Sewage Water Irrigation and Growth Response of Leucaena leucocephala Inoculated With Glomus intrarradices and Application of Organic Matter
The aim of this study is to evaluate the growth response
of Leucaena leucocephala inoculated with Glomus intrarradices
and application of organic matter and the actual level of contamination
with heavy metals Cu, Cr, Zn and Pb in soil irrigated with sewage and
clean water. Sewage water is used for irrigation, which creates both opportunities
and problems. This is an option to reduce the stress on limited fresh
water and help meet the nutrient requirement of crops, but also produces
contamination. In the irrigation District 018, Tulancingo, Hidalgo, Mexico,
forage for cattle has been irrigated with residual water for several years.
To evaluate the level of contamination of two plots, one hectare was irrigated
with residual water and another with clean water. In soil the contamination
of Cu, Cr and Ni are below the established limits for contaminants. Pb
was not found. A comparison of soil irrigated with clean water or sewage
water indicated that Cu is nearly twice as concentrated in soil irrigated
by contaminated water; Ni is slightly greater; Cr is more abundant. Soil
was collected for a greenhouse experiment with Leucaena leucocephala
(guaje) to observe its growth with inoculation of Glomus intrarradices
with different amounts of vermicompost. Later a factorial experiment 7x2
completely randomized design with five replications in the greenhouse
was established. The variables measured were plant height, stem diameter,
root volume, dry weight of biomass and dry weight of roots. The experiment
lasted 180 days from planting until harvesting. It is concluded that the
inoculation with Glomus intrarradices increased the absorption
by Leucaena leucocephala in nutrient adsorption.
to cite this article:
A. Khalil-Gardezi, A. Exebio-Garcia, E. Mejia-Saenz, E. Ojeda-Trejo, L. Tijerina-Chavez, Habibsha -Gardezi and M. Delgadillo-Pinon, 2009. Sewage Water Irrigation and Growth Response of Leucaena leucocephala Inoculated With Glomus intrarradices and Application of Organic Matter. Journal of Applied Sciences, 9: 1373-1377.
Reuse of sewage is one of the best options to reduce the stress on limited
fresh water available and help to meet the nutrient requirement of crops. Sewage
water is either used or disposed of on land for irrigation purposes, which creates
both opportunities and problems (Yadav et al., 2002;
Horswell et al., 2003; Kakar
et al., 2006). The use of raw or untreated sewage can cause accumulation
of heavy metals in the soil and phytotoxicity with an impact on the quality
of soil and in forage (Gradezi et al., 2004; Malla
et al., 2007). Metabolic activity of soil microorganisms has also
been reported to increase when sewage effluent is used for irrigation (Meli
et al., 2002`Ramirez-Fuentes et al., 2002;
Gardezi et al., 2007).
Leucaena leucocephala is a long-lived tropical legume and is a nutritious
forage tree. It has a great variety of other uses: firewood, timber, human food,
green manure, shade and erosion control (Gutteridge and Shelton,
1994; Shelton and Brewbaker, 1994). Leucaena leucocephala
is the mostly widely used species as a valuable fodder shrub for increased
animal production in the tropics (Khamseekhiew et al.,
2001). It is an evergreen forage rich in protein, minerals and B carotene.
The plant can also be grazed directly, is well accepted by livestock, particularly
goats and is quite resistant to heavy, frequent defoliation (Meissner,
1997). Yields of forage vary with soil fertility, rainfall, altitude, density
and cutting frequency from 1-15 t ha-1 year. (Shelton
et al., 1998); the foliage has high nutritive value for ruminant
production. With an edible fraction with 55-70% digestibility, 3-4.5% N, 6-10%
ash, 30-50% N-free extract, 0.8-1.9% Ca and 0.23-0.27% P (Jones
et al., 1992).
The capacity of phytoremediation of Leucaena leucocephala has been evaluated
associated with Glomus and Rhizobium, concluding that this plant
is a good extractor and accumulator of these metals (Habte
and Aziz, 1991; Dahlin et al., 1997; Gardezi,
Leucaena leucocephala and arbuscular endomycorrhiza have been used recently
for bioremediation in soil contaminated with Cu (Gardezi
et al., 2004).
Arbuscular Mycorrhiza Fungi (AMF) give multiple potential benefits to the host
plant growing in practically all soils because its roots are colonized by fungi
(Smith and Read, 1997). These fungi are used in the biofertilization
and as inoculants (Guzman-Plazola and Ferrera-Cerrato, 1990).
