THE QUALITY OF WHEAT VARIETIES FROM THE CENTRAL TRANSYLVANIA AND THE IDENTIFICATION OF MYCOTOXINS CONTAMINATION

Transcription

1 UNIVERSITY OF AGRICULTURAL SCIENCES AND VETERINARY MEDICINE CLUJ-NAPOCA DOCTORAL SCHOOL FACULTY OF AGRICULTURE Eng. Simona Maria AVRAM (MAN) THE QUALITY OF WHEAT VARIETIES FROM THE CENTRAL TRANSYLVANIA AND THE IDENTIFICATION OF MYCOTOXINS CONTAMINATION (SUMMARY OF Ph.D. THESIS) SCIENTIFIC COORDINATOR Prof.univ.Ph.D. Sevastiţa MUSTE CLUJ - NAPOCA 11 XXVIII

2 CONTENT INTRODUCTION...XXX 1. AIM AND OBJECTIVES... XXXI 2. MATERIAL AND METHOD... XXXII 2.1. EXPERIMENTAL FACTORS... XXXII 2.2. THE BIOLOGICAL MATERIAL...XXXIV 2.3. RESEARCH METHODS...XXXV 3. REZULTS AND DISCUSSIONS...XXXV 3.1. RESULTS REGARDING THE INFLUENCE OF THE EXPERIMENTAL FACTORS ON THE QUALITY OF WINTER WHEAT...XXXV The influence of climatic conditions on the quality of wheat...xxxv The influence of variety on the wheat quality...xxxvi The influence of fertilization on the wheat quality...xxxvi The influence of treatment on the wheat quality... XXXVII Interaction factors (years,, variety, agrofond and treatment) on quality parameters of the wheat...xxxix 3.2. RESULTSREGARDING QUALITY THE WHEAT VARIETIESCULTIVATEDIN TWO AREAS DIFFERENT CLIMATIC (TURDA AND TARGU MURES)...XXXIX The influence of culture area on the wheat quality...xxxix The influence of fertilization on wheat quality cultivated in two areas different climatic (Turda and Targu Mures)...XL The influence of variety on wheat quality cultivated in two areas different climatic (Turda and Targu Mures)...XLI Interaction factors (the area of culture, agrofond and variety) on quality parameters of the wheat... XLII 3.3. RESULTS ON THE IDENTIFICATION OF THE FACTORS STUDIED PARTICIPATION RATES FOR THE INTERACTION TO ACHIEVE THE QUALITY OF THE WINTER WHEAT... XLII 3.4. CORRELATIONS BETWEEN QUALITY PARAMETERS OF THE WHEAT UNDER THE INFLUENCE OF THE STUDY FACTORS...XLIV 3.5. REGRESSION ANALYSIS POSITIVELYCORRELATEDBETWEENTHE QUALITY PARAMETERS... XLV The relationship between protein content and the wet gluten...xlvi The relationship between protein content and the gluten index...xlvi The relationship between protein content and the Zeleny index...xlvii The relationship between wet gluten and the gluten index...xlvii The relationship between wet gluten and the Zeleny index... XLVIII The relationship between gluten index and the Zeleny index... XLVIII 3.6. RESULTS OF TOTAL AFLATOXIN CONTAMINATION OF WHEAT APPLYING HIGH PERFORMANCE THINLAYERCHROMATOGRAPHY (HPLC)...XLIX Qualitative determination of total aflatoxins (B1, B2, G1, G2) for winter wheat grown from to SCDA Turda and SCDCB Targu Mures...XLIX 4. GENERAL CONCLUSIONS AND RECOMMENDATIONS... LI SELECTIVE BIBLIOGRAPHY...LIII XXIX

3 INTRODUCTION The special value of the grains for food and diet resulting from advantages they offer over other raw materials of food industry. The main topic of our research is focused on the wheat because of its great importance as food, providing a lot of carbohydrates and proteins necessary to human and release more than half the calories consumed by mankind, being the main raw material in bakery industry. Knowing the biological features of wheat is an essential step both ingenetic and breeding studies and in research related to cultivation technology and processing which aims to achieve quality standards to ensure quality by-products, but also safe for consumption. The potential of the production hence the quality and safety by applying a culture technology which has the starting point for balanced fertilization, disease control (in particular, which are transmitted by seed), keeping the clean chain of weeds, pest control, harvesting at full maturity and proper storage. The clover grown in different climatic conditions, fertilization and treatment are important factors for achieving high yields, quality and safe for consumption, for which reason we conducted research into the. Experience has shown that the storage of grain, grist products and bakery may occur a microflora under certain conditions enhancing the growth and production of mycotoxins. The main mycotoxins that occur in cereals and cereal products, that are regulated in European Community, are aflatoxins, ochratoxins, deoxynivalenol and zearalenone. The recent European studies shown high levels of mycotoxins present in cereals and cereal by-products. These results call requires vigilance and measures to protect consumers and to increase confidence in the products that best exploits the nutrition potential of the grains. The results obtained during these studies can be useful for wheat production in determining the optimal dose for basic fertilization and the ideal combination of treatments applied during the season to obtain high-quality wheat crops. Identification of mycotoxin contamination of wheat processors is important because, although mycotoxins contamination occurs during the vegetation, their levels may increase substantially during storage and industrial processing. STRUCTURE OF THE THESIS The thesis is divided into two main parts, first part including the current state of knowledge in the field, and the second part, own research results and discussion, conclusions and bibliography. FIRST PART: CURRENT STATE OF KNOWLEDGE, has three chapters: Chapter 1. The concept of the wheat quality and its importance, describes the history and importance of wheat, geographical distribution, chemical composition and nutritional value of its, and the importance of ecological, biological and technological factorson the wheat quality. XXX

