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Scientia Horticulturae 111 (2007) 319–325 www.elsevier.com/locate/scihorti Effect of organic and inorganic fertilizers
Scientia Horticulturae 111 (2007) 319–325 www.elsevier.com/locate/scihorti Effect of organic and inorganic fertilizers

Scientia Horticulturae 111 (2007) 319–325

Scientia Horticulturae 111 (2007) 319–325 www.elsevier.com/locate/scihorti Effect of organic and inorganic fertilizers

www.elsevier.com/locate/scihorti

Effect of organic and inorganic fertilizers applied during successive crop seasons on growth and nitrate accumulation in lettuce

Georgios C. Pavlou a, * , Constantinos D. Ehaliotis b , Victor A. Kavvadias a

a National Agricultural Research Foundation (N.AG.RE.F.), Olive and Horticultural Crops Institute, Lakonikis 87, Kalamata 24100, Greece b Agricultural University of Athens, Soil Science and Agricultural Chemistry Lab., Iera Odos 75, Athens 11855, Greece

Received 23 December 2005; received in revised form 10 August 2006; accepted 6 November 2006

Abstract

A romaine-type lettuce ( Lactuca sativa L.) cv. Corsica was cultivated during three successive crop seasons (late-spring, late-autumn and late- winter) in the same soil of an experimental greenhouse in S.W. Peloponnese, Greece. Seven long-term fertilization treatments were tested for their effect on plant growth and nitrate concentration in the external lettuce leaves. Treatments included: three different doses of organic fertilization (composted sheep manure) applied at the start of each crop season, three different doses of inorganic N fertilization applied via fertigation during each crop season, and a control treatment in which no fertilizer was applied. A drip irrigation system was used to water all plants. The highest nitrate levels were observed in the medium and maximum inorganic fertilization treatments (572–664 mg kg 1 ) in all crop seasons. They were significantly higher compared to the respective organic fertilization treatments (253–435 mg kg 1 ) and all other fertilization treatments (148– 435 mg kg 1 ). Crop season affected lettuce growth more than nitrate accumulation in the lettuce leaves: lettuce biomass production was the smallest and most uniform in the late-autumn season and did not respond to the fertilization treatments tested (ranging from 409 to 439 g plant 1 ), while in the late-spring season biomass production was the highest and most variable (561–841 g plant 1 ), it correlated with nitrate concentration in the leaves and in the medium and maximum inorganic fertilizer doses it significantly exceeded production from all other fertilization treatments (827–841 g plant 1 ). Following the three crop seasons the residual availability of N, P and K was clearly enhanced in the soil receiving the organic compared to the inorganic fertilization. Nitrate concentration in lettuce leaves was far below the upper limits set by the European Commission in all fertilization treatments throughout the three crop seasons, a result attributed mainly to the sufficient level of light intensity and duration throughout the year in Southern Greece. # 2006 Elsevier B.V. All rights reserved.

Keywords: Lettuce; Lactuca sativa L.; Fertilization; Organic fertilizers; Manure; Nitrates

1. Introduction

In the temperate zone regions lettuce is cultivated throughout the year, but production and quality characteristics benefit from cool weather and high light intensity. The concentration of nitrates in the edible leaves of lettuce are regulated by the European Commission Regulation No 563/ 2002 which has set upper limits in order to protect consumers from potential toxicological risks following the consumption of nitrate-rich foods (Maynard et al., 1976; Walker, 1990; Bruning-Fann and Kaneene, 1993). Recent studies however, report on beneficial effects of nitrates mainly related to the

* Corresponding author. Tel.: +30 27210 29812; fax: +30 27210 27133. E-mail address: gpavlou.kal@nagref.gr (G.C. Pavlou).

