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Year 2016 , Volume  48, Issue 3
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1
NACL SALINITY-INDUCED CHANGES IN GROWTH, PHOTOSYNTHETIC PROPERTIES, WATER STATUS AND ENZYMATIC ANTIOXIDANT SYSTEM OF NITRARIA ROBOROWSKII KOM
YAN LU, JIAQIANG LEI AND FANJIANG ZENG*

NACL SALINITY-INDUCED CHANGES IN GROWTH, PHOTOSYNTHETIC PROPERTIES, WATER STATUS AND ENZYMATIC ANTIOXIDANT SYSTEM OF NITRARIA ROBOROWSKII KOM
ABSTRACT:
In pot experiments, 1-year-old Nitraria roborowskii Kom plants were planted under control and four levels of NaCl. The plant growth, Na+ and K+ contents, maximal efficiency of photosystem II photochemistry (Fv/Fm), water potential, antioxidative enzyme activities and contents of photosynthetic pigments, relative water, soluble sugars, proline, hydrogen peroxide (H2O2), and malondialdehyde (MDA) in leaves were measured after 90 days of NaCl treatments. N. roborowskii showed tolerance to medium salinity (50 mM NaCl), whereas growth reduction in biomass was observed when ≥ 100 mM NaCl solutions were used for irrigation. Extreme salinity (400 mM NaCl) hardly impacted Fv/Fm, but a marked decreased in chlorophyll content was observed. A gradual decline of the leaf relative water content and water potential with increased NaCl feeding level was accompanied by increased Na+ accumulation. The total proline and soluble sugar contents were not significantly affected by low salinity (50 mM NaCl); however, both of these values decreased gradually at high salinities (≥ 100 mM NaCl). MDA and H2O2 were not markedly affected at low salinity (50 mM NaCl), but both of these values remarkably accumulated at high salinities (≥ 100 mM NaCl). Superoxide dismutase (SOD), guaiacol peroxidase (POD) and catalase (CAT) presented the same variation trends, first increasing with concentrations of 100, 50 and 100 mM NaCl, respectively, and then decreasing compared to the control, whereas APX increased under all NaCl concentrations.

843-851 Download
2
EFFECT OF SEA SALT IRRIGATION ON PLANT GROWTH, YIELD POTENTIAL AND SOME BIOCHEMICAL ATTRIBUTES OF CARISSA CARANDAS.
TAYYAB1,2*, MUHAMMAD AZEEM2, MUHAMMAD QASIM3 AND RAFIQ AHMAD2

EFFECT OF SEA SALT IRRIGATION ON PLANT GROWTH, YIELD POTENTIAL AND SOME BIOCHEMICAL ATTRIBUTES OF CARISSA CARANDAS.
ABSTRACT:
Carissa carandas (varn. Karonda) is an edible and medicinal plant having ability to grow in saline and water deficit conditions, however, little is known about its salinity tolerance. Therefore, the effect of salinity on vegetative (height and volume), reproductive (number of flowers and number, size and weight of fruits) and some biochemical parameters (leaf pigments, ions, soluble sugars, proteins, and phenols) of C. carandas were studied. Plants were grown in drum pot culture and irrigated with non-saline or saline water of 0.6% and 0.8% sea salt concentrations, for a period of 30 months. Results showed that, plant height, and canopy volume decreased with increasing salinity. The chlorophyll contents and chlorophyll a/b ratio followed the similar trend as for growth, however, carotenoids increased at 0.6% sea salt and subsequently decreased in higher salinity. Unchanged soluble sugar and protein content at 0.6% sea salt, as compared to control, could be attributed to leaf osmotic adjustments which decreased with further increase in salinity. Linear increase in soluble phenols and carotenoid/chlorophyll ratio indicating a protective strategy of C. carandas to minimize photo-damage. Besides increasing Na+ and decreasing K+ contents, plant seemed to maintain K+/Na+ ratio (above 1), especially at 0.6 sea salt, which disturbed at higher salinity. Salinity adversely affected reproductive growth of C. carandas where, production of flowers, and fruits were significantly reduced. In addition, fresh and dry weights of fruits decreased with increasing salinity, but salinity did not affect fruit length and diameter. Present study provides basic information related to plant growth, fruit yield and some biochemical attributes, which suggest that C. carandas is moderately salt tolerant plant. This plant showed potential to grow on saline marginal lands using brackish water irrigation and provide biomass for edible and medicinal purposes. However, in-depth analysis of field and greenhouse experiments could be helpful to understand the detailed eco-physiological responses and mechanisms of salinity tolerance of this species.

853-859 Download
3
INDUCTION OF SALT TOLERANCE IN WHEAT (TRITICUM AESTIVUM L.) SEEDLINGS THROUGH EXOGENOUS APPLICATION OF PROLINE
WAJID MAHBOOB*, MUHAMMAD ATHAR KHAN AND M.U. SHIRAZI

INDUCTION OF SALT TOLERANCE IN WHEAT (TRITICUM AESTIVUM L.) SEEDLINGS THROUGH EXOGENOUS APPLICATION OF PROLINE
ABSTRACT:
To appraise the potential role of foliar applied proline as an alternative shotgun approach to ameliorate the adverse effect of salinity on wheat, a pot experiment was conducted under controlled environmental conditions, two wheat genotypes; a salt tolerant strain ESW-9525 and a moderately tolerant cultivar kherman were used in this study. Factorial combination of treatments with three replications was arranged under completely randomized design. Seven days old wheat seedlings were exposed to various levels of salinity (0, 60 and 120 Mm NaCl) for one week and applied with foliar proline (0, 50 and 100 mM) one week later. Salinity stress caused a significant reduction in plant growth, leaf photosynthetic pigments, as well as alterations in ionic balance. Foliar applied proline significantly improved root and shoot length, seedling fresh and dry weight, photosynthetic pigments, K+ contents and K+: Na+ ratio. Both genotypes varied considerably in endogenous level of proline (Pro), glycine betaine (GB), total soluble sugars (TSS) and total phenolic contents (TPC) in response to salinity and foliar proline as well. Foliar applied proline 50 mM and 100 mM were found as a stimulus for plant growth triggering the physiological and biochemical attributes, However, 100 mM proline was the most effective to ameliorate the toxic effects of salinity by improving root and shoot length, seedling fresh and dry weight, chlorophyll a, b contents, TSS, Pro, GB, TPC and K+ contents and K/Na ratio in both genotypes. These findings confirmed the ability of foliar applied proline to stimulate the salt tolerance in wheat plants.

861-867 Download
4
FOLIAR APPLICATION OF ASCORBIC ACID MITIGATES SODIUM CHLORIDE INDUCED STRESS IN EGGPLANT (SOLANUM MELONGENA L.)
SAMIN JAN1, MUHAMMAD HAMAYUN2, SHER WALI1*, ASMAT BIBI2, HUMAIRA GUL2 AND FAZLI RAHIM3

FOLIAR APPLICATION OF ASCORBIC ACID MITIGATES SODIUM CHLORIDE INDUCED STRESS IN EGGPLANT (SOLANUM MELONGENA L.)
ABSTRACT:
The current work was designed to test the effect of sodium chloride on germination, seedling establishment, vegetative growth, yield, chemical contents and ionic composition of eggplant. The consequences of foliar application of ascorbic acid (AA) on mitigation of adverse effects of sodium chloride were also tested. The seeds of Solanum melongena were germinated using NaCl (60 mM, 100 mM) and ascorbic acid (100 and 200 mM). High levels of salinity significantly affected the seed germination and seedling fresh and dry weights. Plants grown under salinity stress with foliar application of ascorbic acid showed significant increase in germination percentage and seedlings growth as compare to control plants. Sodium chloride stress showed adverse effects on plant height, root length, number of leaves, leaf area, fresh and dry biomass, total chlorophyll, carbohydrates and proteins as compared to untreated plants. The relative water content, electrolyte leakage were increased and Na+ and K+ ions balance was disturbed in different plant parts. Ascorbic acid (100 and 200ppm) enhanced all the growth parameters affected adversely by sodium chloride stress.

869-876 Download
5
IMPACT OF ASCORBIC ACID ON GROWTH AND SOME PHYSIOLOGICAL ATTRIBUTES OF CUCUMBER (CUCUMIS SATIVUS) PLANTS UNDER WATER-DEFICIT CONDITIONS
HIRA NAZ1, NUDRAT AISHA AKRAM1* AND MUHAMMAD ASHRAF2,3

IMPACT OF ASCORBIC ACID ON GROWTH AND SOME PHYSIOLOGICAL ATTRIBUTES OF CUCUMBER (CUCUMIS SATIVUS) PLANTS UNDER WATER-DEFICIT CONDITIONS
ABSTRACT:
Cucumber (Cucumis sativus L.) is a very popular vegetable, which is utilized all-over the world. In the current research, two cultivars of cucumber, Local and Hybrid were chosen to look at the effect of foliar-applied ascorbic acid on some key physio-biochemical attributes under varying water regimes. After one week of seed germination, the plants of both cucumber cultivars were subjected to 100% field capacity (control) and 60% field capacity (water deficit conditions. After one month of water stress treatment, ascorbic acid (AsA) at the rate of 0 (control), 50 and 100 mg L-1 was applied foliarly. After 15-day AsA application, the data were recorded which showed that drought stress significantly reduced the plant growth, chlorophyll contents, relative water contents (RWC), rate of photosynthesis (A), stomatal conductance, internal CO2 concentration (Ci) and Ci/Ca, while drought caused an increase in relative membrane permeability (RMP), and proline and glycine betaine contents. AsA improved the shoot fresh and dry weights, chlorophyll a, RWC, Ci, Ci/Ca and proline contents. No change was observed in both cucumber cultivars under water-deficit and AsA treatments. Overall, growth improvement of cucumber plants under dry arid climate could be attributed to AsA-induced stimulation in the chlorophyll a, proline, RWC contents and Ci concentration.

877-883 Download
6
DE NOVO TRANSCRIPTOME SEQUENCEING AND COMPARATIVE ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES IN DRYOPERIS FRAGRANS UNDER TEMPERATURE STRESS
WEN-ZHONG WANG1,2√, WEI-SHUANG TONG1√, YAN LI1, RUI GAO1, LIGUO ZHANG1,3 AND YING CHANG1

DE NOVO TRANSCRIPTOME SEQUENCEING AND COMPARATIVE ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES IN DRYOPERIS FRAGRANS UNDER TEMPERATURE STRESS
ABSTRACT:
Dryopteris fragrans is a species of fern and contains flavonoids compounds with medicinal value. This study explain the temperature stress impact flavonoids synthesis in D. fragrans tissue culture seedlings under the low temperature at 4ºC, high temperature at 35ºC and moderate temperature at 25ºC. By using Illumina HiSeq 2000 sequencing, 80.9 million raw sequence reads were de novo assembled into 66,716 non-redundant unigenes. 38,486 unigenes (57.7%) were annotated for their function. 13,973 unigenes and 29,598 unigenes were allocated to gene ontology (GO) and clusters of orthologous group (COG), respectively. 18,989 sequences mapped to 118 Kyoto Encyclopedia of Genes and Genomes Pathway database (KEGG), 204 genes were involved in flavonoid biosynthesis, regulation and transport. 25,292 and 16,817 unigenes exhibited marked differential expression in response to temperature shifts of 25ºC to 4ºC and 25ºC to 35ºC, respectively. 4CL and CHS genes involved in flavonoid biosynthesis were tested and suggested that they were responsible for biosynthesis of flavonoids. This study provides the first published data to describe the D. fragrans transcriptome and should accelerate understanding of flavonoids biosynthesis, regulation and transport mechanisms. Since most unigenes described here were successfully annotated, these results should facilitate future functional genomic understanding and research of D. fragrans.