4 Chapter 2. The importance of microflora and mycotoxin xontamination of grain quality, describes sources of contamination and structure of microflora contamination, conditions that favor development needs of microorganisms, prevention of mold contamination and inactivation of mycotoxins, and a description of the main fungi producers of aflatoxins. Chapter 3. Mycotoxins, food comtaminants that affect food safety, includes information on the incidence of mycotoxins in cereals, a general description of aflatoxins, as well as legislation issues related to cereals mycotoxins. PART TWO: PERSONAL RESEARCH, is strictured in five chapters, and include: motovation, purpose and objectives, materials and methods, the study of pedoclimatic conditions during conducting research, results and discussions, conlusions and recommendations. Chapter 4. Thesis aim and main objetives, includes motivation and objectives of the research done. Chapter 5. Materials and methods, in which experimental factors are presented, as well as biological material and research methods involved. Chapter 6. Research about the pedo-climatic conditions in that study were carried out, are presented in terms of geographical and pedo-climatic conditions in the region that were taken into study. Chapter 7. Rezults and discussions, describes the results and discussions are presented to each objective. The second part is ending with Chapter 8. Conclusions and recommendations of the study and associated References. 1. AIM AND OBJECTIVES The purpose of the research was the influence of ecological, biological and technological quality of wheat grown in central Transylvania and the identification and assessment of aflatoxin contamination of raw materials used in bakery. The research objectives are: involvement of ecology, biology and technology factors and their interaction on the quality of winter wheat; comparative study on quality of wheat varieties, cultivated in two different pedoclimatic areas (Turda and Targu Mures); participation rates to identify the factors studied and their interaction to achieve the quality indicators of winter wheat; relationships between the parameters studied in order to know the degree of association between them; identification of aflatoxins in wheat, used as raw material to obtain flour and bakery products. XXXI

5 2.1. EXPERIMENTAL FACTORS 2. MATERIAL AND METHOD To achieve the goal of involvement of ecology, biology and technology and their interaction on the quality of winter wheat in 08, 09, 10, was organized at the Agricultural Research and Development Station Turda, an stationary polifactorial type of experience by the subdivided parcels method with four experimental factors in three repetitions (experiment 1). Experience 1: Influence of variety, agrofond and complextreatment with foliar fertilizers, fungicides and insecticides, on the quality of winter wheat in different climatic conditions, between following ranges: Factor A: production year in terms of pedo-climatic conditions, between following ranges: A (Mt.) A A Factor B: variety, with the following ranges: B1 Arieşan (Mt.) B2 Apullum B3 Dumbrava Factor C: base fertilization, with following ranges: C1 N 40 P 40 kg/ha at sowing (Mt.) C2 N 40 P 40 kg/ha at sowing + N P kg/ha at the resumption of vegetation in spring Factor D: complex combination of treatments with foliar fertilizers, fungicides and insecticides at four different phenological moments significant for vegetation with following ranges (table 1). Graduations factor D (vegetation treatments) Table 1 Graduation The resumntion Fenofaza end twin step Fenofaza of Fenofaza of of vegetation and herbicide step bellows flowering D1(Mt.) ÎF + IS ÎF ÎF+FG+IS ÎF+FG+IS D2 - ÎF + IS ÎF+FG+IS ÎF+FG+IS D3 - ÎF + IS - ÎF+FG+IS D4 ÎF + IS ÎF + IS - ÎF+FG+IS ÎF foliar fertilizers Polyfeed 19:19:19 + microelemente 5kg/ha IS-isecidices: Capypso 100 ml/ha in phenophase 1 and 2, Proteus 0,4 1/ha in phenophase 3 şi 4 FG - fungicid Falcon 480 EC 0,6 1/ha in phenophase 3 and Prosaro 11/ha in phenophase 4 The combination of factors and graduations, for each variety result of experience polyfactoriala type 3 x 3 x 2 x 4, with 72 variants, like, presented in Table 2. XXXII

6 Presenting of experimental s variants in first experience Table 2 A1 (07/08) B1 (ARIEŞAN) B2 (APULLUM) B3 (DUMBRAVA) C1 (N 40P 40) C2 (N 60P 60) C1 (N 40P 40) C2 (N 60P 60) C1 (N 40P 40) C2 (N 60P 60) D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 A2 (08/09) B1 (ARIEŞAN) B2 (APULLUM) B3 (DUMBRAVA) C1 (N 40P 40) C2 (N 60P 60) C1 (N 40P 40) C2 (N 60P 60) C1 (N 40P 40) C2 (N 60P 60) D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 A3 (09/10) B1 (ARIEŞAN) B2 (APULLUM) B3 (DUMBRAVA) C1 (N 40P 40) C2 (N 60P 60) C1 (N 40P 40) C2 (N 60P 60) C1 (N 40P 40) C2 (N 60P 60) D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 D1 D2 D3 D4 Previous cropping was the soy beans. The experiments were made each year according to the experimental design, taking into account the best times for cropping, in terms of optimisation. Experimental plot area was of 3m x 7.5 m (22.5 m 2 ) in three repetitions. Due to the experiment 1, complex treatments were carried out at important moments during the vegetation phonological wheat, aiming their influence on quality indices to achieve the goal of identification of aflatoxins in wheat, raw material, to obtain flour and products bread is done in the experimental fields at SCDA Turda and Mures in 09/10 a polyfactorial experiment following the subdivided parcels method in three repetitions (experiment 2). Experience 2: Influence of variety and agrofond contamination with mycotoxins on the quality of winter wheat in different climatic conditions, with the following ranges: Factorul A the area of culture with the following ranges: A1 Turda (Mt.) A2 Targu Mures Factorul B base fertilization, with the following ranges: B1 unfertilized (Mt.) B2 fertilized with N 40 P 40 Factorul C variety, with the following ranges: C1 Arieşan (Mt.) C2 Apullum C3 Dumbrava C4 Turda 00 C5 Exotic C6 Faur C7 Serina C8 Glosa C9 Josef C10 Dropia XXXIII

