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Effects on growth, yield, root quality and anatomy in sugar beet
(Beta vulgaris L.) using a mixture of yeasts and organic manure as
an alternative to mineral -N fertiliser
Ramadan A. Agami
Agricultural Botany Department, Faculty of Agriculture, Fayoum University, 63514-Fayoum,
Egypt
E-mail address: [email protected]
Tel: +201028020544 & +2 01003230804.
Fax: +2 084 6343970
Abstract
The effects of seed inoculation with a mixture of yeasts and farmyard manure (FYM), as an
alternative to mineral -N fertilizer, on the growth, some metabolites, root yield and quality as
well as anatomy of sugar beet on newly-reclaimed soil were investigated. A field trail was
performed in a randomized complete block design with four treatments. The different treatments
were Yeasts + 17.5 metric tons (MT) FYM, Yeasts + 35 MT FYM, and Yeasts + 70 MT FYM
ha-1. In addition, the recommended dose of mineral -N fertilizer was used as a control.
Significant positive effects of Yeasts + 70 MT FYM ha-1 were observed on growth traits, leaf
photosynthetic pigments content, leaf and root nitrite (NO2–) and nitrate (NO3–) contents, total
sugar yield ha–1, root yield ha-1, root sucrose%, purity%, total soluble solids % (TSS) as well as
anatomy of the leaf and the root when compared with the other two Yeasts + FYM treatments.
However, there were no significant differences in most of these parameters between the Yeasts +
70 MT FYM ha-1 and the recommended mineral -N fertilizer treatment except for leaf and root
1
NO2– and NO3– contents, where significant reductions occurred in the Yeasts + 70 MT FYM ha-1
treatment. The results of this study suggest that yeasts and FYM reduced the amount of synthetic
chemical fertilizer needed for sugar beet production, and reduced the negative effects of
chemical fertilizers on the environment.
Keywords: Anatomy, farmyard manure, mineral- N fertilizer, secondary metabolites, sugar beet,
yeasts.
Introduction
Sugar beet (Beta vulgaris L.) is considered the second source for sugar production in many
countries all over the world after sugar cane. Recently, sugar beet crop has an important position
in crop rotation as winter crop not only in the fertile soils, but also in poor, saline alkaline, sandy
and calcareous soils. The great importance of sugar beet crop is not only from its ability to grow
in the newly reclaimed areas as economic crop, but also for producing higher yield of sugar
under these conditions as compared with sugar cane. Demand for sugar beet is increasing, thus
requiring growers to use additional nutrient inputs, especially mineral N to increase yield. Higher
N application may result in NO2 pollution of groundwater (Mytton, 1993; Shrestha and Ladha,
1998), soil acidification (Kennedy and Tchan, 1992), and increased denitrification resulting in
higher emission of N2O to the atmosphere, which may impact global warming (Bronson et al.
1997). These problems have renewed public interest in exploring alternate or supplementary
nonpolluting sources of N for agriculture (Ladha et al. 1997). Attention has therefore focused on
substitute fertilizers, including fungal biofertilizers and organic substances such as farmyard
manure (FYM). There are numerous reports stating the success in the ability of fungi to promote
plant growth as biofertilizers.
Fungal biofertilizers help to minimize the use of synthetic chemical fertilizers. Rhizosphere
yeast strains, however, belonging to several genera, chiefly, Sporobolomyces roseus (Perondi et
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al. 1996), Rhodotorula sp. (Abd El-Hafez and Shehata, 2001), Candida valida, Rhodotorula
glutinis and Trichosporon asahii (El-Tarabily, 2004) have been reported to promote plant
growth. Trichoderma species improve mineral uptake, release minerals from soil and organic
matter, enhance plant hormone production, induce systematic resistance mechanisms, and
induced root systems in hydroponics (Yedidia et al. 1999). Addition of Kluyveromyces walti,
Pachytrichospora transvaalensis and Sacharromycopsis cataegensis as bio-fertilizer to the soil
cultivated with sugar beet significantly increased the yield and enhanced the growth as well as
external and internal structure of the plants (Agamy et al. 2012).
Farmyard manure has been reported to increase soil organic carbon contents and microbial
biomass significantly (Masto et al. 2006), and is considered to be an eco-friendly way to improve
soil structure and fertility (Dauda et al. 2008). FYM was found to increase the growth and yield
of many crops without any undesirable impacts on the environment (Ali et al., 2001; Parray et al.
2007; Akparobi, 2009; Suthar, 2009). Manure applications improve the chemical, physical and
biological characteristics of the soil and increase yield and quality characteristics of crops
(Lithourgidis et al. 2007; Yolcu et al. 2010). The moves towards safe food production and
organic food should increase both biofertilizers and organic manure use and thus result in
environmental and ecosystem savings. Reduction in the use of chemical fertilizers is necessary to
maintain ecosystem function and develop sustainable agriculture. Thus the objective of this study
was to assess the effects of yeasts as biofertilizers and FYM as an alternative to mineral-N
fertilizer on the growth, some metabolites, root yield, and quality as well as anatomy of sugar
beet plants grown on newly-reclaimed soils.
Methodology
Physical and chemical properties of soil and FYM
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The main characteristics of the newly-reclaimed soil used in this research were determined
according to Black et al. (1982) and are shown in Table 1. The main characteristics of the FYM
were: pH 7.81; EC 7.53 dS m–1; organic matter 64.88% (w/w); C/N ratios 31.54; total N 2.25 %(
w/w); total P 0.38% (w/w); and total K 0.41% (w/w), respectively (Black et al. 1982).
