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Iron chlorosis paradox ingrapevineLuigi Bavaresco a , Elisa Giachino a & RuggeroColla aa Istituto di Frutti‐Viticoltura , UniversitàCattolica S. Cuore , Via Emilia Parmense, 84,Piacenza, I‐29100, ItalyPublished online: 21 Nov 2008.

To cite this article: Luigi Bavaresco , Elisa Giachino & Ruggero Colla (1999)Iron chlorosis paradox in grapevine, Journal of Plant Nutrition, 22:10,1589-1597, DOI: 10.1080/01904169909365739

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JOURNAL OF PLANT NUTRITION, 22(10), 1589-1597 (1999)

Iron Chlorosis Paradox in Grapevine

Luigi Bavaresco, Elisa Giachino, and Ruggero Colla

Istituto di Frutti-Viticoltura, Università Cattolica S. Cuore, Via EmiliaParmense, 841-29100 Piacenza, Italy

ABSTRACT

Vitis vinifera L. cv. Pinot blanc was grown in both pot and commercial vineyardunder lime-stress conditions. Dark green to very chlorotic young leaves weresampled in order to analyze chlorophyll and iron (Fe) concentration andcontent, area, fresh and dry weight, specific weight per area. Iron content perindividual leaf was a good parameter to separate dark green leaves fromchlorotic ones in both trials, while Fe concentration on DW basis was not.Chlorosis occurrence always decreased leaf dry weight and specific weightper area.

INTRODUCTION

The complex aspects of lime-induced chlorosis in cultivated plants have beenstudying for a long time, without so far a clear understanding of the phenomenon,in spite of the huge number of experimental papers. A controversial problem is, forinstance, the role of leaf Fe concentration in the chlorosis occurrence of susceptibleplants growing on calcareous soil (Abadia, 1992; Wallace and Jones, 1993; Terryand Zayed, 1995). In some cases lime-induced chlorosis is related to a low Feuptake and translocation to the leaves (Haussling et al., 1985; Bavaresco et al.,1992), in some other cases to an high leaf Fe level, which has to be somehowinactivated inside the leaves (Mengel and Malissiovas, 1981; Mengel et al., 1984;Bavaresco et al., 1993). Crucial aspects in grapevine seems to be whether or notthe plant is grafted and which is the rootstock and the scion. Comparing green and

1589

Copyright © 1999 by Marcel Dekker, Inc. www.dekker.com

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1590 BAVARESCO,GIACHINO,ANDCOLLA

chlorotic potted plants of the same variety (same aerial apparatus), but with differentrootstocks (different root apparatus), Fe concentration of leaf blade and chlorosisrelated (Bayaresco et al., 1992). On the other hand, potted plants of differentscions grafted on the same rootstock and potted ungrafted plants of different Vitisspecies did not show any correlation between leaf Fe concentration and chlorosis(Bayaresco, 1997;Bavarescoetal., 1994). According to Marschner (1995), the lackof correlation between chlorosis and leaf Fe level occurs mostly in field-growingplants, when sometimes chlorotic plants have higher leaf Fe concentration thangreen ones. This latter aspect which has been called "Fe chlorosis paradox"(R8mheld, 1997) is, as a rule, associated with a severe shoot growth inhibition;high Fe concentration in chlorotic leaves is a consequence of the inhibited leafgrowth and it can be observed in spite of a distinct lower Fe content in the individualleaves. Also, Morales et al. (1998) observed lower Fe content in chlorotic pear andpeach individual leaves than in green ones, even though Fe concentration (on thedry weight basis) was similar. The aim of this paper is to further investigate therelationships between chlorophyll and Fe leaf concentrations and contents ofpotted and field-growing grapevines.

MATERIALS AND METHODS

Plant Material

Pot Trial

Three-year-old plants of Vitis vinifera L. cv. Pinotblanc (clone VCR5) grafted onV. ripariaMlCHX. x V. rapes&is SCHEELE 3309 Clime-susceptible hybrid rootstock,were utilized for the trial. The plants were grown in pot (45 L volume) of a calcareousand a non calcareous soil. The main characteristics of the calcareous soil, collectedin a vineyard nearby Piacenza, were silty-loam texture, pH of 8.6, total carbonates,80%, active lime, 19%, organic matter, 0.2%, Fe(extractedbyDTPA5mM+CaCl210mM+triethanolamine 100 mM) 2.4 mg Kg''. The non-calcareous soil was an artificialsoil done by mixing acid peat (pH 4.5), sand and neutral soil in the ratios 1:1:1; themain soil characteristics were sandy-silt texture, pH of 7.3, total carbonates, 3%,active lime, 2%, organic matter, 7.8% , Fe, 50 mg Kg1. The pots were placedoutside on a platform with a hail protection net and soil moisture was kept nearfield capacity by drip irrigation. The plants were trained to 5 bearing shoots,according to a cane pruning system with shoot positioning.

