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Application of Fertilizer Manganese Doubled Yields ofLentil Grown on Alkaline SoilsR. F. Brennan a & M. D. A. Bolland b ca Department of Agriculture of Western Australia, 444 Albany Highway, Albany, WA, 6330,Australiab Department of Agriculture of Western Australia, Bunbury, WA, Australiac Plant Science, Faculty of Agriculture, The University of Western Australia, Crawley,WA, Australia

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To cite this article: R. F. Brennan & M. D. A. Bolland (2003): Application of Fertilizer Manganese Doubled Yields of LentilGrown on Alkaline Soils, Journal of Plant Nutrition, 26:6, 1263-1276

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Application of Fertilizer ManganeseDoubled Yields of Lentil Grown on

Alkaline Soils

R. F. Brennan,1,* and M. D. A. Bolland2,3

1Department of Agriculture of Western Australia,

Albany, WA, Australia2Department of Agriculture of Western Australia,

Bunbury, WA, Australia3Plant Science, Faculty of Agriculture, The University of Western

Australia, Crawley, WA, Australia

ABSTRACT

The yield increase of 45-day-old lentil (Lens culinaris Medik) seedlings

to applications of two sources of manganese (Mn) fertilizer was compared

in a glasshouse experiment using two alkaline soils from southwestern

Australia. The Mn sources were manganese sulfate (24.6% Mn), the usual

source of Mn for crops in southwestern Australia, and Manganese oxide

(MnO; 77.3% Mn). Both sources were finely powdered, applied at equiva-

lent amounts of Mn and were mixed throughout both soils. The effectiveness

of the two Mn fertilizers was compared using yield and Mn content (Mn

concentration multiplied by the dried yield) of dried lentil shoots harvested

*Correspondence: R. F. Brennan, Department of Agriculture of Western Australia,

444 Albany Highway, Albany, WA 6330, Australia; E-mail: rbrennan@agric.wa.gov.au.

JOURNAL OF PLANT NUTRITION

Vol. 26, No. 6, pp. 1263–1276, 2003

DOI: 10.1081=PLN-120020369 0190-4167 (Print); 1532-4087 (Online)

Copyright # 2003 by Marcel Dekker, Inc. www.dekker.com

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45 days after sowing. Applications of Mn about doubled shoot yields on

both soils. For both soils, relative to the Mn sulfate fertilizer, the Mn oxide

was about half as effective for producing dried shoots and about 40% less

effective in increasing the Mn content of dried shoots so about twice as much

Mn as the oxide was required to produce the same yield or Mn content in

shoots as MnSO4 � 4H2O. The concentration of Mn in youngest mature

growth (YMG) and in dried shoots was used to determine critical concen-

trations of Mn in tissue associated with 90% of the maximum yield. The

critical concentration (mg=kg) of Mn was found to be 18 for YMG and about

21 in dried shoots.

Key Words: Manganese sources; Lentil; Alkaline soils.

INTRODUCTION

Lentil (Lens culinaris Medik) is a promising grain legume to grow

in rotation with cereal crops on alkaline loam to clay soils of southwestern

Australia.[1,2] Manganese (Mn) is a common deficiency of cereal crops grown

on some of these soils which occupy about 4.5 million ha (25%) of the

region.[3] Manganese deficient lentil plants have been observed in farmer

paddocks (pale plants with intervenial leaf chlorosis on the youngest mature

leaves), confirmed by low Mn concentration measured in shoots (<10 mg

Mn=kg dried shoots). The Mn requirement of lentil on Mn-deficient alkaline

soils in southwestern Australia is not known. In the region, manganese sulfate

(MnSO4 � 4H2O) is the Mn fertilizer used.[3] However, manganese oxide

(MnO), a by-product of mining operations in Western Australia, is a cheaper

alternative source. It appears that the availability of various Mn sources for

various plant species can vary greatly.[4] The glasshouse experiment described

here compared the yield and Mn content (Mn concentration multiplied by

yield) response of 45-day-old lentil shoots to applications of either

MnSO4 � 4H2O or MnO applied to two alkaline soils suitable for growing

lentil in southwestern Australia.

