIA
F.N. AL-BARAKAH1* AND M.A.U. MRIDHA2

SYMBIOTIC NITROGEN FIXATION IN ALFALFA (MEDICAGO SATIVA L.) BY SINORHIZOBIUM MELILOTI AT AL-QASSIM REGIONS, SAUDI ARABIA
ABSTRACT:
The nodulation status in alfalfa (Medicago sativa L.) plants by Sinorhizobium meliloti under Saudi field condition was assessed in some selected farms in four seasons for two years. In the present study, we also monitored the introduced S. meliloti strains' activity under Saudi soil conditions. The samples were collected at regular seasonal intervals from the selected farms. The total number of nodules, morphology of the nodules and the effectiveness of N2-fixation was assessed. In general, it was revealed that soils in the selected areas in Saudi Arabia have sufficient bacteria of the proper types to nodulate the alfalfa plants. These nodules are high in number, small in size and white in color. The nodules obtained from most of the selected farms are ineffective for nitrogen fixation. Inoculation of alfalfa seeds with imported S. meliloti strains failed to fix the atmospheric nitrogen sufficiently and also the growth improvement of alfalfa plants. There was a wide variation in the occurrence of number of nodules among the four seasons in two years. It was also observed that summer season severely affected the nodulation making it nearly zero. This low number of nodules exerts a very slow recovery of nodule formation in the next year. The introduced strains were always over competing with the native strains but they did not survive because of hot and dry summer. Nitrogenase activity of the nodules collected from both the inoculated and non-inoculated farms were always very low in all the collected samples, which indicates that the ability of fixing nitrogen by S. meliloti strains in alfalfa under Saudi soils conditions is very low.

345-348 Download
45
PRODUCTION AND CHARACTERIZATION OF A THERMOSTABLE BIOPLASTIC (POLY-s-HYDROXYBUTYRATE) FROM BACILLUS CEREUS NRRL-B-3711
ASAD-UR-REHMAN*, ALIA ASLAM, RUSHDA MASOOD, MUHAMMAD NAUMAN AFTAB, RABIA AJMAL AND IKRAM-UL-HAQ

PRODUCTION AND CHARACTERIZATION OF A THERMOSTABLE BIOPLASTIC (POLY-s-HYDROXYBUTYRATE) FROM BACILLUS CEREUS NRRL-B-3711
ABSTRACT:
The poly-β-hydroxybutyrate (PHB) is a thermostable and biocompatible polyester produced by several bacteria under unbalanced nutritional conditions. Among Gram-positive bacteria, which are not well exploited for PHB production on industrial scale, Bacillus appears to be a prospective candidate due to its excellent biopolymer yield and less rigorous fermentation conditions. Batch culture fermentation was carried out for PHB production. The Bacillus cereus NRRL-B-3711 was the most efficient producer of PHB out of five Bacillus species. The optimized production was achieved with 2% glucose, 37°C, pH value of 8 and ammonium sulfate (2.5 g/L) as a nitrogen source. Carbon to nitrogen ratio of 10 significantly affects the PHB accumulation. The selected specie was able to accumulate PHB up to 56% (14.2 }0.07 g/L) on dry cell weight basis after optimization. This corresponds to a 1.87 folds increase in the production. The optical microscopy showed extremely flat surface of bioplastic thin film indicating the brittle failure of PHB under tensile loading. The FTIR analysis revealed C=O and –CH groups, with thermal properties e.g. Tg; 2°C, Tc; 54°C, Tm;162°C and % crystallinity of 51.3 by differential scanning calorimeter (DSC), thus confirming the presence of PHB bioplastic and enhancing its industrial applications.The results surpassed those reported in the literature for PHB production.

349-356 Download
46
KINETIC EVIDENCE OF A NOVEL INVERTASE IN AN ETHYL METHANE SULPLONATE DEREPRESSED MUTANT OF YARROWIA LIPOLYTICA
SIKANDER ALI1* AND MUHAMMAD UMAR HAYYAT2

KINETIC EVIDENCE OF A NOVEL INVERTASE IN AN ETHYL METHANE SULPLONATE DEREPRESSED MUTANT OF YARROWIA LIPOLYTICA
ABSTRACT:
In the present study, we report on the kinetics of an extracellular invertase production by Yarrowia lipolytica in defined medium i.e., sucrose peptone agar yeast extract, pH 6 (SAPY). The wild-type IIB-II was treated with ethyl methane sulphonate (EMS) as a chemical mutagen. Among the six mutants isolated, EMS-IV was found to be the best enzyme producing mutant strain (51±2.4a U/ml). The maximum enzyme production (73±3.1a U/ml) occurred at 48 h of incubation (67±2.7a mg/ml protein). The potential mutant was stabilized at low levels of 2-deoxy-D-glucose (2dg) and the viable mutants were further optimized both culturally and nutritionally. The sucrose concentration, incubation period and pH were optimized to be 30 g/l, 28°C and 6.5, respectively. EMS-IV exhibited an improvement of over 10 folds in enzyme production when 5 g/l ammonium sulphate was used as a nitrogen source. Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) analysis showed that optimal enzyme activity caused a higher hydrolysis rate of sucrose into monosaccharides (a-D-glucose and b-D-fructose). The values for Qp (1.7±0.12c U/ml/h) and Yp/s (3.7±1.24b U/g) of the mutant were considerably higher in comparison to the wild-type or all other yeast strains. The mutant could be used for enzyme production over the temperature range of 26-34°C, which is highly significant (LSD 0.048, HS).