0304-4238/$ – see front matter # 2006 Elsevier B.V. All rights reserved.

control of the gut flora (Lundberg et al., 2004; Addiscott and Benjamin, 2004). Accumulation of nitrates results from an imbalance between the uptake and translocation of nitrates by the xylem, and the reduction of these nitrates to ammonia which is subsequently rapidly incorporated into amino acids (Maynard et al., 1976). However, the internal nitrate concentration in the plant seems to be controlled by a self-regulatory mechanism exerted either by negative feedback control on the net nitrate uptake rate (Cardenas-Navarro et al., 1998) or by passive control on nitrate efflux (Scaife, 1989). The viewpoint that non-structural carbohydrates and nitrates have a complementary role in maintaining cell tugor (Blom-Zandstra and Lampe, 1985; Behr and Wiebe, 1988) offers a credible model for the plant nitrate regulation mechanism suggesting the accumulation of nitrates in the vacuole as an alternative osmoticum under low radiation

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conditions (Blom-Zandstra, 1989; Buwalda and Warmenhoven,

1999).

Accumulation of nitrates in lettuce has been shown to be affected by the soil texture and the source of fertilizer-N (Scaife et al., 1986; Gianquinto et al., 1992; Gunes et al., 1995), the NH 4 -N-to-NO 3 -N fertilizer-N ratio (McCall and Willumsen, 1998; Demsar and Osvald, 2003), the timing of fertilizer-N release (Tesi and Lenzi, 1998), the light intensity and duration (Behr and Wiebe, 1992; Chadjaa et al., 1999; Drews et al., 1995; Gaudreau et al., 1995), crop season (Gianquinto et al., 1992), and lettuce type and cultivar (Blom-Zandstra and Eenink, 1986; Siomos, 2000; Escobar-Gutierrez et al., 2002). In most types of lettuce, including the romaine type, the highest concentration of nitrates is normally observed in the external leaves (Corre and Breimer, 1979; Santamaria et al., 1999; Abu- Rayyan et al., 2004). In countries of the Mediterranean basin, sheep manures are traditionally used as an organic fertilization source. Their recycling into soils low in organic matter, which are widespread in these regions, could benefit soil structure and long term fertility, and is also an alternative to inorganic fertilizers in the growing organic vegetable production business. In this work we compare short and long term effects of organic versus inorganic sources of N on the growth and the accumulation of nitrates in the external leaves of a romaine lettuce. Composted sheep manure is used for organic fertilization whereas standard fertigation applied by farmers in the area is used for the comparison to inorganic fertilization. Fertilization from both sources is applied at a minimum, a medium and a maximum dose during three successive crop seasons in the same soil. Effects of crop season and long term cumulative fertilization on plant and soil are studied.

2. Materials and methods

2.1. Location and design of experiments

The experiments were carried out during three successive crop seasons: late spring (first, 6 April–17 May 2000), late

autumn (second, 26 October–5 December 2000), and late- winter (third, 6 February–28 March 2001), at the N.AG.RE.F.- Olive and Horticultural Crops Institute of Kalamata, S.W.

Peloponnese, Greece (37 8 03 0 N, 22 8 07 0 E, altitude 9 m). At the start of each crop season, a romaine lettuce crop cv. Corsica (Royal Sluis) was established in a polyethylene covered greenhouse (80–85% transparency to visible irradiation), with 60% side wall cover. The greenhouse soil was a sandy clay loam (SCL) with chemical characteristics presented in Table 1. Seven long-term fertilization treatments were compared including: (i) three different organic fertilization treatments ( O-min , O-mid , O-max ) consisting of composted sheep manure (Table 1) applied at the start of each cultivation period, at doses of 1–3 l plant 1 respectively for the first and second crop seasons, and reduced by 33% for the third crop season (0.67, 1.34, and 2.01 l plant 1 , respectively), (ii) three different treatments of inorganic-N fertilization ( I- min , I-mid , I-max ) applied via drip irrigation during each crop season at doses corresponding to 87, 174 and

261 mg N plant 1 fertigation 1 , respectively (Table 2),

and (iii) a control treatment in which no fertilizer was applied ( C ). The nutrient elements provided in total during each crop season by the organic and inorganic fertilization treatments are shown in Table 3. In all three inorganic fertilization treatments (I-min, I-mid, I-max) the soluble commercial fertilizers 15–30–15 and 20–20–20 plus micro- nutrients were applied, each at a rate of 0.25 g plant 1 fertigation 1 . Extra N fertilization was applied for the I-mid and I-max treatments with the inorganic fertilizer 34.5–0–0 (NH 4 NO 3 ) at rates of 0.25 and 0.5 g plant 1 fertigation 1 , respectively. Each experimental plot consisted of 54 lettuce plants placed in three rows (30 cm apart) of 18 plants (25 cm apart). The plants of the two outer rows and the three plants in each end of the middle row were kept as guard plants. There were five replicate plots per treatment and the experiment was set up as a randomized complete block design.