Plant material, yeasts and yeast culture preparation
Seeds of sugar beet (Beta vulgaris L. cv Gloria) were obtained from the Ministry of Agriculture,
Cairo, Egypt and used in this study. Yeast strains; Pachytichospora transvaalensis UFOSY1240, Kluyveromyces walti UFOSY-1175 and Sccaromycopsis cataegensis UFOSY-1365 were
obtained from Professor J.L.F. Kock, University of the Free State, South Africa for research
purpose. Yeast cultures were prepared by growing the yeast strains in yeast extract malt extract
broth (YMB) (yeast extract 3 gL-1, malt extract 3 gL-1, peptone 5 gL-1 and glucose, 10 gL-1) at 25
± 1°C with shaking (150 rpm) for 48 to 72 h. Equal volume of the three strains suspension mixed
together and the yeast cells were pelletized by centrifugation (5000 r.p.m) for 10 min and
resuspended in sterilized tap water containing 0.025% (v/v) Tween-20 to the desired
concentration (~109 cfu ml-1).
Treatments and planting
A field trail was conducted in newly-reclaimed soil at the Experimental Farm of Faculty of
Agriculture, Fayoum (Southeast Fayoum; 29° 17'N; 30° 53'E), during the two successive seasons
of 2011/2012 and 2012/2013. Before sugar beet sowing, seeds were surface-sterilized in 70%
(v/v) ethanol for 2 min, and rinsed six times in sterile water. Surface sterile seeds were
inoculated by immersion in the mixture of yeast strains suspension (at 109 cfu ml-1) for 2 h on a
rotary shaker at 81 rpm, air-dried, and sown immediately. The cell densities in the yeast
suspensions were adjusted to a final density of approximately 108 cfu seed-1. During soil
preparation for sowing, all experimental areas received the complete dose of mineral-P [590 kg
4
ha–1 calcium superphosphate (15% P2O5)] and mineral-K [150 kg ha–1potassium sulphate (48%
K2O)] fertilizers, as recommended by the Ministry of Agriculture and were then divided into
10.5 m2 plots (3.5 m length x 3 m width), with six ridges 50 apart, 3.5 m in length. The control
plots received the recommended dose of mineral-N fertilizer [550 kg ha–1 ammonium nitrate
(33% N)] as outlined by Ministry of Agriculture, but received no yeasts + FYM.
Treatment I consisted of yeasts +17.5 metric tonnes (MT) FYM ha–1 (i.e., yeasts (108 cfu seed-1)
+ 18.37 kg FYM plot–1) spread on the soil surface, then mixed into the soil-surface layer. The
same procedure was carried out for the other two treatments. Treatment II increased the FYM
input to 35 MT ha–1 (i.e., yeasts + 36.75 kg FYM plot–1) Treatment III increased the FYM input
to 70 MT ha–1 (i.e., yeasts + 73.5 kg FYM plot–1). All treatments were applied in a randomized
complete block design with four replicates. Sowing took place on 15th September, 2011 and 15th
September 2012. Seeds were sown in hills 25 cm apart at using 3-4 seeds per hill. Plants were
thinned to one plant per hill after 40 days from planting (at 4-6 leaf stage). Nitrogen fertilizer
with the above mentioned level was added in two equal doses. The first one was applied after
thinning and the other one 21 days later. The other cultural practices were carried out as
recommended for commercial sugar beet production.
Growth traits, yield, TSS, purity and photosynthetic pigments determinations
At maturity stage (160 days from sowing) ten guarded plants were taken at random from each
plot to determine plant height, number of leaves plant-1, root dimensions cm (length and
diameter) and fresh and dry weight of the top plant-1 as well as fresh and dry weight of roots
plant-1 were estimated (g). The tops and roots were separated dried at 70oC for 3 days and at
105oC for 2 h in air forced-draft oven, to determine their dry weight. At harvest (180 days from
sowing), plants were harvested from the four middle rows of each plot to determine the top and
root yields ton/ha. Sugar yield (t/ha) was calculated by multiplying root yield by sucrose
5
percentage. Total soluble solids (TSS %) was determined by using digital referactometer (Atago,
Tokyo, Japan) at 20°C. Apparent purity percentage (%) was determined as a ratio between
sucrose % and TSS % of roots. Leaf chlorophylls and carotenoids contents (mg g-1 FW) were
determined in acetone extracts according to Arnon (1949) using a spectrophotometer (Shimadzu,
Kyoto, Japan).
Sucrose, total soluble proteins, leaf and root NO2– and NO3– determinations
Sucrose was estimated in fresh roots of sugar beet root by using Saccharometer according to the
method described by the Association of Official Agricultural Chemists (1995). Total soluble
proteins content of the fresh leaves and roots was determined according to the method described
by Bradford (1976) with bovine serum albumin as a standard. For leaf NO2– and NO3–
determination samples of 1.0 g dry matter was placed in a 100 ml polyethylene or glass bottle
and 40 ml of distilled water was added, then capped and shaken for 30 min. The mixture was
filtered, and the filtrate was made up to 100 ml in a volumetric flask (Radojevic and Bashkin,
1999). The same steps were followed with fresh root samples. Determinations of the NO3–
contents of each leaf and root sample solution were performed using a spectrophotometer
(Schimadzu, Kyoto, Japan) at a wavelength of 543 nm. The pre-programme for NO3 (64 NO3– N) was selected and the readings were converted to NO3 contents by multiplying using a
conversion factor of 4.4 (LaMotte, 2000). The NO3 contents (in µg g–1) of the samples were
calculated using the formula:
NO3– content (µg g–1) = C × V / M
where, C was the concentration of NO3– in the sample (µg g–1), V was the total volume of the
sample solution (100 ml), and M was the weight of the sample (1.0 g; LaMotte, 2000).
