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LPLA #324620, VOL 31, ISS 09

Critical Ratio of Calcium and Boron in Maize Shoot

for Optimum Growth
Shamsa Kanwal, Rahmatullah, Tariq Aziz, Muhammad Aamer
Maqsood, and Najam Abbas

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Critical Ratio of Calcium and Boron in Maize Shoot for Optimum
Growth
Shamsa Kanwal, Rahmatullah, Tariq Aziz, Muhammad Aamer Maqsood, and
Najam Abbas

0
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Journal of Plant Nutrition, 31: 1–8, 2008

Copyright © Taylor & Francis Group, LLC
ISSN: 0190-4167 print / 1532-4087 online
DOI: 10.1080/01904160802244530

1 Critical Ratio of Calcium and Boron in Maize Shoot

2 for Optimum Growth

3 Shamsa Kanwal, Rahmatullah, Tariq Aziz,

4 Muhammad Aamer Maqsood, and Najam Abbas

5 Institute of Soil and Environmental Sciences, University of Agriculture,

6 Faisalabad, Pakistan

7 ABSTRACT

8 Boron (B) deficiency is widely reported in alkaline calcareous soils of the world including
9 Pakistan. High calcium (Ca) content in such soils can affect the availability and utilization
10 of B by plants. Effect of applied B at different levels of Ca addition on maize was studied
11 in hydroponics. Four maize cultivars (‘EV-5089’, ‘SWL-2000’, ‘EV-6089’, and ‘Sultan’)
12 were grown at three levels of Ca (0.25 mM, 1 mM, and 2 mM) and two levels of B
13 (0 and 25 µM). Application of both the nutrients increased shoot dry matter production.
14 However, application of Ca antagonized the B concentration in shoot of four maize
15 cultivars. A curvilinear relationship existed between Ca/B ratio in shoot and relative
16 shoot dry matter of maize cultivars. Implication of using of Ca/B ratio for managing
17 commonly occurring B deficiency in calcareous soils is suggested.

18 Keywords: Calcium, Boron, Maize, Ca/B ratio, calcareous soils

19 INTRODUCTION

20 Boron (B) is an essential micronutrient required for normal growth and develop-
21 ment of higher plants. World-wide B deficiency is one of the major constraints
22 to crop production (Sillanpae, 1982). Soils of Pakistan are generally alkaline
23 and calcareous in nature and micronutrient deficiencies especially of zinc (Zn),
24 iron (Fe), and B are reported in various parts of the country (Rashid and Rayan,
25 2004). Boron deficiency was reported in 49% of the soil samples collected

Received 30 April 2007; accepted 8 August 2007.

Address correspondence to Dr. Rahmatullah, Institute of Soil & Environmental Sci-
ences, University of Agriculture, Faisalabad 38040, Pakistan. E-mail: rahmat428g@
gmail.com

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2 S. Kanwal et al.