Table 1 Chemical characteristics of soil and sheep manure used in the experiments

 

E.C. (mS cm 1 )

pH

CaCO 3 (%)

O.M. (%)

C.E.C. (cmol (+) kg 1 )

N

Kjeldahl

P

Olsen

K

exch

 

(g

kg 1 )

(mg kg 1 )

(mg kg 1 )

Soil

2.6

7.4

13.4

1.4

8.6

0.15

37.0

118.2

Manure a

21.7

8.0

8.4

72.0

36.0

1.64

 

1.4 (% d.w.)

1.8 (% d.w.)

a Manure dry weight = 201 g l 1 .

Table 2 Nutrient elements provided per fertigation by the inorganic fertilization treatments applied to lettuce

Fertilization

Nutrient elements provided in each crop season

 

treatment

 

N

(g plant 1

P

(g plant 1

K

(g plant 1

Mg (mg plant 1 fertigation 1 )

Fe (mg plant 1 fertigation 1 )

Mn (mg plant 1 fertigation 1 )

Zn (mg plant 1 fertigation 1 )

Cu

fertigation 1 )

fertigation 1 )

fertigation 1 )

I-max

0.261

0.027

0.036

0

0.195

0.081

0.0163

0.0163

I-mid

0.174

0.027

0.036

0

0.195

0.081

0.0163

0.0163

I-min

0.087

0.027

0.036

0

0.195

0.081

0.0163

0.0163

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321

Table 3 Nutrient elements provided per crop season by the fertilization treatments applied to lettuce

Crop season

Fertilization treatment

Macro-elements (g plant 1 )

 

Micro-elements (mg plant 1 )

 
 

N

P

K

Mg

Fe

Mn

Zn

Cu

First and second

O-max

9.840

0.258

4.692

0.102

13.53

13.11

6.69

4.71

O-mid

6.560

0.172

3.128

0.068

9.02

8.74

4.46

3.14

O-min

3.280

0.086

1.564

0.034

4.51

4.37

2.23

1.57

Third

O-max

6.593

0.173

3.144

0.068

9.07

8.78

4.48

3.16

O-mid

4.395

0.115

2.096

0.046

6.04

5.86

2.99

2.10

O-min

2.198

0.058

1.048

0.023

3.02

2.93

1.49

1.05

First

I-max

3.380

0.356

0.473

0

2.54

1.06

0.21

0.20

I-mid

2.260

0.356

0.473

0

2.54

1.06

0.21

0.20

I-min

1.140

0.356

0.473

0

2.54

1.06

0.21

0.20

Second

I-max

2.080

0.218

0.291

0

1.56

0.63

0.13

0.13

I-mid

1.390

0.218

0.291

0

1.56

0.63

0.13

0.13

I-min

0.700

0.218

0.291

0

1.56

0.63

0.13

0.13

Third

I-max

2.860

0.301

0.398

0

2.15

0.90

0.18

0.18

I-mid

1.910

0.301

0.398

0

2.15

0.90

0.18

0.18

I-min

0.960

0.301

0.398

0

2.15

0.90

0.18

0.18

2.2. Cultivation and irrigation practices

Until around 10 days after transplanting, plants of all treatments were irrigated with water only (0.2–

0.4 l plant 1 irrigation 1 ). Afterwards, when plants were well

established in the soil, fertigation was applied with water

quantities of

concentration in the irrigation water ranged between 0.5 % and 1 % . When plants had reached approximately their highest commercial size they were harvested by cutting the stems at the soil level. Twelve plants from each plot were harvested soon after sunrise on a typical for each season day.

