International Journal of Agricultural

Science and Research (IJASR)

ISSN(P): 2250-0057; ISSN(E): 2321-0087
Vol. 6, Issue 5, Oct 2016, 261-268
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INFLUENCE OF SALINE WATER IRRIGATION ON CRUDE PROTEIN AND

AMINO ACIDS IN TWO GENOTYPES OF FINGER MILLET
(ELEUSINE CORACANA GAERTN)
SANJAY AGARWAL 1 , ASHOK KUMAR2 & P. K. SINGH3
1

Department of Botany, Wilsonia Degree College, Moradabad, Uttar Pradesh U.P., India
2
3

Department of Botany, IFTM University, Moradabad, Uttar Pradesh, India

Department of Botany, Hindu Degree College, Moradabad, Uttar Pradesh, India

ABSTRACT
The field experiment was conducted to study the effect of salt stress on protein and amino acid content in two
varieties of finger millet viz. VL-315 and Local Hills. The experimental fields were irrigated with different saline waters
(3, 4, 7.2, 10, 12 and 14 dSm-1). Tube-well water was used in control sets. Leaf protein content was determined at 90th
day stage while grain protein was examined at the final harvest (130 DAS) while amino acids were examined at 60, 90
and 130 DAS. The leaf and grain protein content declined significantly in both cultivars at 3 to 14 dSm-1. The higher
VL-315. Ten amino acids were identified in different samples. Statistical analysis indicates that amino acids in both
cultivars, exhibited a differential response under varying salinity levels. It was also noted that amino acid contents except
thrionine were higher in cv. VL-315 than Local Hills under different salinity regimes at all three stages. Amino acid
accumulation (e.g. arginine and proline) may be considered as a detoxification mechanism of the ammonium produced
in plants subjected to stress. Thus, it is apparent that cv. VL-315 has performed better in terms of protein and amino
acids over Local Hills under saline water irrigation.

Original Article

protein content in leaves and grains may be due to higher photosynthetic rate of plants due to higher total leaf area in

KEYWORDS: Salinity, Protein, Grains, Millet, Irrigation

Received: Aug 26, 2016; Accepted: Sep 22, 2016; Published: Sep 28, 2016; Paper Id.: IJASROCT201630

INTRODUCTION
Soil salinity is one of the most important abiotic stresses (De bez et al., 2006 and Koyro, 2006) for
worldwide plant production especially in arid and semi-arid regions. Soil salinity has caused heavy loss of national
wealth in India. Uttar Pradesh alone has about 1.37 million hectares of sodic and saline soils. In U.P., the salts
affected districts are Allahabad, Aligarh, Etah, Etawah, Hardoi, Mainpuri, Pratapgarh, Raebareli, Sultanpur,
Jaunpur, Azamgarh and Bulandsahar etc. The agricultural areas affected by salts need amendment and
determination of the most suitable plant species able to grow in these areas. Plants need to have special
mechanisms for internal osmotic adjustments and changing of osmotic pressure in the root environment. Stressed
plants diminish osmotic potential by accumulating free amino acids, ions and other dissolvable substances. Protein
content and composition is an important determinant of cereal grain quality and it is sensitive to drought and heat
stress as well as atmospheric CO2 concentration (Wardlaw et al., 2002; Gooding et al., 2003).

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Sanjay Agarwal, Ashok Kumar & P. K. Singh

Jain et al. (2001) suggested that salt stressed plants accumulate low-molecular weight water-soluble metabolites,
like proline and betaines, in their cells, possibly for osmotic adjustment. Proline plays a role as an osmo-protectant to
adjust

osmotic

stability

(Liu

and

Staden,

2000;

Ghoulam

et

al.,

2002)

and

in

finger

millet

(Manikandan and Desingh, 2009; Chaudhary and Srivastava, 2015). Proline accumulation may be a part of the stress
signal influencing adaptive responses. (Maggio et al., 2002). Sarkadi et al. (2002) observed that the salt stress caused
increase in proline, glutamic acid, aspartic acid, arginine, ornithine and -amino-butyric acid levels in seedlings of wheat.
Leucine, isoleucine, valine, tryptophan and lysine contents remained similar in salt stressed and control leaves, supporting
the view that no change occurs in protein synthesis or degradation when salt accumulation is mild (Robinson et al., 1992).
Garthwaite et al. (2005) studied the effects of salt stress on the accumulation of free amino acids in the leaves of Hordeum
species. This study showed that concentrations of glycinebetaine (an N-trimethylated amino acid), asparagine, and proline
all increased in leaves with increased external NaCl concentration.
Finger millet (Eleusine coracana Gaertn.), commonly known as ragi, birds foot millet and African millet, is
grown under varied agro-climatic conditions, mostly in countries in Africa and Asia. India is the major producer of finger
millet in Asia. It is important staple millet after pearl millet (Pennisetum glaucum) in India. It is cultivated mostly as a rain
fed crop in India under diverse environments. In India the crop is grown in an area of 1.6 million ha with a production of
2.1 million tones and productivity of 1.3 tones ha-1 (Krishnappa et al., 2009). Finger millet is highly nutritious as its
grains