from twenty districts of Punjab for an FAO study (Sillanpae, 1982; Rashid 26
et al., 1997). 27
Boron availability in soil is affected by several factors including soil tex- 28
ture, nature of clay minerals, pH, liming, organic matter, interrelationships with 29
other elements, and environmental conditions like moderate to heavy rainfall, 30
dry weather, and high light intensity (Moraghan and Mascagni, 1991). Pres- 31
ence of free calcium carbonate (CaCO3 ) in alkaline calcareous soils affects the 32
availability of B to crop plants. In addition to its effect on soil pH, calcium 33
carbonate also acts as an important B adsorbent in calcareous soils (Goldberg 34
and Forster, 1991). Boron adsorption is more on soils having higher calcium 35
carbonate content (Elrashidi and O’Connor, 1982). 36
Crop plants differ in their B requirement. In general, dicots have more of 37
a B requirement (20–70 mg kg−1 ) than monocots (5–10 mg kg−1 ) (Marschner, 38
1995). Calcium (Ca) is considered important to affect B availability and its 39
requirement for plants. The amount of either B or Ca influences the availabil- 40
ity and requirement of each other for normal plant growth and development 41
(Teasdale and Richards, 1990). An enhanced B deficiency symptoms in plants 42
by increased Ca supply have been reported (Gupta, 1979). Similarly, B de- 43
ficiency altered Ca translocation to the shoot and fruit (Ramon et al., 1990; 44
Yamauchi et al., 1980). It indicates a balanced supply of Ca and B for normal 45
growth and development of plants. The Ca/B ratio in leaf tissue can be used to 46
asses the B deficiency, sufficiency and toxicity in plants. 47
Genotypic variation in crops has also been reported with respect to B 48
Q1 and Ca uptake (Huo -Yan et al., 2003). Their differential requirement and use 49
can influence interactive effect of Ca and B on plant growth. Therefore, B 50
requirement of crop plants should be determined in relation to Ca availability. 51
Therefore, the present study was conducted to determine the critical Ca/B ratio 52
in plants for optimum maize growth at various levels of Ca and B supply in root 53
medium. 54

MATERIALS AND METHODS 55

Seeds of four maize cultivars viz. ‘EV-5089’, ‘SWL-2000’, ‘EV-6089’, and 56

‘Sultan’ were germinated in polyethylene lined iron trays containing pre- 57
washed river bed sand. Sand in the trays was moistened with distilled water 58
for germination. Two-week-old uniform seedlings were transplanted in foam- 59
plugged holes in a thermopal sheet floating in 200 L of Johnson’s modified solu- 60
tion (Johnson et al., 1957) in four polyethylene lined iron tubs. Three levels of Ca 61
(0.25 mM, 1.0 mM, and 2.0 mM) and two levels of B (0 and 25 µM) were devel- 62
oped using calcium chloride and borax, respectively. Potassium nitrate (KNO3 ), 63
ammonium sulfate [(NH4 )2 SO4 ], magnesium sulfate (MgSO4 7H2 O), man- 64
ganese sulfate (MnSO4 ), zinc sulfate (ZnSO4 ), molybdic acid (H2 MoO4 ), and 65
Fe-ethylenediaminetetraacetic acid (EDTA) were used to add 8 mM nitrogen 66
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Calcium Boron Ratio for Optimum Growth 3

67 (N), 3 mM potassium (K), 5 mM magnesium (Mg), 4 mM sulfur (S), 2 µM man-

68 ganese (Mn), 2 µM Zn, 0.5 µM molybdenum (Mo), and 50 µM Fe in solution
69 form to various tubs. The experiment was laid out in completely randomized
70 design (CRD) with nine replications of each genotype. The pH of the solution
71 was monitored daily and maintained at 5.7 ± 0.3 during the entire growth pe-
72 riod. Average temperature in the green house was 30 ± 5◦ C at different times
73 of day and 22 ± 3◦ C during the night for the experimental period. Relative
74 humidity dropped to 35% at mid day and increased to 85% at mid night. Light
75 intensity varied between 300 and 1400 µmol photon m−2 S−1 depending upon
76 the day and cloud conditions. The plants were harvested thirty days after trans-
77 planting, washed with distilled water and separated into root and shoot. Dry
78 weights of shoot were recorded after oven drying them to their constant weight
79 at 70◦ C in a forced air oven. The samples were then ground to 40 mesh with a
80 Wiley mill fitted with stainless steel blades and chamber. The ground samples
81 of shoot were dry ashed at 600◦ C for 12 hours. After dry ashing, samples were
82 digested in sulfuric acid (H2 SO4 ), filtered, and analyzed for B concentration
83 by UV-Visible spectrophotometer (Shimadzu, UV-1201, Shimadzu, Columbia,
84 MD) at 420 nm wavelength. Calcium concentration in plant digest was deter-
85 mined by atomic absorption spectrophotometer (Perkin Elmer Aanalyst 100,
86 Perkin Elmer, Waltham, MA).
87 The data regarding shoot dry matter, B uptake and Ca uptake were statisti-
88 cally analyzed using PC based MSTAT-C (Russel and Eisensmith, 1983). Two Q2
89 factors, ANOVA and LSD, were used to separate treatment means

