Research ArticleReceived: 29 July 2011 Revised: 13 December 2011 Accepted: 14 December 2011 Published online in Wiley Online Library:

(wileyonlinelibrary.com) DOI 10.1002/jsfa.5590

Health-promoting phytochemicals of Italiancommon wheat varieties grown underlow-input agricultural management†

Raffaella Di Silvestro,a Ilaria Marotti,a Sara Bosi,a Valeria Bregola,a

Antonio Segura Carretero,b Ivana Sedej,c Anamarija Mandic,c

Marijana Sakac,c Stefano Benedettellid and Giovanni Dinellia∗

Abstract

BACKGROUND: The increasing interest in organic food products and environmental friendly practices has emphasised theimportance of selecting crop varieties suitable for the low-input sector. Moreover, in recent years the relationship betweendiet and human health has gained much attention among consumers. The aim of this study was to comparatively evaluate theagronomic performance and the nutrient and phytochemical composition of old and modern Italian wheat genotypes grownunder low-input management.

RESULTS: Research highlighted that several old wheat genotypes were comparable to the modern ones in terms of agronomicperformance and nutrient content. Genotype and environmental conditions (growing season), as well as their interaction,significantly affected the phytochemical composition of wheat grains for most of the analysed bioactive compounds. Highvariability was observed among the wheat genotypes for dietary fibre (154.7–183.3 g kg−1), polyphenol (1.94–2.77 mg g−1),tocopherol (9.1–21.2 mg kg−1) and carotenoid (701.4–3243 µg kg−1) content.

CONCLUSION: The comparative study of old and modern wheat varieties highlighted that, under low-input conditions, ancientgenotypes may equal modern ones in terms of agronomic traits and additionally provide nutraceutical value-added wheatgrains. The most promising ancient varieties for the unique phytochemical profiles are Gentil rosso, Marzuolo d’aqui andVerna.c© 2012 Society of Chemical Industry

Keywords: wheat; old varieties; dietary fibre; antioxidants

INTRODUCTIONIn recent years, low input agriculture have gained muchattention in most developed countries, due to increasing interesttowards more ecologically friendly practices and organic foodproducts considered, among consumers, healthier and saferthan conventional ones. Lammerts van Bueren et al.1 recentlyreviewed the lack of crop varieties specifically bred for thelow-input sector, underlining that more than 95% of theorganic products derives from cultivars bred for the conventionalfarming. The current breeding strategies include both directselection under organic farming and indirect selection underconventional conditions.2 Both strategies are significantly affectedby the interaction between genotype and growing system.Consequently, one of the most important concern for the low-inputsector is to obtain varieties with performance and characteristicsrepeatable in time, specifically adapted to particular subregions.3,4

To reach the present purpose, the first requirement is to intensifyinvestigation concerning the comparison of existing varieties forthe agronomic traits and the physiological reaction under low-input growing conditions. As regards common wheat (Triticumaestivum L.), the low genetic variation of selected modern cultivarssuggests that these genotypes may lack traits of crucial interest

for the low-input sector, even if they are able to guarantee a top-yield production in both conventional and organic agriculture.5 – 7

Several unfavourable conditions are typically related to the organicfarming system where synthetic crop protection and chemicalfertilisation are not allowed such as low soil nutrient status

∗ Correspondence to: Giovanni Dinelli, Department of AgroenvironmentalScience and Technology, University of Bologna, via Fanin 44, 40127Bologna, Italy. E-mail: giovanni.dinelli@unibo.it

† Part of this paper was given as abstract and oral presentation at the FirstInternational Conference on Organic Food Quality and Health Research, 18–20May 2011, Prague, Czech Republic.

a Department of Agroenvironmental Science and Technology, University ofBologna, via Fanin 44, 40127 Bologna, Italy

b Department of Analytical Chemistry, University of Granada, C/Fuentenuevas/n, 18071 Granada, Spain

c Institute for Food Technology in Novi Sad, University of Novi Sad, Bulevar caraLazara 1, 21000 Novi Sad, Serbia

d Department of Crop, Soil and Environmental Science, University of Florence,Piazzale delle Cascine 18, 50144 Florence, Italy

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and pressures deriving from weeds, pests and diseases.8 Asreported by Lammerts van Bueren et al.,1 old wheat varieties mayrepresent a valuable start material for breeding programme inthe low-input sector, as they could guarantee a superior nitrogen(N) extraction in low-N environments, good competitiveness onweed control (due to higher plant height) and a general higherresistance to biotic and abiotic stresses. Traditionally, wheatbreeding programmes selected traits to improve grain yield andprotein content, as well as baking quality, neglecting nutritionaland phytochemical characteristics. Among cereals, wheat is oneof the most consumed staple foods all over the world and itplays an important role in human nutrition. In addition to theessential nutrients (i.e. protein, starch, lipid), whole grain containsseveral bioactive compounds that confer to wheat-derivedproducts unique health promoting properties. Epidemiologicalevidence correlated whole-grain consumption with beneficialeffects on human health by preventing chronic and cardiovasculardiseases, in addition to anticancer properties.9 – 12 The overallhealth benefits of wheat grains have been largely attributedto their unique phytochemical profile, which includes dietaryfibre components (insoluble and soluble dietary fibre, resistantstarch) and antioxidant compounds (polyphenols, flavonoids,tocopherols and carotenoids). As regards dietary fibre, wholewheat was found to be the most important dietary fibresource in the human diet, thus significantly contributing to theachievement of recommended daily intake.13 Beyond dietary fibre,the presence of several phytochemical compounds was recentlyreviewed by Fardet14 who described the complex nutraceuticalcomposition of wheat grains and their relative health-promotingproperties. In wheat, most of the antioxidant compounds areconcentrated in the outer layers of the caryopsis (aleurone, branand germ fractions);15,16 therefore the choice of whole-wheatproducts instead of refined flour-based products may providegreater antioxidant benefits. Previous investigations reportedon the comparison of different wheat varieties in terms ofphenolic,17 – 19 tocopherol20,21 and carotenoid content,18,22 as wellas antioxidant activity.17,20 However, no studies described thecomplete phytochemical variability of wheat grains and differentwheat genotypes. The aim of this study was to comparativelyevaluate the nutraceutical properties of old and modern wheatvarieties, grown at the same location (Bologna, Italy) over twoconsecutive growing seasons. The investigation aimed to providea complete characterisation of each genotype by considering the

agronomic performance under low-input management and screenthe nutrient and phytochemical composition of the involved wheatgermoplasm.

