Field diagnostic techniques to determine nitrogen in maize

Técnicas de diagnóstico en campo para determinar nitrógeno en maíz

Eduhin Roacho-Cortés1 , Javier Z. Castellanos-Ramos2‡ , and J. D. Etchevers1

1
Colegio de Postgraduados, Campus Montecillo. Carretera México-Texcoco km 36.5, Montecillo. 56230 Texcoco, Estado de México, México.
2
Fertilab. Poniente 6 #200, Cd. Industrial de Celaya. 38010 Celaya, Guanajuato, México.
‡
Corresponding author ([email protected])

SUMMARY an arithmetic-graphic technique for the crop to reach a

relative yield of 90% for each method. The proposed
Few technologies are available to quantify crop's CC results require further validation before being
nutritional status during their vegetative cycle, such as used in a commercial application. In synthesis, this
in maize. Semiquantitative alternatives could replace work proposes a procedure for diagnosing N nutrition
the laboratory foliar analysis's classical approach for of maize in the f ield that might improve the current
measuring nitrogen (N) in maize (Zea mays L.) in methods used for this purpose.
the f ield, although they require regional calibration
against standard laboratory tests. In this work, we have Index words: ion selective method, nitrate-nitrogen,
calibrated four semi quantitative and two quantitative NDVI, plant sap, SPAD, total nitrogen, Zea mays.
methods for diagnosing maize nutritional status in the
El Bajio region, Guanajuato, Mexico. An experiment RESUMEN
to test the effect of increasing N rates application on
plant sap (cell extract) N was set up during the 2019 Existen pocas tecnologías, aparte del análisis
agricultural cycle. Five N treatments (0, 120, 240, foliar clásico de laboratorio, que pueden emplear
360, and 480 kg of N·ha-1) were tested, and each los productores para dar seguimiento al estado
treatment was replicated f ive times. We have tested nutrimental que guardan los cultivos a lo largo del
the following semi quantitative N methods: plant sap ciclo vegetativo. Este es el caso particular del maíz.
nitrate concentration (N-NO3) of the central midrib of Sin embargo, se dispone de métodos alternativos, de
the recently mature leaf (RML), plant sap N-NO3 of the carácter semicuantitativo para medir la situación del
base stem (in both cases measured by the ion selective nitrógeno (N) en el maíz en campo, pero para que sean
method, using the portable LAQUAtwin® model NO3- efectivos, deben ser calibrados regionalmente contra
11 nitrate ion meter, single-photon avalanche diode el procedimiento estándar de laboratorio. En este
(SPAD) unit record, and the Normalized Difference ensayo se calibraron cuatro métodos semicuantitativos
Vegetation Index (NDVI). Additionally, a quantitative y dos cuantitativos, para diagnosticar la condición
laboratory method for determining N-NO3 in the nutrimental del nitrógeno (N) del maíz en el Bajío
lower part of the maize plant's stem and total N in the guanajuatense. Para ello se estableció un experimento
RML was used. Data were acquired at different plant en el ciclo agrícola 2019. Los tratamientos fueron dosis
phenological stages: V7, V12, R1, and N-NO3 in the crecientes de N (0, 120, 240, 360, 480 kg de N·ha-1).
dry matter of the stem's base at R5. These results were Cada tratamiento se repitió cinco veces. Los métodos
correlated with the relative grain yield and those with a semicuantitativos calibrados fueron: concentración de
correlation coeff icient (r) greater than 0.7 were accepted. nitrógeno nítrico (N-NO3) del extracto celular (EC)
Later, a critical concentration (CC) was estimated using de la nervadura central (NC) de la hoja recientemente

Recommended citation:
Roacho-Cortés, E., Castellanos-Ramos, J. Z., & Etchevers, J. D. (2021). Field diagnostic techniques to determine nitrogen in maize. Terra Latinoamericana, 39:
1-12. e820. https://doi.org/10.28940/terra.v39i0.820

Received: July, 02, 2020. Accepted: March, 25, 2020.