6
NO2– ion contents were determined in a similar manner, except that different reagents were
used. The pre-programme number for NO2– was 67 NO2– -N, and the reaction time was 5 min
compared to 10 min for NO3–. NO2– -N contents were converted to (µg g–1) NO2– by multiplying
by 3.3 (LaMotte, 2000). The NO2– contents (µg g–1) of samples were calculated using the
formula:
NO2– content (µg g–1) = C × V / M
where, C was the concentration of NO2– in the sample (µg g–1), V was the total volume of the
sample solution (100 ml), and M was the weight of the sample (1.0 g; Radojevic and Bashkin,
1999). Finally, the data obtained were converted to mg g–1 DW of leaf, or to mg 100 g–1 fresh
root.
Anatomy of leaf and root
For observation of leaf and root anatomy, samples of 160-days old from the middle of the sixth
leaf (full matured young leaf) from apex and root from 2 cm from base of the main root were
taken. Samples were killed and fixed in F.A.A. solution (50 ml 95% ethyl alcohol + 10 ml
formalin + 5 ml glacial acetic acid + 35 ml distilled water) for 48 h. Thereafter, samples were
washed in 50% ethyl alcohol, dehydrated and cleared in tertiary butyl alcohol series, embedded
in paraffin wax of 54 to 56°C mp. Cross sections with 20 µm thick were cut with a rotary
microtome (Leitz, Wetzlar, Germany), adhered by Haupt’s adhesive and stained with the crystal
violet-erythrosin combination (Sass, 1961), cleared in carbolxylene and mounted in Canada
balsam. The sections observed and documented using an upright light microscope (AxioPlan,
Zeiss, Jena, Germany) Measurements were done, using a micrometer eyepiece and an average of
5 readings were calculated.
Statistical analysis
7
All data were subjected to ANOVA using the SPSS software package, and means comparisons
between the different treatments were performed using the Least Significant Differences
Procedure (LSD) at the P ≤ 0.05 level, and Duncan’s multiple range tests were applied for
comparing the means (Duncan, 1955).
Results and Discussion
Sugar beet plants grown in the Yeasts + 70 MT FYM ha–1 treatment exhibited the highest
increase in plant height, number of leaves plant-1, root length, root diameter, fresh and dry weight
of tops and roots plant–1 when compared to the two other Yeasts + FYM treatments (Table 2).
There were no significant differences in these parameters between the Yeasts + 70 MT FYM ha–1
treatment and the recommended mineral -N fertiliser (control) treatment except number of leaves
plant-1 and root diameter. The increased growth traits fresh and dry weight of tops and roots
plants-1 obtained in the Yeasts + 70 MT FYM ha–1 treatment may be attributed to the positive
combined effects of yeasts and FYM on soils, which leads to increases in the organic matter
content and available nutrients as a result of the reduction in soil pH (Table 1). In addition, yeast
content of macro and micronutrients, growth regulators and vitamins stimulate the plant to build
up dry matters (Mirabal Alonso et al. 2008; Hesham and Mohamed, 2011). Fungal biofertilizers
help to enhance crop yield and promote sustainable agricultural production and are safe to the
environment (Smith and Zhu, 2001). Farmyard manure and crop residues provide a source of
organic matter, and when returned to soil they increase organic C in soil (Benbi et al. 1998;
Banger et al. 2010; Manure contains polysaccharides and aliphatic and aromatic compounds that
can bind to soil particles and create organo-mineral complexes important for flocculating
aggregates <0.2 µm (Tisdall and Oades, 1982). Application of organic waste alone and in
combination with mineral fertilizer enhanced root and shoot biomass, leaf area development, and
yield of maize plants (Ogundare et al. 2012). The interaction effect between yeast and organic
extracts lead to
a significant improve in nutrients uptake of sugar beet plants (Ibrahim and
8
Ibrahim, 2014). Yeasts + FYM increased the availability of nutrients, resulting in a positive
effect on plant growth. These results indicated that yeasts mixed with FYM were beneficial for
newly-reclaimed soils as an alternative to mineral- N fertiliser.
Leaves of sugar beet plants grown under the Yeasts + 70 MT FYM ha–1 treatment had the
highest contents of chlorophyll a, b and carotenoids (0.57, 0.16 and 0.35 mgg-1 fresh leaves,
respectively) when compared to the other two Yeasts + FYM treatments (Table 2). However, the
photosynthetic pigments contents obtained with the above mentioned treatment were almost
equivalent to those obtained in the mineral- N fertiliser control treatment. The beneficial effects
of yeasts inoculation on increased chlorophyll content might have been due to improve mineral
uptake, release minerals from soil and organic matter and enhance plant hormone production.