MATERIAL AND METHODSGrain samplesWheat samples included 17 old, not dwarf and unregistered Italiangenotypes (Andriolo, Autonomia A, Autonomia B, Benco, BiancoNostrale, Canove, Carosello, Frassineto, Gentil Bianco, Gentil rosso,Gentil rosso aristato, Gentil rosso mutico, Inallettabile, Marzuolod’aqui, Marzuolo val pusteria, Sieve, Verna) and six dwarf andsemi-dwarf registered cultivars (Bilancia, Bolero, Eureka, Mieti,Nobel, Palesio) of common wheat (Triticum aestivum L.). Trials werecarried out at the experimental farm of the University of Bologna,Cadriano (latitude 44◦ 33′ N, longitude 11◦ 21′ E, 32 m a.s.l.),Italy, during the growing seasons 2006–2007 and 2007–2008.Data on monthly mean, minimum and maximum temperature ofthe growing seasons 2006/2007 and 2007/2008, along with thetotal monthly rainfall, are shown in Table 1. During the secondyear (2007/2008), lower temperature values were observed ascompared to 2006/2007. In particular, the period from April toJune (2007/2008), which corresponds to the heading, anthesisand maturity stages of the crop, were colder with respect tothe first growing season. As regards the total rainfall, the twogrowing seasons differed mostly during springtime, with lowerrainfall value in May and more abundant rainfall in June of thefirst year as compared to the same months of 2007/2008. Wheatvarieties were managed under low-input conditions. The soil atthe experimental farm of Cadriano is classified as a fine silty, mixed,mesic, Udic Ustochrepts, and has a silty loam texture, with 380,375, and 245 g kg−1 of sand, silt, and clay, respectively. The pH(1 : 2.5 soil to water) is 7.9 and organic carbon is 8.5 g kg−1. Thefield experiments were conducted as a randomised completeblock design with three replicates (plot dimension equal to6 × 5 m). The recorded data included plant height, grain yield(kg ha−1), 1000-kernel weight (g), test weight (kg hl−1) andHarvest Index (ratio of grain yield to grain-plus-straw yield).Whole wheat grains of each variety, obtained from both growingseasons, were ground in a stone mill and the whole flours storedat −20 ◦C until analysis to protect bioactive components fromdegradation.

Table 1. Data on monthly total rainfall (mm), mean, minimum and maximum temperature (◦ C) during the growing seasons 2006/2007 and2007/2008 at the experimental field (Cadriano, Bologna, Italy)

Rainfall (mm) Mean temperature (◦C) Minimum temperature (◦C) Maximum temperature (◦C)

Month 2006/2007 2007/2008 2006/2007 2007/2008 2006/2007 2007/2008 2006/2007 2007/2008

October 11.6 143.8 16.9 14.4 12.9 11.0 21.3 21.0

November 41.8 17.0 10.7 8.7 7.4 5.8 14.3 12.2

December 28.4 41.0 6.4 4.8 3.8 2.1 9.4 8.1

January 10.6 48.6 6.7 6.2 4.0 3.8 10.2 9.1

February 34.6 16.2 8.3 6.9 5.2 3.5 11.9 10.9

March 100.4 54.2 10.9 10.6 6.8 6.6 15.1 14.8

April 19.8 30.8 16.8 13.8 11.3 9.4 22.2 18.5

May 41.8 133.2 20.5 18.4 15.5 13.5 25.8 23.1

June 131.8 87.8 23.2 22.6 18.8 17.8 28.2 27.9

July 1.2 10.4 27.0 26.4 21.0 20.3 32.9 31.0

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ChemicalsFolin–Ciocalteu reagent, gallic acid, catechin, β-carotene, 2,2-diphenyl-1-picrylhydrazyl, α- γ - and δ-tocopherol were purchasedfrom Sigma–Aldrich (St Louis, MO, USA). Lutein and zeaxanthinwere obtained from Extrasynthese (Genay, France). HPLC-grademethanol and acetonitrile were purchased from Carlo Erba (Milan,Italy). All other chemicals and solvents were of analytical grade.

MacronutrientsGrain protein was measured by the Kjeldahl procedure (N × 5.7)(AACC, method 46–12).23 The gluten content was determinedusing the method previously described by Kieffer et al.24 Lipidanalysis was carried out according to the AOAC25 using chloro-form–methanol (2 : 1, v/v) to extract lipid compounds. Starch wasenzymatically hydrolysed with α-amylase and amyloglucosidaseand measured using a Megazyme assay kit (Megazyme Interna-tional Ireland Ltd, Wicklow, Ireland).

Dietary fibre and resistant starchTotal, insoluble and soluble dietary fibre contents were analysedfollowing the enzymatic–gravimetric procedure previously de-scribed by Prosky et al.,26 using a Megazyme assay kit. Briefly,1.00 g of flour was subjected to sequential enzymatic digestion byheat-stable α-amylase, protease and amyloglucosidase to removestarch and protein. Sample solution was then filtered to obtainthe insoluble dietary fibre (IDF) residue and the filtrate was treatedwith 95% heated ethanol to precipitate the soluble dietary fibre(SDF). IDF and SDF residues were dried and corrected for protein,ash and blank for final calculation of dietary fibre content. Resis-tant starch was analysed according to McCleary and Monaghan27

using a Megazyme assay kit. After overnight α-amylase and amy-loglucosidase digestion, soluble starch was removed with 95%and 50% ethanol consecutive washes. The pellet was extractedwith 2 mol L−1 KOH to dissolve resistant starch, hydrolysed withamyloglucosidase and spectrophotometrically quantified using aglucose oxidase–peroxidase (GOPOD) reagent.

Free and bound phenolic compoundsFree and bound phenolic compounds were extracted as previouslydescribed.28 Whole-grain flours were extracted with cold 80%ethanol to dissolve the free soluble compounds, followed byacid and alkaline hydrolyses to release the bound forms. Extractswere analysed for the polyphenol quantification following thecolorimetric procedure based on the Folin–Ciocalteu reagent(Sigma–Aldrich)29 and using gallic acid as a standard. Furthermore,extracts containing free and bound phenolic compounds wereanalysed for flavonoid content following the spectrophotometricmethod previously described28 using catechin as a standard.