885-898 Download
7
MORPHOLOGICAL, PHYSIOLOGICAL AND BIOCHEMICAL RESPONSES OF CAMELLIA OLEIFERA TO LOW-TEMPERATURE STRESS
JUANJUAN HU1¬, WEI WU2¬, ZHIHUA CAO3, JIA WEN1, QINGLONG SHU1** AND SONGLING FU1**

MORPHOLOGICAL, PHYSIOLOGICAL AND BIOCHEMICAL RESPONSES OF CAMELLIA OLEIFERA TO LOW-TEMPERATURE STRESS
ABSTRACT:
Camellia oleifera Abel originates from China and is high healthy effect food oil species. It is also a high additional plant in southern China and can help to keep some people of mountain area out of poverty. In recent years, climate change has been abnormal frequently. Abnormal low temperature in winter and late spring coldness may cause the hard hit to C. oleifera farmers. Freezing injury can be caused by sudden decreases in temperature in winter. However, C. oleifera varieties differ in their hardiness to low temperatures. The paper investigated cold-resistance mechanisms by determining and analyzing the morphological, physiological and biochemical characteristics of C. oleifera from eastern, western and southern Anhui, respectively. Sensitivity to low temperature was assessed via the number of leaves in spring shoots, leaf thickness, the activities of protective enzymes such as CAT, POD and SOD, and the inclusion contents of WSS, FPro, MDA, benzene-alcohol extracts and lignin. The results showed that C. oleifera varieties had different physiological and biochemical, and morphological responses to low winter temperatures. In different regions, the number of leaves, leaf thickness, WSS content, FPro content and MDA content varied from 5.2-7.8, 398.79µm-465.27µm, 23.41mg/g-24.74mg/g, 41.86µg/g-44.18µg/g and 10.08µmol/g-14.51µmol/g, respectively. The varieties from eastern Anhui, the leaf thickness were thicker. Meanwhile, the protective enzyme activities and inclusion contents were relatively higher. The protective enzyme activities and chemical components contents such as benzene-alcohol extract and lignin represented significantly difference (p<0.05) among three regions. In the future, for the abnormal low temperature in winter, a serious of cultivation measures such as improving the contents of WSS, FPro, benzene-alcohol extract and lignin, were taken to enhance the cold resistance of C. oleifera. The result broadens the understanding of cold-resistance mechanisms in C. oleifera.

899-905 Download
8
INDUCTION OF DROUGHT TOLERANCE IN ZEA MAYS L. BY FOLIAR APPLICATION OF TRIACONTANOL
SHAGUFTA PERVEEN*, MUHAMMAD IQBAL, ATTIA NAWAZ, ABIDA PARVEEN AND SAQIB MAHMOOD

INDUCTION OF DROUGHT TOLERANCE IN ZEA MAYS L. BY FOLIAR APPLICATION OF TRIACONTANOL
ABSTRACT:
In the present study, we assessed the effect of foliar application of triacontanol (TRIA) on various growth and physiochemical parameters of two maize (Zea mays L.) cultivars (cv. MMRI-Yellow and cv. Hybrid S-515) under different irrigation levels i.e., normal watering (control) and watering at 60% of the field capacity (drought). Seeds of the two maize cultivars were sown in plastic pots filled with sandy loam soil (2 kg in each). Foliar application of TRIA (0, 2 and 5 µM) was performed after two weeks of drought stress to 28-day-old plants. Data of 58-day-old maize plants was collected for analysis of various growth and physiochemical attributes. Drought stress significantly decreased growth and superoxide dismutase (SOD) activity while increased the activities of catalase (CAT) and peroxidase (POD) and the contents of total phenolics, total soluble proteins, glycinebetaine (GB) and free proline. Foliar treatment with TRIA further increased CAT and POD activities whereas decreased the contents of hydrogen peroxide (H2O2), malondialdehyde (MDA), total phenolics and GB in the maize plants when under drought stress. Of the two maize cultivars, cv. MMRI-Yellow excelled the growth under both normal and drought stress (60% of the field capacity). Overall, TRIA (5 µM) was much more effective in modulating various growth and physiochemical attributes, and thus improving drought tolerance in maize plants.

907-915 Download
9
VARIATIONS IN ADAPTATION STRATEGIES OF WHEAT CULTIVAR REPLACEMENTS UNDER SHORT-TERM OSMOTIC STRESS
JIAKUN YAN1,3, NINGNING ZHANG1,3, NAN WANG1,2, YUPING LI2, SUIQI ZHANG1,2* AND SHIWEN WANG1,2

VARIATIONS IN ADAPTATION STRATEGIES OF WHEAT CULTIVAR REPLACEMENTS UNDER SHORT-TERM OSMOTIC STRESS
ABSTRACT:
Wheat (Tritium aestival L.) production has increased substantially with wheat cultivar replacements, but little is known about how different-year released wheat cultivars maintain whole-plant water balance especially under drought stress. We investigated a series of indices of plant water balance under two water conditions to identify the mechanisms underlying this process in seedlings of four wheat cultivars released in Shaanxi Province, China, from the 1960s to the present decade. The newer cultivars maintained good water status under well-watered conditions by a high uptake of water by roots. The various cultivars released at different times, however, maintained whole-plant water balance differently under osmotic stress. The newer cultivars decreased water loss by decreasing foliar stomatal conductance and severely down-regulating TaPIP1-2, a gene encoding an aquaporin. The expression of root aquaporin genes was higher in the older cultivars than in the newer cultivars. Root hydraulic conductance was significantly and positively correlated with the relative expression of another aquaporin homologue, TaPIP2-1. The older cultivars maintained high levels of transpiration by a relatively high root hydraulic conductance or a large root surface area. The newer cultivars, with lower root hydraulic conductance, maintained water balance by synchronously decreasing foliar water loss. Wheat breeders should seek more effective solutions for improving the capacity of roots to take up water and for reducing foliar water loss to be able to cope with the expected lower availability of water resources for agriculture in the future.

917-924 Download
10
GENETIC BEHAVIOR OF MORPHO-PHYSIOLOGICAL TRAITS AND THEIR ROLE FOR BREEDING DROUGHT TOLERANT WHEAT
SALMAN SALEEM1*, MUHAMMAD KASHIF1, MAKHDOOM HUSSAIN3, ABDUS SALAM KHAN1 AND M. FARRUKH SALEEM2

GENETIC BEHAVIOR OF MORPHO-PHYSIOLOGICAL TRAITS AND THEIR ROLE FOR BREEDING DROUGHT TOLERANT WHEAT
ABSTRACT:
The development of drought tolerant and high yielding varieties/germplasm is the major objective of any wheat breeding program. In the present study genetic architecture of physiological traits, yield and yield related parameters were studied using the generation mean analysis to improve grain yield under drought stress. A drought tolerant line, 9877 and a drought susceptible line, NR371 were crossed to develop six generations (P1, P2, F1, BC1, BC2, and F2). Results revealed additive, dominant and epistatic effects involved in the inheritance of characters which varied with trait and stress. Additive gene action was observed for canopy temperature, Chlorophyll ‘a’ and turgor potential. Although narrow sense heritability estimates for some traits were low but canopy temperature, chlorophyll ‘a’ and turgor potential expressed reasonably high heritability that supports the results of gene action providing an opportunity for early generation selection to use in a breeding program. The estimation of heritability for leaf carotenoids and turgor potential along with gene action for leaf carotenoids is a new work in wheat. The findings of present study suggested that physiological and bio-chemical traits are the indicators of stress tolerance and their utilization in developing high yielding drought tolerant wheat germplasm can expedite the breeding for stress tolerance.

925-933 Download
11
RELATIVE RESPONSE TO MECHANICAL STRESS OF CO-EXISTING AQUATIC SPECIES, FLOATING-LEAVED NYMPHOIDES PELTATA AND SUBMERGED MYRIOPHYLLUM SPICATUM
QIAN-JIN CAO*, NA LIU AND LING WANG

RELATIVE RESPONSE TO MECHANICAL STRESS OF CO-EXISTING AQUATIC SPECIES, FLOATING-LEAVED NYMPHOIDES PELTATA AND SUBMERGED MYRIOPHYLLUM SPICATUM
ABSTRACT:
With the increasing impact of human activities on the environment, the effects of mechanical disturbance on aquatic plants are increasingly evident. Mechanical perturbations at the upper layer and surface of water frequently damage floating-leaved and caulescent submerged plants. Here, we co-cultivated floating-leaved Nymphoides peltata and submerged Myriophyllum spicatum in mesocosms and then subjected leaf laminas of N. peltata and stems of M. spicatum to different levels of mechanical damage. Similar experiments were conducted twice, the second with having an increased intensity and frequency of disturbance. Generally, mechanical disturbances impacted the growth of these species more significantly in the second experiment than in the first. In the second experiment, the total biomass of N. peltata was decreased and the biomass proportion to leaf laminas increased with increasing disturbance intensity; leaf area was maintained across treatments. For M. spicatum, all disturbance treatments decreased total biomass and the total length of stems, compared to non-damage controls. However, in M. spicatum the number of ramets were increased with increasing disturbance intensity. Our results indicate that N. peltata has a high capacity for foliage compensation, and vegetative reproduction of M. spicatum may be improved by intense mechanical disturbance, though severe damage will suppress growth. Mechanical disturbances in upper water layers may change the relative performance of co-existing submerged and floating-leaved species with a repressive effect on submerged species. Management of mechanical disturbance by human activities is urgently required to maintain the function of submerged plants when restoring contaminated wetlands.

935-943 Download
12
EFFECTS OF DIFFERENT CONCENTRATIONS OF CADMIUM ON GROWTH AND MORPHOLOGICAL CHANGES IN BASIL (OCIMUM BASILICUM L.)
HASSAN NAZARIAN*1, DELARA AMOUZGAR2 AND HOSSEIN SEDGHIANZADEH2

EFFECTS OF DIFFERENT CONCENTRATIONS OF CADMIUM ON GROWTH AND MORPHOLOGICAL CHANGES IN BASIL (OCIMUM BASILICUM L.)
ABSTRACT:
This study was conducted on four treatments (0, 3, 5 and 10 milligrams per liter of cadmium nitrate solution) and three replications in a completely randomized design. The morphological characteristics including fresh weight, dry weight, stem length, root length, leaf weight ratio, measuring index of the imposed stress, specific leaf area and water availability per unit leaf area, and the amount of cadmium in stems and roots were measured. By increasing cadmium treatment at 10 ppm concentration, the root growth varied between 3.47 and 1.93 cm and the lowest root growth belonged to the treatment at 10 ppm concentration measured 1.93 ± 0.4 cm. The rising stem growth was also indicated. Fresh and dry weight analysis indicates their descending growth. The growth reduction was observed by increasing cadmium treatment at 10 ppm concentration in the study of the trait of weight and specific leaf area. Relative water content (RWC) of the leaves varies between 81 to 89 percent which represents the greatest amount of stress in the treatment at 10 ppm concentration. A notable decline in transport from the root to stem and a significant reduction in plant tolerance index were observed through the analysis of transfer factor and tolerance index in plants. The process of reducing metal transition from the root to stem has become slower by increasing the substance concentration. Basil can absorb and accumulate cadmium and its root is able to accumulate more quantity of the metal than its stem. The morphological signs of cadmium toxicity are remarkable through discolored roots, shortening the distance between nodes, creating an amorphous spots on the leaves, reducing the leaves extent and a slight pallor of the leaves.

945-952 Download
13
MITIGATION OF CADMIUM INDUCED STRESS IN TOMATO (SOLANUM LYCOPERSICUM L.) BY SELENIUM
ABD_ALLAH E. F.1*; HASHEM ABEER2,3 AND ALQARAWI A. A.1

MITIGATION OF CADMIUM INDUCED STRESS IN TOMATO (SOLANUM LYCOPERSICUM L.) BY SELENIUM
ABSTRACT:
Pot experiments were performed to assess the role of selenium in growth regulation of Solanum lycopersicum and its ameliorative role under cadmium stress. Cadmium (200 μM CdCl2) treatment reduced growth considerably and application of selenium (1.2μmol/L) improved growth and also mitigated the deleterious effects of cadmium. Selenium increased chlorophyll and carotenoid which were however reduced by cadmium stress. Cadmium treated plants showed higher lipid peroxidation and selenium caused improvement in membrane stability by reducing the production of hydrogen peroxide. Activities of antioxidant enzymes assayed increased with selenium treatment providing stability to cellular structures by quick removal of reactive free radicals. GSH and AsA were increased by 14.2% and 21.07% due to selenium application, however along with cadmium, it caused an increase of 40.5% and 45.7% respectively. Proline and phenol content increased with selenium providing osmotic stability to S. lycopersicum for cadmium tolerance. Moreover, selenium treatment declined the uptake of cadmium significantly.

