Published April 19, 2013

Use of a Chlorophyll Meter to Monitor Nitrogen

Status and Schedule Fertigation for Corn
T. M. Blackmer and J. S. Schepers

Research Question Corn production may lead to groundwater contamination by nitrate

when N fertilizers are applied in excess of the crop’s needs. Practices
that will synchronize N fertilization with the crop’s needs could help
reduce the potential for groundwater contamination by nitrate without
reducing yields.

Literature Summary Monitoring leaf N concentration provides a means of identifying crop N

status, but correlation of this measurement to grain yield is confounded
r
by luxury consumption and requires time consuming lab analysis. Leaf
chlorophyll measurements can also detect N deficiencies, but are not as
sensitive to luxury consumption as other N measurements. The SPAD
502 chlorophyll meter provides an instantaneous means of evaluating
chlorophyll content.

Applied Questions Could the chlorophyll meter detect N deficiencies that resulted in
reduced grain yields?

The chlorophyll meter, when used with a reference strip (an area receiv-
ing adequate N), accurately detected N deficiencies that resulted in
reduced grain yields. Using chlorophyll meter readings without an in-
field reference strip resulted in variability from such factors as different
hybrids, soil types, and growth stage.

Did all sites have the same pattern of N availability over the season?

All sites responded differently. Some sites became more N deficient as

the season progressed. In others, more N became available as the season
progressed. Nitrogen availability at one site remained constant over the
season. This variability may apply only to irrigated sites in the area
studied.

Could deficiencies detected by the chlorophyll meter be corrected soon

enough to prevent yield loss?

Treatments that started with adequate fertilizer and then became defi-
cient were corrected without yield loss. Young plants in a deficient state
could not be corrected to full yield potential.

Full scientific article from which this summary was written begins on page 56 of this issue.

J. Prod. Agric., Vol. 8, no. 1, 1995 11

 Use of a Chlorophyll Meter to Monitor Nitrogen Status
and Schedule Fertigation for Corn
T.M. Blackmer* and J.S. Schepers