Antioxidant, free radical scavenging and metal chelatingactivityAntioxidant compounds were extracted from 5.00 g of eachgenotype flour through an overnight extraction with 80%methanol at room temperature. Appropriate dilutions of methanolextracts were used for the subsequent analyses of antioxidant,free radical scavenging and metal chelating activity. Free radicalscavenging capacity of grain extracts was determined accordingto Yu et al.,17 using the stable 2,2-diphenyl-1-picrylhydrazyl radical(DPPH). Freshly made 100 µmol L−1 DPPH solution was added towheat extracts at various concentrations (5.00–40.00 mg mL−1) to

start the radical–antioxidant reaction. The absorbance at 517 nmwas measured against a blank of pure methanol and used toestimate the remaining percentage radical levels. Free radicalscavenging activity of each wheat extract was reported as theconcentration of antioxidant (mg mL−1) required to scavenge 50%of the initial amount of DPPH (IC50). IC50 values were obtained byinterpolation from linear regression analysis. Reducing activityof grain extracts was measured using the method reportedby Oyaizu.30 Extracts were tested at the concentration range1.00–5.00 mg mL−1 and the absorbance of the reaction mixturewas measured at 700 nm. The IC50 value was calculated asthe effective concentration of extract required to obtain the0.5 absorbance value and was obtained by interpolation fromlinear regression analysis. Chelating activity on Fe2+ of grainextracts was determined according to the method previouslydescribed.31 Different concentrations of methanolic extracts(0.01–0.50 mg mL−1) were tested and the absorbance of thereaction mixture was measured at 562 nm. The IC50 value wascalculated as the exact concentration needed to react with 50% ofinitial amount of Fe2+ and obtained by linear regression analysis.

TocopherolsTocopherols were extracted from whole-wheat flours accordingto the method described by Sedej et al.32 Briefly, after alkalinehydrolysis of 5.00 g of flour, tocopherols were extracted with n-hexane. After evaporation to dryness, extracts were dissolved inHPLC-grade methanol, stored under nitrogen stream and filteredthrough a 0.45 µm pore size PTFE filters (Rotilabo–Spritzenfilter13 mm; Roth, Karlsruhe, Germany) before HPLC injection. Thedetermination of tocopherol composition was performed usingan Agilent 1200 series (Agilent Technologies Inc., Santa Clara, CA,USA) equipped with a XDB-C18 analytical column (4.6 × 50 mmcolumn, 1.8 µm particle size) from Agilent Eclipse. Methanol wasused as a mobile phase and the flow rate was set at 1.00 mL min−1.Separated compounds were detected at 295 nm with referencewavelength set at 550/100 nm and the spectra were acquiredat 210–400 nm. Identification of α-, γ - and δ-tocopherol wasachieved by comparing retention time and spectra with those ofstandards.

CarotenoidsCarotenoid content was determined using the methodpreviously described.33,34 Pigments were extracted withmethanol–tetrahydrofuran (1 : 1, v/v) from 1.00 g of flour, pre-viously added of magnesium carbonate to prevent oxidation. Theextraction was repeated until a colourless residue remained. Thecombined organic phases were evaporated to dryness, redissolvedwith methanol–tetrahydrofuran (1 : 1, v/v) and filtered (0.45 µm)before HPLC injection. Carotenoids were analysed using a reverse-phase high-performance liquid chromatography system (RP-HPLC;Beckman Coulter Inc., Miami, FL, USA) consisting of a Gold 126multisolvent pump, photodiode array detector Beckman 168 anda Spark Holland autosampler. For the separation a Waters reverse-phase C18 column (4.6 mm × 250 nm, 5 µm) was employed. Thebinary gradient (acetone–water, initially 75 : 25) elution was run ata flow rate of 1 mL min−1. The gradient was initiated with the ini-tial proportion for 5 min, then linearly increased to 95 : 5 for 5 minand maintained at this level for 10 min. A 3 min re-equilibrationtime was used after each analysis. Carotenoids detected at 450 nmwere identified comparing retention time and spectra with thoseof lutein, zeaxanthin and β-carotene standards.

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Statistical analysisAnalysis of variance (ANOVA, Tukey’s honest significant differencemultiple comparison) was performed using Statistica 6.0 software2001 (StatSoft, Tulsa, OK, USA). The main effects (genotype andgrowing season) and their interaction were tested for significanceby two-way ANOVA. Significance between means was determinedby least significant difference values for P < 0.05. A lineardiscriminant analysis (LDA) was performed using Statistica 6.0software 2001. The supervised technique was applied to thestandardised data matrix of the nutrient and phytochemicalcontent of the investigated varieties from both growing seasons.LDA allowed scoring of the cases as a function of the first two roots(canonical discriminant functions) to visualise similarities amongthe wheat genotypes.

RESULTS AND DISCUSSIONGrain yield and agronomic traitsSeveral factors are known to affect wheat grain yield, includingvariety, environmental and growing conditions.35,36 In our study,a significant influence of wheat genotype on the grain yield

was observed in both growing seasons, while no differenceswere observed between cropping years (Table 2). Higher yieldswere observed for the modern cultivars Bilancia and Bolero andthe old varieties Autonomia A, Autonomia B, Benco, Canove,Gentil rosso aristato and Verna. Mean grain yield (4323.8 kgha−1) aligned with data previously reported for wheat grownunder organic management8 and low N-rate applications.37 1000-kernel weight ranged from 22.3 g (Gentil rosso mutico) and 41.7 g(Nobel) (Table 2) and was generally lower than values previouslyreported.37 – 39 Higher 1000-kernel weights were observed for theold genotypes Gentil bianco, Inallettabile, Marzuolo val pusteriaand Verna, and the modern variety Nobel (Table 2). Harvest Index(HI) ranged from 0.20 (Benco, Frassineto) to 0.35 (Mieti) andresulted in lower values than data previously reported8,37 (Table 2).As expected, HI varied as a function of wheat genotype and highervalues were observed for all the investigated modern varieties.Results agree with previous investigation by Guarda et al.37 whodescribed a significant increase of HI values in modern varietiescompared to the old ones, mainly due to higher spike fertilityand drawft habitus of modern cultivars. The test weight rangedfrom 70.0 kg hl−1 (Bolero) to 82.8 kg hl−1 (Bianco nostrale) and

Table 2. Plant height, grain yield, 1000-kernel weight, test weight and harvest index of the investigated wheat varieties

VarietyPlant

height (m)Yield (kg

ha−1)1000-kernelweight (g)

Test weight(kg hl−1)