357-262 Download
47
MOLECULAR CHARACTERIZATION OF YEAST STRAINS ISOLATED FROM DIFFERENT SOURCES BY RESTRICTION FRAGMENT LENGTH POLYMORPHISM
MUHAMMAD SHAHZAD ALI AND ZAKIA LATIF*

MOLECULAR CHARACTERIZATION OF YEAST STRAINS ISOLATED FROM DIFFERENT SOURCES BY RESTRICTION FRAGMENT LENGTH POLYMORPHISM
ABSTRACT:
Various molecular techniques like analysis of the amplified rDNA internal transcribed spacers (ITS), intragenic spacers and total ITS region analysis by restriction fragment length polymorphism (RFLP) has been introduced for yeast identification but there are limited databases to identify yeast species on the basis of 5.8S rDNA. In this study, twenty nineyeast strains from various sources including spoiled fruits, vegetables, foodstuffs, and concentrated juices were characterized by PCR-RFLP. PCR-RFLP has been used to characterize yeasts present in different spoiled food samples after isolation of the yeasts. By using this technique, the isolated yeast strains were characterized by direct 5.8S-ITS rDNA region amplification. RFLP analysis was applied to each of the amplification products (varied from 400bp to 800bp) detected, and the corresponding yeast identifications were made according to each specific restriction patterns obtained after treatment with two endonucleases TaqI and HaeIII which yielded a specific banding pattern for each species. For further confirmation amplified products of eleven selected isolates were sequenced and blast on NCBI. Both RFLP and sequence analyses of the strains with accession nos. KF472163, KF472164, KF472165, KF472166, KF472167, KF472168, KF472169, KF472170, KF472171, KF472172, KF472173 gave significantly similar results. The isolates were found to belong five different yeast species including; Candida spp., Pichia spp., Kluyveromyces spp., Clavispora spp. and Hanseniaspora spp. This method provides a fast, easy, reliable and authentic way for determining yeast population present in different type of samples, as compared to traditional characterization technique

363-370 Download
48
ANTIBACTERIAL AND ANTIFUNGAL POTENTIAL OF HIMALAYAN MEDICINAL PLANTS FOR TREATING WOUND INFECTIONS
UME HABIBA1, MUSHTAQ AHMAD1*, SHEHLA SHINWARI1 SHAZIA SULTANA1, ZABTA KHAN SHINWARI2,3 AND MUHAMMAD ZAFAR1

ANTIBACTERIAL AND ANTIFUNGAL POTENTIAL OF HIMALAYAN MEDICINAL PLANTS FOR TREATING WOUND INFECTIONS
ABSTRACT:
Many bacterial and fungal strains are involved in wound infectious diseases as most of these strains become resistant to the most commonly used synthetic drugs in Himalayan region. Plant based natural products seem to be an alternative to this problem. The aim of this investigation was to evaluate the In vitro antibacterial and antifungal activities of 30 medicinal plants used in folk recipes by Himalayan people to treat wound infections against multi-drug resistant pathogens. In total of six medically important Myco-bacterial strains Streptococcus pyogenes, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger were tested against methanolic plant extracts at 5 mg/ml concentration using agar disc well diffusion method to determined Minimum inhibitory concentrations (MICs).The plant extracts showed varied levels of MICs against test microorganisms. The strongest antibacterial activity was reported in methanolic extract of Cynadon dactylon (L.) Pers. against Klebsiella pneumoniae with 20.67±1.36 mm MICs, while Candida albicans was considered to be the most resistant pathogen with MICs9.6±0.57 mm. The findings were compared with results obtained using standard antibiotics, amoxicillin, ciprofloxacin, cefotaxime, fluconazole and itraconazole at conc.5mg/ ml. The results provide an evidence of folk medicinal uses of plants among the Himalayan communities to treat wounds. Further research needs to be carried out to identify the active molecules and evaluate the in vivo antibacterial and antifungal activities as well as toxicity level with clinical trials to use full potential of these plants for drug discovery development to control wounds globally.