Hussain et al. (2002) reported that Saccharomyces sp. is among the microorganisms, which
improve crop growth and yield by increasing photosynthesis, producing bioactive substances,
such as hormones and enzymes and controlling soil diseases. The addition of organic fertilizer to
the soil may increase the level of exchangeable, soluble nutrients, thus the uptake of these
nutrients may be increased which, consequently, increases photosynthetic production (Yosifov,
1984). Based on sucrose % of roots (Table 2), the Yeasts + 70 MT FYM ha–1 treatment
produced sugar beet plants that had higher sucrose percent (18.1%) than the two other Yeasts +
FYM treatments. This result was in agreement with the data obtained by Shalaby and El-nady
(2008) who found that the greatest sucrose values of sugar beet were obtained via soil
inoculation with yeast. The enhancing effect of yeast application might be due to secretion of
cytokinins, enhancing the accumulation of soluble metabolites (Entian and Fröhlich, 1984),
increasing levels of endogenous hormones in treated plants which could be interpreted by cell
division and cell elongation (Khedr and Farid, 2002) , increasing the metabolic processes role
and levels of hormones, i.e. IAA and GA3 due to the physiological roles of vitamins and amino
acids in the yeast extract (Chaliakhyan, 1957). FYM improved the chemical, physical and
9
biological characteristics of the soil, resulted in increased sucrose accumulation in roots. The
recommended mineral -N fertiliser dose (i.e., the control treatment) resulted in significant
increase in sucrose percent when compared to the other treatments (Table 2). The positive effect
of N- fertilizer on sucrose values might be due to the increased efficiency of nitrogen fertilization
in building up metabolites translocations from leaves to developing roots thus increases sucrose
accumulation in roots. No clear differences were recorded in TSS and total soluble proteins in
leaves and roots between the Yeasts + 70 MT FYM ha–1 treatment and the control treatment,
although the former recorded purity values lower than control (Table 2). Significantly lower
TSS, purity and total soluble proteins in leaves and roots were obtained from the Yeast + 17.5
MT FYM ha–1 and Yeast + 35 MT FYM ha–1 treatments than the Yeasts + 70 MT FYM ha–1 and
the control treatments. These findings may be attributed to FYM act as a nutrient reservoir and,
upon decomposition, produce organic acids. Thus, the absorbed ions are released slowly over the
entire growth period, leading to higher root yields and quality. The increase in TSS % and purity
% due to Yeasts + FYM, particularly at the highest FYM rate treatment may be backed to its
roles in increasing growth and dry matter accumulation, consequently enhancement TSS % ,
sucrose % and reduction of impurity parameters in roots. The increase in the total soluble
proteins content could be attributed to the growth hormones produced by yeast (Khalil and
Ismael, 2010), direct stimulation of the synthesis of protein (Stino et al. 2009), providing plants
with essential nutrient elements required for protein formation (Hayat, 2007).
Sugar beet plants grown in the Yeasts + 70 MT FYM ha–1 treatment had the lowest leaf and
root NO2– and NO3– contents among all the Yeasts + FYM treatments tested. Farther, all the
Yeasts + FYM treatments caused lower leaf and root NO2 – and NO3 – contents than the control
mineral fertilizer treatment (Table 3). Yeasts + FYM, specially at the highest rate, resulted in
leaves with lower NO2– and NO3– contents, which was positively reflected in the NO2– and NO3–
contents of the roots for human health and nutrition. Increasing the available nitrogen in the soil
10
by increasing the percentage of mineral nitrogen in the control fertilization regime led to obvious
increases in the NO2– and NO3– contents of plant leaves that were also negatively reflected in the
roots. The accumulation of NO2– and NO3– ions in edible plant parts poses a problem which can
be attributed to the supply of readily available NO2– and NO3– to the plants from mineral
nitrogen fertiliser (Mahmoud et al. 2009). By contrast, in the Yeasts + FYM treated plots, the
release of NO2– and NO3– were comparatively slow. In addition, an increase in the percentage of
organic matter in those plots treated with Yeasts + FYM, particularly at the highest rate (Table
3), may control the release and transformation of nitrogen fertilizer to NO2– and NO3–. In this
regard, Gairola et al. (2009) found that the addition of FYM to cultivated soil was effective in
minimizing NO3– toxicity in beet leaves.
Sugar beet plants grown in the Yeasts + 70 MT FYM ha–1 treatment showed the highest top
fresh and dry yield tons ha–1, root fresh and dry yield tons ha–1 and sugar yield tons ha–1 (12.6,
1.78 , 44.8, 9.2 and 8.1 tons, ha-1 respectively) when compared to the two other Yeasts + FYM
treatments (Table 3). There were no significant differences in these parameters between the
Yeasts + 70 MT FYM ha–1 treatment and the recommended mineral- N fertiliser (control)
treatment although the later produced the highest sugar yield tons ha–1 (9.1 tons ha-1) when
compared to other treatments. The reduction in soil pH by Yeasts + FYM improved the solubility
and availability of nutrients for plant roots, which was positively reflected in the growth and
yield of sugar beet plants. Moreover, the optimum nutrient absorption obtained with the Yeasts +
70 MT FYM ha–1 treatment could be explained by the improved availability of essential nutrients
in the root zone, resulting from their solubilisation caused by the organic acids released by
decomposition of the FYM. The increased availability of nutrients in the soil, and the fact that
Yeasts + FYM enhanced their absorption by plant roots, resulted in increased yields and more
stable soil health. Manure applications improve the chemical, physical and biological
characteristics of the soil and increase yield and quality characteristics of crops (Lithourgidis et
11
al. 2007; Yolcu et al. 2010). The promoting effect of yeasts could be due to the biologically
active substance produced by these bio-fertilizers such as auxins, gibberellins, cytokinins, amino
acids and vitamins (Bahr and Gomaa, 2002).