MATERIALS AND METHODS

Soils

The <4 mm fraction of the top 10 cm of two Mn-deficient alkaline soils

was used. The soils were collected from under remnant native vegetation areas

yet to be cleared for agriculture and had never been fertilized with Mn

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fertilizer. Manganese deficiency in cereals is frequently observed in these

soils.[3] One soil was collected near Salmon Gums (33�520 S, 121�400 E) and

the other near Scaddan (33�500 S, 121�300 E), both north of Esperance (33�S,

121�540 E), about 650 km south east of Perth (32�S, 116�E), Western

Australia. The soil from Salmon Gums was a grey powdery clay-loam

(known locally and hereafter called Kopi soil) and the other soil was a

sandy loam (hereafter called Scaddan soil).

Soil classification[5,6] and some properties, are listed in Table 1. Soil pH

was measured in 0.01 M CaCl2 using a 1 : 5 soil to solution ratio, after the soil

suspensions had been shaken at 23�C for 1 h on an end-over-end shaker

(10 rpm).[10] Particle size analysis, for percentage clay (<2 mm), was by the

Table 1. Soil classification, weight of soil for each pot, and some properties ofthe top 10 cm of the <4 mm fraction of the soils used.

Collection site

Soil classification and properties Salmon Gums Scaddan

Classification

Local name Kopi soil Scaddan soil

Soil survey staff[6] Typic Petrocalcic

Eutrochrept Fluventic

Xerochrept

Northcote[5] Gc 1.22 Dy 5.43

Weight of soil per pot (g=pot) 2850 2950

Soil properties

pHa 8.2 8.2

Clay (%) (Day[7]) 14 20

Organic carbon (%)

(Walkley and Black[8])

2.3 2.7

Iron oxide (%)b 1.2 0.4

Aluminum oxide (%)b 0.16 0.16

DTPA extractable Mn

(mg=kg)d

0.3 0.1

Bicarbonate extractable P

(mg=kg) (Colwell[9])

5 4

CaCO3 (%) (Rayment and

Higginson[10])

7.9 5.2

a1 : 5 soil : 0.01M CaCl2 (w=v).bSesquioxides (Hesse[11]).cGupta and McKay.[12]

dDiethylenetriaminepentaacetic acid (Lindsay and Norvell[13]).

Fertilizer Mn and Lentil 1265

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pipette method.[7] Organic carbon content of the soils was measured as

described by Walkley and Black[8] and sesquioxide concentration in the

soils was determined as outlined by Hesse.[11] Extractable soil Mn was

measured using diethylenetriaminepentaacetic acid.[13] Extractable soil phos-

phorus was determined using sodium bicarbonate[9] and percentage calcium

carbonate content of the soil[10] was also measured.

Experiment

The experiment comprised three replications of two soils (Kopi and

Scaddan), two Mn sources, manganese sulfate (MnSO4 � 4H2O; 24.6% Mn),

Mn oxide (MnO; 77.38% Mn) and seven amounts of manganese (mg

Mn=pot): 0 [Mn0], 2.5 [ Mn1], 5.00 [Mn2], 7.5 [Mn3], 10.0 [Mn4], 15.0

[Mn5], and 20.0 [Mn6]. About 3000 g of each soil (actual amounts listed in

Table 1) was placed into a plastic pot (170 mm diameter) lined with a

polyethylene bag. To ensure Mn was the only nutrient element limiting

plant yield, the following solutions were added to the soil in each pot

(mg=pot): NH4NO3, 250; K2SO4, 328; KH2PO4, 1000; MgSO4, 64; CaCl2,

300; CuSO4, 15; ZnSO4, 23; FeSO4, 55; CoSO4, 0.9; Na2MoO4, 0.8; and

HBO4, 1.0. On a surface area basis 227 mg=pot is about 100 kg=ha. To remove

any Mn contamination, the macronutrient salts were purified using dithiozone-

chloroform, as described by Hewitt.[14] After the soils had dried after adding

the solutions, the MnSO4 and MnO sources were added as finely ground

(<0.02 mm) dry powder. The soil in each pot was then thoroughly mixed by

shaking the soil in the plastic bag, disregarding bags between different

amounts of Mn to avoid Mn contamination. The soils were then watered

with deionized water and maintained at 75% of field capacity for 100 days to

allow the applied Mn to incubate with the soil before sowing lentil seed. The

soil incubation technique described here was used to promote Mn deficiency

where soils have been disturbed in collecting from the field, using procedures

previously described for glasshouse experiments.[15]