Article. Volume 39, April 2021.
2 TERRA LATINOAMERICANA VOLUMEN 39, 2021. e820

madura (HRM), N-NO3 del EC en la base del tallo Easy-access alternatives and economic methods to
(BT), observación de las unidades SPAD, Índice de diagnose crop N supply have been developed in the last
Vegetación de Diferencia Normalizada (NDVI) y four decades (Rauschkolb et al.,1974; Schepers, Francis,
dos métodos cuantitativos de laboratorio: N-NO3 en Vigil, and Below, 1992; Withers, 1982). Currently,
la materia seca (MS) de la BT y N total foliar. Estas available semi quantitative methods offer quick results
mediciones se realizaron en las etapas fenológicas V7, to perform adjustments in the fertilization dose plan
V12, R1 del maíz, y en R5 cuando se evaluó el N-NO3 during the agricultural cycle (Justes, Meynard, Mary,
en la BT. Los resultados de las mediciones realizadas and Plénet, 1997; Etchevers, 1999) at a reasonable
con cada método, fueron correlacionados con el cost (Muñoz-Huerta et al., 2013) and straightforward
rendimiento relativo de grano (RR). Se aceptaron interpretation (Rauschkolb et al., 1974). Nonetheless,
aquellas asociaciones que tuvieron un coef iciente they should be previously calibrated for each crop, and
de correlación (r) superior a 0.70. Para cada método possible for each specif ic agroecological zone.
se estimó mediante una combinación de técnicas Diagnostic methods to assess the nutritional
aritmética y gráf ica, una concentración crítica (CC), status consist of sampling a given portion of the plant
def inida como la concentración suf iciente para que biomass to measure an indicator of the response to the
el cultivo alcanzase un rendimiento relativo (RR) de application to an increased nutrient dose. The indicator
90%. El trabajo constituye un modelo de cómo abordar can be the dry matter accumulation or the nutrient
el problema del diagnóstico de la nutrición nitrogenada concentration. Biomass accumulation -the product
en campo. Las CC propuestas en este trabajo para of the increasing fertilizer dose applied- is related to
diferentes estados fenológicos, se deben entender como the specif ic nutrient concentration in plant sap or dry
preliminares y requieren ser validadas a nivel de lotes matter extracts. The nutrient concentration is specif ic
comerciales en campo. to the plant part collected and the phenological stage.
The values for making the diagnostic are given in pre-
Palabras clave: método selectivo de iones, nitrógeno- established protocols. The best results are achieved
nitrato, NDVI, savia vegetal, SPAD, nitrógeno total, when the experimental crop develops in the absence of
Zea mays. restrictive conditions of any growth factor, except for
the factor under study, that is, water, light, temperature,
INTRODUCTION etc. (Mills and Jones, 1996; Smith and Loneragan,
1997; Alcántar-González and Sandoval-Villa, 1999;
To achieve highly qualitative and quantitative yields Etchevers, 1999).
-without harming the ecosystems- agricultural practice Several semiquantitative methods with various
demands management activities to control plant functioning principles -electrochemical, colorimetric,
nutrition. Soil fertility control by chemical methods optic, enzymatic, photo-tele detection, photochemical
and adequate fertilization application are among these activities- to mention some (Sánchez, Molinos,
practices. Nutrient demand varies as the crop develops, Alcántar, and Sandoval, 2016) are used for diagnosing
and therefore it is essential to know the concentration the N status in crops. For example, Reuter, Edwards, and
dynamics of one or several essential plant tissue Wilhelm (1997) and Mills and Jones (1996) suggested
elements at the different phenological stages. This using portable ion meters of specif ic electrochemical
knowledge allows performing management practices principle, to analyze plant N-NO3, including a section
to satisfy the plant nutritional demand by eff icient with interpretation ranges for different crops. These
crop fertilization and simultaneously takes care of the authors also indicated the feasibility of using optical
environment (Dobermann, 2005; IPNI, 2013). chlorophyll readers, such as SPAD 502® (Minolta,
The chemical plant tissue analysis to diagnose Osaka JP) for this purpose. In maize, Reuter et al.
crop nutritional supply -notwithstanding its reliability- (1997) showed that N-NO3 ranges in the base of stem
shows limitations for its massive use (Muñoz- on a dry base is useful to interpret the N status of the
Huerta et al., 2013; Ali, Al-Ani, Eamus, and Tan, plant. Several methods for N diagnosis in crops, their
2017), including inaccurate knowledge of the sample functioning, calibration for different crops, linear or
collection season, high costs, delay in obtaining, and non-linear relationships as compared to other more
diff iculty in interpreting the laboratory results. robust diagnostic procedures, and other aspects for their
 ROACHO-CORTÉS ET AL. NITROGEN DETERMINATION IN MAIZE 3