90 RESULTS AND DISCUSSION

91 Shoot Dry Matter

92 There was a highly significant (P < 0.01) main and interactive effect of addition
93 of Ca and B and cultivars on shoot dry matter production by the four maize
94 cultivars. Shoot dry matter of the four maize cultivars ranged from 0.68 to
95 3.25 g plant−1 (Table 1). There was a significant constant increase in shoot dry
96 matter production of the four maize cultivars by adding Ca and B to the root
97 medium. There was no general trend for increasing shoot dry matter yield in
98 the four maize cultivars at the two levels of B application along with low (0.25
99 mM Ca), medium (1 mM Ca), and high levels (2 mM Ca) of Ca. However,
100 a significant interaction (P < 0.01) among Ca, B, and maize cultivars had
101 a profound effect on relative shoot dry matter production. For example, B
102 application significantly (P < 0.01) increased shoot dry matter production 62
103 to 159% of the maize cultivars at various levels of Ca addition used in this study.
104 Similarly, application of higher level of Ca at different levels of B significantly
105 increased shoot dry matter production 27 to 111% as compared to its lowest
106 level of 0.25 mM Ca application.
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Table 1
Shoot dry matter of maize cultivars (mean of 9 replications)

Shoot dry matter (g plant−1 )

+ Ca at 0.25 mM + Ca at 1.0 mM + Ca at 2.0 mM

+B at 25 +B at 25 +B at 25
Maize cultivars –B µM –B µM –B µM

EV-5089 0.72 1.55 0.96 2.38 1.40 2.78

SWL-2000 0.68 1.33 0.90 2.17 1.34 2.72
EV-6089 0.83 1.64 1.44 2.56 1.64 2.66
Sultan 0.71 1.84 1.16 2.34 1.50 3.25
LSD (0.01) 0.29
Ca × B × Cultivar

Unfortunately, no certain trend of shoot dry matter production was evident 107
in the four maize cultivars grown at the two levels of B applied along with three 108
levels of Ca in the root medium. 109

Calcium and Boron Uptake in Shoot 110

There was a significant (P < 0.01) main and interactive effect of Ca, B, and 111
cultivar on concentration and uptake of Ca in maize shoot (Tables 2 and 3). 112
Addition of Ca to the growth medium significantly (P < 0.01) increased the 113
Ca concentration in shoot. The calcium concentration ranged from 1.91 to 3.78 114
mg g−1 of shoot dry matter in four maize cultivars. Boron application had a 115

Table 2
Concentration of calcium in shoot of four maize cultivars (mean of 9 replications)

Shoot calcium concentration (mg g−1 )

+ Ca at 0.25 mM + Ca at 1.0 mM + Ca at 2.0 mM

+B at 25 +B at 25 +B at 25
Maize cultivars –B µM –B µM –B µM

EV-5089 1.91 2.67 2.42 2.72 3.46 3.64

SWL-2000 2.32 2.34 2.97 3.24 3.62 3.78
EV-6089 2.06 2.31 3.04 3.72 3.62 3.72
Sultan 2.40 2.57 2.67 3.12 3.11 3.65
LSD (0.01) 0.11
Ca × B × Cultivar
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Calcium Boron Ratio for Optimum Growth 5

Table 3
Total uptake of calcium in shoot of four maize cultivars (mean of 9 repeats)

Calcium uptake by shoot (mg plant−1 )