953-961 Download
14
COMPARATIVE ECOPHYSIOLOGY OF SEED GERMINATION IN TWO PAIRS OF CLOSELY RELATED PINES OF MEDITERRANEAN AND HIMALAYAN ORIGINS
MUHAMMAD SAEED1*, COSTAS A. THANOS2, AGHA MUHAMMAD RAZA1, MUHAMMAD MUSHTAQ1 AND MUHAMMAD LUQMAN1

COMPARATIVE ECOPHYSIOLOGY OF SEED GERMINATION IN TWO PAIRS OF CLOSELY RELATED PINES OF MEDITERRANEAN AND HIMALAYAN ORIGINS
ABSTRACT:
In this study we have examined the comparative ecophysiology of seed germination in two pairs of related pines of Mediterranean and Himalayan origins. One pair comprises Pinus canariensis and Pinus roxburghii, and the second Pinus peuce and Pinus wallichiana. Each pair has close taxonomic and morphological affinities. We have observed considerable differences among these pines about their germination behaviour. Seed germination of Pinus canariensis was feasible within a range of 15-20°C both under light and darkness, while the optimum germination of Pinus roxburghii was achievable at 25°C under white light. Germination rate of Pinus roxburghii was significantly enhanced under white light. A brief red light pulse resulted in promoting germination in Pinus roxburghii; however, the enhancing effect of red light was reversed by a brief far-red pulse; which indicated the mediation of phytochrome system in seed germination. Pinus peuce seeds were deeply dormant and required an extended period of stratification in order to come out of dormancy. Pinus wallichiana seeds were less dormant as compared to Pinus peuce, 30 d stratification or incubation under white light resulted in overcoming the dormancy.

963-969 Download
15
MECHANISM OF FLURIDONE-INDUCED SEED GERMINATION OF CISTANCHE TUBULOSA
QING LIANG CHEN1, YUHAI GUO2, YONG JIANG1 AND PENGFEI TU1*

MECHANISM OF FLURIDONE-INDUCED SEED GERMINATION OF CISTANCHE TUBULOSA
ABSTRACT:
Our previous study disclosed that fluridone, a synthesis inhibitor of abscisic acid (ABA), could stimulate seed germination in the holoparasitic plant Cistanche tubulosa. Nonetheless, the underlying mechanisms have not been thoroughly elucidated. In the present study, an attempt was made to reveal the mechanism of fluridone breaking seed dormancy in C. tubulosa and to determine the contribution of hormones in this process. The ABA level in seeds initially decreased following fluridone treatment and was subsequently maintained at a concentration of 31 ng·g-1 DW (dry weight) three days later. The contents of gibberellins (GAs) initially increased and subsequently were maintained at a level of 40

971-976 Download
16
EFFECTS OF PLANT GROWTH REGULATORS ON SEED GERMINATION AND CALLUS INDUCTION OF HYLOCEREUS COSTARICENSIS
WINSON KOE WEI SHENG1, JEEVANDRAN SUNDARASEKAR2, KATHIRESAN SATHASIVAM1 AND SREERAMANAN SUBRAMANIAM3*

EFFECTS OF PLANT GROWTH REGULATORS ON SEED GERMINATION AND CALLUS INDUCTION OF HYLOCEREUS COSTARICENSIS
ABSTRACT:
Dragon fruit (Hylocereus costaricensis) belongs to the family Cactaceae and are climbing vines which have received worldwide attention in recent years. However, there are still lack of information on the protocols for the establishment of In vitro culture system. In the present study, seed germination percentage were determined by culturing seeds on semi-solid Murashige and Skoog medium (MS) supplemented with 1 ppm 6-Benzylaminopurine (BAP) together with either 0, 0.5 or 0.8 ppm of Indole-3-butyric acid (IBA). Germination percentage was the highest by using plant growth regulators (PGRs) combination of 1 ppm BAP and 0 ppm IBA (93.33%). Subsequently, the cotyledons from seedlings of the germinated seeds were used for subsequent callus induction. Small pieces of cotyledons were excised and cultured on MS medium fortified with 0.45, 0.9, 1.8, 2.7, 3.6, and 4.5 ppm of 2,4-Dichlorophenoxyacetic acid (2,4-D) together with either 0, 0.9 or 1.8 ppm of BAP. Callus induction percentage was highest using the plant growth regulators (PGRs) combination of 3.6 ppm 2,4-D and 1.8 ppm BAP (75%). Hence, 3.6 ppm of 2,4-D and 1.8 ppm BAP was the best combination for callus induction of Hylocereus costaricensis.

977-982 Download
17
ECOLOGICAL IMPLICATIONS AND ENVIRONMENT DEPENDENCE OF THE SEED GERMINATION OF COMMON SPECIES IN COLD DESERTS
SUN YUAN-YUAN1, ZHOU JUAN1, LIU TONG1*, LIU ZUN-CHI1, HAO XIAO-RAN1, LIU HUA-FENG1 AND LI RU2

ECOLOGICAL IMPLICATIONS AND ENVIRONMENT DEPENDENCE OF THE SEED GERMINATION OF COMMON SPECIES IN COLD DESERTS
ABSTRACT:
Vegetation is increasingly affected by climate change in cold deserts. Nonetheless, research is limited regarding the natural environmental demands of seed germination in such deserts. This study was conducted in Gurbantunggut Desert as a research base and 17 common species as subjects to investigate the moisture and temperature needs of seed germination in artificial settings, as well as the relationship between characteristics of seed germination and the local distribution of dune and shrubs. Results showed:(1) all tested species generally display low germination percentages that range between 2.9% and 79.6%. Winter snow melt dictates seed germination in cold deserts. Moreover, the subsequent spring rainfall can increase the survival rate of seedlings and significantly affect the process of seed germination. (2) seeds start to germinate only two days after snow melts at the average daily temperature (day/night) of 3.5°C (6.7 °C/-0.5°C) and at a soil volumetric water content of 24.2%. Fifteen days after snow melt, all species germinate when the soil volumetric water content is 6.0% and the average daily temperature is 12.9°C (18.3°C/7.1°C). (3) The seed germination of the tested species can be divided into four patterns: rapid, transitional, slow, and low. Low-pattern plants mainly grow on upper dunes and are significantly associated with shrubs. Rapid- and slow-pattern plants distribute in middle and lower dunes. A few of these plants are significantly associated with shrubs. Transitional-pattern plants generally develop in the low land between hills and middle dunes. This study provides a reference for the actual environmental needs of seed germination in cold deserts and for the temperature and moisture requirements of this process in future experimental settings.

983-992 Download
18
RELATIONSHIP AMONG WATER USE EFFICIENCY, CANOPY TEMPERATURE, CHLOROPHYLL CONTENT AND SPOT BLOTCH (COCHLIOBOLUS SATIVUS) RESISTANCE IN DIVERSE WHEAT (TRITICUM AESTIVUM L.) GERMPLASM
MUHAMMAD JAMIL1*, AAMIR ALI1, KHALID FAROOQ AKBAR2, ABDUL GHAFOOR3, ABDUL AZIZ NAPAR4, SHEHZAD ASAD5, SAJJAD HUSSAIN7, ALVINA GUL KAZI6 AND A. MUJEEB-KAZI7

RELATIONSHIP AMONG WATER USE EFFICIENCY, CANOPY TEMPERATURE, CHLOROPHYLL CONTENT AND SPOT BLOTCH (COCHLIOBOLUS SATIVUS) RESISTANCE IN DIVERSE WHEAT (TRITICUM AESTIVUM L.) GERMPLASM
ABSTRACT:
The food security vision for 2050 has front- lined wheat as the major conduit to feed the global populace estimated at 9.2 billion. Pakistan ranks 6th in world population and among the top ten countries for wheat production, but annual yield productivity appears to be stagnated due to prevalence of various biotic and abiotic stresses. More recently the incidence of a new foliar wheat disease spot blotch (Cochliobolus sativus) in drought and heat stressed areas of Pakistan has necessitated that a new look on both water use efficiency and spot blotch resistance be taken. Hence this study has attempted to establish a relationship between these two stress influencing constraints. One hundred lines were assessed for intrinsic water use efficiency, canopy temperature, chlorophyll concentration index and spot blotch resistance. Rates of photosynthesis (A) and transpiration (E), stomatal conductance (gs) and internal CO2 (Ci) were estimated using the Infrared Gas Analyzer (IRGA). Chlorophyll concentration index and canopy temperature were also measured to determine the extent of physiological changes under disease pressure. Spot blotch presence was estimated using the standard double digit disease scoring scale. Our results have revealed a direct relationship among water use efficiency, canopy temperature, chlorophyll concentration index and spot blotch resistance. Keeping three growth stages (GS: 63, GS: 69 and GS: 77) as a source of variation, data analyses have shown a significant difference amongst the studied attributes. Structural model equation revealed that about 87.8% variability was explained by the studied attributes. Looking at significance of mean square values of % severity, 17 wheat lines were found spot blotch resistant. These lines are valuable breeding stocks for wheat improvement for hot ecological niches within Pakistan and globally where higher incidence of spot blotch prevails.

993-998 Download
19
THREATENED PLANT RESOURCES: DISTRIBUTION AND ECOSYSTEM SERVICES IN THE WORLD’S HIGH ELEVATION PARK OF THE KARAKORAM RANGES
ARSHAD ALI SHEDAYI1,2, MING XU3*, FARRUKH HUSSAIN4, SEHRISH SADIA5, IQNAA NASEER6 AND SEEMA BANO2

THREATENED PLANT RESOURCES: DISTRIBUTION AND ECOSYSTEM SERVICES IN THE WORLD’S HIGH ELEVATION PARK OF THE KARAKORAM RANGES
ABSTRACT:
This study aims to investigate diversity, distribution, status, ecosystem services and threats to the plant resources in the study area based on field survey and ethno ecological knowledge for effective conservation and sustainable ecosystem services. The present study was conducted in the world’s high elevation Khunjerab National Park (KNP) of the Karakoram ranges in Pakistan bordering China. Tremendous ecosystem services are obtained from the park and considered the most important habitat for many plant biodiversity and wildlife species. Field surveys were conducted to collect plants in transect along the road side of seven valleys ranging from 3160m to 4934m altitudinal variation. The names and traditional uses were recorded from the local people of the area by semi structured questionnaires and direct interviews. The data was analyzed by excel spreadsheets, direct matrix ranking, and pair comparison tests. Asteraceae was the dominant family with 15% species followed by Chenopodiaceae 10%, Poaceae 8%, Papilionaceae and Rocaceae 7%each, Brasicaceae 6%. Plant resources contribute direct and indirect ecosystem services such as food, medicine, fuel, timber, thatching, water purification, mineral and soil retention, and most importantly as sink of global carbon stock especially in the high altitude peatlands. Herbs were the dominant species in the area with 89%. Fodder is the most common usage for plants, followed by medicine. Plants with percentages 27% and 39% found to be highly palatable and palatable respectively. Competition for food between wildlife and livestock was high recorded for 60% plants. Plants used to cure various diseases including stomachache, asthma, cancer and tuberculosis etc. Plant resources in KNP are unique and vary with climate and altitude. This floral wealth is under tremendous threats of global climate change and anthropogenic activities like overgrazing, increasing population, and a rapidly declining traditional knowledge for sustainable use. Awareness for sustainable harvesting of plants, reducing grazing pressure, equitable resource sharing, marketing of the products, effective management and policy decisions and research on ecosystem services assessment, valuation and climate change impact on wild plant resources are recommended.

