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Industrial Crops and Products 39 (2012) 135–138
Contents lists available at SciVerse ScienceDirect
Industrial Crops and Products
journal homepage: www.elsevier.com/locate/indcrop
Short communication
Relationship between chlorophyll meter readings and total N in crambe leaves as
affected by nitrogen topdressing
Tiago Roque Benetoli da Silva ∗ , Aline Cristina de Souza Reis, Cleber Daniel de Góes Maciel
Universidade Estadual de Maringá, Departamento de Ciências Agronômicas, Avenida Colombo n.5790, CEP: 87020-900 Maringá, PR, Brazil
a r t i c l e
i n f o
Article history:
Received 1 February 2012
Received in revised form 4 February 2012
Accepted 6 February 2012
Keywords:
Crambe abssynica Hoechst
Chlorophyll
SPAD reading
Mineral nutrition
Leaf diagnostics
a b s t r a c t
Crambe is a crop winter, with high oil content, function as a source of raw material for the biodiesel
production. This crop deserves attention agriculture because the short cycle ranging from 90 to 100 days. A
portable chlorophyll-content meter for proportional estimate determinations (SPAD-502 indices) allows
instant readings, and may be ideal to predict N requirements, since the method used in the laboratory
is time-consuming and destructive. Consequently, this work’s objective was to evaluate the relationship
between chlorophyll meter readings and total N contents in leaves of crambe plants, at the stage prior to
flowering, as affected by topdressing N levels. This work was conducted in Umuarama Municipal District –
Paraná State, Brazil. Variables analyzed were SPAD index values (chlorophyll) in leaves, as measured
with the chlorophyll meter, and total N content. The data were submitted to analysis of variance, means
comparison, polynomial regression, and simple linear correlation analyses. The results showed that:
nitrogen topdressing applied did not affect the chlorophyll and nitrogen in crambe leaves. There was no
significant correlation between chlorophyll and leaf nitrogen meaning on crambe crop, during the full
flowering.
© 2012 Elsevier B.V. All rights reserved.
1. Introduction
Increase of concerns related to global economic environment
issues, and forecasts that non-renewable energy reserves would be
exhausted in the next 50 years, have encouraged the search for new
sources of energy such as solar, wind and biofuels (Oplinger et al.,
1991).
Biodiesel is a liquid fuel, biodegradable, nontoxic, produced
using different raw materials. It is chemical reaction results, which
reacts vegetable oils (or animal fats) and alcohol (ethanol or
methanol) (Brasil et al., 2007). With the biodiesel use can be
reduced by 78% carbon dioxide emissions, about 90% the emission
of smoke and virtually eliminates sulfur oxides emissions (Oplinger
et al., 1991).
Crambe crop (Crambe abssynica Hoechst) is another alternative feedstock for biodiesel production. It is considered a winter
crop, with high drought resistance and a short cycle (between 90
and 100 days). It fits well in hot and cold soils and is pests and
diseases resistant, with oil content of about 35%. Crambe plants
need on average 54 days (range of 42–64 days) between seedling
and flowering. Flowering usually ends about 12–15 days before
∗ Corresponding author. Tel.: +55 4436244558.
E-mail address: [email protected] (T.R.B.d. Silva).
0926-6690/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.indcrop.2012.02.008
physiological maturity. The ideal humidity for crambe crop should
be 14% (Pitol, 2008).
Research conducted by the Mato Grosso do Sul Foundation, on
Maracajú Municipal District, Brazil, indicate yield between 1000
and 1500 kg ha−1 . In tests on the extraction of oil from crushing
presses, we obtained a yield of 25 l of oil to 100 kg of grain (Pitol,
2008). On Brazil, Silva et al. (2011b) verify yield between 1600 and
2000 kg ha−1 .
To maximize the economic and biological yield, the availability
or nutrients supply is essential because species respond to fertilization, such as sunflower (Bajehbaj et al., 2009; Silva et al., 2011a),
especially nitrogen, which is usually the required element in largest
quantity by plants (Malavolta et al., 1997). When plants are deficient in this element, have leaves with pale-green coloration and
yellow. Nitrogen is necessary for synthesis of chlorophyll and as
part of this molecule, is involved in photosynthesis (Epstein and
Bloom, 2006).
Crambe is responsive to fertilizer application. Silva et al. (2011b)
working with phosphorus and zinc, in the Cascavel Municipal District, Parana State (Brazil) concluded that the leaf application of
zinc did not cause modifications in the vegetative and reproductive development of crambe, however, the phosphorus applied on
seedling stage increased oil content and a thousand grain mass and
consequently also increased crambe yield in two cropping years.
Rogério et al. (2011) working with phosphate fertilizers in
crambe on Umuarama Municipal District, Parana State, Brazil,
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found that the use of phosphorus at sowing increased crambe
yield. Santos et al. (2011) in the same municipality noted
that the potassium application of significantly increased crambe
yield.