At the end of the incubation period, 12 seeds of lentil cv. Digger were

sown 1.5 cm deep in each pot before thinning to eight plants per pot about 18

days after sowing. For the first 20 days after sowing, soils were maintained at

75% of water-holding capacity using deionized water. Thereafter, the soils

were maintained at water-holding capacity by frequent watering to weight. The

pots were completely randomized in the glasshouse, and were re-randomized

each day while watering. The glasshouse experiment was conducted from

early April to late August (100 days incubation plus 45 days growing period,

so 145 days total). Temperatures in the glasshouse (air-conditioned) were set at

18�C day and 14�C night, plus or minus 1�C. These are typical temperatures

used to promote Mn deficiency in glasshouse studies.[15]

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During the growth of the plants, nitrogen (ammonium nitrate,

227 mg=pot) was applied every seven days.

Measurements

At 45 days after sowing, the plants were cut at ground level, divided into

youngest mature growth (YMG) and the rest of shoots (ROS), and dried at

80�C for 48 hr before weighing. For each pot, the weight of YMG was added

to the weight of ROS to give the total dried weight of shoots (TWS). After

weighing, the whole shoots were ground and digested in a nitric and perchloric

acid mixture[16] and the concentration of Mn in the digest was measured by

atomic absorption spectrophotometry.

Analysis of Data

Data for the relationship between yield of dried whole shoots and the

amount of Mn applied were adequately described by a Mitscherlich equation:

y ¼ a � b exp(�cx) (1)

where y is the yield (g=pot) of dried whole shoots (young growth plus the

remainder of the dried shoots), x is the amount of Mn applied (mg Mn=pot), and

a, b, c are coefficients. Coefficient a (g=pot) provides an estimate of the

asymptote or maximum yield plateau. The value of the a coefficient was used

as the maximum yield to calculate percentage of the maximum (relative) yield.

Coefficient b (g=pot) estimates the difference between the asymptote and the

intercept on the yield (y) axis at x¼ 0 and so estimates the maximum yield

increase (response) to applied Mn. Coefficient c (pot=mg Mn) describes the

shape of the relationship and governs the rate at which y (the yield response)

increases as x (the amount of Mn applied) increases. The larger the value of c the

more rapidly the response curve approaches the maximum yield plateau (the

steeper the curve) so that less applied Mn is required to achieve the maximum

yield plateau and any yield below the plateau. The larger the value of c the flatter

the response curve and more applied Mn is required to reach the maximum yield

plateau or any yield below the plateau. Mean data were fitted to the equation by

non-linear regression using a computer program written in compiler BASIC.[17]

For each of the two soils, the two Mn sources produced the same yield for

the nil-Mn treatment and similar maximum yield plateaus. Under these

circumstances, the c coefficient of the relationship between yield and the

amount of Mn applied was used to compare how effectively the lentil plants

used the different Mn sources to produce dried shoots.[18,19] For each soil, the

effectiveness of the Mn fertilizers was calculated relative to the effectiveness

of Mn sulfate, to provide relative effectiveness (RE) values for the different

Fertilizer Mn and Lentil 1267

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sources of Mn (REsource). This was done by dividing the c coefficient of each

source of Mn by the c coefficient of MnSO4. Therefore, by definition, the

REsource of MnSO4 is 1.00 (Mn sulfate is the present Mn fertilizer used in

southwestern Australia).

Data for the relationship between manganese content ([Mn concentration

in YMG multiplied by the yield of YMG] plus [Mn concentration in ROS

multiplied by the yield of ROS]) and the amount of Mn applied was curvi-

linear, but was not well described by the Mitscherlich equation. However, all

data for the nil and first two amounts of Mn applied were adequately described

by a linear equation:

Y ¼ A þ Bx

where y was the Mn content in the dried shoots (mg Mn=pot), x was the

amount of Mn applied (mg Mn=pot) and A and B are coefficients. Coefficient

A provides an estimate of the Mn content of the dried shoots derived from the

soil when no Mn was applied (that is, the uptake of indigenous or native soil

Mn). Coefficient B is the slope of the line and estimates the increase in Mn

content in dried shoots of lentil per unit of applied Mn. For each soil, the slope

(B) coefficient for both sources of Mn was divided by the B coefficient of Mn

sulfate to provide REMn uptake values.