999-1012 Download
20
FLORISTIC COMPOSITION AND ECOLOGICAL CHARACTERISTICS OF PLANTS OF CHAIL VALLEY, DISTRICT SWAT, PAKISTAN
ASGHAR ALI1*, LAL BADSHAH2, FARRUKH HUSSAIN3 AND ZABTA KHAN SHINWARI4

FLORISTIC COMPOSITION AND ECOLOGICAL CHARACTERISTICS OF PLANTS OF CHAIL VALLEY, DISTRICT SWAT, PAKISTAN
ABSTRACT:
The present study was carried out during 2012-2014 to enumerate the floristic and ecological characteristics of plants of Chail Valley, District Swat. A total of 463 species belonging to 104 families were recorded. Leading families were Asteraceae (42 Spp), Poaceae (35 Spp), Rosaceae and Lamiaceae (each with 26 Spp), Papilionaceae (25 Spp), Brassicaceae and Boraginaceae (each with 16 Spp), Apiaceae (14 Spp), Solanaceae (13 Species) and Ranunculaceae (12 Spp). Each of the remaining families had less than 12 species. Therophytes with 188 species, 40.60% were dominant. They were followed by hemicryptophytes (77 species, 16.63%). Cuscuta europaea L., C. reflexa Roxb. and Viscum album L. were the three shoot parasites. The leaf spectra was dominated by mesophylls (147 Spp; 31.75%), microphylls (140 Spp.; 30.24%) and nanophylls (136 Spp.; 29.37%). Two species were aphyllous. Majority of the species (305 Spp., 65.87%) had simple lamina. Eight species (1.73%) had spiny leaves.

1013-1026 Download
21
FLORISTIC ANALYSIS OF THE WANDA MOUNTAIN IN NORTH EASTERN CHINA
HONGFRNG WANG1,2, QINGGUI WANG1,2*, LIJIAN XU1, GUOYONG YAN1,2, ZHI ZHANG1, YAJUAN XING1,3*, LIBO ZHAO1, LIHUA WANG4, SHIJIE HAN4 AND JINGHUA YU4

FLORISTIC ANALYSIS OF THE WANDA MOUNTAIN IN NORTH EASTERN CHINA
ABSTRACT:
The plants of the Wanda Mountain area were investigated between 2009 to 2013. The results show that Wanda Mountain has 95 families of seed plants distributed in 334 genera and 705 species. A geographical component analysis shows that in addition to a small number of cosmopolitan species, cold, temperate and tropical species account for 14.9%, 77.3% and 4.4% of the total species, respectively, indicating that the flora of the Wanda Mountains exhibits a significant temperate nature and includes a small number of tropical components and certain cold components. In addition, the Wanda Mountains show a remarkable level of endemism and are geographically related to other regions in East Asia, particularly Japan. Furthermore, the Wanda Mountains present a complicated species composition, with a total of 14 distribution patterns and 10 variants. The coefficient of similarity between the flora of the Wanda Mountain area and the flora of the Changbai Mountain area is 43.1%, and the coefficient of similarity between the flora of the Wanda Mountain area and the flora of the Lesser Xingan Mountain area is 49.2%, indicating that the plants of the Wanda Mountain area are more common to those of the Lesser Xingan Mountain area.

1027-1034 Download
22
THE SEED ATLAS OF PAKISTAN-. PAPAVERACEAE
RUBINA ABID,* AFSHEEN ATHER AND M. QAISER

THE SEED ATLAS OF PAKISTAN-. PAPAVERACEAE
ABSTRACT:
Numerical analysis based on seed morphological characters of 14 taxa belonging to the family Papaveraceae is carried out. Seed macro and micro morphological characters were found useful to strengthen the taxonomic decisions and trace out the phylogenetic relationship within the family Papaveraceae.

1035-1044 Download
23
SPORE MORPHOLOGY OF SOME GRIMMIACEAE ARN. SPECIES BELONGING TO MOSS FLORA OF TURKEY
FILIZ SAVAROGLU1, ISMUHAN POTOGLU ERKARA1* AND FATIH OZCELIK2

SPORE MORPHOLOGY OF SOME GRIMMIACEAE ARN. SPECIES BELONGING TO MOSS FLORA OF TURKEY
ABSTRACT:
This study includes spore morphology of seven Grimmiaceae taxa from Turkey. The spores of Schistidium trichodon (Brid.) Poelt, S. confertum (Funck) Bruch & Schimp., Grimmia ovalis (Hedw.) Lindb., G. pulvinata (Hedw.) Sm., G. trichophylla Grev., G. dissimulata E.Maier and G. decipiens (Schultz) Lindb. were examined by light and scanning electron microscopy. The apertural region forms from a leptoma in all spores. The spore morphology of the examined taxa of the family is verrucate type. The spore shape of all studied species is prolate-spheroid. Spore size ranges from 6 μm to 17 μm in the family of Grimmiaceae. The spore wall of the family Grimmiaceae includes sclerine and intine. The examined species of mosses are belonged to saxicolous habitat type. The taxonomical and ecological implications of the family Grimmiaceae are discussed on the basis of their spore morphology.

1045-1050 Download
24
POLLEN MORPHOLOGY OF THE SUBFAMILY ARECOIDEAE GRIFF. (FAMILY-ARECACEAE) FROM PAKISTAN AND KASHMIR
ABID ARZOO RASHEED1*, ANJUM PERVEEN1, ROOHI ABID1 AND MUHAMMAD QAISER2

POLLEN MORPHOLOGY OF THE SUBFAMILY ARECOIDEAE GRIFF. (FAMILY-ARECACEAE) FROM PAKISTAN AND KASHMIR
ABSTRACT:
Pollen morphological characters are potentially informative in the systematic of monocotyledons including (Arecaceae), at supra and infra familial level. The pollen morphology of 8 species (subfamily Arecoideae) clearly shows that the qualitative pollen characters (such as aperture, exine pattern and shape) are found to be taxonomically important as compared to the quantitative characters (such as size of the grain and exine thickness).Considerable pollen variations have been found within the subfamily Arecoideae with regard to size, shape, aperture type and exine pattern. For instance, 7 out of 8 species have monosulcate pollen (i.e., 87.5%) and a remaining species (viz., Elaeis guineensis Jacq.) have trichotomosulcate pollen. The pollen are frequently elliptical, but in some cases rounded-triangular pollen were also found. Similarly, there is a great diversity in exine pattern such as punctate, reticulate, vermiculate, spinose, rugulate and perforate or combination of these types have also been found, but the most predominant pattern is the reticulate type.

1051-1060 Download
25
GENETIC AND ANATOMICAL ANALYSIS OF NORMAL AND ABNORMAL FLOWERS OF DATE PALM CULTIVAR 'BARHY' DERIVED FROM OFFSHOOT AND TISSUE CULTURE
OMER H. SHAIR1*, EJAZ ASKARI2 AND PERVAIZ R KHAN3

GENETIC AND ANATOMICAL ANALYSIS OF NORMAL AND ABNORMAL FLOWERS OF DATE PALM CULTIVAR 'BARHY' DERIVED FROM OFFSHOOT AND TISSUE CULTURE
ABSTRACT:
Random Amplified Polymorphic DNA (RAPD) analysis between 6 normal flower producing offshoot derived and 6 abnormal multiple carpel, flower producing tissue culture (TC) derived trees of cultivar (cv.) 'Barhy', was performed with the objective to check genetic variation if any at DNA level. DNA samples were extracted from pollinated and un-pollinated flowers from both sets of plants. Amplified RAPD products were clearly detected with 30 primers used in this experiment but only 3 gave a few polymorphic bands which shows low level of genetic variation among the offshoot and TC derived plants. Cluster analysis by the unweighted paired group method of arithmetic means (UPGMA) showed close genomic similarity among the 12 DNA samples with the range of 0.486-0.904 Nei and Li's coefficient in the similarity matrix. The average similarity among the 12 DNA samples was more than 50%. Floral abnormalities in TC derived plants were also studied microscopically. Abnormalities like more than three carpel development, abnormal ovule development and deformities of style and stigma were observed. The results show that the composition and the abnormalities of flowers in TC derived plants of cultivar 'Barhy' may be attributed to epigenetic changes that takes place at different stages of tissue culture and not due to major changes at DNA level.

1061-1065 Download
26
OVER-EXPRESSION OF ZmARG ENCODING AN ARGINASE IMPROVES GRAIN PRODUCTION IN MAIZE
HONG DI1, ROU LI1, YU TIAN1, XIANYU MENG1, PEI ZHANG1, DAN LIU1, XING ZENG1 AND ZHENHUA WANG1*

OVER-EXPRESSION OF ZmARG ENCODING AN ARGINASE IMPROVES GRAIN PRODUCTION IN MAIZE
ABSTRACT:
Arginase, as one of the three key enzymes in nitrogen catabolism, the physiological role of Arg catabolism in cereal crops has not been fully clarified. Studies have shown that arginase-encoding genes play a key role in providing nitrogen to developing seedlings in many plant species.Yield is a primary trait in many crop breeding programs, which can be increased by modification of genes related to photosynthesis, nitrogen assimilation, carbon distribution, plant architecture, and transcriptional networks controlling plant development. In the present study, a maize arginase gene ZmARG was cloned and introduced into maize inbred lines by Agrobacterium tumefaciens- mediated transformation. Putative transgenic plants were confirmed by PCR, Southern blotting RT-PCR analysis. The expression of the ZmARG gene increased arginase activity in several tissues in transgenic lines. Transgenic maize plants had significantly higher ear weight and 100-seed weight as compared with wild-type control. Our results suggested that ZmARG was a potential target gene for crop yield improvement.

1067-1072 Download
27
DISTRIBUTION OF SEVEN GRAIN GENES AND EVALUATION OF THEIR GENETIC EFFECTS ON GRAIN TRAITS
YADONG ZHANG, QINGYONG ZHAO, CHUNFANG ZHAO, TAO CHEN, ZHEN ZHU, LIHUI ZHOU, SHU YAO, LING ZHAO AND CAILIN WANG*

DISTRIBUTION OF SEVEN GRAIN GENES AND EVALUATION OF THEIR GENETIC EFFECTS ON GRAIN TRAITS
ABSTRACT:
Grain size is one of the most important factors determining rice yield. Several major QTLs for grain size control have been molecular characterized, and their roles in the regulation of grain size or weight have been explored but still remained obscure. In this study, we systematically examined the distributions of GW2, GS3, qSW5, qGL3, GS5, GW8, and TGW6 in 240 RIL of TD70 and Kasalath, compared the phenotypic differences between functional (additive) and non-functional alleles of the seven genes by monogenic lines and analyzed the interaction effect of gs3, gw2, and qgl3. The results showed that the 7 genes could be randomly combined, and an individual gene has its specific roles in grain length, grain width, and grain thickness, the seven functional genes regulating grain weight follows the order as qgl3 >gw2>gs3> GS5>qsw5> GW8>TGW6, and the combination effect among gs3, gw2, and qgl3 were revealed. These findings provide novel insight into grain size regulation in rice and are likely to be useful for marker assisted breeding in rice grain size.

1073-1079 Download
28
GENOME-WIDE IDENTIFICATION AND EXPRESSION ANALYSIS OF THE MALATE DEHYDROGENASE GENE FAMILY IN GOSSYPIUM ARBOREUM
MUHAMMAD IMRAN AND JIN-YUAN LIU*

GENOME-WIDE IDENTIFICATION AND EXPRESSION ANALYSIS OF THE MALATE DEHYDROGENASE GENE FAMILY IN GOSSYPIUM ARBOREUM
ABSTRACT:
Malate dehydrogenase (MDH) is a key enzyme that catalyzes the reversible oxidation of malate to oxaloacetate and plays a crucial role in various cellular processes, such as cell expansion, wall thickening and cell elongation. Although individual genes belonging to MDH gene family have been partially identified in various plants, there have been no reports of a genome-wide characterization of the MDH gene family in cotton. Here, we identified a total of 13 MDH genes from the genome of a diploid cotton Gossypium arboreum and designated GaMDH1-13 based on their chromosomal locations. These MDH members were unevenly distributed on 8 of the13 chromosomes. Segmental duplications that played a dominant role in the expansion of the MDH gene family were estimated to have occurred between 19.07 to 20.47 million years ago (MYA), when a recent large-scale genome duplication occurred in cotton. Phylogenetic analyses showed that the putative MDH proteins formed five groups (I to V) in plant species. GaMDH genes within the same group shared similar gene structures and domain constitutions. Furthermore, expression analysis showed that the GaMDH genes were differentially expressed in root, stem, leaf, hypocotyl, petal and anther, with higher expression levels detected during different fiber developmental stages. Notably, GaMDH13 had the highest expression level during the fast fiber elongation stage that ranged from 5 to 15 day post-anthesis (DPA), suggesting that the MDH gene plays a vital role in fiber development. The results of this study will aid functional analyses of the MDH genes in cotton fiber development.