implementation are compiled in extensive literature after sowing, 35% of the corresponding total N dose
reviews (Lemaire, Jeuffroy, and Gastal, 2008; Titolo, as urea; and (iii), 54 days after sowing the remaining
20121; Muñoz-Huerta et al., 2013; Ali et al., 2017; 35% of the N dose as ammonium sulphate. At sowing,
Rhezali and Lahlali, 2017; Colaço and Bramley, 2018). the experimental lot received a basal phosphorus (P)
Therefore, this study's objective was to calibrate f ive and potassium (K) fertilization of 200 kg·ha-1 of P205
semi quantitative f ield methods and two quantitative and 100 kg·ha-1 of K 2O applied as monoammonium
laboratory methods with f ield experiments to diagnose phosphate and potassium chloride, respectively, and
the nutritional N status in maize in the region of El an extra 100 kg ha-1 of K 2O in the second fertilization.
Bajio, Guanajuato, Mexico. When the plants reached the vegetative
phenological stages V7, V12, and at the start of the
MATERIALS AND METHODS reproductive stage R1, plant tissue samples were
collected for analysis using four semi quantitative and
Study Site two quantitative methods to diagnose the nutritional N
status. The exception was the sampling for performing
Experimental plots were established with maize the analysis of N-NO3 at the dry stem base of the plant,
(Zea mays L.) crop and fertilized with increasing where the sample was obtained at R5. Phenological
N doses in a commercial lot of Juventino Rosas stages for sampling were based on selected crop stages
Municipality, Guanajuato, Mexico (20° 35' 19.2" N, provided by Castellanos et al. (2019).
100° 56' 46.8" W). The soil classif ication corresponds
to a clayey pellic Vertisols (Vp) (WRB, 2015). Before Concentration of Plant Sap N-NO3 of the Central
planting the experiment, non-fertilized maize was Midrib of the RML and of the Base of Stem
established to reduce the soil N content. Soil depth
increment (0-30 cm) had a bulk density of 1.1 g·cm‑3, In each experimental unit, 12 central midribs of the
hydraulic conductivity of 0.77 cm·h-1, soil pH = 7.0 RML were collected in the cases previously indicated.
cation exchange capacity of 34 cmol(+)·kg-1, and organic In the R1 stage, the samples were obtained from the
matter content of 2.0%. After the lot was depleted of leaf wrapping the corn ear (feminine maize flower) and
N, the soil contained 89 kg·ha-1 of N-NO3 in the f irst that of the base of the stem (ten 15 cm sections were
0.60 meters. taken from 10 cm above the soil surface) (Binford,
Twenty-f ive experimental units with an area of Blackmer, and El-Hout, 1990).
30.4 m 2 each, f ive seeding furrows distant 0.76 m with Sample collection was conducted between 09:00
8 m length each, were sown on May 23, 2019 and had and 11:00 am and samples were kept in plastic bags
a plant density at harvest time of 95 000 plants ha-1. in a cooler at 5 °C before transferring them to the
The fertilization treatments consisted of f ive N doses laboratory. Central mid ribs of the RML and base
(0, 120, 240, 360, 480 kg·ha-1), and each treatment was steams were compressed with a manual press (Cadahía-
replicated f ive times. The experimental design was a Lopez, 2008) for obtaining the plant sap samples. The
randomized block. plant sap samples were diluted in distilled water (1:10)
The plots received N application at three different and homogenized with a vortex to reduce interferences
opportunities: (a) at sowing; 35% of the total N (Di Gioia et al., 2010). The collected material was
dose corresponding to each treatment applied as maintained refrigerated (5 °C) until their analysis later
monoammonium phosphate and ammonium sulfate, in the day. A portable LAQUAtwin® model NO3-11
except for the control treatment where only triple (Horiba Kyoto, JP) was used to determine the plant
superphosphate was applied to maintain the plot sap, or cellular N-NO3 extract concentration. The
without N. The treatments with N at 120 or more kg·ha-1 N-NO3 determination was carried out in triplicate, and
received: (i) initially 30% of N, using the sources of the coeff icient of variation verif ied to not exceed 15%;
ammonium suphpate and monoammonium phosphate otherwise, the analysis was repeated. The N-NO3 meter
and the rest of the required dose of N as ammonium was calibrated with the manufacturer solutions (150
sulphate and urea in subsequent dates; (ii) 20 days and 2000 mg L-1 of NO3) every two-triplicate series.