These fungi play a central role in nutrient uptake (George
et al., 1994). The importance of arbuscular symbiosis in crop production
and natural ecosystems has been fundamentally linked to its ability to promote
mineral nutrition in deficient soils, especially for nutrients with low mobility,
such as P, Cr and Cu. As a rule, in nutrient-deficient soils with plants colonized
with AMF the uptake of nutrients with low mobility, such as P, is substantially
Gardezi et al. (2007) evaluated the response
of Leucaena leucocephala to inoculation with arbuscular mycorrhiza with
different doses of cow manure. They found a positive effect in the variables
related to plant growth. They found that the best growth of the plant depends
on different treatments of arbuscular mycorrhiza, because the treatments with
Glomus sp. had higher values for the agronomic variables.
MATERIALS AND METHODS
In the irrigated district 018 in Tulancingo, Hidalgo, Mexico (Fig.
1) forage is produced for cattle in irrigated areas with raw sewage
water. The soil is shallow (40 cm) over tepetate (fragipan), and the soil
is saturated most of the time by frequent irrigations. Two plots of one
hectare each, one with pasture and another without pasture were located.
The plot with pasture was irrigated with sewage water for seven years.
The other plot was cultivated for corn (Zea mays) and irrigated
with clean water. For each sub-plot twenty soil samples were taken in
a 25x20 m sub-plot at three depths (0-5, 5-10 and 10-40 cm). For the soil
analysis the 20 samples were homogenized to obtain 3 samples for each
sub-plot. The rest of the soil (about 500 kg) was prepared for a greenhouse
The study lasted 180 days from transplanting to harvesting and was conducted
under greenhouse conditions in the Colegio de Postgraduados, Montecillos,
Estado de Mexico in the spring of 2007.
The seeds were sterilized with sodium hypochlorite and pregerminated
in plastic trays. The plants were transplanted to polyethylene bags that
had been filled with 3 kg of soil.
The inoculation coincided with the transplanting, mixed with 5 g of sand
and 1-2 g of alfalfa roots with 80% colonization of Glomus intrarradices.
In addition 7 levels of vermicompost (V) were applied as a source of organic
||Localization of soil sample sites in the irrigation
district 018, Tulancingo, Hidalgo, Mexico
||Experimental treatments with vermicompost and Glomus
The vermicompost was prepared with 60 kg of cow manure, 15 kg
of residues of melon, and 25 kg of wheat straw, treated for 5 months with
the action of earthworms. Two levels of Glomus were applied, with
and without Glomus inoculation; with 7 doses of Organic Matter
(OM) as follows: OM 0.0 (0 gV), OM 0.5 (15 gV), OM 1.0 (30 gV), OM 1.5
(45 gV), OM 3.0 (90 gV),OM 6.0 (180 gV) OM 12.0 (360 gV) (Table
The variables evaluated were Plant Height (PH), Stem Diameter (SD), Root
Volume (RV) and Dry Weight of Biomass (DWB).
RESULTS AND DISCUSSION
The soils are shallow (40 cm) underlain by fragipan (tepetate), the soil
is irrigated with residual water and saturated with water most of the
time. The texture of the soil irrigated with residual water is clay loam
in the first five centimeters and clay at 5-40 cm. The soil irrigated
with clean water is clay in all the profile.
Table 2 shows the results of the soil sample analysis for
each depth for each plot (residual and clean water) and the chemical characteristics
of the soil pH, Electrical Conductivity (EC), Organic Matter (OM), Total Nitrogen
(TN) and Phosphorus (P) have higher values in the soil irrigated with residual
water. The pH is alkaline in both plots and it increases with depth in both
soils (except for sample 3). The EC increases from 1.8 to 1.98 dm sec-1
from the plot with residual water and increases with depth. This contrast with
the plot irrigated with clean water, were the EC is lower than that of residual
water, and the EC is lower in samples at more than 0-5 cm. This contrasts also
with the plot irrigated with clean water where the EC decreases with depth.