0.5–0.6 l plant 1 fertigation 1 . The fertilizer

The outline of practices and climatic conditions are presented in Table 4. Lettuce seedlings were pre-grown in 190 ml plastic pots filled with the organic substrate Terra Nature (Holland). They were transplanted in the greenhouse at the five to seven true-leaf stage. Organic fertilizer (sheep manure) had already been incorporated into the soil of the respective plots down to a depth of about 20 cm, 2–3 days before transplantation using a milling machine. Inorganic fertilizers were dissolved in 30 l water tanks and injected by an electric pump at a pressure slightly greater than 1.5 atm separately in each of the three 30 m long primary pipes, from where the plants of the three inorganic fertilization treatments were fertigated by 5 m long secondary dripper- pipes. Plants of control and organic fertilization treatments were irrigated through separate pipes with water only. Every plant was irrigated by a labyrinth type dripper (Netafim) supplying about 3 l h 1 at a pressure of 1 atm. The irrigation water used had the following characteristics:

electrical conductivity 686 m S/cm (at 25 8 C), pH 7.29, S.A.R. 2.7, HCO 3 409 mg l 1 , Ca 2+ 52.1 mg l 1 and Mg 2+

2.3. Analytical methods

Just after harvest the fresh weight (f.w.) of the plants was determined; then four external leaves from each plant were removed and their f.w. was also determined. The leaves were then washed off with tap water and blotted dry on absorbing paper. Their dry weight (d.w.) was subsequently determined following drying in a drying chamber at 70 8C for 60–70 h. Dry leaves were ground to 40-mesh size by a centrifugal mill (Tecator—Sweden). Homogenized leaf powder was reinserted to the drying chamber at 70 8 C for about 24 h, and 0.225 g were

12.2 mg l 1 .

Table 4 Cultivation and irrigation practices and climatic conditions during the three successive crop seasons: late-spring (first), late-autumn (second) and late-winter (third)

 

First crop season

Second crop season

Third crop season

Lettuce transplanting Date of harvesting Crop duration (days) Irrigation water (l plant 1 ) total Total number of irrigations Number of fertigations Sunshine duration (h) total Average photoperiod (h) Average air min temperature ( 8C) Average air max temperature ( 8C)

6 April 2000 17 May 2000

26 October 2000 5 December 2000

6 February 2001 28 March 2001

41

39

50

12.2

6.9

7.7

26

12

17

13

8

11

354

229

329

13.53

10.27

11.47

13.3

11.8

8.8

32.7

29.5

27.7

322

G.C. Pavlou et al. / Scientia Horticulturae 111 (2007) 319–325

Table 5 Yield of lettuce (fresh weight) grown under different organic (O) or inorganic (I) fertilization schemes during three successive crop seasons: late-spring (first), late- autumn (second) and late-winter (third)

Fertilization treatment

First crop season (g plant 1 )

Second crop season (g plant 1 )

Third crop season (g plant 1 )

I-max

841 a

439 a

625 a

I-mid

827 a

445 a

624 a

I-min

689 b

441 a

534 ab

O-max

701 b

443 a

569 ab

O-mid

670 bc

432 a

536 ab

O-min

561 cd

409 a

504 b

Control

541 d

335 b

262 c

Means in the same column (crop season) followed by different letters denote significant differences according to Duncan’s multiple range test ( p = 0.05).

used for colorimetric determination of nitrate-N (NO 3 -N) at 410 nm by a Hitachi U-2001 spectrophotometer following the nitration by salycilic acid method described by Cataldo et al. (1975). Five replicate subsamples were measured to obtain each plot-sample value. Randomly selected samples were also cross- checked for NO 3 -N content with the copperized-Cd reduction method (Keeney and Nelson, 1982) to confirm methodological accuracy. Length and width of the biggest leaf on each of the 12 plants under harvest were measured the day before harvest. Air temperature at plants’ height was recorded by a Lambrecht thermograph, while sunshine duration was obtained from the Kalamata Meteorological Station of the National Meteorological Service. Photoperiod was calculated using the approximate local sunrise and sunset hours. Soil available P was estimated by extraction with NaHCO 3 (Olsen-P) and measured colorimetrically at 880 nm (Hitachi U- 2001) following the ascorbic acid molybdate reduction method (Olsen and Sommers, 1982). Soil available K (exchangeable plus water soluble) was estimated following extraction with NH 4 OAc (Thomas, 1982) and determined by flame photometry (Coring 410). The anaerobic incubation index of soil-N availability was estimated by measuring the NH 4 -N production from soil samples incubated anaerobically for 1 week at 40 8 C according to Keeney (1982).