As concerns the root and leaf anatomical structure, the plants grown in the Yeasts + 70 MT
FYM ha–1 treatment exhibited markedly increased in thickness of growth rings of sugar beet root
by increasing the average diameter of the cells (Table 4). Similarly, average diameter of
secondary xylem vessels was also increased when compared to the two other Yeasts + FYM
treatments. The maximum growth of rings thickness (808.3 µm) and average diameter of the
cells (50.0 µm) was obtained by control treatment. The Yeasts + 70 MT FYM ha–1 treatment
increased the thickness of the leaf blade and mid-vein by increasing the average diameter of
parenchyma cells as well as length and width of the vascular bundles (Table 4). The average
diameter of the vessels increased markedly as compared to the two other Yeasts + FYM
treatments. The beneficial effect of yeasts on sugar beet root and leaf structure may be due to the
crucial role of yeasts in improving soil fertility, absorption of nutrients and water, and plant
growth regulators such as auxins, gibberellins and cytokinins which play a role in cell division
and expansion. In addition, the promoting effect of farmyard manure on sugar beet roots and
leaves structure may be due to its effect on increasing macro and micronutrients availability to
plants, which affect plant organs structure. Warring and Philips (1973) stated that yeast is rich in
tryptophan which consider precursor of IAA (Indole acetic acid) which stimulate cell division
and elongation. Agamy et al. (2012) reported that, anatomy of the leaves and the roots of sugar
beet plants showed an increase in thickness of the blade, midvein, dimensions of the vascular
bundles, and number and diameter of xylem vessels as the result of application of yeasts.
12
Conclusion
The substitution of standard mineral -N fertilization regimes with mixture of yeasts combined
with 70 MT FYM ha–1 enabled sugar beet plants to produce higher root yields of higher quality
on newly-reclaimed sandy soils. Yeasts and FYM applications improve the chemical, physical
and biological characteristics of the soil. They also improved the solubility and availability of
nutrients for plant roots, which was positively reflected in the growth and yield of sugar beet
plants. Yeasts + FYM reduced the amount of synthetic chemical fertilizer needed for crop
production, and ameliorated the negative effects of chemical fertilizers on the environment.
References
Abd El-Hafez A.E., Shehata S.F., 2001. Field evaluation of yeasts as a biofertilizer for some
vegetable crops. Arab. Univ. J. Agric. Sci. 9,169–182.
Agamy A., Hashem M., Alamri S., 2012. Effect of soil amendment with yeasts as
bio-
fertilizers on the growth and productivity of sugar beet. Afr. J. Agric. Res. 7, 6613-6623.
Akparopi S. O., 2009. Effect of farmyard manures on the growth and yield of Amaranthus
cruentus. Agricultura Tropica ET Subtropica. 42, 1– 4.
Ali A. H., Abdel-Mouty M. M., Shaheen A. M., 2001. Effect of bio-nitrogen, organic and
inorganic fertilisers on the productivity of garlic (Allium sativum L.) plants. Egyptian J. of
Appl. Sci. 16, 173–188.
Association of Official Agricultural Chemists, 1995. Official methods of analysis, 16th Ed.,
AOAC International, Washington, DC.
Arnon D.I. 1949. Copper enzymes in isolated chloroplast. Polyphenol-oxidase in Beta vulgaris
L. Plant Physiol. 24, 1-5.
13
Bahr A., A., Gomaa A. M., 2002. The integrated system of bio-and organic fertilizers for
improving growth and yield of triticale. Egypt. J. Appl. Sci. 17, 512-523
Banger K., Toor G.S., Biswas A., Sidhu S.S., Sudhir K., 2010. Soil organic carbon fractions
after 16-years of applications of fertilizers and organic manure in a Typic Rhodalfs in
semiarid tropics. Nutr Cycl Agroecosystems. 86, 391–399.
Benbi D.K., Biswas C.R., Bawa S.S., Kumar, K., 1998. Influence of farmyard manure, inorganic
fertilizers and weed control practices on some soil properties in a long-term experiment.
Soil Use Manage. 14, 52–54.
Black C.A., Evans D.D., White J.L., Ensiminger L.E., Clark F.E., 1982. Methods of Soil
Analysis. Amer. Soc. Agron Inc., Ser. 9 in Agron. Madison, Wisconsin.
Bradford M.M., 1976. A rapid and sensitive method for quantitation of microgram quantities of
protein utilizing the principle of protein-dye-binding. Anal. Biochem. 72, 248-254.
Bronson K.F., Singh U., Neu H.U., Abao E.B., 1997. Auto mated chamber measurements of
methane and nitrous oxide flux in a flooded rice soil: Fallow period emissions. Soil Sci.
Soc. Am. J. 61, 988–993.
Chaliakhyan M. K.h., 1957. Effect of Vitamins on growth and development of plants. Dokly
Akad. Nauk. SSSK, 111, 894-897.
Dauda S. N., Ajaua F. A., Ndor E., 2008. Growth and yield of watermelon (Citrullus lanatus L.)
as affected by poultry manure application. J. of Agric. & Soc. Sci. 4,121–124.
Duncan D.B., 1955. Multiple range and multiple F-test Biometrics, II. pp. 1- 42.
El-Tarabily K.A., 2004. Suppression of Rhizoctonia solani diseases of sugar beet by antagonistic
and plant growth-promoting yeasts. J. Appl. Microbiol. 96, 69–75.
14
Entian K.D., Fröhlich K.U., 1984. Saccharomyces cerevisiae mutants provide evidence of
hexokinase PII as a bifunctional enzyme with catalytic and regulatory domains for
triggering carbon catabolite repression. J. Bacteriol. 158, 29-35.