1081-1090 Download
29
MOLECULAR AUTHENTICATION AND QUALITY CONTROL OF CROCUS SATIVUS AND ALOE BARBADENSIS IN RAW MATERIAL SOURCE AND POLYHERBAL MEDICINE EMPLOYING SCAR MARKERS
IRUM GUL, UMARA NISSAR AND MALIK Z. ABDIN*

MOLECULAR AUTHENTICATION AND QUALITY CONTROL OF CROCUS SATIVUS AND ALOE BARBADENSIS IN RAW MATERIAL SOURCE AND POLYHERBAL MEDICINE EMPLOYING SCAR MARKERS
ABSTRACT:
Crocus sativus L. and Aloe barbadensis Mill. are well known functional ingredients in health and food products and are known for their medicinal properties as antioxidant, antidepressant, relaxant and cathartic agents. In this study, reliable quality control markers were developed for the quality assurance of C. sativus and A. barbadensis at raw material source as well as in finished herbal products. DNA based sequence-characterized amplified region (SCAR) markers were developed from randomly amplified polymorphic DNA (RAPD) markers specific for both the species to detect its adulteration in commercial products. Developed RAPD markers were cloned and sequenced and SCAR primers were generated SCAR markers upon PCR amplification with genomic DNA. These specific SCAR markers enabled unequivocal detection of adulterants in the commercially procured polyherbal medicine. C. sativus could not be identified through SCAR markers, instead SCAR amplicon of Carthamus tinctorius was detected suggesting that although being labelled as one of the constituents, C. sativus may not have been used to prepare the polyherbal medicine. This simple analytical strategy could be used for large scale screening of medicinal plants at raw material source as well as in finished polyherbal medicine.

1091-1098 Download
30
EXPRESSION ANALYSIS OF FIBER RELATED GENES IN COTTON (GOSSYPIUM HIRSUTUM L.) THROUGH REAL TIME PCR
NADIA IQBAL1, ASIA KHATOON1, MUHAMMAD ASIF1 AND AFTAB BASHIR2*

EXPRESSION ANALYSIS OF FIBER RELATED GENES IN COTTON (GOSSYPIUM HIRSUTUM L.) THROUGH REAL TIME PCR
ABSTRACT:
Cotton fibers are unicellular seed trichomes and the largest known plant cells. Fiber morphogenesis in cotton is a complex process involving a large number of genes expressed throughout fiber development process. The expression profiling of five gene families in various cotton tissues was carried out through real time PCR. Expression analysis revealed that transcripts of expansin, tubulin and E6 were elevated from 5 to 20 days post anthesis (DPA) fibers. Three Lipid transfer proteins (LTPs) including LTP1, LTP3, LTP7 exhibited highest expression in 10 - 20 DPA fibers. Transcripts of LTP3 were detected in fibers and non fiber tissues that of LTP7 were almost negligible in non fiber tissues. Sucrose phosphate synthase gene showed highest expression in 10 DPA fibers while sucrose synthse (susy) expressed at higher rate in 5-20 DPA fibers as well as roots. The results reveal that most of fiber related genes showed high expression in 5-20 DPA fibers. Comprehensive expression study may help to determine tissue and stage specificity of genes under study. The study may also help to explore complex process of fiber development and understand the role of these genes in fiber development process. Highly expressed genes in fibers may be transformed in cotton for improvement of fiber quality traits. Genes that were expressed specifically in fibers or other tissues could be used for isolation of upstream regulatory sequences.

1099-1106 Download
31
THE DIFFERENTIAL GENE EXPRESSION OF KEY ENZYME IN THE GIBBERELLIN PATHWAY IN THE POTATO (SOLANUM TUBEROSUM) MUTANT M4P-9
JIAN-BIN SHI1, GUANG-JI YE1,2,3,4,5, YONG-ZHI YANG1,2,3,4,5, FANG WANG1,2,3,4,5, YUN ZHOU1,2,3,4,5, HAI-HONG SUN1,2,3,4,5 AND JIAN WANG1,2,3,4,5*

THE DIFFERENTIAL GENE EXPRESSION OF KEY ENZYME IN THE GIBBERELLIN PATHWAY IN THE POTATO (SOLANUM TUBEROSUM) MUTANT M4P-9
ABSTRACT:
In the present study, the expression patterns of the key genes in the gibberellin synthesis pathway in the potato dwarf mutant M4P-9 were detected using quantitative real-time PCR. Using Actin as an internal control, CPS1, KS, KO, GA20ox1, and GA2ox1, genes for key gibberellin synthesis enzymes, were evaluated, along with a gibberellin receptor gene. The standard curves were obtained from dilutions of PCR product; the correlation coefficient for Actin was 0.995, and those for the target genes varied from 0.994 to 1.000. The expression patterns of gibberellin pathway genes in different growth stages and tissues were calculated according to the method of Pfaffl. These genes showed expression patterns that varied based on growth stage and tissue type. The higher expression levels of CPS1 and GA2ox1 in roots, the lower expression levels of GA20ox1 in roots during tuber formation stage; as well as the increased expression of GA20ox1 and GA2ox1 genes in stems during the tuber formation stage, likely play key roles in the plant height phenotype in M4P-9 mutant materials. This article provides a basis for researching the mechanism of gibberellin synthesis in potato.

1107-1117 Download
32
GA4 AND tZR QUANTIFICATION IN PISTILLATE AND STAMINATE PLANTS OF DASYLIRION CEDROSANUM
ERIKA NOHEMI RIVAS-MARTÍNEZ1, RAHIM FOROUGHBAKCHK POURNAVAB1, M. HUMBERTO REYES-VALDÉS2 AND ADALBERTO BENAVIDES-MENDOZA3*

GA4 AND tZR QUANTIFICATION IN PISTILLATE AND STAMINATE PLANTS OF DASYLIRION CEDROSANUM
ABSTRACT:
‘Sotol’, a protected designation of origin alcoholic beverage, is obtained from Dasylirion cedrosanum spp. plants. There is little knowledge concerning the sexual differentiation mechanisms of this species, which leads to a lack of proposals for not only its conservation and reforestation but also mechanisms to account for the dioecious nature of the plant. Phytohormones have been associated with sexual differentiation in dioecious plants because, individually or in combination, these hormones promote masculinization or feminization of their sexual structures. The objective of this study was to quantify gibberellin A4 (GA4) and trans-zeatin riboside (tZR) levels in samples of different organs of staminate and pistillate plants of D. cedrosanum, which were collected at different stages of floral development. The plant material was obtained at three locations in Coahuila at North Mexico. Gibberellin A4 (GA4) and trans-zeatin riboside (tZR) were quantified by HPLC-UV at 205 nm and 268 nm, respectively. During the later appearance of pollen and seeds, the GA4 levels in the crown and leaves were the same but exceeded those in the inflorescence. There were no differences in tZR levels between the plants of different sexes. Among organs, differences were only found during inflorescence emergence and death, stages during which the crown presented the highest levels of tZR. The results for the GA4/tZR ratio were similar to those reported for GA4. GA4 most likely plays a role in sexual determination in D. cedrosanum because its presence is associated with the appearance of staminate flowers.

1119-1127 Download
33
FERTILITY VARIATION IN TWO POPULATIONS OF TAURUS CEDAR (CEDRUS LIBANI RICH.)
HALİL BARIŞ ÖZEL1* AND NEBİ BİLİR2

FERTILITY VARIATION IN TWO POPULATIONS OF TAURUS CEDAR (CEDRUS LIBANI RICH.)
ABSTRACT:
Fertility variation, measured as “half-sib family coefficient”, based on number of one, two and three years cones were investigated in plantation population (PP), and a natural population (NP) of Taurus Cedar (Cedrus libani Rich.) sampled from southern part of Turkey. Fertility variation was higher in PP than NP for one, two and three years. It was the highest in PP for one year cones (2.34), while it was lowest in NP for three years cones (1.73) as shown in Table 2. The effective number of parents were 21.8 (38.4% of census number) for one year cones, 25.7 (47.9% of census number) for two years cones and 29.8 (52.6% of census number) for three cones in PP. On the other hand the effective number of parents were 28.3 (43.4% of census number) for one year cones, 32.8 (51.6% of census number) for two years cones and 36.4 (58.9% of census number) for three years cones in NP. Diameter at breast height and tree crown area had positive and significant (p<0.05) effective on cone production, while effects of tree height and tree age were not significant (NS) on that (Table 3). There were also positive and significant (p<0.05) correlation between years in cone production.

1129-1132 Download
34
QTL MAPPING OF ADULT-PLANT RESISTANCE TO LEAF RUST IN CHINESE WHEAT CULTIVAR LANTIAN 9
HAN LIUSHA1√, LI ZAIFENG1√, WANG XIAODONG1, KANG ZHANHAI1,2, ZHU LIN1, LI XING1,2*, LIU DAQUN1* AND SYED JAWAD AHMAD SHAH2

QTL MAPPING OF ADULT-PLANT RESISTANCE TO LEAF RUST IN CHINESE WHEAT CULTIVAR LANTIAN 9
ABSTRACT:
Chinese wheat cultivar “Lantian 9” showed a stable high yield in the field. Together with its other desirable traits such as tolerance to cold, drought, leaf rust and stripe rust, made it a good source for wheat breading. In our preliminary test, “Lantian 9” showed a typical adult resistance phenotype (susceptible at seedling stage but high resistance at adult stage) to most of the major Chinese leaf rust pathotypes. To clarify the adult-plant resistance (APR) genes in this cultivar, a population with 197 F2:3 lines was generated by crossing “Lantian 9” with susceptible line “Huixian Hong”. The population was phenotyped in the field over three years (year 2012

1133-1137 Download
35
IMPROVEMENT OF EMBRYOGENESIS AND REGENERATION BY AIR DESICCATION IN MAIZE (ZEA MAYS L.)
SELIM MORSHED, ABU BAKER SIDDIQUE AND S.M. SHAHINUL ISLAM*

IMPROVEMENT OF EMBRYOGENESIS AND REGENERATION BY AIR DESICCATION IN MAIZE (ZEA MAYS L.)
ABSTRACT:
Calli derived from mature embryos of four maize varieties viz. Mohar, Khoi bhutta, Barnali and Shuvra were cultured in three basal media for regeneration (MS, N6 and 6N1) which individually supplemented with four hormonal combinations e.g. H1 = BAP 0.5 mg/l + IAA 0.0 mg/l, H2 = BAP 1.0 mg/l + IAA 0.5 mg/l, H3 = BAP 1.5 mg/l + IAA 1.0 mg/l and H4 = BAP 2.0 mg/l + IAA 1.5 mg/l. The highest frequency of regeneration was found with MS + H2 (41.35%) in Mohar, while the lowest was 17.37% in 6N1 + H1 for Barnali. To enhance the capability of regeneration, calli were pretreated by ten groups (6, 12, 18, 24, 30, 36, 42, 48, 54 and 60 h) of desiccation periods. The degrees of desiccation of pretreated calli were determined; and it was ranged as 6.23 to 40.52% where Khoi bhutta showed the maximum value at 60 h desiccation. The callus of Mohar exhibited the highest frequency of regeneration (75.24%) which desiccated for 48 h; and it was around 2 fold higher than the control. The variety Khoi bhutta showed the lowest efficiency (31.80%) when the callus was desiccated for 6 h. All the varieties performed their maximum regeneration at different periods, where 36, 30 and 42 h desiccation were optimal for Barnali (67.23%), Khoi bhutta (68.03%) and Shuvra (73.98%) accordingly. Analysis of variance (ANOVA) showed significant effect of maize genotype and periods of partial air desiccation to enhance regeneration at p<0.05 level.