2.4. Statistical analysis

Analysis of variance was performed to evaluate differences in leaf length, leaf width, plant fresh weight, and nitrate

concentration in the external leaves of lettuce. Treatments were thereafter compared by Duncan’s multiple range test ( p = 0.05). Simple linear regression was also applied using the SPSS v.10 for Windows OS.

3. Results

3.1. Lettuce plant growth

During the first crop season (late-spring) characterized by the greatest temperatures, photoperiod, and sunshine duration (Table 4) high yields were observed in all treatments (Table 5). Lettuce biomass yield in the maximum and medium inorganic fertilization treatments (I-max, I-mid) was significantly greater than in all other treatments (Table 5). The respective organic fertilization treatments (O-max, O-mid) did not differ significantly from the minimum inorganic fertilization treat- ment (I-min) whereas the control resulted in the lowest yield which was similar to the minimum organic fertilization (O-min) treatment only. During the second crop season (late-autumn), characterized by intermediate temperatures and the shortest photoperiod and sunshine duration (Table 4), reduced and uniform yields were observed among treatments (Table 5). Similar lettuce biomass yield was obtained in all treatments apart from the significantly smaller yield of the control (Table 5). During the third crop season (late-winter) characterized by the lowest temperatures and intermediate photoperiod and sunshine duration (Table 4) plant yields were also increased, but did not reach the yields observed in the first (late-spring)

Table 6 Maximum leaf length and width of lettuce plants grown under different organic (O) or inorganic (I) fertilization schemes during three successive crop seasons: late- spring (first), late-autumn (second) and late-winter (third)

Fertilization treatment

First crop season

Second crop season

Third crop season

Length (cm)

Width (cm)

Length (cm)

Width (cm)

Length (cm)

Width (cm)

I-max

32.92 a

21.85 a

31.49 a

16.63 abc

32.30 a

19.40 a

I-mid

32.33 a

21.23 a

32.10 a

17.48 a

32.28 a

19.35 a

I-min

30.41 b

19.42 b

31.53 a

16.94 ab

31.02 ab

17.69 bc

O-max

30.65 b

19.77 b

31.87 a

17.18 ab

31.94 ab

18.59 ab

O-mid

30.16 bc

19.10 bc

31.17 a

16.30 bc

31.22 ab

17.82 bc

O-min

29.24 cd

18.56 cd

30.92 a

15.83 c

30.90 b

17.28 c

Control

29.00 d

18.12 d

29.44 b

14.15 d

28.14 c

12.72 d

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Table 7 Nitrate concentration, on a fresh weight basis, in the external leaves of lettuce plants grown under different organic (O) or inorganic (I) fertilization during three successive crop seasons: late-spring (first), late-autumn (second) and late-winter (third)

Fertilization treatment

First crop season (mg kg 1 )

Second crop season (mg kg 1 )

Third crop season (mg kg 1 )

I-max

623 a

664 a

662 a

I-mid

572 a

588 a

623 a

I-min

314 b

363 bc

372 b

O-max

311 b

435 b

295 b

O-mid

253 bc

412 b

277 b

O-min

186 c

282 c

148 c

Control

71 d

92 d

25 d

Means in the same column (crop season) followed by different letters denote significant differences according to Duncan’s multiple range test ( p = 0.05).

season (Table 5). Similar lettuce biomass yield was obtained in all treatments apart from the minimum organic fertilization (O- min) treatment which resulted in significantly lower yield compared to the maximum and medium inorganic fertilization treatments (I-max, I-mid) but greater yield compared to the control (Table 5). The width and length of the largest leaf in each lettuce plant were measured, as indicators of plant growth. Statistical differences in leaf length among treatments (Table 6) were identical to the lettuce biomass differences (Table 5). Differences in leaf width were analogous to the lettuce biomass differences for the first crop season, but small differentiations were observed in the following seasons (Table 6).