1
Titolo, D. (2012). Using an active optical sensor to improve nitrogen management in corn production. Master’s Theses and Dissertations-
Plant and Soil Sciences. 11. Retrieved from http://uknowledge.uky.edu/pss_etds/11/
4 TERRA LATINOAMERICANA VOLUMEN 39, 2021. e820

N-NO3 Concentration in the Base of the Stem (Dry Harvest

Matter)
When maize reached physiological maturity (R6),
In samplings at V7, V12, R1, and R5, a basal stem corn cobs were harvested manually in 2.0 m 2 in the
fraction from 10 plants was collected. The samples central furrows of each experimental unit, avoiding
were taken to the laboratory and oven dried at 70 °C the edges. The corn cobs were shelled, and the yield
until they achieved constant weight. The plant material was quantif ied adjusted to 155 kg of moisture t-1 of
was grounded and sieved through a 0.42-mm mesh to grain (CIMMYT, 2012).
determine N-NO3 concentration on a dry base weight
(Cataldo, Maroon, Schrader, and Youngs, 1975). Statistical Analysis

Monitoring Recently Mature Leaves with SPAD An analysis of variance (ANOVA) and a
comparison of means test (Duncan, α = 0.05) were
The SPAD instrument measures the relative performed to determine signif icant yield differences
chlorophyll concentration. When plants reached the among the N fertilization treatments. Yield data is
phenological stage V7, 15 plants located in the central expressed as relative grain yield percentage (RY)
rows of each plot, were selected, and marked (tagged) (Equation 1):
to measure them again in V12, and R1 the SPAD
units with the MC-100 Chlorophyll Concentration Grain yield in the experiment plot
RY = ( ) 100
Meter ® (Apogee Instruments, USA). The SPAD units Average grain yield of the treatment with 480 kg of N · ha−1
were measured on the RML between the vein and the
edge of the leave when the plant showed phenological The measurements using each of the described
stages V7 and V12; in R1 the measurement was methods were graphically and statistically related
performed in the leaf wrapping of the corn ear of the to the respective RY obtained at harvest. Data were
plant. An average of f ifteen readings per plot were adjusted with a segmented function model known
used to calculate the variation coeff icient. as the linear plateau (Sprent, 1961), allowing for a
lineal ascendant relationship and then a plateau.
Normalized Difference Vegetation Index (NDVI) This result shows the variable "Y" response to the
increase of the experimental variable "X" axis. The
The NDVI was measured in the phenological Y variable corresponds to the RY and the X variable
stages V7, V12, and R1 using a manual Green Seeker ® to the measurement value that resulted from applying
(Trimble Agriculture, USA). In each plot, readings a diagnostic method. The normal lowered from the
were performed above each plot's central furrow inflection point of this curve (where it starts getting
canopy to avoid the edge effect. The device was parallel to the X-axis) and intercepts the latter is
displaced 0.60 m above the plants' apex at a constant known as critical concentration (CC) or critical value.
speed of 0.5 m s-1, and each reading was made in Data adjustments were performed with signif icance
triplicate. (P < 0.05). Both N-NO3 concentrations in the dry
matter of the base of the stem at R5 were related
Foliar Nitrogen Analysis with relative grain yield (RY) using the Cate-Nelson
procedure, which allows estimating a CC as a function
In the stages V7 and V12, 12 the middle third of the minimum square sum (Cate and Nelson,
RML, and in R1 middle third of the leaves wrapping 1971). This approach classif ies the Y-axis variable
the corn ear were collected in each plot. Total N in two quadrants with a minimum error, determined
content was determined in the samples using the arbitrarily in a concentration range ±5% around the
Dumas method (Horwitz and Latimer, 2005) in an estimated CC. Figures 1 to 6 show the correlation
elemental analyzer (FlashEA 1112® Thermo Fisher coeff icients and the corresponding equations of
Scientif ic, USA). the lineal ascendant part of the linear plateau model.
 ROACHO-CORTÉS ET AL. NITROGEN DETERMINATION IN MAIZE 5