Harvestindex

Andriolo (O) 1.145i 3777.5gh 26.9k 75.8fg 0.22gh

Autonomia A (O) 1.207fg 4567.5cd 30.9h 75.5gh 0.27d

Autonomia B (O) 1.217e – g 5237.5b 28.7ij 75.9fg 0.25ef

Benco (O) 1.438a 5292.5b 34.2f 76.0e – g 0.20i

Bianco nostrale (O) 1.206g 4210.0e 28.6ij 82.8a 0.24fg

Bilancia (M) 0.630q 5672.5a 35.3e 69.4m 0.32b

Bolero (M) 0.767m 5130.0b 33.1g 70.0lm 0.29c

Canove (O) 1.388b 5417.5ab 28.3j 72.2k 0.21hi

Carosello (O) 0.928k 4125.0ef 31.1h 70.4lm 0.26de

Eureka (M) 0.807l 4167.5e 29.2i 70.6l 0.29c

Frassineto (O) 1.331c 3330.0i 23.4l 75.6gh 0.20i

Gentil bianco (O) 1.240e 3530.0hi 39.8b 78.5b 0.25ef

Gentil rosso (O) 1.230ef 3870.0fg 34.1f 78.1bc 0.25d – f

Gentil rosso aristato (O) 1.20g 4595.0cd 34.4f 76.7d – f 0.25ef

Gentil rosso mutico (O) 1.170h 3872.5fg 22.3m 75.5gh 0.25ef

Inallettabile (O) 1.060j 3840.0fg 37.7d 73.4ij 0.25ef

Marzuol’ d’aqui (O) 1.275d 3822.5gh 27.2k 78.1bc 0.22gh

Marzuolo val pusteria (O) 1.230ef 4367.5de 38.6c 77.1c – e 0.21hi

Mieti (M) 0.693o 4180.0e 32.8g 72.0jk 0.35a

Nobel (M) 0.667p 4172.5e 41.7a 75.2gh 0.31b

Palesio (M) 0.729n 4190.0e 23.2l 72.7jk 0.32b

Sieve (O) 1.198g 3290.0i 34.4f 77.2cd 0.26de

Verna (O) 1.195g 4790.0c 38.1cd 74.5hi 0.24fg

Mean value M 0.716 4585.4 32.5 71.7 0.31

Mean value O 1.215 4231.5 31.7 76.1 0.24

Mean all varieties 1.085 4323.8 31.9 74.9 0.26

Genotype 99.8∗∗∗ 89.8∗∗∗ 99.3∗∗∗ 86.8∗∗∗ 94.0∗∗∗

Year NS NS 0.2∗ NS NS

GxY 0.2∗ 10.1∗∗∗ 0.4∗ 12.9∗∗∗ 5.8∗∗

LSD (0.05 0.23 292.0 0.69 1.14 0.02

a,b,c,d,e,f,g,h,i,j,k,l,m,n,o,p Means followed by the same letter are not significant at P < 0.05.Significance level: ∗ , P < 0.05; ∗∗ , P < 0.01; ∗∗∗ , P < 0.001.M, modern cultivar; O, old cultivar, NS, not significant.

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highly statistically significant differences were observed amongthe investigated wheat genotypes (Table 2). The old varietiesBianco nostrale, Gentil bianco, Gentil rosso and Marzuolo d’aquishowed higher test weights, highlighting a well-formed graindevelopment according to literature data.8,40 However, someinvestigated genotypes from both old and modern cultivarsshowed test weights below the recommended for wheat flourtrade (>75 kg hl−1).41 Low specific weight is a common concernof wheat varieties grown under low-input conditions and affectsthe productivity, efficiency and operating costs of flour milling.36

Among the morphological traits, highly significant differenceswere observed between the old and modern genotypes as regardsthe plant height (Table 2). Plant height of old genotypes variedfrom 0.9 m to 1.4 m. As expected, lower values were observedfor modern cultivar (0.7 m) that were subjected to breedingprogrammes aimed to obtain semi-dwarf varieties to preventstem lodging (Table 2).

Macronutrient contentMacronutrient composition of the investigated wheat grains ispresented in Table 3 for protein, gluten, starch and lipid content.In organic wheat production, one of the most important concernsis the grain quality, especially as regards the protein content.

Old wheat varieties have shown a more efficient nutrient use inlow-N environments compared to the modern ones which arestrictly dependent on high-level available N.1 In our study, theprotein content varied from 125.7 g kg−1 (Eureka) to 159.5 g kg−1

(Frassineto) (Table 3). Lower contents were observed for moderncultivars such as Bilancia, Bolero and Eureka, while the oldgenotypes Frassineto, Sieve and Verna showed higher proteinyields (Table 3). The protein content was significantly influencedby wheat genotype, evidencing a probably different nutrient-useefficiency of the investigated varieties. Data obtained here forthe protein content are generally higher than those reported inliterature for wheat grown under low-input management.8,19,35

As regards lipid content, the wheat genotypes involved in thepresent study showed higher values than the crude fat amount ofwhole-wheat flours previously reported42,43 while starch contentagrees with data present in literature.44 Although the investigatedvarieties showed high protein contents, the gluten fractionresulted lower as compared to data previously reported forold and modern wheat varieties grown under organic and low-input conditions.8 The protein content of wheat grain is mainlydetermined by the genetic background; however, environmentalfactors such as nitrogen application, water access and temperaturemay strongly influence protein composition (gliadin, glutenin,

Table 3. Nutrient and dietary fibre content of the investigated wheat varieties expressed as g kg−1 of whole flours

Variety Protein Gluten Starch Lipid SDF TDF RS

Andriolo (O) 149.3cd 101.5b – e 772.4 34.8d – h 18.5hi 154.9j 7.1e – g

Autonomia A (O) 141.6e – g 100.1c – e 742.4 37.1b – e 18.5hi 183.3a 6.8f – h

Autonomia B (O) 146.9de 103.2b – d 736.4 35.2d – h 17.6i 176.3a – d 2.5m

Benco (O) 134.1h – j 85.0jk 762.4 39.1ab 23.2g 165.8d – j 5.1i – k

Bianco nostrale (O) 146.0d – f 100.2c – e 754.9 39.0ab 22.4g 169.6c – i 16.1a

Bilancia (M) 128.0jk 80.9k 727.9 37.5b – e 21.3gh 161.9f – j 2.9lm

Bolero (M) 127.9jk 83.0k 708.1 36.7b – f 15.9i 160.3g – j 13.8b

Canove (O) 139.2f – h 87.4h – k 724.9 41.6a 28.4ef 178.7a – c 12.8b

Carosello (O) 141.1e – h 97.7d – g 724.8 32.2h – j 16.8i 182.8ab 9.0cd

Eureka (M) 125.7k 92.4f – i 725.7 27.6l 18.5hi 163.1f – j 8.5c – e

Frassineto (O) 159.5a 110.8a 717.4 32.5g – j 28.0f 158.7ij 12.6b

Gentil bianco (O) 130.3i – k 90.8g – j 737.9 33.3g – j 21.7g 173.6a – f 5.4h – k

Gentil rosso (O) 139.6e – h 103.4b – d 738.5 28.7kl 33.3bc 173.4a – f 13.6b

Gentil rosso aristato (O) 135.7g – i 87.5h – k 732.9 37.8b – d 33.4bc 176.9a – d 9.1cd