3.2. Nitrate accumulation in leaves

Nitrate concentration in the external leaves did not exceed the 664 mg kg 1 f.w. (Table 7) in all three cultivation periods, even under the maximum fertilization treatments. Similar levels and patterns of nitrate accumulation in the external leaves were observed in all three crop seasons (Table 7). A remarkable increase in nitrate accumulation was observed in the organic fertilization treatments during the late autumn period, characterized by short photoperiod and sunshine duration (Table 4), but specific experimentation is needed to confirm this trend. In all three crop seasons nitrate accumulation in the external leaves of lettuce plants was significantly greater in the

maximum and medium inorganic fertilization treatments (I- max, I-mid) (Table 7); the two respective organic fertilization treatments (O-max, O-mid) followed, together with the minimum inorganic fertilization treatment (I-min). The concentration of nitrates was even lower in the minimum organic fertilization treatment (O-min) except for the O-mid treatment in the first crop season and the I-min treatment in the second crop season (Table 7). Finally the control (non- fertilized) treatment showed the lowest nitrate concentration compared to all fertilization treatments in each season (Table 7). The same nitrate accumulation patterns and treatment differences were obtained by expressing nitrate concentration on a dry weight basis and the data were highly correlated to the per fresh weight data ( r 2 = 0.892, p < 0.01) apparently due to the small variability of the d.w.-to-f.w. ratio at harvest time, indicating that both expressions were valid for analyzing nitrate accumulation data.

3.3. Residual fertilization effects

Following the three successive crop seasons the residual availability of N, P and K in the soil was estimated. Residual N, P and K availability in the soil was generally higher and tended to increase with organic fertilization treatment dose (O-min, O- mid, O-max) compared to control (non-fertilized) plots, but did not respond to inorganic fertilization dose (Fig. 1A–C,

not respond to inorganic fertilization dose ( Fig. 1 A–C, Fig. 1. Residual effect of three

Fig. 1. Residual effect of three different doses of inorganic fertilization (I-max, I-mid, I-min) and organic fertilization (O-max, O-mid, O-min) applied to lettuce plants in three successive crop seasons on the soil-N availability (A), on the soil-P availability (B) and on the soil-K availability (C). Bars with different letters denote significant differences according to Duncan’s multiple range test, p = 0.05.

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respectively). Specifically, the O-max treatment resulted in a

significantly greater index of soil-N availability compared to all

the inorganic fertilization treatments (Fig. 1A), the treatments

O-mid and O-max resulted in a significantly higher P availability compared to all the inorganic fertilization treat-

ments (Fig. 1B), whereas all the organic fertilization treatments significantly exceeded the exchangeable-K values observed in

the inorganic fertilization treatments (Fig. 1C).

4. Discussion

Comparisons between inorganic and organic fertilizer-N sources are hard to perform since there is usually a dramatic difference in N availability from these two sources of N. Comparisons on the basis of similar amounts of total N applied are therefore of limited relevance to agricultural practice, whereas comparisons on the basis of similar N availability are

hindered by the lack of reliable nitrogen release estimates for organic fertilizer sources derived from animal manures (Van Kessel and Reeves, 2002). A general rule of thump assuming 35% of total-N mineralization in the first year following manure application to soil is occasionally used (Klausner, 1997; Van Kessel and Reeves, 2002), but deviations are large and manure-

N mineralization may also significantly depend on soil

temperatures and moisture (Agehara and Warncke, 2005). Our main objective, however, was to compare nitrate accumulation in lettuce cultivated in different seasons under typical organic and inorganic N fertilization regimes. A standard fertilization by inorganic fertilizers or sheep manure

used in the cultivation area was therefore chosen as the standard fertilization treatment, increased by 50% in N content to make the maximum dose treatment and decreased by 50% in N content to make the minimum dose treatment. As sheep manure had a standard elemental synthesis, all other nutrients applied varied in the same order (+50% and 50% for the O-min and O-max, respectively); however, treatment effects on lettuce growth should be attributed mainly to N availability since other nutrients in soil were adequately available and no deficiencies were observed in randomly checked plants (data not presented). General changes in lettuce growth between crop seasons were in line with changes in photoperiod and sunshine duration. This is expected, since photoperiod and sunshine duration differed widely, whereas minimum temperatures differed only slightly and were near optimum during all seasons. High lettuce plant biomass production occurred mainly in the late-spring season associated with long sunshine duration, which is in line with the positive effects that have been observed on the biomass