+ Ca at 0.25 mM + Ca at 1.0 mM + Ca at 2.0 mM

+B at 25 +B at 25 +B at 25
Maize cultivars –B µM –B µM –B µM

EV-5089 1.38 4.14 2.32 6.47 4.84 10.12

SWL-2000 1.58 3.11 2.67 7.03 4.85 10.28
EV-6089 1.71 3.79 4.38 9.52 5.94 9.90
Sultan 1.70 4.73 3.10 7.30 4.67 11.86
LSD (0.01) 0.61
Ca × B × Cultivar

116 synergistic effect on concentration and uptake of Ca in maize shoot of the four
117 cultivars. It significantly (P < 0.01) increased the concentration by 0.86 to 40
118 % and uptake of Ca by more than two fold in maize shoot. Such an increase in
119 Ca content of plants by B application had also been reported by Yamagishi and Q3
120 Yamamoto (1994).
121 There was also significant (P < 0.01) main and interactive effect of Ca,
122 B and cultivars on concentration and uptake of B in shoot of the four maize
123 cultivars (Tables 4 and 5). Application of B significantly (P < 0.01) increased
124 concentration and uptake of B at various levels of Ca addition to root medium.
125 Calcium application had a significant (P < 0.01) antagonistic effect on concen-

Table 4
Concentration of boron in shoot of four maize cultivars (mean of 9 repeats)

Boron concentration in shoot (mg g−1 )

+ Ca at 0.25 mM + Ca at 1.0 mM + Ca at 2.0 mM

+B at 25 +B at 25 +B at 25
Maize cultivars –B µM –B µM –B µM

EV-5089 0.02 0.74 0.02 0.34 0.02 0.08

SWL-2000 0.016 0.71 0.009 0.42 0.017 0.13
EV-6089 0.02 0.65 0.009 0.26 0.006 0.053
Sultan 0.009 0.57 0.012 0.33 0.009 0.098
LSD (0.01) 0.10
Ca × B × Cultivar
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Table 5
Total uptake of boron in shoot of four maize cultivars (mean of 9 repeats)

Boron uptake by shoot (mg plant−1 )

+ Ca at 0.25 mM + Ca at 1.0 mM + Ca at 2.0 mM

+B at 25 +B at 25 +B at 25
Maize cultivars –B µM –B µM –B µM

EV-5089 0.014 1.147 0.809 0.297 0.028 0.222

SWL-2000 0.011 0.944 0.911 0.222 0.023 0.354
EV-6089 0.017 1.066 0.666 0.256 0.010 0.141
Sultan 0.006 1.049 0.772 0.265 0.014 0.319
LSD (0.01) 0.03
Ca × B × Cultivar

tration and uptake of B in maize shoot. Application of Ca at the lowest level 126
of 0.25 mM allowed maximum concentration of B in shoot while the reverse 127
was true at the highest level of 2 mM Ca in the growth medium. Carpena et al. 128
(2000) have also reported a similar result for B concentration in shoot. Increased 129
B requirement of crops by increasing Ca in the root medium have also been 130
discussed by Tisdale et al. (1985). 131
Shoot growth in four maize cultivars had a poor correlation with B con- 132
centration while correlation was significant (r = 0.722, P < 0.03 n = 24) with 133
Ca concentration in plant shoot. Therefore, a curvilinear relationship between 134
Ca/B ratio in shoot and relative shoot dry matter is depicted in Figure 1. The 135
results suggest Ca/B ratio of 30 in shoot for obtaining 95% of the maximum 136
relative shoot dry matter production. 137

Figure 1. Relation between relative shoot dry matter production and Ca:B ratio in
shoots.
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Calcium Boron Ratio for Optimum Growth 7

138 Further work is warranted on utilizing Ca/B relationship for correcting B

139 deficiency found in crops commonly grown on calcareous soils (Rashid et al.,
140 1997)