7 Experiments were placed in strict compliance with three repetitions of wheat cultivation technology for tracking the experimental factors. Soil tillage consisted of stubble-turning with a disc harrow, followed by plowing at cm and two works with disc harrow for seedbed formation, the previous plant used being a spring plant. The combination of factors and graduations, for each kind of results of polyfactorial experiments variants of 10 x 2 x 2, with 40 variants, alike, are shown in the Table 3. Presenting of experimental s variants in the second experience Table 3 A1 (TURDA) A2 (TÂRGU MUREŞ) B1 (N 0 P 0 ) B2 (N 40 P 40 ) B1 (N 0 P 0 ) B2 (N 40 P 40 ) C1 (Arieşan) C1 (Arieşan) C1 (Arieşan) C1 (Arieşan) C2 (Apullum) C2 (Apullum) C2 (Apullum) C2 (Apullum) C3 (Dumbrava) C3 (Dumbrava) C3 (Dumbrava) C3 (Dumbrava) C4 (Turda 00) C4 (Turda 00) C4 (Turda 00) C4 (Turda 00) C5 (Exotic) C5 (Exotic) C5 (Exotic) C5 (Exotic) C6 (Faur) C6 (Faur) C6 (Faur) C6 (Faur) C7 (Serina) C7 (Serina) C7 (Serina) C7 (Serina) C8 (Glosa) C8 (Glosa) C8 (Glosa) C8 (Glosa) C9 (Josef) C9 (Josef) C9 (Josef) C9 (Josef) C10 (Dropia) C10 (Dropia) C10 (Dropia) C10 (Dropia) Throughout the growing season has followed the influence of climatic factors on the development of plant varieties resistancem to stressors with a particular emphasis on the influence of technological factors on the qualitative characteristics, especially for baking ones. Samples were taken from SCDA Turda and SCDCB Mures, both units are located in the centre of Transylvania. Samples collection and training was conducted in compliance with existing standards SR ISO 13690/01, replacing STAS For each experience, at harvest, samples were chosen and retained of foreign bodies, being storage in paper bags at C to determine the physicochemical and backing characteristics. In the study were prepared samples to determine contamination with mycotoxins, which are stored at 3-4 C until the determination in the laboratory THE BIOLOGICAL MATERIAL In order to study the influence of ecology, biology and technology on the quality of bread wheat (one made in SCDA Turda experience, from 07 to 10) were used three winter wheat clovers, namely Ariesan, Apullum and Dumbrava. For experiement 2 on comparative study of wheat quality and identification of aflatoxins contamination were taken into study ten winter wheat varieties domestic and imported, cultivated in the same technological conditions of the same year (09/10), but in different areas at SCDA Turda and SCDCB Targu Mures. XXXIV

8 The studies involved following winter wheat clovers: Ariesan, Apullum, DUmbrava, Turda 00, Exotic, Faur, Serina, Glosa, Josef and Dropia RESEARCH METHODS In order to assess the quality of wheat clover we determined moisture by drying method in the oven, hectolitre mass was assessed by the hectolitice balance, protein content was determined by Kjeldahl method, wet gluten content was assessed by washing with sodium chloride solution with sodium chloride solution 2%, the deformation index gluten was assessed by using the formula, Zeleny sedimentation index, based on particle sedimentation method of flour in the presence of lactic acid and Bromophenol blue, followed by reading the volume of sediment and falling index with the index Falling number equipment. The identification of mycotoxins were done by the method of assessment of all aflatoxins of vegetable origin by high performance thin layer chromatography (HPTLC), based on a method of extraction from Camag with methanol and distilled water. 3. REZULTS AND DISCUSSIONS 3.1. RESULTS REGARDING THE INFLUENCES OF THE EXPERIMENTAL FACTORS ON THE QUALITY OF WINTER WHEAT The influence of climatic conditions on the quality of wheat In 08/09, years characterized as thermally hot and dry excessively; all quality parameters analyzed were favourable influenced by climatic conditions of the year recording significant positive differences compared to07/08, considered as control. Protein content increased by 5.04% in 08/09 (from 9.31% to 14.35%) and to 1.05% in 09/10 (from 9.31% to 10.36%) and wet gluten increased by 18.62% (from 16.11% to 34.73%) in 08/09 and 09/10 increase to 4.11%. In 09/10, a year characterized with normal temperature values, but excessive rain, only the protein content, wet gluten content and quality of gluten index (index of deformation and gluten index) showed significant positive differences towards the control. Thus hectolitric mass increases from kg /hl to 79.50kg /hl (+3.56), in 08/09 and in 09/10 decreased to kg /hl, accounting significantly negative differences (-3.78) compared to the control. The deformation index registered a slight increase from 3.61mm to 4.67mm in 08/09 to 5.11 mm in 09/10 while the gluten index registered a considerable increase from 28.42% to 58.13% (+29.71), in 08/09 and from 28.42% to 39.71% in 09/10. In terms of sedimentation index and drop index we noted in 08/09, a significant positive differences ( for sedimentation index and for index fall), while in 09/10, which was excessively rainy year is very significant negative ( and for sedimentation index for index fall) differences. These values show the importance of climatic conditions on the quality of protein and especially the amylase activity. XXXV

