Scientific Papers Series B Horticulture

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Scientific Papers. Series B, Horticulture. Vol. LVII, 2013Print ISSN 2285-5653, CD-ROM ISSN 2285-5661, Online ISSN 2286-1580, ISSN-L 2285-5653PRELIMINARY STUDY RELATED HIGHLIGHTING THE INHIBITORYEFFECT OF IN VITRO FUNGUS GROWTH MYCOSPHAERELLAGROSSULARIAE (AUERS.) LIND. BY SAPROPHYTIC FUNGIEugenia PETRESCU 1 , Tatiana-Eugenia ESAN 2 , Florica CONSTANTINESCU 3 ,Maria OPREA 3 , Traian MANOLE 3 , Irina IONESCU-MLNCU 41 Carol Davila School, 161-163, Viitorului Street, 020609, Bucharest, Romania2 University of Bucharest, Biology Faculty, 1-3 Aleea Portocalilor Street, Bucharest, Romania3 Research-Development Institute for Plant Protection Bucharest,8 Ion Ionescu de la Brad Street, 013813, Bucharest, Romania4 Faculty of Biotechnology-USAMV, 59 Mrti Blvd., 011464, Bucharest, RomaniaCorresponding author email: petrescu_eugenia@yahoo.comAbstractMycosphaerella grossulariae (Auers.) Lind., one of the most important pathogenic fungi which affect the black currantecological crops cultivated for alimentary supplements and phytotherapeutics drugs production. The present studybrings the new data related to the possibility of „in vitro” vegetative growth inhibition of fungus mycelium usingsaprophytic fungi species. The species of fungi used for experimental trials was Trichoderma viride, Trichotheciumroseum Link, Epicoccum nigrum Link. and Gliocladium roseum Bainier. The fungus was growth on several culturemedia for comparative testing and establishment of the most efficient medium for vegetative growth of the fungi. In thestudy we also follow the comparative approach doing by biometric measurements of the colonies in which work variantthe pathogenic fungi had the high rate of growing. After the testing was made the PDA medium was selected forexperimentation of inhibitory effect of the saprophytes. The method used for trials was the double cultures technique onthe medium of agar which consists in inoculation of one media fragment of mycelium carrier separated from the agarmedium of saprophytic fungi on some distance from pathogenic fungi. The study carried out had allowed the highlightof the fact that in all experimental variant used the saprophytic fungi had inhibits the Mycosphaerella grossulariaepathogen growing. The detailed analysis of the results we could reveal that the most inhibitory effect was done by T.viride followed by T. roseum and E.nigrum.Key words: blackcurrant crop, inhibitory effect, Mycosphaerella grossulariae, saprophytic fungi.INTRODUCTION429The biological resources and the sustainableexploit potential of aromatic and medicinalplants from our country are immense and representan important sustainable component of Romanianagriculture (Manole, 2008). Romaniahas in his flora up to 3,600 species of plantsand more than 1.000 are considered medicinalplants, spontaneous and cultivated (Alexan etal, 1988; Ardelean and Mohan, 2008; Bojor,2003; Pun, 1995). In the classification ofpharmacological industry are under differentforms included – tea, medicinal and cosmeticsproducts – almost 160 medicinal and aromaticspecies, among 110 are spontaneous collectedand up 50 species are cultivated (Pun, 1995).One of the species introduced in the croppingsystem, the blackcurrant crop (Ribes nigrum L.)are extremely economic important. The extensionof ecological crops of this species areimposed the deeply acknowledgement of thepathogens in the purpose of finding rapidly andefficiently ways to biological control.Mycosphaerella grossulariae (Auers.) Lind. isone of the most important pathogenic fungiwhich affect the black currant ecological cropscultivated for alimentary supplements and phytotherapeuticsdrugs production. In plantationof blackcurrant cultivated as medicinal plant inthe south of Romania, Mycosphaerella grossulariaeproduces spots with picnidia on theblackcurrant’s leaves (Petrescu and Oprea, 2012).The present study brings the new data related tothe possibility of „in vitro” vegetative growthinhibition of fungus mycelium using saprophyticfungi species. The fungal saprophyticspecies used for experimentation are alreadycited as antagonists of various pathogens of
cultivated plants (esan, 1997, esan andOprea, 1995). The in vitro antagonistic activityof some fungi against other fungal pathogens ofblackcurrant was previously investigated(Petrescu and esan, 2012; Petrescu et al., 2012).Biological control with antagonistic strains offungi is an alternative and non polluting methodfor control the plant diseases produced by fungalpathogens (Fokkema, 1996).Related to environmentally friendly alternativecontrol methods of fungal pathogens of blackcurrant,a recent study analyzes the effect ofsome plant extracts on the development of theblackcurrant’s pathogen Sphaerotheca morsuvaethat produces American mildew, and onthe pathogenic fungi isolated from phylloplaneof blackcurrant in the South Eastern part ofRomania, such as Botrytis cinerea, Alternariatenuissima and Fusarium oxysporum (Enacheet al., 2011).The field experimental plots were located in theblackcurrant crops of S.C. Export-ImportHofigal S.A., which are playing the role of cofinancingpartner in the research consortium.The mentioned firm is a promoter of thesystems for ecological agriculture in the case ofsome shrubs crops for alimentary supplementsproduction.MATERIALS AND METHODSWe are using for our study the biologicalmaterial provided from reference isolate of theMycosphaerella grossulariae (Auers.) Lind.CBS 235.37 pathogen, which was purchasedfrom CBS culture collection of microorganisms,Utrecht, the Netherlands, and another 4own isolates of saprophytes fungi obtained in2010 from blackcurrant’s phylloplan, whichwere tested for their in vitro effect on thepathogen. These isolates are four strains of thefungi Trichothecium roseum, Trichodermaviride, Gliocladiumroseum and Epicoccumnigrum.In order to saprophytic fungi isolation theblackcurrant leaves were collected andintroduced in plastic bags and brings to theRDIPP (Research Development Institute forPlant Protection) laboratory of Mycology forprocessing and analysis. In laboratory theleaves were divided into the small pieces withthe help of sterile scissors. The leaves pieces430were then placed on the water-agar media forsporulating stimulation and also on CGA mediaand incubated at the room temperature. Thespores of different saprophytic fungi wereobserved and studied at the optical microscope.The saprophytic fungi spores or a little piece offungi mycelium were being transferred on theculture media in sterile conditions. The culturemedia used for fungi growth and multiplicationwere PDA and MEA. The pathogenic strain ofMycosphaerella grossulariae was inoculatedon different media and the colony diameter andcolony characteristics were registereded. Fourculture media were used, PDA (potato dextroseagar, MEA (malt extract agar) Czapek-Dox andCzapek). Each variant had 3 repetitions.The saprophytic fungi and the pathogen weregrown on PDA medium. On the specified timeintervals biometrics measurements of thereverse side of the diameter of the colonieswere performed in view to establish andcompare the growing rate of the pathogenicfungus Mycosphaerella grossulariae withthose of the saprophytic fungi. Looking for invitro testing of the antagonic effect of the foursaprophytic fungi against the strain ofMycosphaerellagrossulariae the double culturemethod were performed which mean theinoculation of both pathogen and saprophyticfungi on the same Petri dish at the samedistance for the dish centre and the samedistance one for another (Juan, 1964, esan andOprea, 1995). The experimental design consistsin 5 variant on 3 replicates each. The controlmedium culture had inoculated only with thesaprophytic fungi. The Petri dishes selected forthe experiment had a small diameter (60 mm)because of the length rhythm of pathogengrowing. The medium used for testing wasPDA. The diameter of the fungal inoculum,both pathogenic and saprophytic was of 3 mmand the distance between Petri dish centre andinoculum was of 10 mm, respectively. Thedistance between the pathogen inoculum andsaprophytic one was of 20 mm. The saprophyticfungus was inoculated later, at the 5days after pathogen inoculation in the momentwhen the colony characteristics of Mycosphaerellagrossulariae are ready formed. Theincubation was performed at room temperature(±24°C). The periodical measurements ofinternal radius of pathogen colony (the radius
