Cover Crop Short-Term Effects on Soil NO3 --N Availability, Nitrogen Nutritional Status, Yield, and Must Quality in a Calcareous Vineyard of the AOC Rioja, Spain

Communications in Soil Science and Plant Analysis, 44:711–721, 2013Copyright © Taylor & Francis Group, LLCISSN: 0010-3624 print / 1532-2416 onlineDOI: 10.1080/00103624.2013.748122

Cover Crop Short-Term Effects on Soil NO3−-N

Availability, Nitrogen Nutritional Status, Yield, andMust Quality in a Calcareous Vineyard of the AOC

Rioja, Spain

EVA PILAR PÉREZ-ÁLVAREZ, JOSÉ LUIS PÉREZ-SOTÉS,ENRIQUE GARCÍA-ESCUDERO,AND FERNANDO PEREGRINA

Instituto de Ciencias de la Vid y del Vino (CSIC), Universidad de la Rioja,Gobierno de La Rioja, Servicio de Investigación y Desarrollo TecnológicoAgroalimentario (CIDA), Logroño, Spain

Cover crop use in vineyards can affect both vine vigor and must and wine qualitybecause of the competition for soil nutrients and water. Our objective was to studythe short-term effects of a cover crop on the nitrate (NO3

−)–nitrogen (N) availabil-ity throughout the grapevine vegetative cycle, the grapevine and cover crop N uptake,and the yield and must quality. By short-term effects we mean the first crop cycle afterplanting the cover crop. The experiment was set in 2009 on a cv. Tempranillo vineyardplanted in a Oxyaquic Xerorthent soil. The soil had not been fertilized with N since2000, and two types of soil management were studied: (1) conventional tillage (CT) and(2) barley (Hordeum vulgare L.) cover crop (B). Soil samples were taken in March (budbreak), June (bloom), July (setting), and August (veraison) of 2009, and the extractableNO3

−-N was determined. At bloom and veraison, N contents in both blade and petiolewere determined. At bloom the grapevine N uptake was estimated using the aerial parts(leaves, shoots, and bunches), and for the cover crop N uptake was determined. Totalyield, bunch, and shoot weight as well as must anthocyanin and polyphenol contentswere determined. Soil NO3

−-N availability decreased in the cover crop from June untilAugust, with the reduction being similar to the cover crop N uptake. Also N contents inboth petiole and blade decreased in the cover crop at veraison. Regarding must qual-ity, the cover crop increased the anthocyanin content. The reduction of soil NO3

−-Navailability throughout the vegetative cycle of grapevine caused a reduction in bothN nutritional status and grapevine vigor, and therefore cover crops could affect mustquality in the first harvest after the cover crop was sown.

Keywords Barley cover crop, NO3−-N availability, vineyard soil

Introduction

The Rioja wine-growing region has approximately 60,000 ha of vineyards. Climate isMediterranean with continental influences; summers are dry although intense storms canbe recorded. Tillage is the usual soil management in Rioja. Generally vineyard soils have

Address correspondence to Fernando Peregrina, Instituto de Ciencias de la Vid y del Vino(CSIC, Universidad de la Rioja, Gobierno de La Rioja), Servicio de Investigación y DesarrolloTecnológico Agroalimentario (CIDA), Logroño, Spain. E-mail: [email protected]

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712 E. P. Pérez-Álvarez et al.

poor organic-matter contents (<1%), their texture is loamy (Peregrina et al. 2010b), andthey present a medium-high risk of overcrusting according to the FAO-PNUMA index(Fao 1980). Consequently, in La Rioja vineyard soils are highly susceptible to erosion. In arecent study Peregrina et al. (2010a) indicated that the use of permanent cover crops invineyards of the La Rioja region could reduce the erosion risk and improve soil quality.

However, cover crops in vineyard can have a potential negative effect as they decreaseboth vine vigor and yield (Tesic, Keller, and Hutton 2007; Ripoche et al. 2011), althoughthese effects can vary according to soil fertility, cultivar, and the cover crop species used.In spite of this, cover crop effects that could be considered drawbacks may be judgedbeneficial; for example, in fertile soils cover crop can reduce the excess vigor unfavorablefor high-quality wine production (Smart et al. 1991). This effect could be caused by waterand nutrient competition. Various authors have reported that cover crops reduced soil Ninorganic availability (Steenwerth and Belina 2008a; Smith et al. 2008; Celette, Findeling,and Gary 2009; Ripoche et al. 2011), and as N is the main nutrient that regulates grapevinevigor (Conradie 2001) this effect in the soil N availability can explain the reduction invigor reported in vineyards with cover crops. The reduction of soil N availability can bedue to various factors such as the N uptake by the cover crop, the changes that covercrops produce in the conditions affecting organic matter mineralization (Celette, Findeling,and Gary 2009), and the increment of the NO3

−-N recycled by the soil microorganisms(Steenwerth and Belina 2008a).