Gairola S., Umar S., Suryapani S., 2009. Nitrate accumulation, growth and leaf quality of
spinach beet (Beta vulgaris Linn.) as affected by NPK fertilisation with special reference to
potassium. Indian J. Sci. Techn. 2, 35–40.
Hayat A. E. H., 2007. Physiological studies on Hibiscus sabdariffa L. production in new
reclamated soils. M.Sc. thesis, Faculty of Agriculture, Zagazig University.
Hesham A. L., Mohamed H., 2011. Molecular genetics identification of yeast strains isolated
from Egyptian soils for solubilization of inorganic phosphates and growth promotion of
corn plants. J. Microbiol Biotechnol. 21, 55–61.
Hussain T., Anjum A.D., Tahir J., 2002. Technology of beneficial microorganisms. Nature Farm
Environ. 3, 1-14.
Ibrahim Heba A.K., Ibrahim S. M., 2014. Effect of Some Organic Extracts on Essential Nutrients
Uptake of Sugar Beet under Saline Conditions. Res. J. Agric. & Biol. Sci. 10, 53-64.
Kennedy I.R., Tchan Y.T., 1992. Biological nitrogen fixation in non-leguminous field crops:
Recent advances. Plant Soil. 141, 93–1.
Khalil S. E., Ismael E., G., 2010. Growth, Yield and Seed Quality of Lupinus termis as Affected
by Different Soil Moisture Levels and Different Ways of Yeast Application. J Am Sci. 6,
141-153.
Khedr Z.M.A., Farid S., 2002. Response of naturally virus infected tomato plants to yeast extract
and phosphoric acid application. Annals of Agric. Sci. Moshtohor, Egypt. 38, 927-939.
15
Ladha J.K., de Bruijn F.J., Malik K.A., 1997. Introduction: Assessing opportunities for nitrogen
fixation in rice-A frontier project. Plant Soil. 194, 1–10.
La Motte 2000. Smart Spectro Water and Waste Water Procedure Analysis Manual. La Motte
Inc.,Washington,DC, USA: PP. 68–180.
Lithourgidis A.S., Matsi T., Barbayiannis N., Dordas C.A., 2007. Effects of liquid cattle manure
on corn yield, composition, and soil properties. Agron J. 99, 1041- 1047.
Mahmoud E., Abd EL-Kader N., Robin P., Akkal-Corfini N., Abd EL-Rahman L., 2009. Effects
of different organic and inorganic fertilizers on cucumber yield and some soil properties.
World J Agric Sci. 5, 408–414.
Masto R. E., Chhonkara P. K., Singh D., Patra A. K., 2006. Changes in soil biological and
biochemical characteristics in a long-term field trial on a subtropical in ceptisol. Soil Biol
Biochem. 38, 1577–1582.
Mirabal Alonso L., Kleiner D., Ortega E., 2008. Spores of the mycorrhizal fungus Glomus
mosseae host yeasts that solubilize phosphate and accumulate polyphosphates. Mycorrhiza
18,197-204.
Mytton L., 1993. Nitrogen fixation, in Institute of Grassland and Environmental Research Rep.
Inst. Grassland and Environ. Res., Aberystwyth, UK., PP. 46–50.
Ogundarem, K., Agele S., Aiyelari P., 2012. Organic amendment of an ultisol: effects on soil
properties, growth, and yield of maize in Southern Guinea savanna zone of Nigeria.
International Journal of Recycling of Organic Waste in Agriculture. 1,11.
Parray, B. A., Ganai A., Fazili K. M., 2007. Physiochemical parameters and growth yield of
tomato (Lycopersicon esculentum): role of farmyard manure and neemcake. American-
16
Eurasian J. of Agric. & Environ. Sci. 2, 303–307.
Perondi N.L., Luz W.C., Thomas R., 1996. Microbiological control of Gibberella in wheat.
Fitopatol Bras. 21, 243–249.
Radojevic M., Bashkin N. V., 1999. Practical Environmental Analysis. Royal Society of
Chemistry Thomas Graham House, Cambridge, UK: PP. 180 – 430.
Sass J.A. 1961. Botanical Microtechnique. 3rd Ed. The Iowa State University Press, Ames.
Shalaby M.E., El-Nady M.F., 2008. Application of Saccharomyces cerevisiae as a biocontrol
agent against Fusarium infection of sugar beet plants. Acta Biol Szeged. 52, 271-275.
Shrestha R.K., Ladha J.K., 1998. Nitrate in groundwater and integration of nitrogen-catch-crop
in rice–sweet pepper cropping system. Soil Sci. Soc. Am. J. 62, 1610–1619.
Smith S.E., Zhu Y.G., 2001. Application of arbuscular mycorrhizal fungi: Potentials and
challenges, In: Stephen B.P and K.D. Hyde, (eds) Bio-Exploitation of Filamentous Fungi.
Fungal Diversity Research Series. 6, 291-308.
Stino R. G., Mohsen A. T., Maksouds M. A., Abd El- Migeed M. M. M., Gomaa A. M., Ibrahim
A. Y., 2009. Bioorganic fertilization and its Impact on Apricot young trees in newly
reclaimed soil. American- Eurasian. J. Agric. & Environ. Sci. 6, 62-69.
Suthar S. 2009. Impact of vermicompost and composted farmyard manure on growth and yield
of garlic (Allium sativum L.) field crop. Int. J. of Plant Prod. 3, 27–38.
Tisdall J.M., Oades J.M., 1982. Organic matter and water-stable aggregates in soils. J. Soil Sci.
62, 141–163.
