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 2285-5653STUDY ON THE FREE AMINO ACID PROFILE OF LEAVES FOR GRAPESVARIETIES OF GALBEN DE ODOBETI SORTOGROUPMarioara BOSOI 1 , Constantin ÂRDEA 21 Research and Development Station for Viticulture and Oenology Odobeti, Stefan cel Mare no. 61,625300, Odobeti, Romania2 University of Agricultural Sciences and Veterinary Medicine Iai, Mihail Sadoveanu Avenue no.3,Iai, RomaniaAbstractCorresponding author email: mioara_bosoi@yahoo.comResearches on amino acid profile carried out so far in the genus Vitis showed its great variability, suggestingimportance of the genetic involvement in this variability This paper presents preliminary results of investigating theamino acid profile from leaves to the grape varieties of sortogroup Galben de Odobeti. The grape varieties taken inthe study were: Galben de Odobeti, Btut neagr, Zghihar de Hui, Negru moale and Negru vârtos. For extractionof free amino acids from the leaves of the vine has been used the method Bieleski & Turner (1966) adjusted forgrapevine, and separation technique was performed by thin-layer chromatography (TLC). Identification andquantitative estimation of free amino acids was performed using analysis and image processing software - ImageJver.1.46. The amount of free amino acids ranged between 18.76 mg / mg fresh leaf at Galben de Odobeti variety and14.33 mg / mg for Negru moale variety. In the varieties Galben de Odobeti, Btut neagr and Zghihar de Hui themost abundant free amino acids were: aspartic acid (Asp), glutamic acid (Glu), serine (Ser), threonine (Thr), proline(Pro) and glycine (Gly). The predominant amino acids for Negru moale variety were: aspartic acid (Asp), serine (Ser),glycine (Gly), glutamic acid (Glu), threonine (Thr), proline (Pro) and arginine (Arg). Amino acids predominantly toNegru vârtos variety are: aspartic acid (Asp), glutamic acid (Glu), serine (Ser), threonine (Thr), proline (Pro) andarginine (Arg). Aspartic acid (Asp) is found in large quantities compared with other amino acids, representing 31.7%of total amino acids identified for Negru vârtos variety, 31.5% at Zghihar de Hui variety, 30.5% fpr Negru moalevariety, 27. 5% for Galben de Odobeti variety and 25.3% for the variety Btut neagr. Statistical ratio Pro / Argand amino predominant ratio (Asp / Ser), genetically differentiate the Btut neagr variety the other varietiesbelonging to the sortogroup Galben de Odobeti, with a confidence interval of 95%. These preliminary data provide abasis for further research that can demonstrate that amino acid profile of the leaves can be used as method indiscriminant analysis of grape varieties.Key words: amino acid profile, Galben de Odobeti, sortogroup, thin-layer chromatography.INTRODUCTIONincreases during aging reaching values between200 and 6500 mg/l of each amino acidThe fund management in grape germplasm isvariations from one year to another, and fromone of the most important issues is the growingone variety to another (ârdea, 2007). Shareconcern of researchers from all countries.amino acids in grapes is high and is 20 -30% ofClassical identification methods based onthe total nitrogen compounds (Poux andcharacteristics ampelographic / botanical notOurnac, 1970). Total amino acid contentthe most accurate, with certain restrictions dueespecially predominant ratio of amino acidsto instability morphological characteristicsdiffer significantly from one variety to another,influence environmental conditions.so knowing the amino acid spectrum ofResearch on amino acid profile made so far invegetative organs and grapes can be a means ofthe genus Vitis showed its great variability,differentiation of vine varieties in terms ofsuggesting importance of the geneticinvolvement in this variability (Kliewer et al.,genetically (Hernández-Orte et al., 1990).1966, Kliewer 1969; Klub et al., 1978; MarcyStudies on free amino acid profiles of grapeset al. 1981; Notsuka et al., 1984, Huang andmade so far reported variations in the geneticOugh, 1991). On vines, the total amino acidsmaterial and suggests new biochemicaldescriptors (Shiraishi, 1996).163
The characterization of the grapevine varietiesbased on free amino acid profile shows a greatimportance to complement the modern methodson investigation of the vine varieties(isoenzymatic analysis and DNA analysis).MATERIALS AND METHODSThe biological material was represented by fivelocal varieties to Galben de Odobetisortogroup (Galben de Odobeti, Zghihar deHui, Btut neagr, Negru moale and Negruvârtos), belonging ampelographic collection ofResearch and Development Station forViticulture and Winemaking Odobeti.Samples for analyses were the young leafobtained by forcing cuttings - eye. The leafsamples (2 or 3 leaves, ~ 5 g ) were collectedfrom each variety in plastic bags, labeled,stored on ice and brought to laboratory foranalyses.For extraction of free amino acids from theleaves of the vine has been used the methodBieleski & Turner (1966) adjusted forgrapevine. This method ensures the extraction,separation and quantitative estimation of aminoacids from small samples of vegetal freshtissue, by thin layer chromatography technique.For extraction of free amino acids in biologicalmaterial and for the preparation of standardsolutions of amino acids was used a mixturesolution with methanol, chloroform and waterin a ratio of 12:5:3 v/v/v. For each grapevariety was weighed a sample of 0.5 gbiological material. The fresh leaves werelyophilized and crushed in a mortar with liquidnitrogen until was obtained a fine powder.0,200 g was transferred for each sample in aEppendorf tube 1.5 ml. Over powder was added200 ml extraction solution (methanol:chloroform: water - M/C/W) in ratio of 12/5/3(v/v/v), which allowed the removal of pigmentsand lipids from plant material in chloroformlayer. Cell suspensions were treated with sonicfrequencies for 1 hour at 4 ° C, causing rupturecell membranes. After sonication the sampleswere stored in a refrigerator at 4 º C for at least1 h. To obtain the supernatant containing theamino acids, the sample was centrifuged at13,500 rpm for 15 minutes at 4 ° C.The chromatographic separation of amino acidswas achieved on TLC plates with silica gel164stationary phase 60 on aluminum foil 20 x 20(TLC Silicagel 60), manufactured by MerckKGaA, Germany. TLC plates were made withsize 10 x 7 cm and have been marked the areasof application for samples. Using standardamino acids were produced by Merck KGaA,Germany, in