The SPAD 502 chlorophyll meter' (Minolta Camera Co., Excessive levels of available N can result in luxury con-
Osaka, Japan) is a new instrument that has been introduced sumption, but usually do not increase the meter readings
as a tool to improve N management. This study was conducted (Schepers et al., 1992a). The plateau in chlorophyll meter
to evaluate the ability of the chlorophyll meter to detect plant readings at high fertilizer N rates is thought to reflect the
N deficiencies in corn (Zeu mays L.) by identifying when it fact that nutrients other than N are limiting chlorophyll
would be appropriate to supply N fertilizer in irrigation water. production. As such, luxury consumption is not detect-
Nitrogen response studies were conducted on five irrigated sites ed by the meter, which makes it ideal for detecting an
in central Nebraska in 1991. Crop N status was monitored dur- N deficiency. Plants treated the same except for N avail-
ing the growing season and additional N was added to simu- ability should show differences only if an N deficiency
late fertigation when apparent N deficiencies were detected with exists. Because chlorophyll meters are not sensitive to lw-
the meter. Changes in N status over the season were determined ury consumption, one need not know the exact level of
relative to an adequately fertilized in-field reference plot. Early- N required for maximum yield as long as a slight excess
season (V6) N deficiencies were poorly correlated with yield be-
is maintained in a reference area of the field. Well ferti-
cause factors such as nitrate leaching, organic matter minerali-
lized reference areas can be established within a field and
zation, and nitrate present in irrigation water modified the crop weekly comparisons of chlorophyll meter readings can
N supply during the growing season. Nitrogen deficiencies de-
tected late (R4-R5) in the season were more highly correlated be made to determine if deficiencies are present. If a defi-
with yield than early season N stresses. Treatments that started ciency is detected, fertilizer can be applied with the irri-
with adequate fertilizer and then became deficient were corrected gation water. This strategy permits fertilizer to be applied
without yield loss. Young plants in a deficient state could not only when needed. This N management approach ac-
be corrected to full yield potential. Chlorophyll meters can be counts for fluctuations in seasonal N availability result-
a valuable tool for N management of irrigated corn produc- ing from parameters difficult to estimate before the
tion when used to assess crop N status in the irrigated Great growing season, such as N mineralization or leaching loss-
Plains. es. For this strategy to be practical, however, deficien-
cies must be detected early enough to be corrected by
supplemental fertilizer N before the yield potential
declines.
L EAF N CONCENTRATION at silking has been shown to
be highly correlated with grain yield in corn (Tyner
and Webb, 1946). Chlorophyll concentration (leaf green-
The use of chlorophyll meters offers several advantages
over conventional tissue testing procedures for detecting
N deficiencies. The most obvious advantages are porta-
ness) in corn has been found to be positively correlated bility and rapid assessment of N status in the field without
with leaf N concentration (Wolfe et al., 1988; Lohry, destroying plant tissue. In a practical sense, chlorophyll
1989; Wood et al., 1992) and N sufficiency (Zelich, 1982; meters have been shown to be an effective tool for iden-
Girardin et al., 1985; and Lohry, 1989). It follows that tifying sites that are responsive and nonresponsive to side-
leaf chlorophyll concentration reflects relative crop N sta- dressed N (Piekielek and Fox, 1992). Chlorophyll meters
tus and yield level. The SPAD 502 chlorophyll meter pro- have also been shown to effectively quantify N status dur-
vides a convenient means of assessing relative leaf ing the reproductive stages of corn growth (Schepers et
chlorophyll concentration and therefore offers a promis- al., 1992b; Wood et al., 1992). These observations raise
ing tool for evaluating the N status of corn during the the possibility of using this tool to identify sites that
growing season. To measure relative chlorophyll concen- would be responsive to N fertilizers and where N could
tration, the meter is clamped on the corn leaf and light be injected into irrigation water during the growing sea-
transmittance through the leaf is determined at 650 and son to correct an N deficiency. Such use of the meter
940 nm. The transmittance at 940 nm is used as a refer- offers potential economic and environmental advantages
ence to compensate for factors such as leaf moisture con- because it enables producers to reduce rates of insurance
tent and thickness while the 650 nm source is sensitive N with minimal risk of yield reduction. However, the con-
to chlorophyll concentration. Chlorophyll meter readings ceptual use of chlorophyll meters presented above has not
provide a relative indication of leaf chlorophyll concen- been evaluated under field conditions. The objective of
tration (Yadava, 1986; Marquard and Tipton, 1987; this report is to evaluate the chlorophyll meter as a tool
Dwyer et al., 1991). for detecting N deficiencies and scheduling N fertiliza-
tion during the growing season of irrigated corn.
T.M. Blackmer, Dep. of Agronomy, Univ. of Nebraska, and J.S.
Schepers, USDA-ARS and Dep. of Agronomy, Univ. of Nebraska, Lin-
coln, NE 68583. Joint contribution of USDA-ARS and the Nebraska
Agric. Res. Div., J. Series no. 10472. Received 20 Aug. 1993.
'Corresponding author ([email protected]). I Mention of trade names or proprietary products does not indicate
endorsements by USDA, and does not imply its approval to the exclu-
Published in J. Prod. Agric. 856-60 (1995). sion of other products that may also be suitable.