In the absence of supplied nitrogen to the soil, the plant degrades
the chlorophyll molecule, nitrogen going to regions of active
growth, where it performs it is functions (Furlani Júnior et al., 1996),
thus persisting N deficiency and there is a reduction in chlorophyll
content, the plants will not use sunlight as an energy source to
carry out essential functions such as other nutrients absorption and
carbohydrates for their growth and development (Potafos, 1998).
Traditionally methods employed for the chlorophyll content
determination in leaves are destructive and labor intensive processes of extraction and quantification were done in the laboratory,
which precludes its use for predicting N deficiency quickly and
routinely (Argenta et al., 2001; Godoy, 2002). With the invention of a portable direct reading which estimates the proportion
of chlorophyll content (Minolta, 1989), measuring the green coloration intensity of the leaf (the amount of light absorbed by the
pigment), it is possible to obtain values in the field, so instantaneously and nondestructively (Furlani Júnior et al., 1996; Malavolta
et al., 1997; Godoy, 2002).
Nitrogen is the macroelements for which there have been no
effective method to assess its needs on the ground (Furlani Júnior
et al., 1996) and has an index of content in the soil that serves
as an indicator for fertilizer recommendation. It is known that
oilseed plants respond to nitrogen, as in the case of castor bean
(Silva et al., 2007), but the complex dynamics of this nutrient
(leaching, immobilization–mobilization, denitrification, mineralization, etc.) and can vary in their content field after a rain and
even packed samples (Godoy, 2002). Green color determination
on leaves using a portable colorimeter can estimate the chlorophyll content, and indicate the level of N in plants crambe, since,
according to Malavolta et al. (1997), chlorophyll and nitrogen are
correlated positively.
Furlani Júnior et al. (1996), Chapman and Barreto (1997), Booji
et al. (2000), Argenta et al. (2001) and Godoy (2002) observed in
their experiments that chlorophyll was positively correlated with
N content in shoots of plants. In this context, the chlorophyll is
being used to predict the need for nitrogen fertilization in some
species, like rice (Stalin et al., 2000), maize (Argenta et al., 2001;
Godoy, 2002) and bean (Furlani Júnior et al., 1996), for the crambe
plants has not been verified. It is noteworthy that results of research
aimed at “balancing” this equipment is more practical importance,
as its use is becoming more widespread, especially among farmers
more technical.
It is important to correlation establish between the chlorophyll
meter readings and the total content of N from leaves to Brazilian conditions, with specific species such as crambe plant, carrying
out necessary changes in the methodology of using the chlorophyll meter (Godoy, 2002). In this sense, logically, inaccuracies of
the equipment must be taken into account as in the case where
there is excessive absorption of N (luxury consumption), without
an increase in the intensity of green leaves color, because the second
Crusciol et al. (2001), the increased availability and consumption of
the nutrient in question do not cause the plant to produce chlorophyll than you need. Fact converse should also be observed when it
detects a yellowing of leaves, is theoretically considered as an indication of N deficiency, but that actually may be caused by many
other factors like deficiency of other minerals or even the attack
pests and diseases.
Crambe an unfamiliar plant in the Brazil country, there is a
need to develop appropriate cultivation techniques, where the use
of chlorophyll in relation to nitrogen brings something new for
this species. The present work aimed to evaluate the relationship
between the readings provided by the chlorophyll index (SPAD) and
the total nitrogen content in the crambe leafs in the full flowering
stage, with different levels of nitrogen topdressing applied.
2. Materials and methods
This work was conducted under field conditions in 2010 agricultural year, at Farm School from Universidade Estadual de Maringa
College, in Umuarama Municipal District, Paraná State, Brazil.
Climate is mesothermal subtropical with average annual temperatures around 19 ◦ C. The local soil is an Oxisol Dystrophic (Embrapa,
2006), with sandy texture.
Soil samples were collected before the establishment of the
experiment for chemical characterization, presenting at 0–20 cm
depth: 5.2 mg dm−3 of phosphorus (Mehlich 1 extractor); 7.8 g kg−1
of organic matter; 4.3 pH (CaCl2 ); 0.4; 1.5; 0.75 and 8.1 cmolc dm−3
of K, Ca, Mg and Cation Capacity Change respectively, bases saturation of 33.1%.
The experiment was conducted in accordance with the randomized block design with four replications. Nitrogen rates were 0, 30,
60, 90 and 120 kg ha−1 . The font used for was urea fertilizer, in topdressing application at 20 days after emergency. All plots received
an uniform sowing fertilization of 20 kg ha−1 by nitrogen (urea),
60 kg ha−1 of phosphorus (super phosphate simple) and 60 kg ha−1
of potassium (potassium chlorate).
Crambe seeds were provided by Mato Grosso do Sul Foundation
– FMS. The plots consisted of six rows with five meters long, with
spacing between them of 0.25 m, totaling 20 plots. Sowing area
was fenced and dried, removing debris from previous crops and
weeds. Sowing was done manually on September 10, 2010 with
the Crambe FMS-Brilhante cultivar, targeting a population of 750
plants per plot and fertilizer was added beside the seed furrow.
After the 45 days after emergency, was held to hand harvested
10 plants of each plot to determinate the chlorophyll meaning and
Leaf Nitrgen (Total N).