9 The influence of variety on the wheat quality The variety is one of the most important factors in wheat culture technology. In terms of genetics, there are quite high differences between varieties of wheat depending on the content of protein substances. For wheat the protein and gluten content are two hereditary characteristics that are strongly influenced by the environmental factors and culture conditions. The three studied, wheat varieties, Arieşan, Apullum and Grove, are clearly different regarding thir quality for bread. Thus, the variety Arieşan used as a blanck in the analyzed experiences is the largest variety from the area production, being known for its high production potential associated with a good crop quality. For all the analyzed quality parameters, this variety has proven to have a better quality than the varieties Apullum and Dumbraba, with values of kg / hl for hectolitrical weight, 11.80% for protein content, 25.45% for wet gluten content, 5.38 for deformation index, % for falls index and ml for the Zeleny index. Apullum and Dumbrava wheat varieties have shown very significant negative differeces for all the quality parameters, except the hectoliter weight, which showed very significant positive differences compared to the blanck, by increasing the hectoliter mass from kg/hl to 78. kg/hl ( 3.65%) from the Apullum variety, and from kg/hl to kg/hl (0.30%) for the Dumbrava variety. A decreased of the α-amylase activity was also noticed for the Dumbrava variety, that has shown very significant negative differences (-32.43) by raport to the Arieşan variety, considered as blanck The influence of fertilization on the wheat quality An important factor for wheat quality is the fertilization which by differential amounts of nitrogen and phosphorus given to plants in various stages of vegetation can affect the gluten proteins accumulation in the wheat grains, with repercussions on their qualitative characteristics. The fertilization variety for seeding N40P40 kg/ha, was chosen as blank in the three-year experience achieved at ARDS Turda. Significant positive values were noticed for all the analyzed parameters, except the drop index which shown very significant negative values by report to the dose of fertilization applied (-18.0). Applying the nitrogen and phosphorus dose during seeding and in the spring, when the vegetation in the spring (N60P60) an increase of the quality parameter values has been noticed depending on the studied variety. The hectoliter weight increased by kg / hl (figure 1), the protein by % (figure 2), the wet gluten by % (figure 3),the gluten index by % (figure 4) and the sedimentation index by ml (figure 5). Also it was noticed that all the quality parameters, regardless of fertilization applied, the Arieşan wheat variety has showed the highest values, except the hectoliter mass, which recorded the highest values regardless the fertilization applied for Apullum variety. XXXVI

10 DUMBRAVA (+0.29) DUMBRAVA (+0.97) APULLUM (+ 0.57) ARIEŞAN (+0.89) N60P60*** N40P40(Mt.) APULLUM (+1.17) ARIEŞAN (+0.75) N60P60*** N40P40(Mt.) Masa hectolitrică/weight test (kg/hl) Fig. 1 The evolution of weight test depending on fertilization at winter wheat varieties (SCDA Turda) Conţinut de proteină/protein content (%) Fig. 2 The evolution of protein content depending on fertilization at winter wheat varieties (SCDA Turda) DUMBRAVA (+4.09) DUMBRAVA (+6.83) APULLUM (+3.98) ARIEŞAN (+3.67) N60P60*** N40P40(Mt.) APULLUM (+4.92) ARIEŞAN (+4.08) N60P60*** N40P40(Mt.) Gluten umed/wet gluten (%) Fig. 3 The evolution of wet gluten depending on fertilization at winter wheat varieties (SCDA Turda) Indice glutenic/gluten index (%) Fig 4 The evolution of gluten index depending on fertilization at winter wheat varieties (SCDA Turda) DUMBRAVA (+6.67) APULLUM (+5.56) ARIEŞAN (+4.28) N60P60*** N40P40(Mt.) Indice Zeleny/Zeleny index (ml) Fig. 5 The evolution of Zeleny index depending on fertilization at winter wheat varieties (SCDA Turda) The influence of treatment on the wheat quality The success of wheat crops is often conditioned by the appearance of different diseases and pests which by the realized attack helps to the production potential and varieties quality decreasing below the provided agrotechnical conditions. During the growing season the wheat crop monitoring is necessary, by using in time the insecticides and fungicides that are necessary to fight diseases and pests, and also by foliar fertilizer application when the vegetation is regained in order to get a better crop quality. The graphical representation of the quality parameters evolution, for each studied variety was realized depending on the treatments applied during wheat growing season. The treatment variety D1, for which complex treatments have been applied to all four major phenological moments, was considered as blank. XXXVII

11 For all the experimental variants by applying treatments in different stages of vegetation, significant positive or negative differences was noticed for all the analyzed quality parameters except the deformation and failure index where insignificant differences were noticed in most variants. Regarding the quality parameters: protein content (figure 7), wet gluten (figure 8), indicating gluten (figure 9) and Zeleny sedimentation index (figure 10), very significant negative differences were observe compared to the blank for all the studied wheat varieties. The treatment variety D1, for which were applied vegetation treatments in all four major phenological times, the highest values of 11.33% % for protein, 23.38% % for wet gluten, 43.33% - 46% for index gluten, ml ml for sedimentation index have been recorded. Concerning the hectoliter weight (figure 6), very significant positive differences have been noticed compared to the blank (D1) for all wheat varieties, for treatment variants D3 and D4, which suggest an increase of grains filling capacity by not applying treatments during skin phase. Masa hectoliotrică/weight test (kg/hl) Arieşan Apullum Dumbrava D1 (Mt.) D2 (ooo) D3 (***) D4 (***) Gluten umed/wet gluten (%) Arieşan Apullum Dumbrava D1 (Mt.) D2 (-) D3 (ooo) D4 (ooo) Fig. 6 The evolution of weight test depending on treatment Fig.8 The evolution of wet gluten depending on treatment Conţinut de proteină/protein content (%) Arieşan Apullum Dumbrava D1 (Mt.) D2 (ooo) D3 (ooo) D4 (ooo) Indice glutenic/gluten index (%) Arieşan Apullum Dumbrava D1 (Mt.) D2 (ooo) D3 (ooo) D4 (ooo) Fig. 7 The evolution of protein content depending on treatment Fig. 9 The evolution of gluten index depending on treatment Indice Zeleny/Zeleny index (ml) Arieşan Apullum Dumbrava D1 (Mt.) D2 (ooo) D3 (ooo) D4 (ooo) Fig. 10 The evolution of Zeleny index depending on treatment XXXVIII