371-375 Download
49
ALGAL GROWTH AND WASTE STABILIZATION PONDS PERFORMANCE EFFICIENCY IN A SUB-TROPICAL CLIMATE
AAMIR ALAMGIR, MOAZZAM ALI KHAN* AND S. SHAHID SHAUKAT

ALGAL GROWTH AND WASTE STABILIZATION PONDS PERFORMANCE EFFICIENCY IN A SUB-TROPICAL CLIMATE
ABSTRACT:
Both irrigation and potable water are in diminutive supply in most of the developing countries particularly those situated in tropical and subtropical regions where, often untreated wastewater is utilized for the purpose of irrigation. Treated wastewater has proved to be a potential asset serving as an alternate source for the expansion of irrigated agriculture. Waste stabilization ponds (WSP) are considered as less costly and effective substitute for the wastewater water treatment in tropics. The principle of wastewater treatment in waste stabilization pond is based on the symbiotic relationship between bacteria and various algal species. In this study, an attempt was made to relate algal growth and different extrinsic factors using multiple regression models. The predominant algal species found in WSP systems were Chlorella, Euglena, Oscillatoria and Scenedesmus. The growth of individual algal species and overall algal growth was principally governed by temperature, total sunshine hours and Total Kjeldhal Nitrogen (TKN). The study suggested that algal bacterial symbiotic relationship works well and the dissolved oxygen production through algal photosynthesis was optimum to decompose heavy organic load resulting in oxygen-rich effluent (liquid fertilizer) which could be successfully exploited for unrestricted irrigation.

377-385 Download
50
ADDITIONS TO THE POLYPORALES OF PAKISTAN
ABDUL RAZAQ1 AND SALEEM SHAHZAD2

ADDITIONS TO THE POLYPORALES OF PAKISTAN
ABSTRACT:
During a survey of macrofungi, nine species belonging to order Polyporales of Phylum Basidiomycota viz., Bjerkandera adusta, Fomes fomentarius, Heteroporus biennis, Laetiporus sulphureus, Phaeolus schwinitzii, Polyporus squamosus, Rigidoporus ulmarius, Royoporus badius and Trametes versicolor were collected for the first time from Gilgit-Baltistan. Of these, Bjerkandera adusta, Fomes fomentarius, Heteroporus biennis, Phaeolus schwinitzii, Rigidoporus ulmarius, Royoporus badius and Trametes versicolor appeared to be new records from Pakistan not hitherto reported.

387-391 Download
51
THE EVALUATION OF DIFFUSIVE GRADIENTS IN THIN FILMS (DGT) AND CaCI2 EXTRACTION ON PHOSPHORUS-ZINC INTERACTION IN SUDAN GRASS
OSMAN SONMEZ1*, GARY PIERZYNSKI2, CENGIZ KAYA3 AND SALIH AYDEMIR3

THE EVALUATION OF DIFFUSIVE GRADIENTS IN THIN FILMS (DGT) AND CaCI2 EXTRACTION ON PHOSPHORUS-ZINC INTERACTION IN SUDAN GRASS
ABSTRACT:
Diffusive gradients in thin films (DGT) has been used for the assessment of Zn availability and offers some advantages over routine batch extractions of soil samples. To evaluate the effectiveness of DGT compared with CaCl2-extraction to P additions in Zn contaminated soil, a greenhouse study was established. Zinc was added at the rate of 0, 150, 300, 600 and 1200mg Zn kg-1 as ZnSO4 in order to create various levels of Zn phytoavailability. Soil was amended with KH2PO4 at 0, 100 and 200mg P kg-1. Increasing application of Zn to soil elevated Zn adsorption by DGT, plant Zn concentrations, CaCl2 extractable and water-soluble Zn and decreased soil bioavailable P and pH. Internal plant concentrations and CaCl2-extraction were influenced by P additions, whereas DGT Zn concentrations were not affected. Phosphorus application decreased plant Zn concentrations and increased plant biomass yields.

393-397 Download
52
ANTI-RHEUMATIC POTENTIAL OF PAKISTANI MEDICINAL PLANTS: A REVIEW
MUSTAFA KAMAL1, MUHAMMAD ADNAN1*, WAHEED MURAD1, HAMEEDA BIBI2, AKASH TARIQ1, HAZIR RAHMAN3 AND ZABTA KHAN SHINWARI4

ANTI-RHEUMATIC POTENTIAL OF PAKISTANI MEDICINAL PLANTS: A REVIEW
ABSTRACT:
Present review aimed to provide a comprehensive documentation of plants used as anti-rheumatic ethnomedicines in Pakistan and to suggest future recommendations. Data on anti-rheumatic plants was collected from published scientific papers, reports and thesis using online searching engines such as Google Scholar PubMed and Science Direct. Five distinct zones in the country were classified on the basis of geography, humidity and rainfall. We used Sorenson similarity index for plants and their parts used between different zones. A total of 137 anti-rheumatic plant species representing 55 families and 104 genera are used in Pakistan. Herbs (87 plants) were the primary source of anti-rheumatic medicinal plants, while leaves (22 % plant species) were the most frequently used part in the preparation of ethnomedicinal recipes. Highest number of 52 medicinal plant species were found in Zone A having high mountains and cold climate where the prevalence of rheumatism was more common. Solanum surattense was found with highest conservation concerns as it was using in 13 different areas against rheumatism. Results of Sorenson index revealed that there is a similarity of plants and its parts uses between different zones. In conclusions, geography and climate have an important role in causing rheumatic disease. Pakistan has a number of anti-rheumatic plants that are used by the local populations through their traditional knowledge. Moreover, inter zonal similarities among plants and its part uses indicate higher pharmacological potency of these medicinal plants. Further, the review will also provide an insight regarding the conservation status of reported plants.

399-413 Download
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