contain

65-75%

carbohydrates,

5-8%

protein,

15-22%

dietary

fiber

and

2.5-3.5%

minerals

(Chetan and Malleshi, 2007). The total dietary fiber (22.0%) of finger millet grain is relatively higher than that of most of
other cereal grains (eg. 12.6%, 4.6%, 13.4% and 12.8% in wheat, rice, maize and sorghum, respectively)
(Klopfenstein and Malleshi, 1998). Ragi is also rich in proteins, sulphur containing amino acids and because of its low
glycemic index with high fiber, it is recommended specially for diabetic patients. Therefore, it is considered an especially
wholesome food for diabetics.
The aim of the present study was to assess the impact of salt stress on leaf and grain protein content, amino acids
in leaves of two cultivars of ragi (one tolerant and one sensitive).

MATERIAL METHODS
The pure line seeds of finger millet (VL-315 and Local Hills) were sown in experimental plots (11 m2 each).
Plots were irrigated with saline water of different EC levels (3, 6, 7.2, 10, 12 and 14 dSm-1) which was prepared by mixing
the salts of NaCl, Na2SO4, NaHCO3 and CaCl2 in tube well water as described by U.S. Salinity Laboratory Staff
Handbook (1954). Control sets were irrigated with equal amount of tube well water whose electrical conductivity was
0.6 dSm-1. All sets were marked in triplicates. Protein content in leaves and grains of ragi were estimated by the method of
Lowery et al. (1951). For this purpose second and third leaves from the top of 90 days old plants were collected from
different sets of salinity. Sampling was done in triplicate for all estimating protein and amino acids. These fresh leaves of
each variety were dried in hot air oven and each dried sample was ground separately in pastel and mortar. At harvesting,
grains were also collected to evaluate the protein content from each sets (non salinized and salinized). The protein was
estimated using a standard curve prepared from bovine serum albumin (B.S.A.). Amino acids have been extracted,
identified and estimated from dry powder of leaves following the procedure of Block, Durrum and Zwieg (1985) with
some modification.

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Influence of Saline Water Irrigation on Crude Protein and Amino Acids

in Two Genotypes of Finger Millet (Eleusine Coracana Gaertn)

263

RESULTS & DISCUSSIONS

The effect of saline water irrigation on protein of leaf and grains of two cultivars of ragi are presented in figures 1
and 2. Leaf protein content declined significantly in both cultivars at 3 to 14 dSm-1 at 90 DAS. Data indicate 7.4 to 63.7%
and 32.3 to 72.8% reductions in leaves of cv. VL-315 and Local Hills at all salinity regimes at 90 DAS respectively. It is
also clear from data that leaf protein was higher in cv. VL-315 than Local Hills except in controls. The range of reduction
was 8.14 to 50.7% and 21.16 to 60.41% in grain protein of genotypes VL-315 and Local Hills respectively at different
salinity regimes at 130 DAS. Leaf and grain proteins were higher in cv. VL-315 than Local Hills at different salinity levels,
however, the crude protein was higher in grains than leaves. Inhibitory effects of salinity on leaf protein were also reported
by El-Sayad et al. (1995) in cotton and maize and Xu et al. (2010) in bentgrass. The present findings are confirmed by
Tammam et al. (2008) in wheat, Ayala- Astroga (2010) in Paulownia species and Kumar et al. (2010) in oat.
The reduction in protein contents may be due to decreased availability of amino acids and denaturation of the enzymes
involved in the amino acid and protein synthesis under saline-alkali conditions (Levitt, 1972).