Chlorophyll meaning was determined using a portable Chlorophyll Meter, model SPAD-502 (Soil and Plant Analysis Development) of Minolta Co. Ltd., Osaka, Japan (1989). The device has two
LEDs (LED) positioned at the tip of the meter, which emit light in the
range of 600–700 nm and 860–1060 nm range, in sequence, when
closed (Godoy, 2002). The light passes through the emission and the
part that crosses the sheet reaches a receiver (photodiode silicon)
and is converted into electrical signals, amplified and converted
into digital signals and is used by a microprocessor to calculate the
SPAD values, which are shown on a display. The values obtained
are proportional to the amount of chlorophyll present in leaves
(Argenta et al., 2001), because the wavelength bands used in equipment were based on two absorbance peaks of chlorophyll in vitro
(Godoy, 2002).
It is noteworthy that the precision of the device is a unit SPAD
values between 0 and 50 SPAD units (Godoy, 2002), and that their
use requires, among some conditions, that the only variable influencing the concentration of chlorophyll should be N content by the
fact that, according Malavolta et al. (1997), almost all the deficiencies and excesses in some cases, cause chlorosis.
Readings were taken in the morning, in the shade to avoid direct
incidence of sunlight on the chlorophyll, always in the middle third
of the plants, avoiding necrotic areas by the attack of pests and
diseases, and the edges of the leaf midrib. The developmental stage
of sampling was in full flowering.
Nitrogen leafs content was carried out was collected immediately after reading the SPAD index values, the same leaves used in
this determination which estimates direct proportion the amount
of chlorophyll, which are then packed in paper bags and placed
in a greenhouse at 60 ◦ C with forced air for 48 h. Then there was
the grinding of the material in Willey mill type to be analyzed
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T.R.B.d. Silva et al. / Industrial Crops and Products 39 (2012) 135–138
137
Fig. 1. SPAD index values (estimated proportion of chlorophyll content) (a) and the total content of N (b) in crambe leaves as a function of nitrogen applied topdressing.
Umuarama (PR-Brazil), 2010. n.s., not significant.
N content, according to the method described by Malavolta et al.
(1997).
Statistical analysis was performed followed by analysis of variance model, through Sisvar program, by using the 5% significance
level. The mean nitrogen rates were compared by regression with
the same significance level. The chlorophyll and leaf nitrogen was
comparing by simple linear correlation analyses.
3. Results and discussion
As can be seen in Fig. 1a and b, there was no significant influence
of nitrogen topdressing on chlorophyll content and nitrogen. This
result does not corroborate those obtained by Crusciol et al. (2001),
where the highest levels of available soil N and greater consumption of this element did not make the plants produce chlorophyll
than they needed. Therefore, the portable meter SPAD-502 proves
to be ineffective to the luxury consumption of N, because the device
measures the intensity of green coloration and nitrogen that is not
embedded in the chlorophyll molecules do not reflect the variation of the staining intensity (Godoy, 2002). Lack of response fact
of nitrogen is due, probably, the dynamics in the soil and rain lack
during the application of the element in question.
Nitrogen critical levels for various species were established after
years of research, due to peculiarities. However, as no literature was
found in the appropriate content in the crambe leafs, was used as a
comparison the critical level (leaf analysis) for the castor bean and
sunflower (Malavolta et al., 1997). In Fig. 1b, was observe that the
mean value content 29 g kg−1 it is considered below for castor bean
(40–50 g kg−1 ) and sunflower (33.0–35.0 g kg−1 ) to Malavolta et al.
(1997). In this context the N critical level determination for crambe
crop, can be important goal of future work study, and subsequently
also possible the development of another survey to determine the
needs of N, apply as from the use of chlorophyll.
Analysis was performed using simple correlation (r) between
these two variables, being highlighted the significant correlation
lack (Fig. 2). This result disagree with Furlani Júnior et al. (1996) and
Godoy (2002), who reported the using possibility the chlorophyll
in the indication of nitrogen deficiency in bean and maize, respectively. Argenta et al. (2001), in an experiment correlating the N
content of corn leaf chlorophyll and reading, go further, concluding
that this method can be well spent, depending on the developmental crop development stage, because in the early stages the values
of the readings do not are very accurate.
It is necessary to standardize the reading of the chlorophyll
meter SPAD-502 in different climate conditions, for several genotypes, different developmental stages and in various cultural
practices, for not controlling these factors may limit the reliability of the statement of possible N deficiency and in diagnosing the
content of this element in leaf (Costa et al., 2001). With that, who
knows, in the near future, using the chlorophyll meter to calculate how much N fertilizer applied, correcting an imminent failure,
during the crop cycle, immediately. New research should be conducted in crambe plants, aiming to calibrate chlorophyll in relation
to nitrogen in the leaf.
4. Conclusion
Nitrogen topdressing applied did not affect the chlorophyll and
nitrogen in crambe leaves.
There was no significant correlation between chlorophyll and
leaf nitrogen meaning on crambe crop, during the full flowering.
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