56 J. Prod. Agric., Vol. 8, no. 1, 1995

Table 1. Site description for N fertilizer response trials conducted in central Nebraska.
Water Initial
Site Soil type nitrate-N Previous crop Residual N t Hybrid$ N rate Fertigationss

PPm lb Nlacre lb Nlacre no. lb Nlacre

1 Leshara silt loam (fine-silty, mixed, mesic 2 Sweetclover [Melliotus officinalis 65.1 3394 0 4 120
Typic Haplaquolls) (L.)Lam.] 40 3 90
80 1 30
120
160
200
2 Wood River silt loam, (fine, montmorillonit- 1 Soybeans [Glycine max (L.) 52.2 3417 0 4 120
ic, mesic Typic Natrustolls) Merr.] 40 3 90
80 2 60
120 1 30
160
3 Hord silt loam (fine-silty. mixed, mesic Cu- 17 Corn (Zea mays L.) 47.4 3162 0 2 60
mulic Haplaquolls) 40 1 30
80
120
160
4 Hord silt loam 6 Sorghum [Sorghum bicolor (L.) 26.2 Unknown 0 1 30
Moench] 40
80
120
160
5 Hord silt loam 31 Corn 49.0 3379 0 2 60
40 1 30
80
120
160
t Calculated from available N (ammonium and nitrate) present in the top 24 in. of soil.
$ Pioneer brand hybrid.
8 Fertigation applied rates of 30 lb Nlacre, and values in parenthesis represent the total N applied.

Table 2. Linear regression parameters for relationships between

MATERIALS AND METHODS grain yields and chlorophyll meter readings.
V-6 stage R4-R5 stage
This study was conducted on five N-response irrigat-
ed corn sites in central Nebraska (Table 1). Preliminary Site Intercept Slope r2 Intercept Slope r2
residual soil samples were collected by gathering 16 cores 1 -46.0 5.5 0.48 -53.3 4.8 0.88
(0.70 in. diameter, to a depth of 24 in.) with a hand probe. 2 -2.6 2.9 0.06 64.1 1.6 0.89
3 29.8 4.6 0.67 -87.2 5.4 0.89
The samples were oven dried, ground to pass a 14 mesh 4 -835.9 25.6 0.31 4.9 2.0 0.79
sieve, and extracted using 2 N KCl. Nitrate and ammon- 5 139.6 0.8 0.55 5.6 3.1 0.55
ium-N concentrations were determined using a Lachat Combined -25.9 4.3 0.25 -155.4 6.3 0.84
flow-injection analyzer (Lachat Instruments, Mequon,
WI). At each location, plots were either 30 by 40 ft (12 recent fully collared leaf). First readings were collected
rows, 30-in. spacings) or 48 by 40 ft (16 rows, 36-in. spac- when corn plants were at V6 (12 in. tall), as discussed
ing) arranged in randomized complete-block designs with by Ritchie et al. (1992). Measurements were taken mid-
four replications. Each study was planted with a differ- way between the stalk and the tip of the leaf, and mid-
ent corn variety between 21 Apr. and 14 May 1991. Am- way between the margin and the mid-rib of the leaf from
monium nitrate fertilizer was broadcast at various initial 30 representative plants randomly selected from the center
N rates (Table 1) and incorporated with a rototiller shortly two rows of each plot. Plants unusually close together
after planting. At sites 1,2, 3, and 4, soils were cultivat- or far apart, or those that were damaged were not sam-
ed to form furrows for irrigation before the corn was 18 pled. After silking, readings were taken using similar
in. tall. Site 5 required no furrows because irrigation was methods, except the ear leaf was sampled. Chlorophyll
applied with a sprinkler system. Other than fertilizer ap- meter data were collected until leaf senescence increased
plication and grain harvest, all plots were managed us- plant-to-plant variability (approximately R5).
ing practices common to corn production in the Central Treatments were characterized as N deficient at the site
Platte River Valley. when the sufficiency index was < 0.95 for two successive
To monitor the N status of the crop, N sufficiency in- weeks. When such deficiencies were detected, the plots
dexes were calculated as the ratio of chlorophyll meter were split and N was applied to one of the subplots to
readings for the treatment receiving lower amounts of N simulate fertigation. Nitrogen was applied to the split
fertilizer (all treatments except the highest N rates) to the plots as ammonium nitrate (dissolved in water) at a rate
treatment receiving the highest amount of N (the refer- of 30 Ib N/acre (a typical rate of N that could be applied
ence area). through irrigation systems without injuring corn plants).
Chlorophyll meter readings were collected using the These fertilizer applications were then incorporated with
SPAD 502 at approximately weekly intervals during the typical irrigation for the site.
growing season. Prior to silking, readings were collected The center two rows of each plot were hand harvested
from the most recent fully expanded leaf (i.e., the most using 20-ft segments from each plot. A mechanical sheller