Figure 1: Effect of Salinity on Total Protein in Leaves of Finger Millet

(Eleusine Coracana Gaertn.) in cv. VL-315 and Local Hills at 90 Days after Sowing

Figure 2: Effect of Salinity on Total Protein in Grains of Finger Millet

(Eleusine Coracana Gaertn.) in cv. VL-315 and Local Hills at Final Harvest Stage
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Sanjay Agarwal, Ashok Kumar & P. K. Singh

Amino acids play an important role in salt tolerance of crop as well as in wild plant species. Data on amino acids
in finger millet at different growth stages (60, 90 and 130 DAS) are presented in tables (1& 2). Ten amino acids were
identified in different samples which are cystine, iso-leucine, leucine, lysine, methionine, phenylalanine, threonine,
tryptophan, valine and proline. Statistical analyses indicate that amino acids exhibited a differential response under varying
salinity levels in both varieties. Leucine, methionine, threonine and valine were declined significantly as salinity raised
from 3 to 14 dSm-1, but gradually increased with the advancement of plant age (60-130 DAS). On the other hand, cystine,
iso-leucine, tryptophan, lysine, phenylalanine and proline increased significantly with increasing salt concentrations
(3-14 dSm-1) as well as with increase in plant. It is interesting to note that the amino acid contents were higher in cv.
VL-315 than Local Hills under different salinity regimes at 60th, 90th and 130th day stages except threonine. Cv. VL-315
registered significant increase in proline content under various salinity regimes at different durations and the accumulation
was higher in this variety than Local Hills. Among all individual free amino acids, proline (Pro) content in salt-treated
leaves increased significantly in finger millet cultivars. Proline serves as an important compatible osmolyte, and its
accumulation is believed to reduce cellular water potential and avoid deleterious toxicity of high ionic strength, has also
been proposed to serve as reactive oxygen species scavenger (Verbruggen and Hermans, 2008) and its accumulation can
stabilize the structure of membranes and proteins to minimize the damage of cells under salt stress. The present findings
also confirmed the work of Prasad et al. (1980) in finger millet and Parihar and Bajal (1982) in berseem. Accumulation
of proline under salt stress has also been reported by Mishra et al. (2006) in green gram, Ghoulam et al. (2002) and
Monreal et al. (2007) in sugar beet. These findings also confirmed by Rai et al. (2004) in Ocimum tenuiflorum, Kasim
(2006) in Vicia faba, Fayek et al. (2010) in jojoba, Amirjani (2010) in soybean and Kader et al. (2011) in Zea mays.
Amino acid accumulation (e.g. arginine and proline) may be considered as a detoxification mechanism of the ammonium
produced in plants subjected to stress (Slocum and Weinstein, 1990). The changes in accumulation of free amino acid
content induced by salt stress have an important role, since these relations were obtained several times in a relation-ship
with stress tolerance by Livia et al. (2002) in cereal plants. Survival and growth of plants in saline environments is the
result of adaptive processes such as ion transport and compartmentation of osmotic solute, synthesis and their accumulation
lead to the osmotic adjustment and protein turnover for cellular repair (Munns and Termaat, 1986).
Table 1: Effect of Saline Water Irrigation on amino Acids (g/100mg Dry Weight)
in Leaves of Finger Millet cv. VL-315 at Different Growth Stages

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Influence of Saline Water Irrigation on Crude Protein and Amino Acids

in Two Genotypes of Finger Millet (Eleusine Coracana Gaertn)

265

Table 2: Effect of Saline Water Irrigation on amino acids (g/100mg dry weight)
in Leaves of Finger Millet cv. Local Hills at Different Growth Stages

CONCLUSIONS
Salinity disturbed the metabolism and slowdown the activity of nitrate reductase enzyme with lesser nitrogen
contents and crude protein, cv. Local Hills proved to be lesser tolerant to salinity over cv. VL-315. It is concluded from the
findings that accumulation of proline and other amino acids function as a protector of macromolecules in plants subjected
to hyper-osmotic stress and its accumulation was assumed to be a metabolic adaptation which confers survival value by
relieving stress and the levels of proline in the plants is mainly designates their ability to tolerate or to adapt to saline
conditions. Due to better performance and superiority of cv. VL-315, this variety could be recommended for cultivation
under saline condition or where ground water is saline in arid and semi-arid regions.

ACKNOWLEDGEMENTS
We are thankful to the principal, Hindu College for providing all lab facilities and Dr. D. Sharma, Principal,
Wilsonia Degree College, Moradabad for giving us the valuable suggestion during research. We acknowledge the help of
Dr. Vijay Kumar Yadav for providing the pure line seeds of finger millet for present experiment.
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in Two Genotypes of Finger Millet (Eleusine Coracana Gaertn)

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