Gentil rosso mutico (O) 144.7d – f 87.3i – k 739.5 38.9a – c 33.1bc 172.6a – f 5.0jk

Inallettabile (O) 151.0b – d 105.9a – c 739.5 34.2e – i 37.1a 175.1a – e 3.0lm

Marzuolo d’aqui (O) 150.2b – d 97.2d – g 730.1 37.8b – d 35.0ab 159.6h – j 7.3e – g

Marzuolo val pusteria (O) 149.4cd 111.2a 762.3 30.2j – l 32.4bc 182.1ab 4.4kl

Mieti (M) 136.7g – i 86.5i – k 739.1 27.0l 31.3c – e 171.8a – g 6.1g – j

Nobel (M) 150.9b – d 106.2a – c 695.5 33.4f – j 32.4bc 171.0b – h 6.6f – i

Palesio (M) 146.8de 94.5e – h 750.0 31.5i – k 31.5cd 154.7j 7.9d – f

Sieve (O) 156.9ab 99.4c – f 751.8 36.6b – f 28.7d – f 163.6e – j 10.1c

Verna (O) 154.5a – c 108.3ab 714.1 35.7c – g 32.1bc 177.6a – d 7.6d – g

Mean value M 136.0 90.6 724.4 32.3 25.1 163.8 7.6

Mean value O 145.3 98.6 740.1 35.6 27.1 172.0 8.1

Mean all varieties 142.9 96.5 736.0 34.7 26.6 169.9 8.0

Genotype 55.3∗∗∗ 28.6∗∗∗ NS 26.9∗∗∗ 59.7∗∗∗ 44.1∗∗∗ 55.2∗∗∗

Year 12.1∗∗∗ 50.9∗∗∗ NS 38.1∗∗∗ 30.6∗∗∗ 12.3∗∗∗ 3.6∗∗∗

GxY 32.7∗∗∗ 20.5∗∗∗ NS 35.1∗∗∗ 9.7∗∗∗ 43.6∗∗∗ 41.1∗∗∗

LSD (0.05) 7.40 7.12 – 3.29 3.01 11.98 1.62

a,b,c,d,e,f,g,h,i,j,k,l,m Means followed by the same letter are not significantly different at P < 0.05.SDF, soluble dietary fibre; TDF, total dietary fibre; RS, resistant starch; M, modern cultivar; O, old cultivar.

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albumin, globulin content) and therefore the gluten relativeamount.8,35

Dietary fibre and resistant starch contentThe bran layer of the wheat kernel is particularly rich in dietaryfibre (DF), including insoluble (IDF) and soluble (SDF) dietary fibreand resistant starch (RS). The 23 investigated wheat varieties werecharacterised for fibre component content, as reported in Table 3.In recent years, consumers have increased their awareness on therelationship between food and health, raising the consumption ofwhole-grain products as major source of fibres in the human diet.Several epidemiological studies largely demonstrated the healthbenefits of dietary fibres.45 – 50 In particular, IDF has direct beneficialeffects on the colon by preventing constipation and cancer, whileSDF and RS are known to modulate blood glucose and insulinlevels and exert high prebiotic activity. Gebruers et al.13 comparedthe dietary fibre content of several cereal crops, highlighting thatwholemeal wheat flour contained the highest fibre amount. Thisfinding underlines the important contribution of whole-wheatconsumption to reaching the daily dietary fibre intake in humandiet. Moreover, in the abovementioned study, high significant dif-ferences for fibre content were observed among the genotypes,suggesting the importance of screening and selection of highdietary fibre-containing varieties that may contribute to the main-tenance of human health. In addition, environmental conditionsmay affect dietary fibre content of wheat grain; for instance, Shewryet al.51 showed a positive correlation with the mean temperaturebetween heading and harvest. In our study, dietary fibre contentwas strongly influenced by wheat genotype and the interactionbetween genotype and growing season (G×Y) (Table 3). Indeedthe total dietary fibre content varied greatly among the 17 old andsix modern wheat varieties. It ranged from 154.7 g kg−1 (Palesio)to 183.3 g kg−1 (Autonomia A) and the soluble components variedfrom 15.9 g kg−1 (Bolero) to 37.1 g kg−1 (Inallettabile) (Table 3). Re-sistant starch is defined as the starch fraction that escapes digestionin the small intestine and is generally classified as a component ofdietary fibres. As outlined above, RS possesses prominent health-promoting properties (i.e. hypocholesterolaemic, hypoglycaemicand prebiotic activities) and additionally RS can be fermented bythe microflora of the large intestine, liberating short chain fattyacids that exert a positive impact preventing colon cancer.48,49

Statistically significant variability was observed among the in-vestigated genotypes for resistant starch content, ranging from2.9 g kg−1 (Bilancia) to 16.1 g kg−1 (Bianco nostrale) (Table 3). Theobtained average dietary fibre and resistant starch contents ofwhole grains are in agreement with data previously reported forseveral wheat varieties.13,52 However, it is to note that some of theinvestigated old genotypes showed higher contents of IDF, SDFand RS than those reported in literature data, thus highlighting anintriguing phytochemical composition.

Polyphenol and flavonoid contentPolyphenols are the most representative class of antioxidantsin the wheat kernel, mainly concentrated in the outer layer ofthe caryopsis.16 In wheat, phenolic compounds exist in the soluble(free) and insoluble (bound) forms. Both phenolic fractions possessvaluable health-promoting properties, acting as radical scavengersand preventing several chronic diseases.14,53 Moreover, boundpolyphenols cross-linked with cell wall components, may resistupper digestive process and reach the colon where, after intestinalmicroflora digestion, directly exert their health benefits reducing