J. Prod. Agric., Vol. 8, no. 1, 1995 57

 Table 3. Fertilizer rate comparisons that resulted in differences son. This site had been depleted of N by previous crop-
in R-5stage chlorophyll meter readings and corresponding N ping without fertilization as reflected by low residual N
fertilizer effects on grain yield. Comparisons include only non-
fertigated treatments.t in the soil (Table 1). No starter fertilizer was applied at
site 4 and wet weather caused a delay in application of
Probability of a greater F value
the initial N treatments until the plants were about 4-in.
Site Comparison$ For yield For chlorophyll meter tall.
1 0 vs. 80 0.006 0.039 Relationships between grain yields and chlorophyll
1 0 vs. 120 0.004 0.016 meter readings taken when the corn plants were at V6
1 0 vs. 160 0.011 0.011
1 0 vs. 200 0.002 0.006 showed a trend for increased yields as chlorophyll meter
1 40 vs. 120 0.049 0.050 readings increased within locations, but variation among
1 40 vs. 160 0.126 0.019 locations resulted in poor predictability (Fig. 2). When
1 40 vs. 200 0.026 0.034
2 0 vs. 40 0.267 0.017 data from all sites were combined, the relationships be-
2 0 vs. 80 0.075 0.002 tween chlorophyll meter readings and grain yields had
2 0 vs. 120 0.014 0.001 poor predictability, even though it was statistically sig-
2 0 vs. 160 0.008 0.001
2 40 vs. 120 0.064 0.038 nificant (r2 = 0.25, P < 0.01). Poor relationships (poor
2 40 vs. 160 0.114 0.002 predictability) at V6 were also observed when data from
2 80 vs. 160 0.236 0.013 several of the individual sites were analyzed separately
3 0 vs. 120 0.138 0.044
3 40 vs. 120 0.171 0.015 (Table 2). These observations indicate that, early in the
4 0 vs. 40 0.226 0.044 season, chlorophyll meters have limited potential as a
4 0 vs. 80 0.090 0.001 general tool for predicting yield responses to N fertilizer
4 0 vs. 120 0.428 0.009
4 0 vs. 160 0.223 0.003
unless the readings can be adjusted to account for other
environmental differences.
t Fertigation treatments were not significantly different than the reference
plots. Relationships between grain yields and chlorophyll
$ Initial N treatments in lb Nlacre. meter readings at R4 (dough stage) or R5 (dent stage) were
was used to separate the grain from the cob. Grain yields better than V6 relationships both within and across lo-
were adjusted to 15.5% moisture content. cations (Fig. 3). When data from all sites were combined,
The regression analyses (Table 2) were performed on a statistically significant linear correlation (r2 = 0.84, P

....
the relationship between grain yields and chlorophyll
meter readings at the V6 growth stage and the R4-R5 3 2501 SITE 1
growth stage. To detect treatment differences for Table b
3, Fisher's LSD approach was used on the nonfertigated c?
3 200-
AAA #
plots. All comparisons found statistically different ( P e SITE 3 Q3
SITE 5
< 0.05) for either the grain yield parameter or the chlo- 9 150-
rophyll meter parameter at the R4 to R5 stage are report-
ed. Analysis of variance was used to determine grain yield u-l
SITE 2
response to N.
z
100-
t
RESULTS AND DISCUSSION $ 50- SITE 4
(3
Corn grain yields observed for the highest N rates
ranged from 72 to 225 bu/acre (Fig. 1). Analysis of vari- 30 35 40 45 50 55 60
ance showed a significant ( P < 0.05) effect of N rate at CHLOROPHYLL METER READINGS
sites 1 and 2 only. Relatively low yields at site 4 may have Fig. 2. Relationships between corn grain yields and early season chlo-
rophyll meter readings at the V6 growth stage.
been caused by an extreme N deficiency early in the sea-
- 250- SITES 1 8 3