The TN is greater in the irrigated soil with residual water at all depths with
differences up to 0.09%, with the principal accumulation in the upper horizon
for irrigation with residual water. The OM is 4.5% up to 2.87% greater than
in clean water, the principal accumulation in clean water is in the upper layer
with 2.0%. The horizon of 10-40 cm has lower differences up to 1.3% to those
irrigated with clean water. With respect to phosphorus there is a large difference
among the two soils, up to 7.75 times greater for the depth of 5-10 cm. The
amount of phosphorus is from 140 to 170 mg kg-1 and soil irrigated
with residual water; in contrast the soil irrigated with clean water had only
20-30 mg kg-1.
With respect to the characteristics of the soil, there is an increase
in pH, EC, TN, OM and P (Table 2) which would probably
continue to increased with time. The farmer now considers that irrigation
with residual water is beneficial because it promotes the high yields
of grass. All the metals were more highly concentrated in the upper five
centimetres of the soil and decline with depth. However, when in time
the increase of organic matter and pH could have toxic effects on grassland
due to high pH and inadequate balance of nutrients.
In general there is no contamination with the heavy metals copper (Cu),
chromium (Cr), lead (Pb) and nickel (Ni) but their concentration has increased
by irrigation with residual water, except for chromium which has a higher
level in soil irrigated with clean water (Table 2).
The levels of the soils irrigated with residual water showed differences
between residual and clean water. Cu is almost twice the concentration
in residual water, Ni is slightly higher, and Cr is less abundant.
Soil analysis shows the presence of Cu, Cr and Ni in low concentrations,
and that Pb is absent in the soil.
Table 3 shows the result of analysis of variance: Significant
differences were observed among all four variables. When the individual variable
was analyzed using factorial analysis, there were significant differences from
the principal effects (OM and GI). For the studied variables the behaviour was
similar to that found by Gardezi et al. (2007).
The plants inoculated with endomycorrhizae fungi (Glomus intraradices) were
taller than those not inoculated at all levels of organic matter (Fig.
2). However, the curves had differences that are explained by the significant
interactions. However, in the inoculated plants there was a positive response
to the increase of organic matter up to 6 t ha-1, reducing the highest
content in plants not inoculated. The height of the plant was greater when the
content of organic matter increased in the total range studied.
|| Soil analysis for the three depths (0-5, 5-10, 10-40
|| Analysis of variance for four variables evaluated in
|MSD: Minimum Significant Difference, CV: Coefficient
||Effect of arbuscular endomycorrhizae fungi and different
levels of organic matter in plant height of Leucaena leucocephala
||Effect of arbuscular endomycorrhizae fungi and different
levels of organic matter in stem diameter (cm) of Leucaena leucocephala
with Erythrina americana Miller, Sesbania emerus Aubul, Dodonea
viscosa, Dodonea angustifolia L.f. (Gardezi et
al., 1995, 1999, 2000)
reported that the arbuscular endomycorrhizae fungi stimulate the growth of the
plant more than phosphorus fertilization.
There were different tendencies in stem diameter. For plants inoculated
with endomycorrhiza (Glomus intraradices) there was an increase
in stem diameter. With addition of 0 and 0.5 t ha-1 there was
no increase, but in 1 to 12 t ha-1 of organic matter the stem
diameter increased progressively (Fig. 3).
The volume of roots of Leucaena leucocephala increased in the
presence of Mycorrhizae Glomus intraradices (Fig.
4). The inoculated and not inoculated plants show a well defined tendency
from 0 to 1 t ha-1 of OM. The increase of organic material
also increases the root volume, especially in inoculated plants up to
6 t ha-1. The root volume was the same at 6 and 12 t ha-1.
However, the root volume without inoculation decreased.
Effect of arbuscular endomycorrhizae fungi and different
levels of organic matter
in root volume (cm3
) of Leucaena
In general the levels of Copper (Cu), Chromium (Cr), Lead (Pb) and Nickel
(Ni) are higher in the soil irrigated with sewage water, almost twice
that of the other soil, except for chromium which has a lower level in
soil irrigated with clean water.
However, in time the increase of organic matter and high pH in the soil
could have toxic effects on grassland and inadequate balance of nutrients.
This situation that is not now considered as a problem by the farmers
of this area.
The inoculation with Glomus intraradices and the application of
6 t ha-1 of OM produced the higher value for all evaluated
variables (plant height, stem diameter, root volume) of Leucaena leucocephala.
Previous evidence showed that this plant has higher dependence on this
mycorrhizal fungus, because the treatment with this fungus produces the
highest values for all evaluated variables.
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