Furthermore, N availability can affect must quality even when other aspects such asvine vigor, yield, and/or berry size are not significantly affected. Various authors havereported a reduction in the concentrations of anthocyanins and/or polyphenols in mustsfrom plants that had been fertilized with N (Keller and Hrazdina 1998; Hilbert et al. 2003;Delgado et al. 2004) or plants with higher N status (Choné et al. 2001; Tregoat et al. 2002).

On the other hand, most studies have not taken into account the agrosystem’s dynamicsthroughout the transition phase (i.e., the first crop cycle with cover crop). This transition isimportant because yield is influenced by the conditions occurring during both the currentand the previous crop cycle (Boss et al. 2002). Thus, very scarce information exists aboutthe short-term effect of the cover crops on yield and must quality.

Finally, a great majority of the studies on the effect of cover crops in vineyards havebeen carried out in regions where the climatic conditions were different than those in theregion of La Rioja. Besides this, very little research has been done on the effect of per-manent cover crops on the Tempranillo cultivar, which is the most important in the RiojaAOC.

Within the general objectives of optimizing cover crop utilization and improving thequality of the wine produced in the Rioja AOC, this study assesses the effect, in the firstgrapevine cycle, of the cover crop on the soil NO3

−-N availability levels, the N grapevinenutritional status, yield, and must quality.

Materials and Methods

Experimental Design and Vineyard Plot Description

The experiment was set up in 2009 in a vineyard plot located on a terrace of the RiverNajerilla in the municipality of Nájera (La Rioja, Spain) (latitude 42◦ 26´ 34.18´´ N;longitude 2◦ 43´ 32.31´´ W). The field slope was about 0.2% with west–east orientation.The soil was classified as Oxyaquic Xerorthent (Soil Survey Staff 2006). Additional detailsfor soil characteristics are shown in Table 1. The climate in the area is semi-arid according

Cover Crop Short-Term Effects 713

Table 1Soil characteristics of the Ap horizon

Parameter Value

Soil classification(Soil Survey Staff 2006) Oxyaquic XerorthentAp horizon depth (cm) 15pH (H2O, 1:5) 8.5EC 1:5 (dS m−1) 0.11Organic matter (%) 1.24Carbonates (%) 4.2Color Munsell (dry) 10 YR 5/4

Particle size distribution (%)Sand (2000–50 µm) 38.3Silt (50–2 µm) 43.2Clay (<2 µm) 18.5

to the UNESCO aridity index (UNESCO 1979). Meteorological data were recorded at ameteorological station installed near the experimental plot (latitude 42◦ 27′ 42.75´´ N; lon-gitude 2◦ 42′ 45.66′′ W at altitude of 465 m above mean sea level). For 2009, the annualprecipitation was 409.6 mm, average annual temperature was 12.6 ◦C, and annual potentialevapotranspiration (FAO-Penman) was 1003.9 mm.

The selected vineyard had been planted in 1999 with Vitis vinifera L., cv. Tempranillo,grafted on 110-Ritcher rootstock. Vine and row spacings were 1.3 and 2.7 m, respectively,rows had a west–east orientation, and the planting density was 2,850 plants ha−1. Vineswere trained to a vertical shoot position trellis system on a Double Cordon Royat and werespur pruned to 12 buds per vine. All water shoots were removed in the spring.

The experimental design was a randomized complete block with two treatments andthree replications per treatment. Each replicate (plot) had four rows with 60 vines in each.Two types of soil cover management were studied: CT, conventional tillage between rows,and B, cover crop of barley Hordeum vulgare L. cv. Naturel, sown with a rotary seed drillin February 2009.

The treatments had not received any N fertilization since 2000. In CT treatment, soiltillage was carried out approximately once every 4 to 6 weeks, at 0–15 cm deep.