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SCIENTIFIC PAPERSSERIES B. HORTICUL
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SCIENTIFIC COMMITTEE Bekir Erol AK
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Fructification - Florin Constantin
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Phenological Studies on Some Variet
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Scientific Papers. Series B, Hortic
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Table 2. The organoleptic appreciat
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processes become even slower and th
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- usingselectedbacteriafromm Leucon
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To organoleptic analysis of the win
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works were pickling the cucumbers i
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days after removal of weeds by burn
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it appears that the culture has gro
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Maintainance of the genetic structu
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Table 3. Interaction variety of dwa
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Experimental scheme is situated in
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improvement especially laboratory s
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Table 1 shows the type 2x3x2 trifac
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Neamtu G., Gheorghe Campeanu, Carme
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Experience consisted of the followi
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Table 7. Total production on plantV
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vineyard, several methods were used
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Table 4. Structure and number and r
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gets serious role through the ferme
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Table 3. Element content of the ber
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Figure 4. The length of internodes
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The process of fruit forming was de
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Compomist F1 hybrid reaches value o
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Figure 2. Experimental results repr
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and total number of tubers per plan
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and quality of organic potatoes for
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Petri dishes, the pathogenic fungi
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Variant 4R 2 11.0 4.0 15.0 7.0 22.0
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Analyzing the dynamics of the total
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Table 2. Indicative data on volumes
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Table 3. The Influence of the level
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Table 8. The Influence of irrigatio
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Figure 1. The aspect of apples from
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Conclusions on changes in chemicalc
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RESULTS AND DISCUSSIONSRaspberry is
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CONCLUSIONSAs a result of scientifi
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observations were made on phenologi
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Figure 1. Flowering period and blac
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content, A-acidity content) to the
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sugar accumulated in grapes and 1,0
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Figure 2. DryerFigure 3. Programmer
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Appearance of the apricots after 5
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where: P F represent the coordinate
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Table 2. The production obtained at
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As shown in figures 5 and 6 the vit
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At two weeks from planting it was a
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If we take into account the both fa
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F1 - + infectedF2 - + infectedF3 +
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HORTICULTURALBIODIVERSITY ANDGENETI
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RESULTS AND DISCUSSIONSFruit charac
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The chorology maps of Artemisia alb
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Figure 5. Chorology of Artemisia le
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Molecular differentiation showed a
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The characterization of the grapevi
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Figure 4. Amino acid profile of the
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Table 4. The significance of differ
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The studied phenophases (table 1),
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REFERENCESCapusan Janina, 2013. Rez
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Neoaliturus fenestratus Herrich-Sch
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occurrence in western part of Roman
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Pop 3 = Population 3 obtained by cr
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well adapted in our country, and th
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Variations in acidity of the variet
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grape. Analele Universitatii din Cr
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Flow cytometry has proved to be an
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cell nuclei, using chromosome count
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Figure 1. Prekos F1Figure 4. Prekos
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Interpretation of the results conce
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Photo 1- Redhaven (Control)The Roma
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Dry matter (determinate refractomet
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was evaluated by using a large-size
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During the growing season have been
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Likewise, vitamin C content of ‘G
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Figure 2. Air temperature (°C) in
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REFERENCESAnconelli S., Antolini G.