1139-1144 Download
36
EVALUATION OF POLYPHENOL CONTENT IN DIFFERENT PARTS OF PHYSALIS IXOCARPA
WAJID KHAN1,2 JEHAN BAKHT1* AND MOHAMMAD SHAFI3

EVALUATION OF POLYPHENOL CONTENT IN DIFFERENT PARTS OF PHYSALIS IXOCARPA
ABSTRACT:
In the current study extracts of leaf, stem, fruit and calyx with different polarity was investigated for their phenolic content using high performance liquid chromatography and spectrophotometric assay. Among different parts, stem contain high concentration of total polyphenol and gallic acid. The effect of extraction solvent on polyphenol quantification was observed in both assays. Spectrophotometric analysis of the data regarding polyphenol content indicated that among different extracts from the stem, leaf and fruit tissues; ethyl acetate extracted fraction of stem measured maximum polyphenol content of 110.376 mgGAE/g of dry extract. The ethyl acetate extracted sample of leaf showed high polyphenol (Gallic acid) content of 95 mg GAE/g of dry extract using high performance liquid chromatography assay. The amounts of phenolic content (Gallic acid) extracted from the parts of the plant with the different solvent ranged from 0.0354- 95 mg GAE/g of the dry extract using HPLC, however, spectrophotometric assay indicated total polyphenol ranged from 38-110.37 mgGAE g-1 of the dry extract. The current study suggested that ethyl acetate is an effective solvent for the extraction of polyphenol in different parts of P. ixocarapa.

1145-1151 Download
37
MICROPROPAGATION OF GINGER (ZINGIBER OFFICINALE VAR. RUBRUM) USING BUDS FROM MICROSHOOTS
ZURAIDA A.R.1*, MOHD SHUKRI M.A.2, ERNY SABRINA M.N.3, AYU NAZREENA O1 , CHE RADZIAH C. Z.4, PAVALLEKOODI G.5 AND SREERAMANAN S.5

MICROPROPAGATION OF GINGER (ZINGIBER OFFICINALE VAR. RUBRUM) USING BUDS FROM MICROSHOOTS
ABSTRACT:
Zingiber officinale var. Rubrum (ZOR) is cultivated for its medicinal value despite the constraints of longer life cycle. The study has established an efficient and reproducible protocol to micropropagate ZOR using buds generated on the surface of the ginger. Surface sterilized young buds of 0.5-1 cm and 2-4 cm cultured on Murashige and Skoog (MS) supplemented with BAP showed the highest survival rate (55-65%) and produced the highest average number of microshoots per explant (3.2±0.06) respectively. MS medium supplemented with different concentrations and combinations of auxin and cytokinin were used to evaluate shoot multiplication and root induction. BAP concentrations between 3.0-5.0 mg/L was very effective in promoting microshoots and resulted in 100% of microshoot propagation. Microshoots cultured on MS medium supplemented with 3 mg/L BAP and 0.5 mg/L NAA produced the highest number of shoots while 0-0.5 mg/L BAP enhanced shoot length and 3mg/L NAA in combination with BAP produced highest number of roots. Microshoots maintained on MS medium supplemented with 4.5% sucrose produced the highest number of plantlets (23±2.5) and roots per explants (15.4±2.4) meanwhile reducing the length of lateral roots (2.6±0.2).

1153-1158 Download
38
ETHNOMEDICINAL UTILIZATION OF WILD EDIBLE VEGETABLES IN DISTRICT HARNAI OF BALOCHISTAN PROVINCE-PAKISTAN
NIAZ MOHAMMAD TAREEN1,2,3, SAEED-UR-REHMAN2, MUSHTAQ AHMAD1*, ZABTA KHAN SHINWARI4 AND TAHIRA BIBI1,5

ETHNOMEDICINAL UTILIZATION OF WILD EDIBLE VEGETABLES IN DISTRICT HARNAI OF BALOCHISTAN PROVINCE-PAKISTAN
ABSTRACT:
Wild edible plants have a tremendous influence on human being even before civilization. These plants contain considerably high nutritional value. Present survey was conducted to explore edible wild vegetables species and their ethnomedicinal uses by the inhabitants of district Harnai, Balochistan, Pakistan. Information was obtained through informed free listing interviews with randomly selected informants and field interviews with key informants selected after free listing. A total of 59 wild edible vegetables belonging to 41 genera, 59 species and 20 plant families are used not only as vegetables and salads but treatment of various diseases The most common plant families in terms of the number of species are the Brassicaceae (10 species), Apiaceae (9 species) and Asteraceae (6 species). The most common parts of the plants used as vegetables and medicine are their leaves (44.45%) and whole plant (22.22%). Plants are often used as decoction (34%), powder (26%). Highest plants species are used for gastrointestinal diseases (45 species). Highest ICF value (0.4) was recorded for dermatological disorders category. 100% fidelity level was found for two plant species i.e., Descurainia sophia, and Caralluma tuberculata. The highest use value was reported for the Lepidium sativum (0.63). Highest RFC value was calculated for Caralluma tuberculata (0.14). Highest use report was calculated for three species Apium graveolens Lepidium sativum and Mentha longifolia, (7 UR for each). The highest FIV was calculated for family Brassicaceae (14 FIV).Our study reveals that plants are still used as a major source of food like vegetables as well as medicine for the local people. Too little or no information is available on their uses, cooking methods and nutritional and physiotherapeutic values. Necessary steps should be taken to perform phytochemical and pharmacological studies to explore the potential nutritional values and herbal drug discovery of such plants.

1159-1171 Download
39
EFFECT OF VARIOUS TILLAGE PRACTICES ON SOIL PROPERTIES AND MAIZE GROWTH
NAIMATULLAH LEGHARI1, A.Q. MUGHAL2, K. QASIM LEGHARI3, W. FARHAD4*, M.S. MIRJAT1 AND H. MOHKUM HAMMAD5

EFFECT OF VARIOUS TILLAGE PRACTICES ON SOIL PROPERTIES AND MAIZE GROWTH
ABSTRACT:
Appropriate tillage practices are vital for good tilth that is pre-requisite for aggregate formation, soil aeration, better root development and plant growth. A field experiment of maize was carried out at the experimental site of Sindh Agriculture University Tandojam during two consecutive growing seasons 2009 and 2010. A randomized complete block design with three treatment conventional tillage (CT), reduced tillage (RT) and no tillage (NT) was used in the study. Significant differences between tillage treatments were observed in the soil properties, growth and root development of plants. The NT treatment retained higher soil water contents (15.8 and 16.0%) measured at 0-20 cm depth during 2009 and 2010, respectively. Likewise, the soil bulk density (1.4 and 1.4 cm-3) was higher at this depth consequently; it resulted in greater soil strength (81 N m-2 and 79 N m-2) during 2009 and 2010, respectively. The negative and significant correlations were recorded between root dry weight and soil strengths. On the other hand, positive and significant relationship of root dry weight with mean total dry matter production and LAI was observed. Moreover, the root development related observations were significantly enhanced under CT as compared to RT and NT treatments. The results indicate that conventional tillage improve maize growth and root development by improving soil properties.

1173-1182 Download
40
CHARACTERIZATION OF FOUR SALSOLA SPECIES AND THEIR GENETIC RELATIONSHIP BY AFLP
AMAL M.E. ABDEL-HAMID1, 2

CHARACTERIZATION OF FOUR SALSOLA SPECIES AND THEIR GENETIC RELATIONSHIP BY AFLP
ABSTRACT:
Amplified length polymorphism (AFLP) technique was used to characterize and detect molecular genetic markers for four Salsola species collected from Al Jouf region in the northern of Saudi Arabia and to shed light on their genetic relationships. Three primer combinations were used for AFLP analysis of the four Salsola species, they generated a total of 181 fragments of which 133 were species specific markers scored across Salsola species. The dendogram produced by Jaccard's coefficient and the UPGMA clustering method showed one main cluster, subdivided into two subclusters. The first sub cluster included Salsola schweinfurthii and Salsola tetrandra. The second sub cluster included Salsola villosa and Salsola cyclophylla. It is worth mentioning that this is the first study to use AFLP markers to characterize and detect molecular genetic markers for the four Salsola species and their genetic relationships.

1183-1187 Download
41
ALLELOPATHIC ACTIVITY OF LEONURUS SIRIBICUS L. ON SEED GERMINATION AND SEEDLING GROWTH OF WHEAT AND IDENTIFICATION OF 4- HYDROXY BENZOIC ACID AS AN ALLELOCHEMICAL BY CHROMATOGRAPHY
M.A. SAYED1, 2*, RAHSED IMAM1, M.N. SIDDIQUI3, S.M. RAIHANUN-NABI4, S. AKTAR1, AND S.R. DAS1

ALLELOPATHIC ACTIVITY OF LEONURUS SIRIBICUS L. ON SEED GERMINATION AND SEEDLING GROWTH OF WHEAT AND IDENTIFICATION OF 4- HYDROXY BENZOIC ACID AS AN ALLELOCHEMICAL BY CHROMATOGRAPHY
ABSTRACT:
The aim of this study was to investigate the allelopathic effects of L. siribicus extract on seed germination and seedlings growth of wheat as well as to identify potential allelochemical. The different concentration (5, 10 and 15%) of aqueous extract were applied during the time of sowing and at 5 days after sowing of wheat seed. L. siribicus extract showed concentration and time - depending activity. Different concentration of aqueous extract inhibited seed germination, seedlings growth, when extracts were applied during the time of seed sowing. The stimulatory effect of seedlings growth were found for 5 % aqueous extract, in contrast 10 and 15% extract inhibited seedlings growth, when extracts were applied at 5 days after sowing. Apart from, 4-hydroxy benzoic acids affected seedlings growth irrespective of application time. The weight of dry matter of wheat seedlings were increased for 5% than 10 and 15% extracts. Thin layer chromatography suggested that the presence of 4-hydroxy benzoic acid including other allelopathic and growth regulatory compounds inhibited germination and seedlings growth. Mineral composition was determined and its might have some stimulatory effect on seedlings growth. It was interesting that 5% extract inhibited germination and seedlings growth, when it was applied during the time of seed sowing, but stimulated seedling growth, when it was applied at 5 days after sowing. The extract of this plants can be used either for bioherbicide as well as growth stimulatory agents for the organic farming system. To find out molecular mechanism behind it, further research is to be done.

1189-1195 Download
42
ALLELOPATHIC SORGHUM WATER EXTRACT HELPS TO IMPROVE YIELD OF SUNFLOWER (HELIANTHUS ANNUUS L.)
SABIR HUSSAIN SHAH1, 2*, EJAZ AHMAD KHAN2, HUSSAIN SHAH3, NADEEM AHMAD4, JABBAR KHAN5 AND GHAZANFAR ULLAH SADOZAI2

ALLELOPATHIC SORGHUM WATER EXTRACT HELPS TO IMPROVE YIELD OF SUNFLOWER (HELIANTHUS ANNUUS L.)
ABSTRACT:
Allelopathy provides eco-friendly environment in managing weeds by reducing the use of synthetic herbicides that cause environmental pollution and herbicide resistance problems. Therefore, weeds have been controlling by plant derived organic compounds as an alternative of inorganic herbicides since the last two decades. In this study, sorghum aqueous extracts were applied individually as well as accumulatively with reduced levels of Dual Gold® (S-Metolachlor) as foliar sprays in sunflower at 50, 70 and 90 DAS. For comparison, standard level of S-Metolachlor was also applied as foliar sprays along with weedy check. The highest reduction of total weed density (93.7%) was recorded by three sprays of sorghum aqueous extracts at rate of 15 L/ha mixed with 1/3rd S-Metolachlor at 1.6 L/ha as foliar applications. This reduction rate was statistically similar to one that was obtained by standard level of S-Metolachlor (1.6 L/ha). The highest achene yield was achieved by applying three foliar sprays of aqueous sorghum extracts along with reduced doses of S-Metolachlor, which was almost similar to full recommended dose of S-Metolachlor. These findings demonstrate that allelopathy offers environment friendly and economical opportunity for weed control in sunflower reducing the dependence and cost of herbicides.