12 Interaction factors (years,, variety, agrofond and treatment) on quality parameters of the wheat The complexity of wheat quality parameters is even more obvious if is taken into account the interaction of these elements with the ecological, biological and technological factors, by improving the quality of the crops. The interaction between the studied factors years, variety, agrofond and treatments on wheat quality parameters is different depending on the analyzed parameter. Thus, the factors interaction was statistically analyzed by the polifactorial analysis of the variance in subdivided plots, with which combinations between variants and positive or negative significations were obtained. For the interaction between year and variety, regardless the cultivated variety, the achievement of the analyzed quality parameters is influenced by the climatic conditions of the experimental years, with a specific reference to the index of fall whose values are strongly influenced by the climatic conditions, especially by precipitations. Regarding the interaction between agrofond and year, regardless of climatic conditions of the culture, the applied agrofondul favorably influence all the quality parameters. If treatment interactions with year, regardless of climatic conditions of culture, treatments can influence the quality parameters, especially the hectoliter weight by not applying the treatments during the skin phase, but it cannot influence the index of fall, which is stronger influenced by the climatic conditions RESULTS REGARDING QUALITY THE WHEAT VARIETIES CULTIVATED IN TWO AREAS CLIMATIC DIFFERENT (TURDA AND TARGU MURES) The influence of culture area on the wheat quality The representative ecological factors, regarding the different pedo-climatic conditions of the culture areas can influence the accumulation of protein and gluten in wheat grain, and also their quality, by changes of temperature and precipitation, especially during grain filling. The experience made at ARDS Turda and SCDCB Targu Mures, in the year 09/10, considered as thermal normal, but excessive in rains in both culture areas, all the quality parameters analyzed showed significant and significant positive values by raport to the culture area from Turda, considered as blank (table 4). This proves that with the favorable influence of climatic conditions of the area can also interact other ecological factors which have t been studied. Thus the protein content increased by 0.36% (11.94% of the ARDS Turda, from 12.30% to SCDCBB Targu Mures), wet gluten content increased by 1.26% (26.51% of the ARDS Turda, from 27.77% to SCDCB Targu Mures ) and Zeleny sedimentation index increased from ml in ARDS Turda, to ml (+2.03) in Targu Mures SCDCB. XXXIX

13 Table 4 The influence of the culture on the quality parameters of wheat, in Area of the culture (A) Specification Protein content, % Wet gluten, % Zeleny index, ml TURDA (A1) Mt. Value (Mt.) TÂRGU MUREŞ (A2) Value % Dif. ± Semnif. * ** ** Characterization the year Normal heat Excessive rain Normal heat Excessive rain 5% DL 1% % The significance of effect: NS not significant, * significant pozitive, ** significant distinct pozitive, *** very significant pozitive; 0 significant negative, 00 significant distinct negative, 000 very significant negative; The influence of fertilization on wheat quality cultivated in two areas climatic different (Turda and Targu Mures) An important factor on the quality of wheat is the agrofondul, which by its different amounts of nitrogen and phosphorus given to the plants can influence the accumulation of gluten proteins from the wheat grains with repercussions on their qualitative characteristics. In the experiments realized at ARDS Turda and SCDCB Targu Mures in the years 09 10, ten varieties of wheat have been cultivated on the fertilized and nonfertilized agrofond, with a dose of nitrogen and phosphorus N40P40 kg / ha. From the statistical data analyze presented in Table 5 can be noticed very significant positive changes for all the analyzed quality parameters. The presence of nitrogen and phosphorus doses is strongly felt by the protein content which is increased from 11.01% to 13.23% (+2.22) and the wet gluten which increases from 23.29% to 30.98% (+7.69), and the Zeleny sedimentation index which increases from ml to ml (+16.85). Table 5 Influence of factor fertilization on the wheat quality parameters, in SCDA Turda and SCDCB Targu Mures(09/10) Dose of fertilizer Protein Wet gluten, Specification (kg/ha) (B) content, % % Zeleny index, ml N 0 P 0 (B1) Mt. Value (Mt.) Value N 40 P 40 (B2) % Dif. ± Semnif. *** *** *** DL 5% % % The significance of effect: NS not significant, * significant pozitive, ** significant distinct pozitive, *** very significant pozitive; 0 significant negative, 00 significant distinct negative, 000 very significant negative XL

14 The influence of variety on wheat quality cultivated in two areas climatic different (Turda and Targu Mures) The variety is one of the most important factors in wheat culture technology, because its quality depends in a relatively high genetic potential of each variety. The wheat variety Arieşan was used as a blank in the analyzed experiences being the most cultivated variety in the research areas. The analyzed quality parameters values were 12.25% protein, 27.38% wet gluten and ml index, sedimentation. Based on the statistical data analysis was noticed that for the majority of wheat varieties taken into study, significant negative or insignificant differences were noticed reported to Arieşan variety considered blank. An exception was made by Josef wheat variety which shown very significant positive differences for all the analyzed qualitative parameters, with an increase of +0.83% for protein content (figure 11), +3.60% for wet gluten content (figure 12) and ml for Zeleny sedimentation index (figure 13). Josef wheat variety was followed by Turda wheat variety in 00, which showed significant differences in protein content (+0.11 compared to the blank) (figure 11), very significant positive differences in wet gluten content (+0.52% to of blank) (figure 12), and Zeleny sedimentation index (+1.85 ml compared to the blank) (figure 13). Conţinut de proteină/protein content ( %) Apullumº Dumbravaººº 11.9 Turda 00 (NS) (+0.11) Exoticººº Faur(NS) Serinaººº (-1.05) Glosaººº Josef*** 13.1 (+0.83) (+0.30) Dropia** Gluten umed/wet gluten ( %) Apullumººº Dumbravaººº (+0.52) Turda 00*** Exoticººº Faur(NS) Serinaººº (-3.37) Glosaººº (+3.60) Josef*** (+0.75) Dropia*** Fig.11 The variation of protein content to ten varieties of winter wheat (SCDA Turda, SCDCB Targu Mures, 09/10) Fig. 12 The variation of wet gluten to ten varieties of winter wheat (SCDA Turda, SCDCB Targu Mures, 09/10) Indicele Zeleny/Zeleny index (ml) (+1.85) (-6.57) 44.2 (+7.28) (+0.55) Apullum** Dumbrava(NS) Turda 00*** Exoticººº Faurººº Serinaººº Glosaººº Josef*** Dropia(NS) Fig. 13 The variation of Zeleny index to ten varieties of winter wheat (SCDA Turda, SCDCB Targu Mures, 09/10) XLI