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Taking into the account these and t
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postharvest decays in fruits and ve
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manure semifermentated. In vegetati
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Figure 1. Prima Cl. 1022 varietyThe
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temperature required for a differen
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MATERIALS AND METHODSTo accomplish
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climate (IH4), becomes for this yea
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quantities of sugar accumulated in
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ibosomal to investigate 13 species
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Biogeography of Genus MangiferaTwo
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productionand uses. Center for trop
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analysis of variance to express the
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highlighted by a correlation coeffi
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without heat and in open field. Cro
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ut the fruit had a reduced number o
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Genetic autochthonous heritage wasi
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Table 1. The main characteristics o
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Data were subjected to statistical
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Also, the dry biomass and root leng
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“Clapp’s Favorite” in 2011 wh
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2012 the order was “Conference”
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microspore to mature pollen, determ
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microspores at certain date. Micros
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viin utilizate ca genitori potentia
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The objective of this paper, is to
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Table 1. Relationship between germi
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Figure 2. Tassels lengthIn case of
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ORNAMENTAL PLANTS,DESIGN ANDLANDSCA
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of Chisinau, also the old parks. Fo
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y cambium on the very center - meth
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flower is fully colored. Postharves
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Results from table 8 shows that Mar
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Pinus sylvestris ‘Gold Coin’, R
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Figure 4. Excelsa (H 18/1)Descripti
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Figure 1. Plants in cellular trays
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espectively 69%, followed by hybrid
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observations made on the plants of
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The values of pH diminished in all
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Figure 5. Content of total soluble
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Figure 1. Plan of Bran domain (Ion
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Carol the second, her brother, on h
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Figure 14. Tea house in the natural
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Cults, and in conformity with the c
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consult the scientific novelties an
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,,The restoration is the methodolog
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MATERIALS AND METHODSThe chorology
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Merce, 2011; Zamfirescu, 2010) (IAS
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experiment because it shows and ill
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among all the administrations, exce
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solution known as AKN. Content: pot
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As shown in the figure, Chrysal is
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MotivationsOfferprospectingCustomer
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conclude the presented case, with k
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Also, comparative studies, concerni
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done before. Also the speed of the
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as Marcellus theatre or Trajan foru
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ian regime, by huge contradictions
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Pyongyang, the hidden cityThe capit
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“ordinary” buildings as the gen
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- The Writer’s Rotunda“The cons
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Figure 4. Vegetation planRegarding
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Figure 9. BenchesVasesThere are 4 k
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REFERENCESPanoiu A., 2011, Evolutia
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RESULTS AND DISCUSSIONSResults of t
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Figure 5. Average growth rate of
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In 2012 there were higher onset tem
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Page 387 and 388: REFERENCESArchibugi F., 1997. The E
Page 389 and 390: Figure 1. Optimized version of comp
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Page 394 and 395: Scientific Papers. Series B, Hortic
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Page 401 and 402: RESULTS AND DISCUSSIONSFor this pur
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Page 409 and 410: Figure 2. The „Eco-leaf” logo o
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Page 418 and 419: Table 1. Sampling procedures applic
Page 420 and 421: Table 2. Example of sampling proced
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Page 426 and 427: One of the first and simplest condi
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Page 434 and 435: Figure 5. Macroscopic view of the c
Page 436 and 437: Related to the process of the evolu
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Page 442 and 443: Imposition of the pot is done every
Page 444 and 445: Figure 8. Test of Strength Vetiver
Page 448 and 449: Table 3 defines LSD test for genoty
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Scientific Papers Series B Horticulture

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