In all the treatments, a 0.8-m-wide strip was kept using herbicide beneath the vinesthus allowing a cover crop width of 1.7 m. Other management practices were similar in alltreatments; pest and disease control and grapevine canopy management were carried outmechanically except for harvesting, which was done manually.

Nitrogen Grapevine Nutritional Status

At bloom (17 June) and veraison (9 August), 30 leaves were sampled from nodes oppositethe first bunch at bloom and opposite the second bunch at veraison. In each leaf, blades andpetioles were separated, washed with tap water, and rinsed with distilled water. The plantmaterial sample was dried at 60 ◦C in a forced-air oven for 72 h, ground, and sieved to0.5 mm in an ultracentrifugal mill (Retsch ZM1; Haan Germany). Nitrogen concentrationin leaf blades and petioles was determined with a CNS analyzer elemental (TruSpec


CN, Leco Inc., St. Joseph, Mich.). Concentrations were expressed on a dry-weight basis(Romero, García-Escudero, and Martín 2010).

Grapevine Agronomic Performance

Grapevine performance was assessed by measuring yield, bunch number, and their totalweight per vine at harvest (all treatments were hand harvested on 7 October). In eachtreatment 20 grapevines were hand pruned in December, and their canes were collected todetermine the shoot weight.

Grapevine and Barley Nitrogen Uptake

Grapevine N uptake was determined in the aerial parts produced during each growingseason. Five shoots from five grapevines were sampled at random in each plot, and eachsample was divided into three parts: stems, leaves, and fruits. Plant material was washedwith tap water, rinsed with distilled water, and dried at 60 ◦C for 72 h to calculate its drybiomass. The samples of dry mass of shoots, leaves, and bunches were used to measure theN content determined by combustion analysis with CNS elemental analyzer (TruSpec CN,Leco Inc., St Joseph, Mich.). The total N uptake by grapevine was expressed as N kg ha−1

considering the plant density.The N uptake of barley cover crop was estimated in the aerial organs. Aboveground

cover crop and weed biomasses were collected from randomly placed quadrants (0.5 m ×0.5 m; n = 2 per plot). Plant material was cut at soil level and then taken to the laboratorywhere it was washed with tap water, rinsed with distilled water, and dried at 60 ◦C for 72 h.The dry biomass was weighed and ground through a 0.5-mm sieve with an ultracentrifugalmill (Retsch ZM1). Nitrogen concentration was determined by combustion analysis withCNS elemental analyzer. Nitrogen uptake by cover crop was expressed as N kg ha−1.

Nitrate-N Content throughout the Growing Season

In each plot, soil samples were collected at 0–15 and 15–40 cm deep in the interrow. Foursamplings were carried out on 22 April (bud break), 18 June (bloom), 16 July (setting), and10 August (veraison) of 2009. The composite samples of three subsamples were collectedwith an Edelman-type auger.

Samples were air dried and the soil was ground and sieved to 2 mm. Soil coarsefragments (>2 mm) percentage was determined in each sample. NO3

−-N was extractedwith a calcium sulfate saturated solution and determined by the second derivative method(Sempere, Oliver, and Ramos 1993). Nitrate-N content in kg ha−1 was estimated using thesoil bulk density at 0–15 and 15–45 cm deep as determined by the core method (Grossmanand Reinsch 2002) and considering the soil percentage of coarse fragments (>2 mm).

Must Quality Analysis

At harvest (7 October), fruit composition was evaluated using a randomized sample of500 berries in each plot. Must was obtained from 200 berries that were weighed andcrushed. Total polyphenol index (TPI) was determined by spectrophotometry measuringultraviolet absorption at 280 nm (Ribéreau-Gayon et al. 1972). An additional 300 berrieswere treated for 30 min with 1% hydrochloric acid (HCl) at 60 ◦C. In this extract, berry


skin anthocyanins were determined using the sodium bisulfite discoloration method andwere expressed as berry weight (Ribéreau-Gayon and Stonestreet 1965).

Statistical Analysis

Treatment effects on measured variables were tested using ANOVA (univariate linearmodel), and comparisons between treatment means were made using the least significantdifference (LSD) multiple range test calculated at P < 0.05. Statistical procedures werecarried out with the software program Statgraphics Plus for Windows (1998) (StatgraphicsPlus, Rockville, Md.).