The statistical analyses were managed with the of the response expressed in RY (Sprent, 1961). The
program R vs. 3.5.1 (The R Foundation for Statistical correlation coeff icients (r) of this association were
Computing, 2018). greater than 0.86 (Figure 1).
The CC and its adequate range of concentration in
RESULTS AND DISCUSSION V7 was 700 ± 35 mg·L-1 of N-NO3. This result agrees
with the CC proposed by Withe et al. (1996) for a yield
Grain Yield of 15 Mg ha-1 of fresh corn. Reed and Hageman (1980)
reported that approximately 20% of the total nitrate
Maize grain yield responded signif icantly ions in V7 were found in the leaves' central midrib,
(P < 0.001) to the application of increasing nitrogen whereas in V12, the plant sap N-NO3 concentration
fertilization doses, which were 0, 120, 240, 360, in the central midrib and its respective range was
and 480 kg·ha-1 producing 5.8, 11.2, 13.9, 15.7, and 450 ± 22 mg·L-1. When the CC and their respective
18.3 Mg·ha-1, respectively. The relationship between ranges of adequate concentration -estimated in V7 and
N doses and grain yield was adjusted to a quadratic V12- were compared, a decrease of the second one was
function (P < 0.01) with an adjustment coeff icient (R 2) observed caused by the dilution effect described by
of 0.86, that is, 86% of the yield was explained by the Plénet and Lemaire (1999). This result may be because
N dose applied. the expression base of the N-NO3 concentration, e.g.,
Table 1 shows the CC estimated with each the dry leaf matter, increases with the plant age. At
diagnostic method for the three phenological stages the start of the reproductive stage R1, the CC and
(V7, V12, R1), and in the case of the diagnostics that the range considered acceptable ranges between
used the base of the stem, one CC estimated in R5 was 230 ± 11 mg·L-1 of N-NO3. This result contrasted with
included. Additionally, a percentage of deviation of the the CC suggested by Reisenauer (1978), 500 mg·L-1
means was included, which considered an arbitrary for the same phenological stage of the sampling leaf
range where the N concentration of the plant would be -the wrapping of the corn ear, but in sweet corn. This
adequate. difference could be because sweet corn accumulates
less biomass than f ield corn grown for grain, showing
Plant Sap N-NO3 Concentration in the Central less dilution of the N concentration. The hybrid used
Midrib in this experiment was crystalline white corn and
had an average height of ~2.80 m at the start of the
The plant sap N-NO3 concentration in the central reproductive stage, and the differences in critical
midrib of the RML measured in the phenological stages concentration between sweet and crystalline corn are
V7, V12, R1 was associated to the lineal ascendant greater as the cycle advances.

Table 1. Critical values in different maize phenological stages as estimated in the RML by several diagnosing methods at V7, V12
and R1 by using the Linear-plateau procedure proposed by Sprent (1961) and at R5 by using the procedure proposed by Cate and
Nelson (1971)†.