Warring P. E., Phillips I. D. G., 1973. The control of growth and differentiation in plants. E L B
S ed., Pub by Pergamon Press Ltd. VK.
17
Yedidia I., Benhamou N., Chet I., 1999. Induction of defense responses in cucumber plants
(Cucumis sativus L.) by the biocontrol agent Trichoderma harzianum. Appl Environ
Microbio. 65, 1061-1070.
Yolcu H., Günes A., Dascı M., Turan M., Serin Y., 2010. The effects of solid, liquid and
combined cattle manure applications on yield, quality and mineral concentrations of
common vetch and barley intercropping mixture. Ekoloji. 19, 71-81.
Yosifov M. A., 1984. Photosynthetic activity in watermelon inrelation to nutrition. Sel`skohozy
aistvennaya Biologiya. 1: 36–39
18
Table (1): Physical and chemical properties of the experimental soil before treatment and sowing (BS) and 160 days
after sowing (AS).
Soil property
BS
AS
Sand [% (w/w)]
91.35
91.21
Silt [% (w/w)]
4.45
4.35
Clay [% (w/w)]
4.20
4.44
Sandy
Sandy
pH
7.71
7.21
EC (dS m–1)
7.76
7.45
Organic matter [% (w/w)]
0.58
0.81
CaCO3 [% (w/w)]
6.48
6.21
Available N (mg kg–1 DW)
56
98
Available P (mg kg–1 DW)
7
22
Available K (mg kg–1 DW)
56
111
Available Fe (mg kg–1DW)
16
29
Available Mn (mg kg–1DW)
10
16
Available Zn (mg kg–1 DW)
4
6
Soil texture
19
Table (2): Effect of yeasts, FYM and mineral - N fertilisers on growth traits, photosynthetic pigments, sucrose%, TSS, purity and total soluble proteins of
sugar beet plants grown on a sandy reclaimed soil. Mean values (n = 5) ± SD in each column followed by different lower-case letters are significantly
different (P ≤ 0.05) by Duncan’s multiple range test.
Treatments
Control
†
Plant height (cm)
No. of leaves
-1
plant
a
29.0 ± 1.7
b
22.3 ± 1.5
a
23.6 ± 0.6b
a
61.2 ± 0.4
Root length
(cm)
a
23.3 ± 0.6
c
20.0 ± 0.9
Root diameter
2
(cm )
a
14.0 ± 0.3
c
7.7 ± 0.7
d
195.1 ± 6.4
b
9.4 ± 0.3
c
233.3 ± 9.5
a
11.2 ± 0.9
Yeasts + 17.5 MT FYM
-1
ha
45.5 ± 2.2
Yeasts + 35 MT FYM
-1
ha
58.0 ± 1.7
Yeasts + 70 MT FYM
-1
ha
59.0 ± 1.7
26.0 ± 1.7
Chlorophyll a
−1
(mgg F.W.)
Chlorophyll b
Carotenoids
-1
(mgg F.W.)
Treatments
c
21.6 ± 1.1
b
23.6 ± 0.6
(mgg F.W.)
Control
†
a
0.17 ± 0.00
c
0.14 ± 0.00
b
0.16 ± 0.00
a
0.16 ± 0.01
0.58 ± 0.00
Yeasts + 17.5 MT FYM
-1
ha
0.48 ± 0.01
Yeasts + 35 MT FYM
-1
ha
0.54 ± 0.01
Yeasts + 70 MT FYM
-1
ha
0.57 ± 0.00
†
a
Fresh weight of
-1
top plant
a
37.4 ± 1.5
c
25.5 ± 1.1
b
34.3 ± 0.6
a
35.5 ± 1.1
259.9 ± 7.1
b
251.1 ± 3.5
Sucrose%
Dry weight of
-1
top plant
T.S.S
a
921.0 ± 19.1
c
633.6 ± 23.4
b
827.4 ± 31.1
ab
889.8 ± 13.5
0.36 ± 0.00
a
20.1 ± 0.4
b
0.28 ± 0.01
a
a
a
20.8 ± 0.3
c
16.1 ± 0.7
0.34 ± 0.00
a
0.35 ± 0.00
a
c
18.4 ± 0.3
17.5 ± 0.3
b
19.5 ± 0.3
18.1 ± 0.3
b
20.5 ± 0.3
-1
Recommended dose of mineral N fertiliser [550 kg N ha ammonium nitrate (33%N)].
Yeasts: Kluyveromyces walti, Pachytichospora transvaalensis and Saccharomycopsis cartaegensis
FYM: farmyard manure
20
a
96.9 ± 1.6
a
9.5 ± 0.4
c
87.0 ± 4.9
b
8.3 ± 0.2
b
88.4 ±2.8
b
a
89.6 ±2.6
b
Dry weight of
-1
roots plant
a
190.5 ± 9.8
c
146.1 ± 5.1
b
172.2 ± 3.8
a
184.8 ± 8.5
Proteins in leaves
-1
(mgg F.W.)
Purity
-1
a
Fresh weight of
-1
roots plant
a
c
b
ab
Proteins in roots
-1
(mgg F.W.)
a
6.4 ± 0.3
a
b
5.2 ± 0.2
9.2 ± 0.5
a
5.8 ±0.3b
9.2 ± 0.7
a
6.1 ± 0.1
c
ab
-
-
Table (3): Effect of yeasts, FYM and mineral - N fertilisers on top yield, root yield, sucrose yield and nitrite (NO2 ) and nitrate (NO3 ) contents ion
of sugar beet plants grown on a sandy reclaimed soil. Mean values (n = 5) ± SD in each column followed by different lower-case letters are
significantly different (P ≤ 0.05) by Duncan’s multiple range test.