15 Interaction factors (the area of culture, agrofond and variety) on quality parameters wheat The interactions between the agrofond culture area and the variety culture area has significantly influences the quality of wheat, although there were noticed also insignificant values. It can be concluded that regardless of culture area, the agrofond by applying nitrogen and phosphorus fertilizers and the cultivated variety, by its genetic potential, can contribute to wheat quality parameters improvement RESULTS ON THE IDENTIFICATION OF THE FACTORS STUDIED PARTICIPATION RATES AND THEIR INTERACTION TO ACHIEVE THE QUALITY OF WINTER WHEAT The specific objectives pursued in our research were to: study the influence of ecological factors (temperature and precipitation years experience and areas of culture), the variety and fertilization with different doses of fertilizers and foliar fertilizer complex treatments, fungicides and insecticides on a seven specific parameters, the quality of wheat grain and flour quality of bread made from wheat. For this purpose we used statistical calculations namely variance analysis to determine the cause variability of each factor on the technological characteristics of wheat and wheat flour. Of the statistical parameters that characterize the best variability of factors on quality indices such studies have been taken into account the sum of squares, degrees of freedom, and average square factor F. For a more suggestive illustration of the influence of complex factors and their interaction on the quality parameters was analyzed using analysis of variance estimation percentage share of each factor involved in the implementation characteristics of wheat bread. The coefficient of participation of the climatic conditions of the three experimental years, the experience made in Turda polifactorială is very high for protein content, 88% (figure 15), wet gluten, 89% (figure 16), indicating gluten, 88% (figure 17), sedimentation index, 85% (figure 18) indicate a fall, 82% (figure 19) and mass per storage volume, 71% (figure 14). Share in the climatic conditions in the year in Targu Mures Turda and influences but in a very small proportion of protein and wet gluten. From the analysis participation rates of the factors to achieving quality parameters, that variety is the greatest contribution to the hectoliter weight, 22% (figure 14). The action fertilizer for three years experience influenced both share similar protein content, 4% (figure 15), wet gluten content, 5% (figure 16) and index gluten, 4% (figure 17) and index Zeleny sedimentation, 17% (figure 18), but the action of fertilizers for agricultural year experience in hand, but in different areas of culture, influences a large proportion of protein content, 78% (figure ), wet gluten, 78% (figure 21) and sedimentation index, 82% (figure 23). The complex treatments on vegetation, influenced in a small proportion of qualitative parameters, respectively, 2%, mass per storage volume (figure 14), gluten XLII

16 index (figure 17), sedimentary index (figure 18), 1%, protein content (figure 15) and wet gluten (figure 16). The share of interactions was higher if the index falls 11% (figure 19), interaction effect on other quality parameters were much lower. Very significant for research conducted in the climatic conditions of the years in ARDS Turda is striking influence on the quality of the harvest, followed by a variety of climatic conditions, respectively agrofond., And for research in two different areas in Turda and Targu Mures a high percentage had agrofondul, followed by variety and growing area. Masa hectolitrică/weight test (kg/hl) Conţinut de proteină/protein content (% ) 71% A (ani)/ A(year) B (soi)/ B(variety) 88% A (ani)/ A(year) B (soi)/ B(variety) 4% 2% 1% 22% C (agrofond)/ C(fertilization) D (tratamente)/ D(crop tratament) I (interacţiuni)/ I(interactions) 4% 1% 4% 3% C (agrofond)/ C(fertilization) D (tratamente)/ D(crop tratament) I (interacţiuni)/ I(interactions) Fig.14 Participation rates of climatic conditions (A), variety (B), fertilization (C) and the crop treatments at weight test Fig. 15 Participation rates of climatic conditions (A), variety (B), fertilization (C) and the crop treatments(d) at protein content Gluten umed/wet gluten (%) Indice glutenic/gluten index (%) 89% A (ani)/ A(year) 88% A (ani)/ A(year) B (soi)/ B(variety) B (soi)/ B(variety) 3% 1% 5% 2% C (agrofond)/ C(fertilization) D (tratamente)/ D(crop tratament) I (interacţiuni)/ I(interactions) 5% 2% 4% 1% C (agrofond)/ C(fertilization) D (tratamente)/ D(crop tratament) I (interacţiuni)/ I(interactions) Fig. 16 Participation rates of climatic conditions (A), variety (B), fertilization (C) and the crop treatments(d) at wet gluten Indice de sedimentetre Zeleny/Zeleny index (ml) Fig. 17 Participation rates of climatic conditions (A), variety (B), fertilization (C) and the treatments at gluten index Indice de cădere/falling number (sec) 85% A (ani)/ A(year) B (soi)/ B(variety) 82% A (ani)/ A(year) B (soi)/ B(variety) 5% 2% 7% 1% C (agrofond)/ C(fertilization) D (tratamente)/ D(crop tratament) I (interacţiuni)/ I(interactions) 6% 11% 1% 0% C (agrofond)/ C(fertilization) D (tratamente)/ D(crop tratament) I (interacţiuni)/ I(interactions) Fig. 18 Participation rates of climatic conditions (A), variety (B), fertilization (C) and the treatments at Zeleny index Conţinut de proteină/protein content (%) Fig. 19 Participation rates of climatic conditions (A), variety (B), fertilization (C) and the treatments at Falling number Gluten umed/wet gluten (%) 78% A (zona de cultură) / A (area of the culture) 78% A (zona de cultură) / A (area of the culture) B (agrofond)/ B (fertilization) B (agrofond)/ B (fertilizarion) C (soi)/ C (variety) C (soi)/ C (variety) 2% 5% 15% I (interacţiuni)/ I (interactions) 2% 4% 16% I (interacţiuni)/ I (interactions) Fig. Participation rates of area of the culture (A), fertilization (B) and variety (C) at protein content Fig. 21 Participation rates of city (A), fertilization (B) and variety (C) at wet gluten XLIII