Diagnostic method Measurement unit V7 V12 R1 R5

CC ±5%‡ CC ±5%‡ CC ±5%‡ CC ±5%‡

N-NO3 in central midrib sap mg·L-1 700 35 450 22 230§ 11 -
N-NO3 in base of stem sap mg·L -1
330 16 460 23 370 18 350† 17
N-NO3 in base of stem dry matter mg·kg -1
8500 425 8300 415 7000 350 2647 †
132
NDVI units NDVI 81 4 83 4.1 80 4 -
SPAD units SPAD 43 2 39 2 47 §
2 -
Total foliar N in RML %N 3.4 0.17 2.2 0.11 2.6§ 0.13 -
‡
Concentration ranges were set considering ±5% of each estimated critical concentration. § Samples from the leaf that envelops the jilote.
6 TERRA LATINOAMERICANA VOLUMEN 39, 2021. e820

Relative yield %

N-NO3 (mg·L-1)


Figure 1. Modeling plant sap N-NO3 concentration in the central midrib of the RML at V7 and V12 and of the leaf
wrapping the corn ear at R1 of its relationship with the relative grain yield, estimated with the Linear-plateau model
(Sprent, 1961).

Plant Sap N-NO3 Concentration of the Base of the et al. (1997). These authors assessed the relationship
Stem between nitrogen nutrition index -dimensional value
between 0 and 1.8 obtained by dividing the total N
The plant sap N-NO3 concentration of the maize concentration in the biomass by the total N critical
base of the stem in different phenological stages (V7, concentration- and plant sap N-NO3 concentration
V12, R1) was associated positively (r > 0.83) with RY from the base of the stem of the phenological stages
in the linear-plateau model (Sprent, 1961) and were from V7 to R1. These authors also proposed a general
adjusted to Cate-Nelson model in R5 data (Figure 2). CC of 337 mg·L-1 N-NO3, estimated by using the Cate-
The CC results estimated in the phenological Nelson technique. This value represents the minimum
stages (V7, V12, R1, and R5) ranged from 330 to concentration of plant sap N-NO3 in the base of the
480 mg·L-1 of N-NO3, and at least the lowest limit stem, to obtain a nitrogen nutrition index similar or
of this range agreed with the CC reported by Justes greater than one.

Figure 2. Modeling plant sap N-NO3 of base of stem and its relationship with relative grain yield for the
phenological stages V7, V12, R1, and R5, as estimated with the linear-plateau method (Sprent, 1961) and Cate
and Nelson (1971) model at R5.
 ROACHO-CORTÉS ET AL. NITROGEN DETERMINATION IN MAIZE 7

N-NO3 Concentration in the Base of Stem Dry Matter Notwithstanding, this coincidence should be taken
with a certain reserve since these authors obtained
The N-NO3 concentration in the base of stem dry samples from freshly harvested sweet corn at R2 and R3
matter in the stages (V7, V12, R1) was associated with stages. According to Dechorgnat et al. (2018) each corn
the relative grain yield adjusting data to the linear- hybrid has specif ic requirements in acquiring N (ion
plateau model (Sprent, 1961), and in R5 adjusted by NH4 or NO3) influenced by the soil-climate condition
using the Cate-Nelson model (Figure 3). in which the maize develops. Nitrate concentration in
The correlation coeff icient of the lineal ascendant the dry matter of the stem's base undergoes influence
model in stages V7 and V12 was higher than 0.80 but of the prevailing solar radiation and soil humidity
lower in R1 (r = 0.64). The CC in stage R5 obtained before sampling (Fox, Roth, Iversen, and Piekielek,
from adjustment by Cate-Nelson procedure yield better 1989). Thus, CC could show some variation among
results. According to Ta (1991), as much as 40% of sites and years (Sainz-Rozas et al., 2001). Due to the
N in corn plants were in the stem during R1. In V7, above, this experiment's results should be considered
CC of N-NO3, the base of the stem and its respective preliminary and require several years of additional
range considered as adequate was 8500 ± 425 mg·kg-1, and validated experimentation. Consequently, the CC
concentrations in agreement with the results reported reported with this and other methods for the vegetative
by Sainz-Rozas, Echeverria, Herfurth, and Studdert stages and start of flowering may only be extrapolated
(2001) for corn in stage V6 with a yield of 12 Mg·ha-1. regionally while more research generates new results.
The results of V7 contrasted with the CC range Extrapolation to other ecosystem conditions must be
reported by Reuter et al. (1997) with values of 9000 to conducted with reserve.
17 800 mg·kg-1 of N-NO3 – values that corresponded To evaluate if the N application was moderate or
to 95% of relative yield. In V12, the CC and its range excessive, during the f inal reproductive cycle, Binford
resulted in being 8300 ± 415 mg·kg-1 of N-NO3 and et al. (1990) proposed performing the N-NO3 analysis
in R1 7000 ± 350 mg·kg-1 of N-NO3. These results in the dry matter of the base of the stem sample when
agree with the CC proposed by Heckman, Samulis, and the crop is close to stage R6 (physiological maturity).
Nitzsche (2002). This N-NO3 measurement has been calibrated for