The relationship between yield of TWS, expressed as relative yield, as

the dependent variable (y-axis), and the concentration of Mn in dried shoots

of YMG or ROS, as the independent variable (x-axis), was used to define

critical Mn concentration in tissue. The critical value is the Mn concentration

that was related to 90% of the maximum dried yield of shoots (TWS).[20]

The value of the a coefficient of the Mitscherlich equation fitted to the

relationship between yield of dried shoots and the amount of Mn applied

was used as the maximum yield to calculate relative yield. Data for the Mn

concentration in whole shoot of lentil did not fit a Mitscherlich equation, so

hand-drawn curves were fitted to the data and critical Mn was determined

from the hand-fitted curves.[21]

RESULTS AND DISCUSSION

Symptoms of Manganese Deficiency

Five weeks after emergence of seedlings, visual symptoms of Mn

deficiency were observed for plants growing on the Mn0 and Mn1. Manganese

deficient plants were pale with interveinal leaf chlorosis on the youngest

mature leaves. Three weeks after sowing (two weeks after emergence), the

deficient lentil plants were stunted.

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Dry Matter Production of Whole Shoots

The deficiency reduced growth at about three weeks after sowing and

growth progressively worsened. Yield of shoots of lentil increased

with increasing amounts of Mn application from both Mn sources.

However, the yield response varied with Mn source and soil type (Fig. 1).

Figure 1. Relationship between yield of dried shoots (TWS) of lentil and the amount

of Mn applied (mg Mn=pot) for (a) Kopi soil and (b) Scaddan soil. Key: In each case:

(r) MnSO4 and (j) MnO.

Fertilizer Mn and Lentil 1269

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For the Kopi soil, maximum yield increases of dried lentil shoots due to

applications of Mn were about a 50% [Fig. 1(a)]. For the sulfate source,

increases in yield occurred up to the Mn2 treatment, with the Mn3 to Mn6

treatments being on the maximum yield plateau [Fig. 1(a)]. Both Mn sources

reached the same maximum yield plateau (see ‘a’ values, Table 2). However,

for the oxide source the maximum yield was not reached until the highest

amount of Mn was applied (Mn6).

For the Scadden soil, additions of Mn increased yield of lentil shoots by

about 50% [Fig. 1(b)]. For the sulfate source, increases occurred to the fourth

level of Mn application (Mn3), whereas the oxide source reached the same

maximum yield plateau at the fifth level of Mn addition (Mn4).

For both soils, the REsource values suggests that MnO was about half as

effective as the sulfate source, so that, relative to the sulfate source, about

twice as much Mn as the oxide needed to be applied to produce the same yield

(Table 2).

Manganese Content of Dried Whole Shoots

The sources differed in their ability to supply Mn to increase the Mn

content of whole shoots of lentil (Fig. 2, Table 3). For each amount of Mn

Table 2. Values of the coefficients of the Mitscherlich equationa fitted to therelationship between yield of dried whole shoots (g=pot) and the amount of Mnapplied (mg Mn=pot) to the Kopi and Scaddan soil, and the relative effectiveness(REyield) of Mn sources.

Source a b c r2

Yield

response (%)c REyield

Kopi soil

Sulfate 2.64 1.34 0.4 0.99 51 1.00b

Oxide 2.59 1.36 0.2 0.98 52 0.50

Scaddan soil

Sulfate 2.63 1.08 0.41 0.99 41 1.00

Oxide 2.66 1.09 0.21 0.97 41 0.51

aEquation fitted: y¼ a� bexp(�cx), where y is the yield of dried whole shoots

(g=pot), x is the amount of Mn applied (mg Mn=pot), a provides an estimate of the

maximum yield plateau (g=pot), b is the yield response to added Mn (g=pot), and c

describes the shape of the relationship by estimating the rate at which y approaches the

maximum yield plateau as the amount of Mn applied is increased.bCalculated for each source by dividing c of each source by c for MnSO4, so that, by

definition, REsource for MnSO4 is 1.00.cYield response, b=a � 100%, calculated using values of a and b from above equation.