17 Indice de sedimentare Zeleny/Zeleny index(ml) 82% A (zona de cultură) / A (area of the culture) B (agrofond)/ B (fertilizarion) C (soi)/ C(variety) 1% 4% 13% I (interacţiuni)/ I (interactions) Fig. 22 Participation rates of city (A), fertilization (B) and variety (C) at Zeleny index 3.4. CORRELATIONS BETWEEN WHEAT QUALITY PARAMETERS UNDER THE FACTORS INFLUENCE STUDY For the association value of quality parameters analysis from three wheat breeds harvested at SCDS Turda in the experimental years and also for ten wheat breeds harvested in the same conditions and in the same experimental year (09-10), but in different areas at SCDA Turda and at SCDCB Targu Mures, it was calculated correlation coefficients ( r ). Studying the existent relationships between the analysed quality parameters has a great impact in mill and bakery products industry, by allowing the selection of wheat breeds with several optimal quality parameters for bakery, but it also gain attention of how difficult it`s to accomplish this tasks. The results regarding the correlation coefficients between studied quality parameters from those three wheat breeds harvested at SCDA Turda in and from those ten autumn wheat breeds from SCDA Turda and SCDCB Targu Mures are illustrated in table 6 and 7. It can be observe that gained correlation coefficients shows the existence of tight and significant links between the studied characteristics. From the experiment realized at SCDA Turda in the years (table 46), the strong positive relationship between proteins and gluten content (r = ***) is stated, a natural correlation regarding that most of the proteins are represented by glutenic proteins, such as glyadine and glutenine, components fractions of gluten. Protein content is also positively correlated with glutenic index (r = ***), which characterize the quality of gluten, it was also correlated with the sedimentation index (Zeleny index) (r = ***), which characterize the quality of protein. Hectolitric mass has been positively correlated with Zeleny index ( r = ***), with falling index (r = ***), with protein (r = ***), with wet gluten (r = ***) and with glutenic index (r = ***), and it was not correlated with deformation index (r = **). From the others positive correlations gained between 07 and 10 at SCDA Turda, it`s shown the strong positively correlation between glutenic index and sedimentation index (r = ***), sedimentation index and falling index ( r = ***) and the significant correlation between deformation index and glutenic index (0.2723*). From the insignificant correlation gained at SCDA Turda, we state the relationship between deformation index and Zeleny sedimentation index (r = NS ) and the relationship between deformation index and falling index (r = NS ). XLIV

18 Table 6 The correlation coefficients between quality parameters at wheat varieties in polifactorial experience with 4 factors in 3 repetitions (07 10) Quality parameters Weight test, kg/hl Protein content, % Wet gluten, % Deformation index, mm Gluten index,% Zeleny index, ml Falling number, sec Weight test, kg/hl 1 Protein content, % *** 1 Wet gluten, % *** *** 1 Deformation index, mm ** *** ** 1 Gluten index,% *** *** *** * 1 Zeleny index, ml *** *** *** N.S *** 1 Falling number, sec *** ** *** N.S N.S *** 1 r 5% = 0.24 r 1% = 0.31 Results analysis shows that correlation coefficient between protein content and wet gluten from SCDA Turda and from SCDCB Targu Mures, gained a high positive value, r = *** at SCDA Turda and r =0.9801*** at SCDCB Targu Mures. In the same time it`s shown a high positive correlation between protein and sedimentation index at SCDA Turda (r = 0,9574***) and (r = ***) at SCDCB Targu Mures, which suggest the importance of protein content on quality index, giving the possibility to use the Zeleny sedimentation index as a selection criterion for wheat quality in identifying the breeds with very good qualities for bread manufacturing. The correlation between sedimentation index and wet gluten is also worth to be stated here, (r = ***) at SCDA Turda and (r = ***) at SCDCB Târgu Mureş, which suggest once again the importance of protein and its quality on wet gluten. Table 7 The correlation coefficients between quality parameters at 10 wheat varieties grown in Turda and Targu Mures(09/10) Quality parameters Protein content % Wet gluten, % Zeleny index, ml r 5% = 0.44 r 1% = 0.56 Protein content, % TURDA Wet gluten, % Zeleny index, ml Protein, % *** *** 1 TARGU MURES Wet gluten, Zeleny index, ml % *** *** *** *** REGRESSION ANALYSIS POSITIVELY CORRELATED BETWEEN THE QUALITY PARAMETERS For a more precise examination of correlations found in autumn wheat, between bread manufacturing quality parameters, regressions were made for the most important attributes. For parameters analysed in different cities (Turda and Targu Mures) the XLV

19 regressions were illustrated on the same graph for an easier comparison of existent relationship between quality parameters, in different haverst zones The relationship between protein content and the wet gluten The association of protein content with wet gluten from the experiment realized at SCDA Turda in 07-10(figure 23) and at SCDCB Targu Mures in 08/09(figure 24), it can be seen that for autumn wheat breeds the association is described by an regression line with a strong ascending slope (b=3.66) at SCDA Turda and (b=3.45) at SCDCB Targu Mures, which shows that the wet gluten is direct proportional with protein content. The determination coefficient (R 2 ) calculated show that the total variation of wet gluten content 98% (SCDA Turda, 07 10) and 96% (SCDA Turda and SCDCB Targu Mures, 10) is determined by the variation of protein content Gluten umed/wet gluten (%) y = x r = R 2 = Conţinut de proteină/protein content (%) Fig. 23 The relationship between protein content and wet gluten at three varieties of winter wheat (Turda, 07 10) Gluten umed/wet gluten (%) TARGU MURES: y = x r = R 2 = TURDA: y = x r = R 2 = Conţinut de proteină/protein content (%) TURDA TARGU MURES Linear (TURDA) Linear (TARGU MURES) Fig. 24 The relationship between protein content and wet gluten at ten varieties of winter wheat (Turda and Targu Mures, 09/10) The relationship between protein content and the gluten index In figure 25 is graphically represented the relationship between protein content and glutenic index for autumn wheat breeds from SCDA Turda between Correlation coefficient with a value of r = *** indicates the existence of a very strong positive relation, suggesting the importance of this indicator for quality appreciation. The determination coefficient (R 2 = ) indicates that for analyzed breeds, from total variation of glutenic index only approximately 91% can be attributed to protein content variation. XLVI