(d
3 200-
3 200
SITE 3
e SITE 2
BP
e SITE 5 n
-I
150-
0 .
e m
SITE 5
e
SITE 2
II!
100-
A .AA
z A
z SITE 4 50- SITE 4
(3
(3 04 . ' .
- .
' I 1 . 1 . I '

", . I . I . . 30 35 40 45 50 55 f D
0 40 ao 120 160 200 240
N RATE (Ib N/acre) CHLOROPHYLL METER READINGS
Fig. 3. Relationships between corn grain yields and late season chlo-
Fig. 1. Corn grain yield responses to initial fertilizer N rates at five sites. rophyll meter readings at R4-RS growth stage.

58 J. Prod. Agric., Vol. 8, no. 1, 1995

< 0.001) between chlorophyll meter readings and grain 2). The lack of relationship between grain yields and
yields was observed. The practical significance of this early-season chlorophyll meter readings at site 2, for ex-
relationship for diagnosing N deficiencies is questionable, ample, should be expected because yield differences were
however, because the trend was largely related to differ- caused by deficiencies that developed later in the season.
ences between sites instead of differences due to N ap- The lack of a significant relationship between early sea-
plied within sites. The overall trend shown in Fig. 3 was son meter readings and grain yield at site 5 resulted from
caused by differences in leaf greenness related to factors an N deficiency observed early in the season that was ap-
such as hybrids, rotation, and locations rather than N parently corrected by nitrate present in the irrigation
status. These observations indicate that chlorophyll meter water or N supplied by mineralization.
use in midseason probably has limited potential as a tool Leaf greenness can be influenced by a number of fac-
for predicting yield responses to N fertilizer unless the tors (e.g., hybrid, stage of growth, and some nutrients),
readings are interpreted for a specific location. but soil N availability probably has the greatest effect
Seasonal trends in N sufficiency indexes observed on within a field. Changes in N sufficiency indexes through-
unfertilized plots were different at each site (solid circles) out the growing season reflect the initial N status of the
in Fig. 4. The severity of N deficiency tended to decrease soil and N additions from various sources (Fig. 4). Be-
with time at sites 3 and 5, but tended to increase with cause fertigation treatments were applied only when defi-
time at sites 2 and 4. Increasing N deficiency over the ciencies were detected, the number of fertigated plots
season could be explained by depletion of available N as varied among the sites. In general, plots receiving lower
a result of plant uptake and low N content in irrigation fertilizer application rates at planting required more fer-
water. Situations showing increasing N sufficiency over tigation, while sites with greater N fertilizer inputs and
time could be explained by mineralization of available N from irrigation water (sites 3 and 5) required less ferti-
N from organic matter, more extensive extraction of N gation (Table 1). Situations with low concentrations of
from volumes of soils containing available N, or avail- nitrate in the water or a high leaching potential had the
ability of nitrate in irrigation water. Nitrates in irriga- greatest need for fertigation (site 2). Prescribed fertiga-
tion water offer a reasonable explanation for the tion treatments were able to maintain minimal N defi-
correction of deficiencies at sites 3 and 5 because large ciencies as indicated by the sufficiency index (Fig. 4).
amounts of nitrate were contained in the water (Table Evidence that the chlorophyll meter had adequate sen-
1). The failure of nitrate in irrigation water to correct defi- sitivity to detect N deficiencies that could significantly
ciencies at site 4 may be explained by visual observations reduce yields is provided in Table 3. This table contains
of poor early season root development and lower levels only comparisons between fertilizer N rates that were sig-
of residual N resulting in a visual early season N defi- nificantly different for either yield or chlorophyll meter
ciency that lowered the yield potential. Grain N concen- readings at the R5 growth stage. Results show that eight
trations for this site were high for the higher N treatments grain yield comparisons and 20 chlorophyll meter read-
suggesting that N availability was not a problem (data ing comparisons were statistically different. Each of the
not presented). eight cases that was statistically different for yield was
Seasonal trends in N sufficiency indexes illustrated in also statistically different for chlorophyll meter reading
Fig. 4 help to explain why late-season chlorophyll meter comparisons. This concurrence of observations is a strong
readings resulted in better relationships with grain yields indicator that the chlorophyll meter can detect N defi-
than did early season chlorophyll meter readings (Table ciencies that could result in yield reductions.