the solution of methanol:chloroform: water (12:5:3 v / v / v). Samples (2l) were applied with an automaticmicropipette with volume control, from left toright at the bottom of the plate, in the areasmarked for each cultivar. The chromatographicseparation or development of plates was donein mono dimension, was used the developmentsystem based on n - butanol / glacial acetic acid/ water (ratio 4:1:1 v/v/v). Afterchromatographic separation, the TLC plates areready for viewing and identification of aminoacids separated. To ensure reproducible results,the tests were repeated three times forpredominant amino acids, to all five varieties.View amino acids was performed usingspecific reagents – ninhydrin (a solution inconcentration of 0,25%). Amino acids appearas separate spots colors. The TLC platesviewed ninhydrin solution, sensitive to lightwere scanned or photographed for thequalitative and quantitative evaluation of freeamino acids separated from each sample. Toidentify amino acids separated in samples wascalculated the retention factor (Rf) comparedwith each standard amino acid. Quantitativedetermination of free amino acids in thesamples was performed using image analysisprogram - ImageJ ver 1.46 r, and the valuesobtained were expressed in g free amino acidfor 1 mg fresh leaf tissue.The data obtained were processed statisticallyfor analysis of variance (ANOVA test) and hasbeen determined significance of differences forthe total free amino acids content and the ratiobetween the predominant amino acids (Studenttest). Analysis of variance and Student test wasperformed using the Microsoft Excel menuTools - Data Analysis.RESULTS AND DISCUSSIONS14 amino acids were identified. In Figure 1 and2 are presented TLC plates with amino acidsseparated and visualized with ninyidrin solutionfor the five cultivars studied. Standard aminoacids used: alanine (Ala) and histidine (His). In
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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
Page 113 and 114: Figure 1. Flowering period and blac
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Page 118 and 119: content, A-acidity content) to the
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Page 124 and 125: Figure 2. DryerFigure 3. Programmer
Page 126 and 127: Appearance of the apricots after 5
Page 130 and 131: where: P F represent the coordinate
Page 134 and 135: Table 2. The production obtained at
Page 136 and 137: As shown in figures 5 and 6 the vit
Page 140 and 141: At two weeks from planting it was a
Page 142 and 143: If we take into account the both fa
Page 146 and 147: F1 - + infectedF2 - + infectedF3 +
Page 148: HORTICULTURALBIODIVERSITY ANDGENETI
Page 151 and 152: RESULTS AND DISCUSSIONSFruit charac
Page 156 and 157: The chorology maps of Artemisia alb
Page 158 and 159: Figure 5. Chorology of Artemisia le
Page 162: Molecular differentiation showed a
Page 167 and 168: Figure 4. Amino acid profile of the
Page 169 and 170: Table 4. The significance of differ
Page 171 and 172: The studied phenophases (table 1),
Page 173 and 174: REFERENCESCapusan Janina, 2013. Rez
Page 175 and 176: Neoaliturus fenestratus Herrich-Sch
Page 177 and 178: occurrence in western part of Roman
Page 179 and 180: Pop 3 = Population 3 obtained by cr
Page 181 and 182: well adapted in our country, and th
Page 186 and 187: Variations in acidity of the variet
Page 188: grape. Analele Universitatii din Cr
Page 191 and 192: Flow cytometry has proved to be an
Page 193 and 194: cell nuclei, using chromosome count
Page 195 and 196: Figure 1. Prekos F1Figure 4. Prekos
Page 197 and 198: Interpretation of the results conce
Page 199 and 200: Photo 1- Redhaven (Control)The Roma
Page 201 and 202: Dry matter (determinate refractomet
Page 203 and 204: was evaluated by using a large-size
Page 208 and 209: During the growing season have been
Page 212 and 213: 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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fact that root injuries is the majo
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We recommend that the species and c
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and parameters of quality. The phys
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AESTHETICALLEVEL OFAPPROACHLEVEL OF
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REFERENCESArchibugi F., 1997. The E
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Figure 1. Optimized version of comp
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CONCLUSIONSThe optimized version (F
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etween 182-212 g/l, the lowest valu
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CONCLUSIONSThis study showed that 2
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RESULTS AND DISCUSSIONSFor this pur
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according to quality class.The calc
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third party countries who wish to b
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Stakeholder comments concerned nota
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Figure 2. The „Eco-leaf” logo o
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identified to facilitate its tracea
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- Analysis reports;This document ma
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Table 1. Sampling procedures applic
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Table 2. Example of sampling proced
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should be 5 kg.” According to thi
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One of the first and simplest condi
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of RIN transcription factor in ethy
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oriented towards the centre of the
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Figure 5. Macroscopic view of the c
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Related to the process of the evolu
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efore it is printed using the main
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Imposition of the pot is done every
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Figure 8. Test of Strength Vetiver
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Table 3 defines LSD test for genoty
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