the incidence of colon cancer. During kernel development, thebiosynthesis and accumulation of phenolic compounds vary de-pending on the wheat genotype and the environmental and grow-ing conditions. In our study, the accumulation of free polyphenolsand flavonoids was equally affected by both variety and growingseason, whereas the insoluble bound phenolics varied as a functionof the wheat genotype and the interaction G×Y (Table 4). The vari-ance components of G×Y interactions showed high values for allthe phenolic fractions, highlighting a possible different metabolicreaction of the wheat genotypes to changing growing conditions(temperature, rainfall). The obtained results confirm previous in-vestigations concerning the variability of phenolic content amongseveral common wheat varieties and different cropping location,especially in relation to environmental conditions and the agricul-tural management.15,17,54 – 56 Biotic and abiotic stresses are knownto induce phenolic biosynthesis in plants, consequently organiccrops are often thought to contain more phenolic compounds.A comparative study on polyphenol content among differentwheat varieties cropped under organic and conventional condi-tions showed that organic wheat had higher polyphenols contentwith respect to the conventional one.56 However, this result is incontrast with previous investigation that showed a greater im-pact of the environmental factors on the phenolic amounts ratherthan agricultural management (organic/conventional).55 Furtherstudies will be necessary to understand the existing variability ofphenolics in wheat grain and the metabolic reaction of differentvarieties to environmental and growing conditions. Results onpolyphenol and flavonoid content (both free and bound fractions)of the 17 old and 6 modern wheat genotypes are shown in Table 4.The total polyphenol content ranged from 1.93 g kg−1 (Benco) to2.77 g kg−1 (Gentil rosso) and the insoluble phenolics accountedfor 1.40–2.15 g kg−1. The higher values were obtained for theold varieties Bianco nostrale, Carosello, Gentil rosso and Verna.On average, the bound fraction contributed to 76% of the totalamount, confirming that in wheat polyphenols primarily exist inthe bound forms. Among the different phenolic classes, flavonoidis one of the most abundant in wheat kernel. The total flavonoidcontent varied from 0.40 g kg−1 (Sieve) to 0.65 g kg−1 (Gentil rossomutico) with the bound fraction contributing to 57–72% of thetotal content (Table 4). The obtained polyphenol and flavonoidamounts are comparable with data previously reported for freephenolic content,20,57 while higher values were observed for thebound fraction of the investigated genotypes.18,22

Antioxidant activityThe antioxidant properties of the investigated wheat varieties werecomparatively assessed using three different tests: DPPH, reducingpower and chelating activity assays. Results are presented in Ta-ble 4 and expressed as IC50 values. In the DPPH assay, the old geno-type Verna showed the lowest IC50 value (15.3 mg mL−1) that cor-responds to the highest antioxidant capability (Table 4). The strongantioxidative activity of this genotype might be attributed to theelevated content of free and bound phenolics (Table 4) as theycan donate H-atoms from the OH-groups and stabilise the DPPHradical.58 Along with Verna, high antiradical activity against DPPHwas observed for other ancient varieties as Canove, Gentil rosso,Marzuolo d’aqui and Sieve (Table 4). A statistically significant vari-ability was also observed among the wheat samples for the reduc-ing power. The IC50 values ranged from 4.9 (Bolero) to 7.8 mg mL−1

(Mieti) and the strongest reducing activity was observed for onemodern (Bolero), followed by five old (Frassineto, Gentil rosso,Gentil rosso aristato, Gentil rosso mutico, Inalletabile) varieties

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Table 4. Polyphenol, flavonoid content and antioxidant activity of the investigated wheat varieties

VarietyBPC

(mg g−1)TPC

(mg g−1)BFC

(mg g−1)TFC

(mg g−1)DPPH

(IC50 mg mL−1)RP

(IC50 mg mL−1)CA

(IC50 mg mL−1)

Andriolo (O) 1.67h 2.16i – k 0.41a 0.64a 18.9i – k 6.85cd 17.7a

Autonomia A (O) 1.84ef 2.32fg 0.33c – e 0.48ij 20.3gh 7.68ab 9.9l

Autonomia B (O) 1.53i 2.04lm 0.27ij 0.43l 24.2bc 7.58ab 13.8e – g

Benco (O) 1.48ij 1.93n 0.41a 0.61b 21.9d 6.88cd 12.8f – j

Bianco nostrale (O) 2.15a 2.65bc 0.39b 0.55d 19.1ij 6.20fg 13.1f – i

Bilancia (M) 1.54i 2.08kl 0.27j 0.46jk 21.7de 6.58e 13.9d – g

Bolero (M) 1.73gh 2.27gh 0.33c – e 0.51f – h 18.1lm 4.88k 14.9c – e

Canove (O) 1.83ef 2.50de 0.30f – h 0.47jk 16.7n 6.40ef 15.8bc

Carosello (O) 1.94cd 2.64bc 0.42a 0.58c 20.9fg 7.73a 14.7c – e

Eureka (M) 1.90c – e 2.47e 0.32d – f 0.52e – f 23.5c 6.58e 14.0d – f

Frassineto (O) 1.40j 1.94mn 0.29g – i 0.45kl 23.6bc 5.78i 12.4h – k

Gentil bianco (O) 1.89d – f 2.45e 0.35c 0.54de 19.5hi 7.73a 14.0d – f

Gentil rosso (O) 2.05b 2.77a 0.34cd 0.52e – g 17.5mn 5.23j 13.1f – i

Gentil rosso aristato (O) 1.79fg 2.50de 0.32e – g 0.50f – i 21.4d – f 4.95k 15.1b – d

Gentil rosso mutico (O) 1.98bd 2.60cd 0.43a 0.65a 18.3kl 5.80hi 13.6e – h

Inallettabile (O) 2.04b 2.58cd 0.35c 0.54de 18.5j – l 5.90hi 16.4b

Marzuolo d’aqui (O) 1.98bd 2.61c 0.41a 0.60bc 15.9o 6.00gh 11.7jk

Marzuolo val pusteria (O) 1.71gh 2.19h – j 0.33c – e 0.50g – i 26.2a 7.03c 12.1i – k

Mieti (M) 1.91c – e 2.41ef 0.29h – j 0.45kl 21.9de 7.78a 12.6g – j

Nobel (M) 1.71gh 2.26g – i 0.32d – f 0.49hi 24.3b 7.48b 13.0f – j

Palesio (M) 1.71gh 2.23g – j 0.27j 0.44kl 21.1ef 6.80d 11.1kl

Sieve (O) 1.47ij 2.14jk 0.28ij 0.40hi 17.3n 6.33f 11.9i – k

Verna (O) 1.99bc 2.74ab 0.39b 0.60b 15.3o 6.95cd 12.7f – j

Mean value M 1.75 2.29 0.30 0.48 21.8 6.68 13.2

Mean value O 1.81 2.40 0.35 0.54 19.7 6.53 13.6

Mean all varieties 1.79 2.37 0.34 0.52 20.3 6.57 13.5

Genotype 38.4∗∗∗ 48.3∗∗∗ 42.5∗∗∗ 50.7∗∗∗ 64.6∗∗∗ 58.2∗∗∗ 28.9∗∗∗

Year 13.2∗∗∗ 1.9∗∗∗ 0.3∗ 5.4∗∗∗ 7.4∗∗∗ 31.6∗∗∗ NS

GxY 48.5∗∗∗ 49.9∗∗∗ 57.2∗∗∗ 43.8∗∗∗ 28.1∗∗∗ 10.2∗∗∗ 70.5∗∗∗

LSD (0.05) 0.096 0.101 0.022 0.022 0.750 0.220 1.310

a,b,c,d,e,f,g,h,i,j,k,l,m,n,o Means followed by the same letter are not significantly different at P < 0.05.BPC, bound phenolic compounds; TPC, total phenolic compounds; BFC, bound flavonoid compounds; TFC, total flavonoid compounds; RP, reduncingpower; CA, chelating activity; M, modern cultivar; O, old cultivar.