Field Trial

Twenty-seven-year-old plants of Vitis vinifera L. cv. Pinotblanc grafted on V.berlandieri* Vitis riparia Kober 5BB hybrid rootstock were utilized for the trial.The vineyard was located in Pigazzano (300 m elevation, Piacenza province) on thesame calcareous soil type utilized for the pot trial. The vines were trained doublecane-pruning with about 20 shoots per plant and vine spacing 2.5 mx2 m. The

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IRON CHLOROSIS PARADOX IN GRAPEVINE 1591

vineyard was not treated with Fe chelates and chlorosis symptoms changed alongthe time depending on the climatic factors and fruit load. During 1997 growingseason some vines showed chlorosis symptoms while other ones remained green.

Plant Tests

Twenty-nine young leaves (4th and 5th leaf beginning from the shoot tip) weresampled from the potted plants growing in both soils at fruit set (June 6, 1997)when the average shoot length of the plants growing in the calcareous soil(chlorotic and some pale green leaves) and in the other one (pale green and darkgreen) was respectively 60 cm and 130 cm; the age of the two leaf sets was different,the leaves from the calcareous soil being older than those from the other soil.

Twenty young leaves (4th and 5th from shoot tip) were sampled from chloroticand green plants growing in the vineyard, at veraison (July 27,1997); shoot lengthwas not recorded.

The ieaves were weighed in order to know the individual fresh weight (FW) andarea was recorded by the gravimetric method according to Kvet and Marshall(1971), ari outline of the leaf was traced on a paper which had a uniform weightdistribution with area. The leaf shape was cut from the paper and weighed and leafarea was then calculated from the weight to area relationship of the paper, establishedby weighing pieces of paper of known area.

Chlorosis symptoms were rated using the scale of Pouget and Ottenwaelter(1978), ranking from 0 (dark green leaves) to 5 (severe chlorosis with more than10% of the blade surface with necrosis).

Chlorophyll (Chi) concentration was indirectly calculated by using the portableChlorophyll Meter SPAD 502 (Minolta Corp., Ramsey, NJ). The leaves were testedin 5 positions of the interveinal tissue corresponding to the leaf lobes and thevalues were averaged to get one datum per individual leaf. SPAD units weretransformed into Chi concentrations (mg 100'g "' DW, mg g "' FW, ug cm2),according to equations calculated in a previous work (Bavaresco, 1995). Chlorophyllcontent per individual leaf was obtained multiplying leaf area and weight by Chiconcentrations, and it was expressed as ug leaf"1.

Iron leaf concentration was assayed after wet destruction of the oven-driedmaterial, by atomic absorption spectrometry (AAS) and expressed as ug g"' DWand FW, ug cm ~2. Iron content per individual leaf was obtained multiplying leafweight by Fe concentration, and it was expressed as ug leaf'1.

Specific leaf weight per area (SLW) was calculated dividing leaf dry weight byleaf area and expressed as mg cm2 .

Statistical Analysis

Simple regressions were calculated as follows: Fe concentrations versus Chiconcentrations; Fe content versus Chi content; Chi concentration versus leaf dryweight (LDW), fresh weight (LFW), area (LA) and specific leaf weight per area(SLW).

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1592 BAVARESCO, GIACHINO, AND COLLA

TABLE 1. Correlations between chemical and morphological leaf parameters of pottedcv. Pinotblanc.