of lettuce by supplementary light (McCall and Willumsen,

1999) and long photoperiod (Chadjaa et al., 1999). Fertilizer effects on growth were most clearly differentiated in this spring season; on the contrary a poor response of plants to either organic or inorganic N fertilization was observed in the late autumn season indicating that suboptimal environmental conditions (short photoperiod and sunshine duration) limited plant growth independent of N availability. Seasonality affected plant growth more than nitrate concentration in leaves. Apparently, Mediterranean climatic

conditions led to adequate photosynthetic activity and nitrate reduction rates in planta during all seasons and therefore the accumulation of nitrates in the lettuce leaves were far bellow the maximum limits set by the E.C. 563/2002 Regulation for lettuce fresh product (3500 and 4500 mg kg 1 for the cultivation periods April-to-September and October-to-March, respectively). Indeed raised nitrate reductase activity has been observed at increased light levels (Gaudreau et al., 1995) and a close negative correlation between photosynthetic activity and nitrate content has been reported for various lettuce cultivars (Behr and Wiebe, 1992) and was used for modeling plant nitrate accumulation (Scaife and Schloemer, 1994). Nitrate accumulation in lettuce leaves was roughly doubled in all crop seasons in the maximum and medium inorganic fertilization treatments compared to the following treatments (maximum and medium organic, minimum inorganic). This is despite the higher total N in the respective sheep manure treatments and is apparently explained by the higher N availability in the inorganic fertilizer applications. However, a significant growth advantage was obtained for those two inorganic fertilization treatments in the late-spring crop season only, with the highest sunlight intensity and duration (20% and 23% yield increase compared to the respective maximum and medium organic fertilization treatments). Sheep manure or low- dose inorganic N fertilizer applications may therefore be successfully used under suboptimal environmental conditions to avoid unnecessary costs and nitrate accumulation in lettuce leaves. Reduced nitrate accumulation results have been reported for farm yard manure fertilized lettuce for one crop season only (Gianquinto et al., 1992; Stopes et al., 1989). Our results show that composted sheep manure applications were particularly safe in terms of nitrate accumulation in lettuce leaves in the long-term. They remained significantly lower than in the respective inorganic fertilization treatments, even after sub- sequent high level applications in different cultivation seasons (for example, I-max caused 100%, 53% and 124% increase in nitrate concentration compared to O-max in the respective first, second and third crop season, but a significant yield benefit was observed in the first season only). Moreover, sheep manure applications contributed to the built up of soil fertility at the end of the experiment, since they resulted in significantly increased N, P and K availability, contrary to the inorganic fertilization treatments that showed residual fertilizer effects in soil not different from the control. The effects of fertilization treatments on lettuce yield were similar to their effects on lettuce growth indicators (largest leaf length and width). However, leaf length showed limited variability between treatments (28–33 cm) and was not a sensitive indicator of lettuce growth ( r 2 = 0.218, p = 0.03); on the contrary leaf width varied widely (13–22 cm) and was strongly correlated to lettuce growth ( r 2 = 0.938, p < 0.01).

5. Conclusions

Nitrate concentration in romaine lettuce leaves depends on fertilizer type and dose rather than on the crop season and may

G.C. Pavlou et al. / Scientia Horticulturae 111 (2007) 319–325

325

remain in Southern Greece far below the upper limits set by the European Commission Regulation, even under high and repeated fertilization treatments, a result attributed mainly to the sufficient sunlight intensity and duration throughout the year in this region. High application rates of sheep manure may be needed to reach the lettuce growth obtained by inorganic fertilizers under optimal climatic conditions only. Increased inorganic fertilizer doses should be avoided since, especially under suboptimal light conditions, they might result in significant rise in nitrate accumulation in leaves but marginal yield increase; on the contrary composted sheep manure may be safely applied even at high doses. High residual availability of N, P and K may be obtained in the soil, following the sheep manure applications, which relates to manure dose; on the contrary no residual availability of N, P and K was obtained by inorganic fertilization applied via fertigation.

Acknowledgement

This

work

was

financially

supported

by

the

National

Agricultural Research Foundation.

References

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