Results

Conventional tillage treatment had greater soil NO3−-N content than B from bloom (June)

to veraison (August) (Figure 1). Soil NO3−-N content in CT showed an increasing trend

throughout the growing cycle and especially in the months July and August; in contrastB did not show the same pattern. Various authors have reported decreases in the soilNO3

−-N content under cover crop in Californian Mediterranean vineyards (Smith et al.2008; Steenwerth and Belina 2008a); also Celette, Findeling, and Gary (2009) foundlower N inorganic in a French Mediterranean vineyard. In addition, Ripoche et al. (2011)reported a reduction of soil N inorganic at setting in the first season with a cover crop in aMediterranean vineyard of France.

Regarding the N uptake, the B treatment agreed with the results reported by Celette,Findeling, and Gary (2009) in a Mediterranean vineyard. The N uptake by B was similar tothe reduction in the soil NO3

−-N (Table 2). Furthermore, N uptake by the cover crop wastwofold less than grapevine N uptake at bloom. Therefore, in our conditions one of causes

Figure 1. Soil NO3−-N contents in the 0- to 45-cm soil depth during the vegetative cycle (bud break,

bloom, setting, and veraison) in the conventional tillage (CT) and barley cover crop (B) treatments.Different letters indicate significant differences between treatments with LSD test (P < 0.05). Barsrepresent standard error.


Table 2Barley cover crop nitrogen uptake at bloom and fruit setting

Parameter Bloom (22 June) Setting (22 July)

N uptake (kg ha−1) 13.80 ± 4.72a 15.90 ± 4.72a

aStandard error.

for the grapevine N uptake reduction could be the competition of the barley cover crop forthe soil NO3

−-N. Various factors could explain the reduction of soil NO3−-N availability

under the cover crops. Celette, Findeling, and Gary (2009) suggested that the soil waterreduction due to the cover crop water consumption can reduce the microbial activity andtherefore the organic-matter mineralization. Steenwerth and Belina (2008b) indicated alsothat the increment of soil microbial activity under the cover crop can immobilize NO3

−-Nwhereas under conventional tillage the soil microorganism activity is limited by the lowercontent of soil organic C.

With respect to the grapevine N nutritional status, at veraison the N content in petioleand blade decreased in the B treatment with respect to CT. At bloom in B treatment, Ncontent in both blade and petiole tended to decrease (Figure 2). Tesic, Keller, and Hutton(2007) and Ingels et al. (2005) found that N content in leaf tissues decreased after variousseasons with cover crops in an Australian and a Californian vineyard, respectively. In thisstudy, at veraison, the N reduction was greater than in bloom because the grapevines haddeveloped actively in this period and therefore the N uptake increased.

The N uptake by aerial grapevine parts at bloom showed a decreasing trend in Brespect to CT (Figure 3). Ripoche et al. (2011) found a decrease in the N uptake in thefirst season with a cover crop. These results agreed with the reduction in the N nutritionalstatus in the cover crop treatment.

Nitrogen nutritional status and N uptake of the grapevine are important because theyare related to the potential quality of the berries. Choné et al. (2001) and Tregoat et al.(2002) showed that vines with lower N status under similar water uptake conditions

Figure 2. Blade and petiole N content at bloom and verasion, in conventional tillage (CT) and barleycover crop (B) treatments. Different letters indicate significant differences between treatments usingthe LSD test (P < 0.05). Bars represent standard error.


Figure 3. N uptake by aerial grapevine parts at bloom in conventional tillage (CT) and barley covercrop (B) treatments. Different letters indicate significant differences between treatments using theLSD test (P < 0.05). Bars represent standard error.

have more anthocyanin and polyphenols than vines with greater N status in cv. Merlotgrapevines.

Therefore the lower soil NO3−-N availability in the soil from bloom to veraison in B

treatment caused a lower N nutritional status and N uptake of the grapevine.Regarding grapevine performance, the shoot weight (Figure 4) tended to decrease in

the B treatment (significantly at P = 0.88 with Tukey’s test). Various authors found lower

Figure 4. Shoot weight, grapevine yield, and bunch weight in conventional tillage (CT) and barleycover crop (B) treatments. Different letters indicate significant differences between treatments usingthe LSD test (P < 0.05). Bars represent standard error. Asterisks indicate significant differencesbetween treatments using the LSD test (P < 0.12).


grapevine vigor after various seasons with cover crops, including Ingels et al. (2005) ina California vineyard with cv. Merlot, Wheeler, Black, and Pickering (2005) in a NewZealand vineyard with cv. Cabernet Sauvignon, and Tesic, Keller, and Hutton (2007) in anAustralia vineyard with cv. Chardonnay. As soil NO3

−-N decreased in B, this result agreeswith Conradie (2001), who found that grapevine vigor is stimulated principally by the soilN availability.