Figure 3. Modeling N-NO3 concentration in the base of stem dry matter estimated with the
linear-plateau model from V7 to R1, and the critical concentration estimated with the Cate-
Nelson model at R5.
8 TERRA LATINOAMERICANA VOLUMEN 39, 2021. e820

different soil and climate regions: northern China that NDVI measurements showed variation among
(Huang, 2013); southeastern Buenos Aires, Argentina seasons and sites due to the characteristics of each
(Sainz-Rozas et al., 2001), and Pennsylvania, USA hybrid (Hatf ield, Gitelson, Schepers, and Walthall,
(Fox, Piekielek, and Macneal, 2001). 2008), as well as population density and space between
In the present experiment, we estimated a CC of plantation rows (Maddonni, Otegui, and Cirilo, 2001).
N-NO3 in the dry matter of the stem base for the stage Finally, the present study results showed that NDVI
R5 of 2647 mg·kg-1 N-NO3: this value corresponded explained -on average- only 53% of the relative yield
with 95% of the relative grain yield index. This value variability, an exceptionally low value, that contrast
would be the closest one to the high limit of optimum with the results reported in wheat by Raun et al. (2001)
concentration since other authors (Blackmer and and Santillano, López, Ortiz, and Raun (2013), 83%.
Mallarino, 2000; Isla and Blackmer, 2007; Beegle and
Rotz, 2009) situated this value in a range from ~700 SPAD Units in Recently Mature Leaf
to ~2000 mg·kg-1 of N-NO3. We can speculate that the
sum of fertilization dose and N contribution to soil The measurements of SPAD units made in the
were higher than N demand to reach a relative grain RML in the phenological stages (V7, V12, R1) were
yield (RY) of 95%, which is not strange in the area. associated with RY and adjusted to the linear-plateau
model (Figure 5).
NDVI The correlation coeff icient (r) was higher than 0.78
in all cases. In stage V7, a CV of 43 SPAD units was
The NDVI units performed on the maize plant estimated, slightly lower than the calculated critical
canopy in the phenological stages (V7, V12, R1) were value proposed by Edalat, Naderi, and Ega (2019), i. e.
associated with RY and adjusted to the linear-plateau 48 SPAD units. In V12, the CV, 39 SPAD units, had a
(LP) model. In the lineal part of the function, the lower value than those def ined by those authors of 51
correlation coeff icient was higher than 0.70 (Figure 4). SPAD units. In stage R1, the measurement performed
Since we did not measure N-NO3 concentration on the leaf wrapping the corn ear was 47 SPAD units
with this method, we proposed to call the parameter – a lower value than that def ined by Shapiro, Attia,
calculated critical value instead of CC. The calculated Ulloa, and Mainz (2016) of 57 SPAD units. The
critical value of the estimated NDVI in V7 (81 units) of contrast in the results reinforces the conclusion that
maize was lower but close to 83 units proposed by Yin other reports (Schepers et al., 1992; Xiong et al., 2015)
and McClure (2013). These authors estimated this value documenting that SPAD measurements were an indirect
for V12 of 80 NDVI, which is lower than the calculated measurement of N supply affected by environmental
in the present study (84 units), for the same stage. When factors, such as changes in light intensity in short
the crop reached the R1 stage, the calculated value was periods, specif ic green color characteristics of each
80 units. The discrepancy in the result agrees with that hybrid and N fertilization regime. This value must be
pointed out by Inman, Khosla, Reich, and Westfall related to the SPAD index of one well-supplied N band
(2007) and Colaço and Bramley (2018), who reported to make sense.