1197-1202 Download
43
EFFECT OF SPATIAL ARRANGEMENT AND DENSITY ON WEED INFESTATION AND YIELD OF MAIZE (ZEA MAYS L.)
MUHAMMAD SAEED1, ZHAOFENG HUANG1, HONGJUN HUANG1, SHOUHUI WEI1, CHAOXIAN ZHANG1*, SAIMA HASHIM2 AND KHAN BAHADAR MARWAT3

EFFECT OF SPATIAL ARRANGEMENT AND DENSITY ON WEED INFESTATION AND YIELD OF MAIZE (ZEA MAYS L.)
ABSTRACT:
Field experiments were conducted during summer season 2012 and consequently repeated in 2013 to assess the efficacy of row and plant spacing on weed infestation and yield of maize crop. The experiments were carried out in Randomized Complete Block (RCB) design with split plot arrangements. Three row spacings i.e. 60, 75 and 90 cm were assigned to main plots while different plant spacings i.e. 10, 15, 20, 25 and 30 cm were allotted to subplots, respectively. The results showed that for both the year’s narrow row and plant spacing effectively suppressed weeds while wider row and plant spacing resulted in higher weed density. The data showed that the maximum weed density (202.07 and 218.70 m-2) was recorded in 90 cm row spacing in 2012 and 2013, respectively. However, among plant spacing highest weed density of 214.89 m-2 and 219.83 m-2 was recorded in 30 cm plant spacing during 2012 and 2013, respectively. The data regarding biological yield showed maximum biological yield in 60 cm row spacing while among plant spacing the highest biological yield was resulted in narrow plant spacing of 10 cm during both years. Furthermore, highest grain yield of 4928.9 kg ha-1 in 2012 and 5063.9 kg ha-1 in 2013 was recorded in 75 cm row spacing while lowest grain yield of 3026 kg ha-1 in 2012 and 3989 kg ha-1 in 2013 was observed for 90 cm row spacing. Among plant spacing highest grain yield of 4474.8 kg ha-1 and 5228.5 kg ha-1was recorded in 15 cm plant spacing whereas lowest grain yield of 3554 kg ha-1 and 4010.6 kg ha-1 was observed for 30 cm row spacing in 2012 and 2013, respectively. The regression analysis also showed highest grain yield form 15-20 cm plant spacing during both years. Similarly the correlation data showed that with increase in weed density the grain yield decreases accordingly. The two years of research showed that narrow spacing (15-20 x 75 cm) enhanced the competitive ability of maize crop and suppressed weed growth.

1203-1207 Download
44
GLOMUS ETUNICATUM ROOT INOCULATION AND FOLIAR APPLICATION OF ACETYL SALICYLIC ACID INDUCED NACL TOLERANCE BY REGULATION OF NAC1 & LeNHX1 GENE EXPRESSION AND IMPROVED PHOTOSYNTHETIC PERFORMANCE IN TOMATO SEEDLINGS
BUSHRA GHAZANFAR, CHENG ZHIHUI*, CUINAN WU, HANQIANG LIU, HEZI LI, RANA NAVEED UR REHMAN, IMRAN AHMAD AND ABDUL REHMAN KHAN

GLOMUS ETUNICATUM ROOT INOCULATION AND FOLIAR APPLICATION OF ACETYL SALICYLIC ACID INDUCED NACL TOLERANCE BY REGULATION OF NAC1 & LeNHX1 GENE EXPRESSION AND IMPROVED PHOTOSYNTHETIC PERFORMANCE IN TOMATO SEEDLINGS
ABSTRACT:
Salinity stress hampers plant growth and cause significant yield losses thus induction of salinity stress tolerance in crop plants is one of major goals of agriculture research. Arbuscular mycorhizae fungi Glomus etunicatum and acetyl salicylic acid were tested for induction of NaCl stress tolerance in tomato seedlings, cultivar No. 4. The seedlings were inoculated with Glomus etunicatum and exogenously sprayed with acetyl salicylic acid (0.30 mM) followed by salinity stress (150 mM). It was observed that both Glomus etunicatum and acetyl salicylic acid (singly or in combination) were significantly effective to minimize the injurious effects of salinity by improving root morphological parameters (length, diameter, surface area, volume and number of tips, nodes, bifurcations and connections), photosynthetic parameters (net photosynthesis Pn, stomatal conductance Gs) and chlorophyll contents compared to sole salinity treatment. The bio-inoculant Glomus etunicatum and chemical ameliorator acetyl salicylic acid also notably improved vegetative (fresh and dry weights) and reproductive growth (percent seedlings with flower buds and opened flowers, number of flower buds and opened flowers per seedling) of the plants as compared to the sole salinity treatment. The studied salt responsive genes (LeNHX1 and NAC1) were also regulated to different extents in seedling roots and leaves which was consistent with enhanced salinity stress tolerance. From these observations it is suggested that the individual or synergetic use of the AMF (Glomus etunicatum) and acetyl salicylic acid can be useful for tomato cultivation in the marginally salinity effected soils and warrants further investigations.

1209-1217 Download
45
AGROBACTERIUM-MEDIATED TRANSFORMATION OF VERTICILLIUM DAHLIAE WITH GFP GENE TO STUDY COTTON-PATHOGEN INTERACTION USING A NOVEL INOCULATION METHOD
FENG LI1, NOREEN BIBI1,2, KAI FAN1, MI NI1, SHUNA YUAN1, MING WANG1 AND XUEDE WANG1*

AGROBACTERIUM-MEDIATED TRANSFORMATION OF VERTICILLIUM DAHLIAE WITH GFP GENE TO STUDY COTTON-PATHOGEN INTERACTION USING A NOVEL INOCULATION METHOD
ABSTRACT:
Verticillium dahliae is a soil-born fungal pathogen which causes Verticillium wilt in economically important crops including cotton. We conducted a study to monitor the interaction between the fungus and cotton. V. dahliae was transformed with the gene encoding green fluorescent protein. The gene can be constitutively expressed and fluorescence was clearly visible in both hyphae and spores. Due to heterogeneous gene insertion, the growth rate, colony morphology and pathogenicity of fungus transformants showed differences compared with corresponding wild type. Similarly, quantitative real-time PCR analysis also indicated significant differences in the gene expression among different V. dahliae transformants. To study cotton-pathogen interaction, we devised a novel inoculation method and developed a successful infection by keeping GFP-expressed mycelial plug alongwith aseptic cotton seedlings. After 6-day inoculation, the LSM microscopic image showed that the fungus rapidly formed a mycelial network on the surface of the stems and colonized into plant tissue, displayed an intercellular infection pattern. The early events during cotton colonization by V. dahliae can be successfully observed in 10 days including the plant growth period. Besides, pathological changes of seedlings like tissue discoloration, wilting, stem dehiscence and necrosis can be clearly observed without the influences of soil and other microbes. This inoculation method provides a rapid, effective and environmental friendly technique for the study of cotton-pathogen interaction and identification of resistant plant cultivars.

1219-1227 Download
46
ASSOCIATIONS AMONG HALOTOLERANCE, OSMOTOLERANCE AND EXOPOLYSACCHARIDE PRODUCTION OF AUREOBASIDIUM MELANOGENUM STRAINS FROM HABITATS UNDER SALT STRESS
BENJAWAN YANWISETPAKDEE1, PONGTHARIN LOTRAKUL1*, SEHANAT PRASONGSUK1, TOSAK SEELANAN2, JAMES F. WHITE JR.3, DOUGLAS E. EVELEIGH4, SEUNG WOOK KIM5* AND HUNSA PUNNAPAYAK1*

ASSOCIATIONS AMONG HALOTOLERANCE, OSMOTOLERANCE AND EXOPOLYSACCHARIDE PRODUCTION OF AUREOBASIDIUM MELANOGENUM STRAINS FROM HABITATS UNDER SALT STRESS
ABSTRACT:
Associations among halotolerance, osmotolerance and exopolysaccharide (EPS) production of 50 Aureobasidium melanogenum strains isolated from various habitats along the coasts of Thailand were compared. Using Fisher’s Exact Test, significant associations were found between halotolerance vs osmotolerance (P = 0.004), halotolerance vs EPS production (P = 0.049) and osmotolerance vs EPS production (p0.001). Highly to moderately halotolerant strains were found to be moderately osmotolerant, but not vice versa. Tolerant strains against either salt or sugar produced moderate to low EPS yield. Strains intolerant to salt and/or sugar varied widely in EPS production. The effect of osmotic stress on the growth and EPS yield were investigated with three strains different in halotolerance, osmotolerance and EPS production by comparing their cultures in media with increasing sucrose concentrations. As sucrose concentration increased, a significant reduction in conversion efficiency was observed. Both moderately halotolerant (PBUAP13) and osmotolerant (PBUAP50) strains with moderate EPS production lost their conversion efficiency more drastically than the relatively stress intolerant, high EPS producing strain (PBUAP34). The reduction in EPS production at high osmotic stress was apparently not the result of growth inhibition for both moderately tolerant strains. Cellular accumulation of mannitol was detected in all strains tested.

1229-1239 Download
47
ASSESSMENT OF FUNCTIONAL TRAITS IN THE ASSEMBLAGE OF ENDOPHYTIC FUNGI OF ANACARDIUM OTHONIANUM RIZZINI
PAULA SPEROTTO ALBERTO FARIA1, JAQUELINE ALVES SENÁBIO2, MARCOS ANTÔNIO SOARES3, FABIANO GUIMARÃES SILVA4, ANA PAULA ALVES CUNHA5 AND EDSON LUIZ SOUCHIE6

ASSESSMENT OF FUNCTIONAL TRAITS IN THE ASSEMBLAGE OF ENDOPHYTIC FUNGI OF ANACARDIUM OTHONIANUM RIZZINI
ABSTRACT:
Plants maintain symbiotic relationships with microorganisms as a strategy to withstand adversities. From this exchange, organisms receive photoassimilates and provide benefits to the plant. Anacardium othonianum Rizzini, locally known as ‘caju-de-árvore-do-cerrado’ (tree cashew ofthe cerrado), is a tree species of the family Anacardiaceae nativeto the Midwest region of Brazil. The objective of this study was to characterize the culturable endophytic fungal community, its functional traits and its association with the roots of A. othonianum. The roots of A. othonianum were fragmented (1 cm) and inoculated in medium for the isolation of endophytic microorganisms. The molecular identification of the isolates was performed through the partial sequencing of the internal transcribed spacer (ITS). The endophytic isolates were tested for the synthesis of indole acetic acid (IAA) and phosphate solubilization through the colorimetric method.The root fragments were cleared, stained and examined under a microscope. Structures characteristic of endomycorrhizal and endophytic microorganisms were found on the slides analyzed. A total of 67 fungal strains were isolated and identified in 12 species: Fusarium oxysporum, Bionectria ochroleuca, Periconia macrospinosa, Phomopsis lagerstroemiae, Penicillium kloeckeri, Eupenicillium shearii, Phomopsis asparagi, Penicillium pinophilum, Agaricomycetes sp., Diaporthe sp., Cladosporium cladosporioide sand Paecilomyces lilacinus. All the genera found have been reported in the literature as endophytic species. It can be concluded that A. othonianum maintains associations with endomycorrhizal and endophytic fungi. Twelve endophytic strains were isolated from A. othonianum Rizzini, seven of which have potential for phosphate solubilization and IAA synthesis.

1241-1252 Download
48
CHARACTERIZATION OF SULFUR-OXIDIZING BACTERIA ISOLATED FROM ACID MINE DRAINAGE AND BLACK SHALE SAMPLES
WASIM SAJJAD1, TARIQ MEHMOOD BHATTI2, FARIHA HASAN1, SAMIULLAH KHAN1, MALIK BADSHAH1, ABBAS ALI NASEEM3 AND AAMER ALI SHAH1*

CHARACTERIZATION OF SULFUR-OXIDIZING BACTERIA ISOLATED FROM ACID MINE DRAINAGE AND BLACK SHALE SAMPLES
ABSTRACT:
Acid mine drainage (AMD) and black shale (BS) are the main habitats of sulfur-oxidizing bacteria. The aim of this study was to isolate and characterize sulfur-oxidizing bacteria from extreme acidic habitats (AMD and BS). Concentration of metals in samples from AMD and BS varied significantly from the reference samples and exceeded the acceptable limits set by the Environmental Protection Agency (EPA) and the World Health Organization (WHO). A total of 24 bacteria were isolated from these samples that were characterized both morphologically as well as through biochemical tests. All the bacteria were gram-negative rods that could efficiently oxidize sulfur into sulfate ions (SO4-2), resulted into decrease in pH up to 1.0 when grown in thiosulfate medium with initial pH 4.0. Out of 24, only 06 isolates were selected for phylogenetic analysis through 16S rRNA sequencing, on the basis of maximum sulfur-oxidizing efficiency. The isolates were identified as the species from different genera such as Alcaligenes, Pseudomonas, Bordetella, and Stenotrophomonas on the basis of maximum similarity index. The concentration of sulfate ions produced was estimated in the range of 179-272 mg/L. These acidophiles might have various potential applications such as biological leaching of metals from low-grade ores, alkali soil reclamation and to minimize the use of chemical S-fertilizers and minimize environmental pollution.