20 Indice glutenic/gluten index (%) y = x r = R 2 = Conţinut de proteină/protein content (%) Fig. 25 The relationship between protein content and gluten index at three varieties of winter wheat (Turda, 07 10) The relationship between protein content and the Zeleny index A very tight relationship between protein content and sedimentation index is described by the regression line with a slope b = 4.15% (SCDA Turda )(figure 26), b=6.48 and b=8.18 ( SCDA Turda and SCDCB Targu Mures, 10) (figure 27), a natural relationship stating that sedimentation index characterizing protein quality. The determination coefficient R 2 has high values, (SCDA Turda, 07 10), and (SCDA Turda and SCDCB Târgu Mureş, 10), which means that the sedimentation index variation is determined in proportion of 80%, 91% and 98% by the protein content variation. Indice Zeleny/Zeleny index (ml) y = 4.181x r = R 2 = Conţinut de proteină/protein content (%) Fig. 26 The relationship between protein content and Zeleny index at three varieties of winter wheat (Turda, 07 10) Indice Zeleny/Zeleny index (ml) TARGU MURES: y = 8.183x r = R 2 = TURDA: y = x r = R 2 = Conţinut de proteină/protein content (%) TURDA TARGU MURES Linear (TURDA) Linear (TARGU MURES) Fig. 27 The relationship between protein content and Zeleny index at ten varieties of winter wheat (Turda and Targu Mures, 09/10) The relationship between wet gluten and the gluten index The liniar relantionship between wet gluten content and glutenic index is showed in figure 28. Association of wet gluten content with glutenic index is described by a regression line with an ascendant slope b= 1.47, which suggest the tight link between and gluten quality, represented by glutenic index and gluten quantity. Determination coefficient R 2 = ), shows that 92% of glutenic index variation is influenced by wet gluten content XLVII

21 Indice glutenic/gluten index (%) y = x r = R 2 = Gluten umed/wet gluten (%) Fig. 28 The relationship between wet gluten and gluten index at three varieties of winter wheat (Turda, 07 10) The relationship between wet gluten and the Zeleny index In figures 29 and 30 are graphically represented the relationships between wet gluten content and Zeleny index for autumn wheat breeds from SCDA Turda in years of and from SCDA Turda and SCDCB Târgu Mureş from 09/10. The determination coefficient (R 2 ) calculated indicates that from total variation of Zeleby index 80% (SCDA Turda, 07-10), 92% and 88% (SCDA Turda and SCDCB Târgu Mureş, 10) is determined by the variation of wet gluten content. Indice Zeleny/Zeleny index(ml) y = x r = R 2 = Gluten umed/wet gluten (% ) Fig. 29 The relationship between wet gluten and Zeleny index at three varieties of winter wheat (Turda, 07 10) Indice Zeleny/Zeleny index (ml) TARGU MURES: y = x r = R 2 = TURDA: y = x r = R 2 = Gluten umed/wet gluten (% ) TURDA TARGU MURES Linear (TURDA) Linear (TARGU MURES) Fig. 30 The relationship between wet gluten and Zeleny index at ten varieties of winter wheat (Turda and Targu Mures, 09/10) The relationship between gluten index and the Zeleny index The relationship between glutenic index and Zeleny index is represented in figure 31. The linear regression equation shows that a relationship exist between these quality indexes (b=0.67), but not as tight as in between protein and wet gluten, which suggest that besides the technologically factors applied in favor of quality and quantity of proteins and of wet gluten, in a large part the climate condition of experimental year influence the quality of protein and gluten. The determination coefficient (R 2 = ) indicates that, for analyzed breeds, from total variation of Zeleny index only 66% can be attributed to glutenic index variation. XLVIII

22 70 Indice Zeleny/Zeleny index(ml) y = x r = R 2 = Indice glutenic/gluten index (%) Fig. 31 The relationship between gluten index and Zeleny index at three varieties of winter wheat (Turda, 07 10) 3.6. RESULTS FOR TOTAL AFLATOXIN CONTAMINATION OF WHEAT, BY THIN LAYER CHROMATOGRAPHY HIGH PERFORMANCE (HPLC) Qualitative determination of total aflatoxins (B1, B2, G1, G2) of winter wheat grown from SCDA Turda and SCDCB Targu Mures The mycotoxins followed in this study are: total aflatoxins (AFL B 1, AFL B 2, AFL G 1, AFL G 2 ), were chosen because of their massive presence in cereals In table 8 are represented the results for 40 version, which consider the presence or absence of aflatoxins Table 8 Area of the culture TURDA TÂRGU MURES Identifying the presence of aflatoxins in winter wheat varieties grown in Turda and Targu Mures Sample code NOT FERTILIZED Variety of Type of wheat aflatoxin Sample code FERTILIZER P1 Ariesan AFL B 2 P11 Ariesan ND P2 Apullum ND P12 Apullum ND P3 Dumbrava ND P13 Dumbrava ND P4 Turda 00 ND P14 Turda 00 ND P5 Exotic ND P15 Exotic ND P6 Faur ND P16 Faur ND P7 Serina ND P17 Serina ND P8 Glosa ND P18 Glosa ND P9 Josef ND P19 Josef ND P10 Dropia ND P Dropia ND P21 Ariesan ND P31 Ariesan ND P22 Apullum ND P32 Apullum ND P23 Dumbrava ND P33 Dumbrava ND P24 Turda 00 ND P34 Turda 00 ND P25 Exotic AFL B 1 P35 Exotic ND P26 Faur ND P36 Faur ND P27 Serina ND P37 Serina ND P28 Glosa ND P38 Glosa ND P29 Josef AFL B 1 P39 Josef ND P30 Dropia ND P40 Dropia ND ND- unidentified XLIX