1270 Brennan and Bolland

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applied, the Mn content in shoots was always larger for the sulfate source of

Mn (Fig. 2). The REMn uptake values for Mn oxide were about 60% the REMn

uptake values for Mn sulfate, indicating that about 40% more Mn as oxide

needed to be applied to produce the same Mn content in dried shoots as Mn

applied as sulfate (Table 3). This suggests plant roots were better able to access

Mn from the sulfate than the oxide source.

Figure 2. Relationship between the Mn content in dried shoots (mg Mn=pot) of lentil

and the amount of Mn applied (mg Mn=pot) for the (a) Kopi and (b) Scaddan soils.

Key: In each case: (^) MnSO4 and (j) MnO.

Fertilizer Mn and Lentil 1271

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Critical Manganese Concentration in Tissue

Different yields for the nil-Mn treatment were produced for each of the

two soils so relative yield was used to determine critical Mn tissue test values

[Fig. 3a), (b)]. Note that when relative yield was used, for each species and

plant tissue (YMG or ROS), data for the two sources and soil types were

similar, so there are 28 data points (14 for each soil type) shown for each

species and plant tissue in Fig. 3. That is, data for the relationship between

relative yield and Mn concentration in dried shoots was similar for YMG and

ROS for both the soils. The Mitscherlich equation fitted to all YMG data for

both soils was y¼ 101.3� 20046.9exp(�0.43x), r2¼ 0.89 for YMG

[Fig. 3(a)].

For ROS, the relationship between relative yield and Mn concentration in

dried shoots showed that the plants from the nil-Mn treatment had higher Mn

concentration than plants from soil to which the two lowest amounts of Mn

fertilizer (Mn1 and Mn2 treatments for both MnO and MnSO4 � 4H2O) had

been applied [Fig. 3(b)]. This phenomenon, first described by Steenbjerg[22]

was not observed for YMG [Fig. 3(a)].

The critical Mn concentration (mg=kg) for ROS was 21 and for YMG 18.

We could find no data in the literature for the critical concentration of Mn in

either YMG or whole shoots.

Table 3. Values of the coefficients of the relationship between Mncontent in dried whole shoots (mg Mn=pot) and the amount of Mnapplied (mg Mn=pot) and the relative effectiveness (REMn uptake) ofMn sources calculated using Mn content in whole shoots.

Source Aa B r2 REuptakeb

Kopi soil

Sulfate 19.24 4.89 0.98 1.00

Oxide 17.65 2.91 0.98 0.60

Scaddan soil

Sulfate 33.16 4.36 0.98 1.00

Oxide 31.24 2.48 0.97 0.57

aEquation fitted: y¼AþB � x, where y is the Mn content of dried

whole shoots (mg Mn=pot), x is the amount of Mn applied (mg

Mn=pot), B is the slope coefficient and estimates rate of change in

Mn content per unit of applied Mn (g=pot), and A is the intercept and

provides an estimate of the Mn uptake from indigenous Mn.bCalculated for each source by dividing B (slope) of each source by B

for MnSO4, so that, by definition, REuptake for MnSO4 is 1.00.

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Figure 3. Relationship between percentage of the maximum (relative) yield of dried

shoots of lentil and the concentration of Mn in dried (a) young tissue (YMG) and (b)

whole shoots (ROS). Data are for both Mn sources, so there are 28 data points for YMG

or whole shoots for each species (14 for each soil type). In each case: (r) Kopi soiland (j) Scadden soil.

Fertilizer Mn and Lentil 1273

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CONCLUSIONS

On both soils, compared to the standard sulfate Mn source used in

Western Australia, the MnO source was about 50% less effective in

producing dried shoots and about 40% less effective in increasing the Mn

content of the dried shoots of lentil. That is, about twice as much Mn as the

oxide source was required to produce the same relative yield as the sulfate

source. Although MnO is only slightly soluble in water, its effectiveness for

plant uptake has been found to increase if applied as a finely ground

powder.[23,24] We did not study the effect of particle size of Mn fertilizer in

this work. Our results support the results of various workers who found

MnO to be less effective than MnSO4 for a range of crop species.[25–28] In

some situations, MnO did not alleviate Mn deficiency.[26,29] However, in the

present study, the oxide source at the highest amount applied did fully

alleviate Mn deficiency in lentil.

ACKNOWLEDGMENTS

The Chemistry Centre (WA) measured soil properties and Mn concentra-

tions in tissue. The Department of Agriculture of Western Australia provided

funds and facilities.

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