V

Chl(mglOOr'DW)

Chi (nig- g-1 FW)

Chi (ngcnr1)

Chl(|igleaf')

LeafDWfeleaf')

LeafFWfeleaf"1)

Leararea(cmMear')

SLWCmgcnr2)

X1

FeOig-.g-'DW)

Fefrgg-'FW)

Fe (ngcmJ)

Fe(ngleaf')

ChlOigcnr2)

ChlOig-cnr*)

ChlOigcnr1)

Chi (pgcnr2)

n'

29

29

29

29

29

29

29

29

r4

-0.62"

ns

ns

0.56"

0.88"

0.81"

0.73"

0.85"

chlorotic

leaves

81+231

0.20+0.64

2.60+10.28

91+391

0.10+0.16

0.51+0.71

30.9+43.5

2.96+4.45

Grange

green

leaves

249+448

0.70+1.36

11.27+22.80

557+1288

0.20+0.37

0.69+1.24

43.9+73.0

3.47+6.35

X range

chlorotic

leaves

79+182

14.4+32.7

0.26+0.54

9.8+19.2

2.60+10.28

2.60+10.28

2.60+10.28

2.60+10.28

green

leaves

37+96

8.6+27.4

0.13+0.54

7.6+26.9

11.27+22.80

11.27+22.80

11.27+22.80

11.27+22.80

'Dependent variable,independent variable.3Number of tested leaves.••Correlation coefficient."Significance at P=0.01.

RESULTS

Pot Trial

The sampled leaves ranged from SPAD value 4 (corresponding to rating 4 of theabove mentioned scale, which means yellow to whitish leaves with little necroses)to 28.7 (corresponding to rating 0, which is dark green leaves); the correspondingvalues as Chi concentrations are reported in Table 1 (Y range). Iron concentrationsranged according to X range of Table 1, with average values of 102.2±33.6 ppm and62.4±17 ppm for chlorotic and green leaves respectively (data not shown);correlation between Chi and Fe concentrations on the DW basis was indeednegative (r=-0.59), while no significant correlations were calculated between thesame parameters on FW and area basis (Table 1). A significant positive regression(r=0.56) was calculated between Chi and Fe contents per individual leaf, but,according to Figure 1, there is not a clear separation between Fe content of chloroticand pale green leaves. Chlorotic and pale green leaves, in fact, had similar Fecontent (12.5±3.8 ug leaf"1 and 15.5±4.4 ug leaf"1, respectively), while only the darkgreen leaves had an higher Fe content (24.9±1.7 ug leaf, data not shown). Leafdry and fresh weight, area and specific leaf weight per area, were positively correlatedwith Chi concentration (Table 1).

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IRON CHLOROSIS

1,400-

1.200-

1,000-

leaf

-

O)

^ 600-O

400-

200-

J

+

+

X

PARADOX IN GRAPEVINE

a

+ + + / ^

+ ± ^^^ +

X

* X

X

1 1 1 1

10 15 20 25Fe (ug leaf -1)

1592

r=0.56*«

• dark green

+ pale greenx chlorotic

FIGURE 1. Pot trail: correlation between Chi and Fe content per individual leaf.

Field Trial

The sampled leaves ranged from SPAD value 4.4 (corresponding to rating 3 ofthe above mentioned scale, which means yellow leaves with green veins) to 35.3(corresponding to rating 0, which means dark green leaves); the correspondingvalues as Chi concentrations are reported in Table 2 (Y range). The maximum Chivalues are higher than those of the pot trial, as well as Fe concentrations (X rangeof Table 2). Chlorophyll and Fe concentrations on DW basis did not significantlycorrelate (Table 2), while the same parameters on FW and area basis did correlate(r=0.78 and 0.76, respectively). Iron average concentrations of chlorotic and greenleaves were 112±34 ppm and 143±27 ppm, respectively. A positive and significantcorrelation (r=0.82) between Chi and Fe content per individual leaf occurred (Figure2); chlorotic leaves, pale green leaves and dark green leaves had an Fe content of27.5±10.5 ug leaf', 39.0±2.6 ug leaf' and56.4±13.1 ug leaf1, respectively (datanotshown). Leaf dry weight and specific leaf weight per area, were positively correlatedwith Chi concentration (Table 2).

DISCUSSION

The Fe paradox was evident for potted plants, especially when Chi and Fe wereexpressed on DW basis; chlorotic leaves had higher Fe levels because ofconcentration effect, meaning that high Fe values are the consequence, and not

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1594 BAVARESCO, GIACHINO, AND COLLA

TABLE 2. Correlations between chemical and morphological leaf parameters of cv. Pinotblanc growing in the field.

Y1

Chl(mglOOg'DW)

Chi (mg- g-1 FW)

CM fog-cm1)

Chi Gig-leaf •')

LeafDW(gleaf ' )

LeafFWfeleaf ' )

Leaf area (cmMeaf ' )

SLW(mgcm*)

X!