Figure 4. Relationship of NDVI with the relative grain yield of the phenological stages V7, V12, and R1, estimated with
the linear-plateau model.
 ROACHO-CORTÉS ET AL. NITROGEN DETERMINATION IN MAIZE 9

Figure 5. Relationship SPAD units with the relative grain yield in the phenological stages V7, V12 in the RML, and in
R1, taken in the leaf wrapping the corn ear, estimating the CC with the linear-plateau model.

Total N in the RML CONCLUSIONS

Total N concentration of the RML collected at Diagnostic methods for characterizing the N
the phenological stages V7, V12, R1 was associated nutritional status in maize crop in El Bajio, Guanajuato,
with relative grain yield when they were lower than Mexico with potential use under f ield conditions were
the critical concentration. In all the cases a correlation successfully calibrated for three different phenological
coeff icient higher than 0.70 was obtained (Figure 6). development stages. The critical N-NO3 concentration
In stage V7, we estimated a CC of 3.4% ± 0.17 N. measured under f ield conditions with semi quantitative
This result agrees with the CC reported by Plénet and and with quantitative methods conducted in the
Lemaire (1999), Campbell (2013), and Castellanos laboratory presented an acceptable association with
et al. (2019). At the stage V12, the CC was 2.2% ± the relative yields of maize fertilized with increasing
0.11% of N, which is slightly lower than that proposed N doses. The plant sap N-NO3 of the central leaf
by Castellanos et al. (2019) of 2.5 at 3.5% of N. This midrib -usually the recently mature leaf- and the
difference is probably due to the ability of the hybrid to N-NO3 measured in the plant sap of the stem base were
acquire N, which depends on competence (population associated with relative grain yield with a coeff icient of
density, intrinsic development characteristics of the root correlation higher than 0.80. The measurement obtained
system, and soil conditions of the site and N availability with optical sensors (SPAD and GreenSeeker) showed
in the soil. In stage R1, the CC was 2.6%. This result a lesser association with the relative yields although
agreed with the CC proposed by Reuter et al., 1997), with coeff icients of correlation higher than 0.77. The
Plénet and Lemaire (1999), and Castellanos et al. CC of total N measured in the recently mature leaf in
(2019). the phenological stages (V7, V12, R1) in general agreed

Figure 6. Relationship between total N in the RML and the relative grain yield at the phenological stages V7, V12, R1.
In R1 the sample was taken from the leaf wrapping the feminine flower, and CC was estimated with the linear-plateau
model.
10 TERRA LATINOAMERICANA VOLUMEN 39, 2021. e820

with those suggested by other authors. Consequently, and tables, and wrote and approved the f inal manuscript
maize producers and technical advisors from the Bajio draft: E.R.C. Conceived and designed the experiments,
region in Mexico now have references for diagnosing analyzed the data, prepared the f igures and tables,
N maize status by different analytical techniques and supervised the writing, and edited the f inal draft: J.D.E.
various development stages. These references allow Conceived, designed, and supervised the experiments,
them to verify the nutritional N status maintained by analyzed the data, prepared the f igures and tables,
the plant and take the corrective measurements, so this corrected, and edited the manuscript: J.Z.C.R.
element does not constitute a limiting factor for the
crop to reach the maximum yield possible, allowed by ACKNOWLEDGMENTS
the agroecosystem and at the same time gives better
information for not to exceed the N fertilization with The authors are thankful to CONACYT for funds
adverse effects in the environment. to carry out Master studies. Special thanks to Fertilab®
More studies are required to conf irm the CC and PROAIN®, Mexican enterprises for their f inancial
established and to identify variations among year, support in this research, to the seed company Aspros®
environments, and commercial corn hybrids, as well as for the facilities offered to perform sampling of cellular
to validate relative grain yield prediction. extract in their commercial lots and the donation
of the seed “Samurai” for the establishment of the
ETHICS STATEMENT experiment, to Miguel Peña for helping in performing
whitening nitrogen work of the lot, and to Dr. Elhadi
Not applicable. Yahia and Diana Fischer for reviewing and editing the
manuscript.
PUBLICATION CONSENTMENT
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