Treatments
Fresh top yield
tons ha
Dry top yield
-1
tons ha
a
Control†
13.1 ± 0.3
-1
Yeasts + 17.5 MT FYM ha
9.8 ± 0.3
c
Yeasts + 35 MT FYM ha
-1
11.7 ± 0.4
Yeasts + 70 MT FYM ha
-1
12.6 ± 0.1
Treatments
Leaf No2
-1
a
9.5 ± 0.4
a
9.1 ± 0.3
a
1.2 ± 0.1
c
31.9 ± 1.1
c
7.3 ± 0.3
c
5.1 ± 0.3
c
41.6 ± 1.5
b
8.4 ± 0.3
b
7.5 ± 0.3
b
44.8 ± 0.7
a
9.2 ± 0.4
ab
8.1 ± 0.1
b
1.78 ± 0.0
Leaf No3
b
ab
–
–
Root No2 mg 100g
FW
-1
-1
–
Root No3 mg 100g
1
FW
mgg DW
a
3.0±0.1
a
38.7±0.1
a
5.9±0.1
a
0.18± 0.00
b
1.4±0.1
bc
31.2±0.1
c
5.2±0.1
b
c
1.5±0.1
b
37.8±0.1
b
5.2±0.1
b
1.3±0.1
c
30.8±0.1
d
5.1±0.1
b
0.26 ± 0.01
Yeasts + 17.5 MT FYM ha
46.4 ± 0.9
a
-1
Yeasts + 35 MT FYM ha
-1
0.15± 0.01
Yeasts + 70 MT FYM ha
-1
0.13± 0.01d
†
Sucrose yield tons
-1
ha
a
1.7 ± 0.0
-
Dry yield of roots
-1
tons ha
1.8 ± 0.6
b
mgg DW
Control†
-1
Fresh yield of roots
-1
tons ha
-1
Recommended dose of mineral N fertiliser [550 kg N ha ammonium nitrate (33%N)].
Yeasts: K. walti, P. transvaalensis and S. cartaegensis
FYM: farmyard manure
21
-
Table (4): Effect of yeasts, FYM and mineral - N fertilisers on anatomical structure of root and leaf blade of sugar beet plants grown on a sandy
reclaimed soil. Mean values (n = 5) ± SD in each column followed by different lower-case letters are significantly different (P ≤ 0.05) by
Duncan’s multiple range test.
Treatments
Characters
Growth rings
Average
(μm)
Control
†
Yeasts + 17.5 MT FYM ha
-1
Secondary xylem
thickness Average diameter Average number Average diameter of
of cells (μm)
of vessels/row
vessels (μm)
808.3 ± 8.3
a
50.0 ± 2.0
a
6 ± 0.5
a
33.8 ± 0.8
b
583.3 ± 3.3
c
33.3 ± 3.3
c
4 ± 0.2
c
30.6 ± 0.6
c
Yeasts + 35 MT FYM ha
-1
641.0 ± 6.0
b
40.0 ± 3.0
b
5 ± 0.4
b
33.1 ± 1.1
Yeasts + 70 MT FYM ha
-1
650.0 ± 5.0
b
41.7 ± 1.7
b
5 ± 0.3
b
43.7 ± 0.7
Treatments
a
Characters
Midvein thickness
(μm)
Control
b
†
Yeasts + 17.5 MT FYM ha
-1
Blade
thickness
(μm)
Average diameter of
parenchyma
cells(μm)
2937.0 ± 17
a
290.0 ± 7
a
150.1 ± 4.9
2187.5 ± 19
d
260.0 ± 5
c
83.2 ± 3.2
a
d
Dimensions of vascular bundles
Length (μm)
Average diameter of
vessels(μm)
Width (μm)
626.0 ± 6.0
b
490 ± 7
b
51.3 ± 1.3
a
445.0 ± 5.0
d
405 ± 5
c
40.8 ± 1.8
c
Yeasts + 35 MT FYM ha
-1
2700.0 ± 12
c
270.0 ± 6
bc
100.0 ± 5.0
c
500.0 ± 8.0
c
540 ± 4
a
39.4 ± 0.4
c
Yeasts + 70 MT FYM ha
-1
2750.0 ± 18
b
280.0 ± 8
ab
125.2 ± 5.2
b
666.7 ± 6.7
a
550 ± 8
a
45.0 ± 2.0
b
†
-1
Recommended dose of mineral N fertiliser [550 kg N ha ammonium nitrate (33%N)].
Yeasts: K. walti, P. transvaalensis and S. cartaegensis
FYM: farmyard manure
22
Fig. (1): Transections of sugar beet root as affected by application of yeasts, FYM and mineral- N
-1
-1
fertilisers. A) Control; B) Yeasts +17.5 MT FYM ha ; C) Yeasts + 35 MT FYM ha ; D) Yeasts + 70 MT
-1
FYM ha ; gr, growth ring; Sxv, secondary xylem vessels and sp, storage parenchyma.
23
Fig. ( 2): Transections of sugar beet leaf blade as affected by application of yeasts, FYM and mineral -1
-1
N fertilisers. A) Control; B) Yeasts + 17.5 MT FYM ha ; C) Yeasts + 35 MT FYM ha ; D) Yeasts + 70 MT
-1
FYM ha ; b, blade; pc, parenchyma cells and vb, vascular bundles.
24