141 REFERENCES

142 Carpena, R. O., E. Esteben, M. J. Sarro, J. Penalosa, A. Garate, J. J Lucena,

143 and P. Zornoza. 2000. Boron and calcium distribution in nitrogen-fixing
144 pea plants. Plant Science 151: 163–170.
145 Elrashidi , M. A. and G. A. O’Connor. Boron sorption and desorption in soils. Q4
146 Soil Science Society of America Journal 46(1): 27–31.
147 Goldberg, S. and H. S. Forster. 1991. Boron sorption on calcareous soils and
148 reference calcites. Soil Science 152: 304–310.
149 Gupta, U. 1979. Boron nutrition of crops. Advances in Agronomy 31: 273–307.
150 Hu , H. and P. H. Brown. 1994. Localization of boron in cell walls of squash Q5
151 and tobacco and its association with pectin. Evidence for a structural role
152 of B in the cell wall. Plant Physiol. 105: 681–689.
153 Huo-yan, W., W. Yun-hua, Du. Chang-wen, Xu. Fang-sen, and Y. Yu-hua. 2003.
154 Effects of boron and calcium supply on calcium fractionation in plants and
155 suspension cells of rape cultivars with different boron deficiency. Journal
156 of Plant Nutrition 26 (4): 789–806.
157 Johnson, C. M., R. R. Strout, T. C. Broyer, and A. B. Carlton. 1957. Comparative
158 chlorine requirement of different plant species. Plant Soil 8: 327–353.
159 Marschner, H. 1995. Mineral Nutrition of Higher Plants. San Diego, CA: Aca-
160 demic Press.
161 Moraghan, J. T. and H. J. Mascagni. 1991. Environmental and soil factors
162 affecting micronutrient deficiencies and toxicities. In: Micronutrients in
163 Agriculture. 2nd ed, eds. J. J. Mortvedt, F. R. Cox, L. M. Shuman, and R.
164 M. Welch, pp. 371–425. Madison, WI: SSSA.
165 Ramon, A. M., R. O. Carpena, and A. Garate. 1990. The effect of short term
166 deficiency of boron on K, Ca and Mg distribution in leaves and roots of
167 tomato (Lycopersicum esculentum) plants. In: Plant Nutrition Physiology
168 and Applications, ed. M. L. van Beusichem, pp. 287–290. Dordrecht, The
169 Netherlands: Kluwer.
170 Rashid, A., E. Rafique, and N. Bughio. 1997. Micronutrient deficiencies in
171 rainfed calcareous soils of Pakistan. III. Boron nutrition of sorghum. Com-
172 munications in Soil Science and Plant Analysis 28: 444–454.
173 Rashid, A. and J. Rayan. 2004. Micronutrient constraints to crop production in
174 soils with Mediterranean-type characteristics: a review. Journal of Plant
175 Nutrition 27(6): 959–975.
176 Russel, D. F. and S. P. Eisenmith. 1983. MSTAT-C. Crop Soil Sci. Dept., Michi-
177 gan State Uni., USA, East Lansing, MI.
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LPLA˙A˙324620 601.cls July 3, 2008 4:3

8 S. Kanwal et al.

Sillanpae, M. 1982. Micronutrients and nutrients status of soils: a global study. 178
FAO Soils Bulletin No. 48, Rome, Italy: FAO. 179
Teasdale, R. D. and D. K. Richards. 1990. Boron deficiency in cultured pine 180
cells: quantitative studies of the interaction with Ca and Mg. Plant Physi- 181
ology 93: 1071–1077. 182
Tisdale, S. L., W. L. Nelson, and J. D. Beaton. 1985. Soil Fertility and Fertilizers. 183
New York: MacMillan Publishing Company. 184
Yamagishi, M. and Y. Yamamato. 1994. Effects of boron on nodule development 185
and symbiotic nitrogen fixation in soybean plants. Soil Science and Plant 186
Nutrition 40: 265–274. 187
Yamauchi, T., T. Hara, and Y. Sonoda. 1980. Distribution of Ca and B in the 188
pectin fraction of tomato leaf cell wall. Plant Cell Physiology 27: 727–73. 189