Grapevine yield and bunch weight (Figure 4) presented no differences between B andCT. Similar results were reported after various season with cover crop by Wheeler, Black,and Pickering (2005) in a New Zealand vineyard of cv. Cabernet Sauvignon, Smith et al.(2008) in a California vineyard of cv. Chardonnay, Lopes et al. (2008) in a Portuguesevineyard of cv. Cabernet Sauvignon. Also, Bahar and Yasasin (2010) found similar resultsafter the first season with cover crop in a cv. Cabernet Sauvignon Turkish vineyard andRipoche et al. (2011) for cv. Cabernet Sauvignon with a reduction of yield only afterthe second season with a cover crop. However, the competition between cover crop andgrapevine has also resulted in a reduction in yield after various seasons with a cover crop(Tesic, Keller, and Hutton 2007; Ripoche et al. 2011). According to Ripoche et al. (2011),the absence of effect on the yield could be due to the floral initiation that occurs during theprevious season in latent buds. Therefore, the effect in the yield after the first season withcover crop would not be a good estimator of the yield in the next season with a cover crop.

Regarding must quality, the B treatment increased the anthocyanin content withrespect to CT (Figure 5). Furthermore, the polyphenol content tended to increase in Bthough there were no significant differences between treatments (Figure 5). These resultsagree with the increments in berry anthocyanin and polyphenol content with cover cropreported by Wheeler, Black, and Pickering (2005), Lopes et al. (2008), and Bahar and

Figure 5. Anthocyanin content in berries and total polyphenol index (ABS 280 nm) for conventionaltillage (CT) and barley cover crop (B) treatments. Different letters indicate significant differencesbetween treatments using the LSD test (P < 0.05). Bars represent standard error.


Yasasin (2010). They also agree with the results presented by Xi et al. (2010), who foundincrements of phenolic compounds with cover crops in a Chinese vineyard. The covercrop competition for soil N could explain these results; in fact high N supply inhibitedthe synthesis of anthocyanins in cv. Merlot grapevines (Hilbert et al. 2003). A similarresult was reported by Keller and Hrazdina (1998) on continuously irrigated potted cv.Cabernet Sauvignon plants, and in cv. Tempranillo, Delgado et al. (2004) found that Nsupply decreased the phenolic compound accumulation in grape skin. Also, consideringdifferent vineyard soils, low vine N nutritional status induced high berry anthocyanin andpolyphenol content in cv. Cabernet Sauvignon (Choné et al. 2001) and cv. Merlot (Tregoatet al. 2002) in the Bordeaux region (France). Therefore, in this study the increment ofanthocyanin and polyphenol could be due to the reduction in soil NO3

−-N availability inthe B treatment.

Conclusions

The soil NO3−-N availability under conventional tillage increased throughout the

grapevine growing cycle, with a maximum at verasion. Therefore, conventional tillage hada high capacity for N mineralization.

In the first season with a barley cover crop, the soil NO3−-N availability decreased

throughout the vegetative growing season of the grapevines with respect to the conventionaltillage; this effect could be due principally to the barley cover crop N uptake.

Consequently, because of the reduction of the soil NO3−-N in the barley treatment,

both the grapevine N nutritional status and the grapevine vigor decreased. However, theyield parameters were not affected and must quality improved as a result of the greatercontent in the anthocyanin and polyphenols.

Therefore, barley cover crops sown in calcareous vineyard of the Rioja region can beused to improve the must quality in the short term.

Acknowledgments

We thank Juan Antonio Leza and Ricardo Leza for providing the vineyard site to conductthis study. Also, we thank the staff of the CIDA and Laboratorio Regional, especially ClaraLarrieta and Mikel Colina. This study was supported by a predoctoral contract for E. P.Pérez-Álvarez funded by the INIA and the European Social Fund, postdoctoral contract forF. Peregrina funded by the INIA and the European Social Fund), and by Grants R-07–08(from Autonomous Government of La Rioja) and RTA 2009-00101-00-00 (from INIA andFEDER funds).

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Cover Crop Short-Term Effects on Soil NO3 --N Availability, Nitrogen Nutritional Status, Yield, and Must Quality in a Calcareous Vineyard of the AOC Rioja, Spain

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