Fefogg'DW)

Fe fog- g-1 FW)

Fe fog-cm')

Fe fog-ieaf •')

Chi fogcnr2)

Chi fog-cm"1)

Chi (tig cm'2)

Chi fogcm1)

n1

20

20

20

20

20

20

2d

. 20

r4

ns

0.78"

0.76"

0.82"

0.76"

ns

ns

0.88"

chlorotic

leaves

88+222

0.22+0.61

2.92+9.79

203+1049

0.17+0.44

0.88+1.86

38.5+105.7

2.59+4.41-

Y range

green

leaves

307+536

0.89+1.67

14.54+28.20

895+2486

0.25+0.46

0.98+1.53

52.2+76.4

- 4.02+6.60

chlorotic

leaves

68+65

12.4+31.9

0.23+0.73

14.3+47.8

2.92+9.79

2.92+9.79

2.92+9.79

2.92+9.79

X range

green

leaves

114+198

30.5+59.3

0.51+1.12

36.5+81.2

14.54+28.20

14.54+28.20

14.54+28.20

14.54+28.20

'Dependent variable,independent variable.3Number of tested leaves.••Correlation coefficient."Significance at P=0.01.ns-Not significant.

3,000-

2,500-

2,000-

- 1;500-

o 1,000-

500-

r=0.82**

* dark green

+ pale green

* chlorotic

10 20 30 40 50 60 70 80Fe{ugleaf-')

FIGURE 2. Field trail: correlation between Chi and Fe content per individual leaf.

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IRON CHLOROSIS PARADOX IN GRAPEVINE 1595

the cause, for chlorosis. Chlorosis, in fact, was related to small leaves and lowspecific leaf weight per area. On the other hand a positive correlation between Chiand Fe leaf contents per individual leaf occurred, meaning that Fe content per leafwas a better Fe index in relation to chlorosis occurrence, as reported also for field-growing peach and pear (Morales et al., 1998); data from this pot trial showed thatonly dark green leaves differed from chlordtic ones as regarding Fe content. UnderFe stress conditions, therefore, Chi synthesis is impaired because Fe was initiallypoorly translocated into the leaves which remain small; this is likely due to the lowphotosynthesis rate occurring in chlorotic plants, which causes also the slow andshort shoot growth. The low Fe uptake per leaf (arid per plant) in chlorotic vines isdue to the effect of soil bicarbonate (Mengel et al., 1984) which impaires Fe uptakemechanisms of the root system (Marschrier et al., 1986), and reduces root growth(Bavaresco et al., 1993). The high Fe concentrations in yellow leaves means that ithas been accumulated in an unavailable form (Mengel, 1995), but this is, accordingto Romheld (1997), a secondary event occurring in the leaf. It is still unclear themechanism for Fe to be inactivated in chlorotic leaves, even though somehypotheses have been done (Terry and Low, 1982;Koleshetal., 1987; Mengel andGeurtzen, 1988). Attempts to correlate active Fe, as Fe (II) extracted byphenantroline, with chlorosis occurrence of grapevine potted plants gavecontroversial data (Bavaresco et al., 1993; Bavaresco et al., 1995; Bavaresco andFogher, 1996).

Leaves from field-growing plants showed higher maximum values for Chi andFe. Also, in this experiment Fe paradox was evident, since some chlorotic leaveshad higher Fe concentrations (DW basis) than green ones. Unlike potted plants,Chi and Fe concentrations expressed on FW and area basis were significantlyrelated, as well as the contents per individual leaf. As already observed in the pottrial, Fe content per individual leaf did separate well only dark green leaves fromchlorotic ones. In this case Fe deficiency seems to be the cause for leaf chlorosissince chlorotic leaves had a lower Fe concentration and content. Chlorotic leaveshad also a low specific leaf weight per area, due to the low photosynthesis rate,providing poor nutrient supply to leaf tissue. Under field conditions, according tothe data of this experiment, Fe concentration per leaf fresh weight and area providea better way to express Fe as related to chlorosis.

ACKNOWLEDGMENTS

The authors want to thank MURST (Ministry for University and Scientific andTechnological Research) for financial support, and Mr. G. Bruzzi (lab crew) for hiscontribution to the project.

REFERENCES

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