(Table 4). Results showed that wheat flours could be a rich sourceof dietary antioxidants and may prevent oxidative damage in thebiological system, reacting with peroxyl radicals and thereby termi-nating the auto-oxidation chain.59 Moreover, the 23 wheat extractswere tested for their chelating activity, namely the reduction andstabilisation of transition metals (i.e. Fe2+) which are the causeof the free radical formation and consequently of the oxidativechain inception.54 Highly significant differences were observed forchelating activity among the investigated wheat varieties, with IC50

values ranging from 9.9 (Autonomia A) to 17.7 mg mL−1 (Andriolo)(Table 4). The variability of antioxidant and chelating activity variedas a function of the wheat genotype and the interaction G×Y (Ta-ble 4). This finding supports previous investigations that comparedthe radical scavenging capacities and chelating activity of differentwheat varieties and the influence of cropping locations.17,20,54

Tocopherol contentAlthough cereals are a modest source of lipids, whole-grainderived products are a good source of vitamin E (lipid soluble com-pounds, namely tocopherols and tocotrienols) in the human diet.Previous investigations demonstrated that wheat tocopherols

possess higher bioaccessibility with respect to other vitamin Efood sources,60,61 implying to the important role of whole-grainconsumption in meeting the recommended dietary allowance(RDA) (15 mg per day). In wheat kernel, tocopherols occur in theα, β , γ and δ forms, depending on the number and localisationof the methyl groups of the chromanol ring. Vitamers possessstrong antioxidant activity as they can reduce and stabilise lipidperoxyl radicals by donating hydrogen atoms.20,62 Few studies in-vestigated the effects of environmental conditions on tocopherolcontent and the quali-quantitative variability of tocopherolsamong different wheat cultivars.21,63 Recently, Lampi et al.21 re-ported on the tocopherol content of 26 wheat varieties (includingold and modern cultivars) and showed that the environmentalfactors may affect the accumulation of this class of compounds inwheat kernel. Interestingly some of the old genotypes tested byLampi et al.21 resulted less sensitive to the environmental changes(year, location), showing a greater stability with respect to themodern ones. In our study, the observed variability of vitamer con-tent resulted equally influenced by genotype, growing season andinteraction G×Y (Table 5). α-Tocopherol was the most abundantin all the investigated wheat samples, ranging from 7.5 mg kg−1

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Table 5. Tocopherol (α, γ , δ) and carotenoid (lutein, zeaxanthin, β-carotene) content of the investigated wheat varieties

Tocopherol (mg kg−1) Carotenoid (µg kg−1)

Variety α γ δ Lutein Zeaxanthin β-Carotene

Andriolo (O) 9.250fg 1.025h 0.850a – c 726.250gh 297.025no 247.500j – l

Autonomia A (O) 11.825b – g 2.350c – h 0.800a – d 612.325h 318.250m – o 519.975e – g

Autonomia B (O) 10.225e – g 3.300a – d 1.000ab 640.250h 313.000m – o 542.500e – g

Benco (O) 13.450a – f 2.300c – h 0.275e – g 1048.575c – e 208.025p 329.125hi

Bianco nostrale (O) 9.950e – g 1.675gh 0.325d – g 1581.950a 437.850ij 802.475b

Bilancia (M) 16.275ab 2.350c – h 0.525b – f 316.725i 288.450no 196.100l

Bolero (M) 15.400a – c 4.325a 0.975ab 1136.650b – d 275.350o 593.750de

Canove (O) 16.200ab 3.900ab 1.100a 1176.625b – d 713.675b 571.125e – g

Carosello (O) 12.850a – f 2.425c – h 0.400c – g 704.850gh 376.250kl 279.850i – k

Eureka (M) 11.850b – g 3.200a – f 0.100fg 703.375gh 415.825i – k 227.050kl

Frassineto (O) 16.900a 3.050a – g 0.300d – g 1032.350c – e 514.350fg 357.850h

Gentil bianco (O) 14.050a – e 3.025a – g 0.675a – e 597.450h 562.175ef 306.650h – j

Gentil rosso (O) 11.475c – g 3.575a – c 0.225e – g 1179.600b – d 604.400de 583.150d – f

Gentil rosso aristato (O) 11.175c – g 3.050a – g 0.225e – g 1195.225bc 779.650a 836.425ab

Gentil rosso mutico (O) 10.650d – g 2.650b – g 0.300d – g 1148.325b – d 656.500c 882.300a

Inallettabile (O) 13.025a – f 2.025d – h 0.250e – g 1243.025b 596.850de 699.800c

Marzuolo d’aqui (O) 11.650c – g 1.675gh 0.275e – g 723.150gh 354.625lm 524.675e – g

Marzuolo val pusteria (O) 7.450g 1.675gh 0.000g 902.875ef 460.225hi 542.100e – g

Mieti (M) 15.150a – d 2.400c – h 0.200e – g 1019.650de 409.475jk 514.050fg

Nobel (M) 10.125e – g 1.850e – h 0.125fg 1017.325de 498.775gh 688.600c

Palesio (M) 11.500c – g 3.250a – e 0.150fg 297.275i 334.700l – n 219.950k – l

Sieve (O) 11.150c – g 2.875b – g 0.925ab 837.250fg 638.925cd 500.725g

Verna (O) 10.250e – g 1.825f – h 0.575b – f 1013.975de 535.600fg 655.275cd

Mean value M 13.383 2.896 0.346 748.500 370.429 406.583

Mean value O 11.854 2.494 0.500 962.591 492.199 540.090

Mean all varieties 12.253 2.599 0.460 906.741 460.433 505.262

Genotype 36.3∗∗ 33.0∗∗ 40.7∗∗∗ 91.2∗∗∗ 74.3∗∗∗ 76.9∗∗∗

Year 30.1∗∗∗ 41.1∗∗∗ 20.6∗∗∗ 4.1∗ 16.2∗∗∗ 13.7∗∗∗

GxY 33.6∗ 25.9∗ 38.7∗∗∗ 4.6∗ 9.5∗∗∗ 9.4∗∗∗

LSD (0.05) 4.53 1.42 0.50 174.90 49.60 73.90

a,b,c,d,e,f,g,h,i,j,k,l,m,n,o,p Means followed by the same letter are not significantly different at P < 0.05.M, modern cultivar; O, old cultivar.