1.1 -r I
I

I=== n
I I

I I I
I
--
I

I Ir
I

r w
I

ar1
I
- NITROGEN RATE
(Ib N/acre)
INITIAL FERTIGATION
1.0 - 0 0 NO
5 0 0 YES

z 0.9 - -- v 40 YES
80 YES
>-
O 0.8 - SITE 1 -- SITE 2 \ 120 YES
Z
w
I I I I I I I

-I --
I I I I I I I I I I I

g 1.1
LL r I = I I 1111 1 1 r1
2
LL
1.0 - -- r..-.-&jz=-%.
0’ .A=*
0.9 -
0.8 - SITE 3
- SITE 4 SITE 5
I I I I I 1 I I 1 I I I

160 180 200 220 160 180 200 220 160 180 200 220
CALENDAR DAY
Fig. 4. Nitrogen sufficiency indexes for plots receiving various combinations of initial N rates and fertigation treatments.

J. Prod. Agric., Vol. 8, no. 1, 1995 59

 Reasons for the greater number of significant compar- ability of the chlorophyll meter to schedule fertigation
isons with the chlorophyll meter than with yield could is promising because it offers the possibility of conserv-
be a result of at least two factors. First, a higher vari- ing fertilizer and protecting the environment. More
ance for the yield measurements than for the chlorophyll research is needed to identify the best N sufficiency in-
meter readings could result in detecting fewer yield com- dex to initiate fertigation and to determine if this strate-
parisons that were truly different (type II error) from gy has application outside of the Great Plains.
chlorophyll meter readings. When using the chlorophyll
meter, damaged plants were not sampled and the same
number of plants (30) were always sampled for each plot.
In contrast, yield measurements were made on an area
basis. Yield variability is inevitable because damaged
plants were included in the measurements, and plant
stands may vary from plot to plot. This leads to the poten-
tial for greater variability for yield data than from chlo-
rophyll meter measurements.
Secondly, the detection of luxury consumption by the
meter could result in more significant comparisons for
the chlorophyll meter readings than grain yield. However,
because the chlorophyll meter was able to detect differ-
ences between 0 and 80 Ib N/acre, but usually was not
able to detect differences between 80 and 160 Ib N/acre,
suggests that luxury consumption does not present a
problem when interpreting the data. If luxury consump-
tion was a significant problem, we would expect more
significant comparisons involving higher N rates.
Neither the chlorophyll meter nor yield response meas-
urements were able to detect differences between the
highest initial N treatments (adequately fertilized) and the
fertigation treatments (Table 3). This observation presents
preliminary evidence that use of the chlorophyll meter
is an effective way to schedule fertigation in the irrigat-
ed Great Plains Region.

CONCLUSIONS

Chlorophyll meters were able to distinguish between

fertilizer N treatments that resulted in N deficiencies that
reduced corn grain yields. Chlorophyll meter data fol-
lowed trends similar to grain yield. Calculating an N suffi-
ciency index relative to chlorophyll meter readings from
a non-N-limited area made it possible to compare crop
N status across fields, hybrids, and sampling dates. The

60 J. Prod. Agric., Vol. 8, no. 1, 1995