1253-1262 Download
49
PRODUCTION AND CHARACTERIZATION OF TANNASE FROM A NEWLY ISOLATED BACILLUS SUBTILIS
MUHAMMAD NAUMAN AFTAB, HAMID MUKHTAR AND IKRAM-UL-HAQ

PRODUCTION AND CHARACTERIZATION OF TANNASE FROM A NEWLY ISOLATED BACILLUS SUBTILIS
ABSTRACT:
The work describes the production and characterization of tannase from a newly isolated Bacillus subtilis. The strain was isolated from the garden soil and was capable of producing tannase at particular temperature (41ºC) and pH (5) in 24 h. Addition of 10 % glucose as a carbon source and 12% tannic acid as an inducer resulted in the improved rate of enzyme production. The enzyme was purified up to 4.86 fold with 96.25% yield. It exhibited optimal temperature and pH tolerance of 45ºC and 5, respectively. However, the enzyme was found to be notably more functional in a broad range of temperature (20-80ºC) and pH (3-10). Furthermore it remained remarkably stable at wide range of pH (3-8) and at a higher salt concentration (~3M). The shelf life of enzyme was also prolonged and remained stable up to a maximum of 8 months.

1263-1271 Download
50
GENETIC DIVERSITY OF MUNGBEAN GENOTYPES IN RELATION TO RESISTANCE AGAINST MUNGBEAN YELLOW MOSAIC VIRUS
RANA BINYAMIN1*, MUHAMMAD ASLAM KHAN2, AZEEM IQBAL KHAN3, FAISAL SAEED AWAN4, NASIR AHMAD KHAN2 AND SOHAIL AKHTAR1

GENETIC DIVERSITY OF MUNGBEAN GENOTYPES IN RELATION TO RESISTANCE AGAINST MUNGBEAN YELLOW MOSAIC VIRUS
ABSTRACT:
Mungbean Yellow Mosaic virus (MYMV) caused by single stranded DNA begomovirus is the most potential and prevailing threat to mungbean worldwide. This study was conducted to determine genetic variation in mungbean genotypes to know whether these genotypes, which differ in field screening results, also vary on a genetic basis or not. One hundred and twenty seven genotypes were screened against MYMV under the field condition. RAPD analysis showed significant amount of genetic diversity within mungbean germplasm. Twenty six primers produced 66% of polymorphism with average 5.36 bands per primer. Minimum similarity was observed among genotypes 8008 and Azri-06, which is 61.15%, while the maximum similarity was observed among genotypes NM-54 and NM-92, which is 91.37%. Three genotypes belonging to resistant category remained unclustered i.e., NM-2011, NM-2006 and Azri-06.Genetic differentiation among mungbean genotypes was similar to the field screening results. RAPD analysis revealed an extensive amount of variation, which could be used for cultivar identification. This information will be helpful in recognition of resistant germplasms and will be utilized for mungbean improvement program.

1273-1277 Download
51
IN VITRO INVESTIGATIONS ON HOST SPECIFICITY OF RALSTONIA SOLANACEARUM AMONG SOLANACEOUS CROPS AND ITS BIOLOGICAL CONTROL IN TOMATO
SADIA HASSAN1, M. INAM-UL-HAQ1*, FARAH NAZ1, M. IBRAHIM TAHIR1 AND ZAHID ALI2

IN VITRO INVESTIGATIONS ON HOST SPECIFICITY OF RALSTONIA SOLANACEARUM AMONG SOLANACEOUS CROPS AND ITS BIOLOGICAL CONTROL IN TOMATO
ABSTRACT:
Ralstonia solanacearum, the causal organism of bacterial wilt of solanaceous crops is a major limitation on the production of solanaceous crops worldwide. The present studies were carried out to explore the prevalence, severity, virulence and host specificity range of R. solanacearum in chili, tomato, eggplant and potato plants while biocontrol of this pathogen was studied on tomato. The isolation and the initial identification of bacterium were done on TTC and 523 media. Out of 32 isolates, 30 showed positive hypersensitive reaction (HR) with variable response and were confirmed as R. solanacearum by performing biochemical tests i.e. Gram staining, KOH, catalase oxidation, Kovacs oxidation, levan production from sucrose, lipase activity on tween 80 agar, production of fluorescent pigment, and oxidation/fermentation of glucose. Race identification studies showed that all the isolates belonged to Race 3 while biovar confirmation tests revealed that 4, 3 and 23 isolates belonged to biovar 1, biovar 2 and biovar 3, respectively. The isolates belonging to distinct biovar class were tested for their wideness of host range by challenging them to chili, tomato, eggplant and potato plants. It was observed that biovar 3 is the most aggressive and has widest host range as compared to counterparts. Biocontrol studies through antagonistic rhizobacteria resulted four antagonistic isolates; PRB10, PAP5, PAT1 and PTR6 having the highest biocontrol activity with 98.75%, 97.5%, 93.75% and 91.25% respectively.

1279-1287 Download
52
SEASONAL DYNAMICS OF EPIPHYTIC MICROALGAE AND THEIR HOST SEAWEEDS FLORIDEOPHYCEAE AT JEDDAH COAST, THE RED SEA, SAUDI ARABIA
SALIM MARZOOG AL- HARBI AND MD. ABU AFFAN*

SEASONAL DYNAMICS OF EPIPHYTIC MICROALGAE AND THEIR HOST SEAWEEDS FLORIDEOPHYCEAE AT JEDDAH COAST, THE RED SEA, SAUDI ARABIA
ABSTRACT:
Algae play a very important role as feed and shelter for fish, crustacean and gastropods. Thus, this study was carried out to know dynamics of epiphytic microalgae and their host macroalgae of Florideophyceae at Jeddah coast, the Red Sea. The water temperature, salinity, pH and dissolved oxygen varied from 21.6 to 32.30°C, 36.40 to 39.70 ppt, 8.04 to 8.60, and 3.00 to 8.90 mg/L, respectively. A total of 10 seaweeds species of Florideophyceae was found, and among them Gracilaria corticata was found throughout the year. Gracilaria tikvahiae and Digenea simplex occurred from spring to autumn, and Acanthophora muscoides occurred from winter to summer. A total of 108 taxa of epiphytes were identified; 95 belong to Bacillariophyceae, 9 Cyanophyceae, 2 Dinophyceae, 1 Chlorophyceae and 1 Raphidophyceae. The epiphytes on A. muscoides varied from 9.50 to 66.10´105 cells/100 g of A. muscoides, and the highest cell abundance was in spring. Epiphytic cell abundance on Amphiroa sp., fluctuated from 5.38 to 40.78´105 cells/100 g of Amphiroa sp. Similarly, for Digenea simplex, epiphytes varied from 27.56 to 51.69´105 cells/100 g of D. simplex. Epiphytic cell abundance on Gracilaria corticata varied from 7.28 to 74.21´105 cells/100 g of G. corticata, and the lowest and highest cell abundance were in winter and summer, respectively. Among the microalgae, the contribution of Bacillariophyceae was highest (87.96%), followed by Cyanophyceae (8.33%) and Dinophyceae (1.85%). A total of 32 species contributed above 10% in epiphytic microalgae, and Licmophora sp., Navicula sp., Leptocylindrus sp., Nitzschia sp., Tabellaria sp., and Thalassionema sp., were the dominant species.

1289-1298 Download
53
EVALUATION OF BIOCONTROL POTENTIAL OF EPIPHYTIC FLUORESCENT PSEUDOMONAS ASSOCIATED WITH HEALTHY FRUITS AND VEGETABLES AGAINST ROOT ROT AND ROOT KNOT PATHOGENS OF MUNGBEAN.
HABIBA1, RUBINA NOREEN2, SYED ABID ALI3, VIQAR SULTANA4, JEHAN ARA1 AND SYED EHTESHAMUL-HAQUE2

EVALUATION OF BIOCONTROL POTENTIAL OF EPIPHYTIC FLUORESCENT PSEUDOMONAS ASSOCIATED WITH HEALTHY FRUITS AND VEGETABLES AGAINST ROOT ROT AND ROOT KNOT PATHOGENS OF MUNGBEAN.
ABSTRACT:
Endophytic and rhizospheric fluorescent Pseudomonas have widely been used as biological control agents against soilborne plant pathogens. In this study, fifteen epiphytic fluorescent Pseudomonas isolated from the surfaces of citrus (grapefruit, orange and lemon) melon and tomato fruits were characterized for their in vitro activity against root rotting fungi viz., Macrophomina phaseolina, Fusarium solani, F. oxysporum and Rhizoctonia solani and nematicidal activity against the second stage juveniles of Meloidogyne javanica. Out of fifteen Pseudomonas isolates HAB-16, HAB-1 and HAB-25 inhibited the growth of all the test fungi and showed maximum nematicidal activity against second stage juvenile of M. javanica. Based on their effective in vitro activity nine epiphytic fluorescent Pseudomonas were evaluated for their growth promoting ability and biocontrol activity in screen house on mungbean. Pseudomonas isolates (HAB-13, HAB-2, HAB-4, HAB-1, HAB-14, HAB-9, HAB-7 and HAB-25) used as soil drench greatly reduced the root rot-root knot infection and thereby enhanced plant growth, root nodulation and yield in mungbean. Besides, rhizospheric and endophytic, epiphytic fluorescent Pseudomonas associated with healthy fruits may be used as biocontrol agent against root rotting fungi, besides, using for the mangemnet of postharvest diseases.

1299-1303 Download
54
COMBINED EFFECT OF SOIL AMENDMENT WITH OIL CAKES AND SEED PRIMING IN THE CONTROL OF ROOT ROT FUNGI OF LEGUMINOUS AND NON-LEGUMINOUS CROPS
HIRA RAFI1, SHAHNAZ DAWAR1* AND MARIUM TARIQ2

COMBINED EFFECT OF SOIL AMENDMENT WITH OIL CAKES AND SEED PRIMING IN THE CONTROL OF ROOT ROT FUNGI OF LEGUMINOUS AND NON-LEGUMINOUS CROPS
ABSTRACT:
Organic amendments of soil help in proper aeration, rising of temperature and water holding capacity which results in better uptake of nutrients with root system gets extensive establishment. In this study, effects of soil amendment with oil seed cakes including mustard (Brassica campestris L.), cotton (Gossypium hirsutum L.), almond (Prunus amygdalus L.) and black seed (Nigella sativa L.) cakes @ 0.1 and 1% w/w and priming of seeds with Acacia nilotica (L.) Willd. ex Delile and Sapindus mukorossi (L.) leaves extracts and microbial antagonists (Trichoderma harzianum and Rhizobium melilotii) was observed on the growth of plants and in the suppression of root infecting fungi. The results obtained showed that combined effect of bio-priming of seeds with T. harzianum spore suspension and amendment of soil with mustard cake @ 1% was found to be most effective for the growth of leguminous and non-leguminous crop plants (peanut, chickpea, okra and sunflower) and for the reduction of root infecting fungi like Macrophomina phaseolina, Fusarium spp followed by R. meliloti primed seeds in combination with cotton, almond and black seed cakes amendment respectively as compared to control (non treated seeds and soil).

1305-1311 Download
55
NEW RECORDS OF ORDER BOLETALES FROM PAKISTAN
ABDUL RAZAQ1 AND SALEEM SHAHZAD2

NEW RECORDS OF ORDER BOLETALES FROM PAKISTAN
ABSTRACT:
During a survey of basidiomycetous fungi, eight species belonging to order Boletales were collected from different parts of Gilgit-Baltistan, Pakistan. The collected members of phylum Basidiomycota included Aureoboletus gentilis, Boletus reticulatus and B. subtomentosus belonging to family Boletaceae, and Suillus bovinus, S. granulatus, S. luteus, S. placidus and S. viscidus belonging to family Suillaceae. Of these, Aureoboletus gentilis, Boletus reticulatus, B. subtomentosus, S. bovinus, S. luteus and S. viscidus appeared to be new records from Pakistan not hitherto reported.

1313-1317 Download
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