(Marzuolo val pusteria) to 16.9 mg kg−1 (Frassineto) (Table 5). Theother vitamers detected were γ and δ tocopherols, the first ina smaller extent (1.0–4.3 mg kg−1) and the second not presentin significant amount (0.1–1.1 mg kg−1) (Table 5). Tocopherolcontent of the investigated wheat genotypes is comparable withdata previously reported for whole wheat grains.20,21,64

Carotenoid contentCarotenoids are plant secondary metabolites that play animportant physiological role in wheat kernels as they canscavenge free radicals and protect grains against disruptionand deteriorative changes.65 Lutein is a carotenoid present inthe highest amount in whole grains, followed by zeaxanthinand β-carotene.18,20,22 When adequately introduced in thediet, carotenoids may act as powerful antioxidants protectinghuman health against several chronic disease.66 In particular,epidemiological studies have correlated high dietary intakesof carotenoids with a lower incidence of age-related maculardegeneration. Indeed, lutein and zeaxanthin are the mostabundant pigments in human macula which protect eyes against

oxidative damage by screening out blue light and quenchingfree radicals.67,68 In our study, carotenoid content varied greatlyamong the investigated old and modern wheat varieties foreach detected compound (Table 5). The growing season partiallyaffected the pigment amounts, but the observed variabilityresulted strongly dependent by the genetic characteristics.This finding agrees with previous investigations reporting highsignificant differences among several wheat varieties.18,20,22 Theelevated genetic variation for pigment content highlights theimportance of screening the existing varieties to identify thosewith higher carotenoid concentration and possibly developa breeding programme to select these traits, as was donebefore for durum wheat varieties improved for yellow colour.In our study, lutein was found to be the most representativecarotenoid in the majority of investigated varieties, rangingfrom 297.3 µg kg−1 (Palesio) to 1582.0 µg kg−1 (Bianco nostrale)(Table 5). Moreover, the 17 old and 6 modern varieties showedthe presence of significant amount of zeaxanthin and β-carotene,with the ranges 275.4–779.7 µg kg−1 and 882.3–196.1 µg kg−1,

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Figure 1. Scatterplot of 6 modern (solid symbols) and 17 ancient (empty symbols) wheat varieties from both growing seasons according to the nutrientand phytochemical content defined by the first two canonical functions (Root 1 and Root 2). Square and circle symbols refer to the first (2006/2007)and second (2007/2008) growing season, respectively. AN, Andriolo; AA, Autonomia A; AB, Autonomia B; BE, Benco; BN, Bianco nostrale; BI, Bilancia; BO,Bolero; CA, Canove; CR, Carosello; EU, Eureka; FR, Frassineto; GB, Gentil bianco; GR, Gentil rosso; GRA, Gentil rosso aristato; GRM, Gentil rosso mutico; IN,Inallettabile; MA, Marzuolo d’aqui; MP, Marzuolo val pusteria; MI, Mieti; NO, Nobel; PA, Palesio; SI, Sieve; VE, Verna.

respectively (Table 5). The obtained results for pigment contentagree with those previously reported.18,20,22

Discriminant analysisThe multivariate techniques of data analysis have been usedto explain the observed variability concerning the nutrient andphytochemical content of the investigated wheat genotypesharvested in the first (2006/2007) and second (2007/2008) growingseasons. LDA allowed to obtain more information as regardsthe variables that mainly influence the sample similarities anddifferences.69 Fig. 1 shows the plot of the wheat samples on thespace defined by the first two canonical functions. The calculatedWilks’ lambda value of 0.0094 (significant at P < 0.00001) isindicative of high discrimination power of the applied model. Aclear differentiation between the two growing seasons has beenobserved. The separation of the samples deriving from 2006/2007and 2007/2008 was affected by the nutrient (gluten, lipid), dietaryfibre and antioxidant (flavonoid, tocopherol, DPPH and RP activity)content, as revealed from the values of canonical functionsstandardised within variances for each variable on Root 1 (valuesof canonical discriminant function 1 equal to −0.85, −0.77, −0.67,−0.88, −0.91, −1.0, −0.70, −0.50, −1.10, −0.77 for gluten, lipid,TDF, % of SDF, RS, total flavonoid, % of free flavonoids, DPPH, RPand total tocopherols, respectively). Moreover, the old and modernvarieties grouped into separated clusters within each year, exceptfor the ancient genotype Frassineto and the modern cultivar Mietiof the second growing season (Fig. 1). The described clusterisationof the cases along second canonical function (Root 2) resultedstrongly influenced by the starch, carotenoid, free polyphenoland flavonoid content (values of canonical discriminant function2 equal to 0.95, 0.74, 0.95 and −1.02, respectively). Accordingly tothe results obtained, the year of cropping induced strong effectson kernel composition of primary and secondary metabolites.However, in each year the different physiological response of

modern and ancient varieties to environmental inputs is reflectedin peculiar phytochemical content of wheat grains.

CONCLUSIONSAs a result of current and foreseeable expansion of the low-input farming (i.e. organic agriculture) the development ofbreeding programmes to select wheat varieties suitable for low-input practices has become a key requirement. The aim of thepresent study was to comparatively evaluate the agronomicand physiological response of old and modern wheat varietiesto low-input growing conditions. A complete description ofthe involved germoplasm was given in terms of agronomicperformance, nutrient content and phytochemical composition.The research highlighted that several ancient wheat genotypesshowed agronomic traits comparable with those of modernvarieties as regards grain yield, 1000-kernel weight and test weight.Genotype and environmental conditions (growing season), aswell as their interaction, significantly affected the phytochemicalcomposition of wheat grains for most of the analysed bioactivecompounds. Some of the investigated old varieties, namelyGentil rosso, Marzuolo d’aqui and Verna, showed an intriguingphytochemical profile, with the higher amounts of dietaryfibre and antioxidant compounds (polyphenol, flavonoid). Thepresent findings highlight that ancient wheat genotypes maybe successfully used in breeding programmes aimed to selectvarieties suitable for low-input farming and rich in health-promoting compounds.

ACKNOWLEDGEMENTSThe authors are grateful to the Emilia Romagna region, Italy(Project ‘‘BioPane’’, CUP J31J09000430002) and the Italian Ministry

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of Agriculture (Project ‘‘Pane della salute’’ MiPAF-OIGA, CUPJ31J09000370001) for financially supporting the research.

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