Biological and Clinical Sciences Research Journal
ISSN:
2708-2261
www.bcsrj.com
DOI:
https://doi.org/10.47264/bcsrj0201001
Biol. Clin.
Sci. Res. J., Volume, 2021: e001
Original Research
GENETIC
EVALUATION OF LEGUME SPECIES UNDER HEAVY METAL AND BIOGAS WASTEWATER TREATMENTS
*Kamran, Ali Q, Malik A
Institute
of Molecular Biology and Biotechnology, The University
of Lahore, Lahore Pakistan
Corresponding
author email: kamrankhan002121@gmail.com
Abstract
The
legumes are very important food crops, called pulses grown throughout the world
for their grain which contains essential vitamins, carbohydrates, protein, fat,
minerals, and dietary fiber. Chickpea (Cicer arietinum L.), pea (Pisum sativum L.), green mung bean (Vigna radiate L.), and black mung
bean (Vigna mungo L.) is
important pulse crops that belong to the family Fabaceae
or Leguminosae. The present experiment was conducted
at the greenhouse of Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore during the season of July
to August of 2020 to study the effects of different treatments of ZnSO4
and biogas wastewater on the growth of chickpea, pea, green bean, and black
bean. The experiment consisted of two treatment combinations comprising of two
levels of ZnSO4 at 0.5M and 0.25M along with the two levels of
biogas wastewater at 500ml and 250ml along with including the control group. It
was observed from results that the performance of Chickpea, pea, green mung bean, and black mung bean
genotypes were variable under heavy metal zinc sulfate treatment. The results
suggested that the treatment of a higher concentration of 0.5M ZnSO4
was toxic as compared with 0.25M ZnSO4. The application of biogas
wastewater was found relatively fit for the seedling growth of all of four
pulse crop species. The results showed that there was a significant correlation
among root length, shoot length, and leaf length under the application of
different treatments. A higher genetic advance was reported for shoot length
and root length which revealed that the selection of legumes may be fruitful to
improve yield under stress conditions.
Keywords:
chickpea, pea, green mung bean, black mung bean, stress, ZnSO4
Chickpea
(Cicer arietinum L.),
pea (Pisum sativum L.),
green mung bean (Vigna radiate L.) and black mung bean (Vigna mungo L.) are an
important pulse crops belong to family Fabaceae or Leguminosae. Chickpea has been cultivated in the Middle
East while green bean and black bean have been cultivated in East Asia and
Southeast Asia from ancient times of old civilizations. Chickpea and pea are
sensitive to abiotic stress such as heat, drought,
cold and heavy metals. The introduction and use of high input fertilizer
varieties has caused micronutrient depletion in soils, mainly zinc (Zn). About
60 % of the world’s soil is deemed insufficient for crops production of some
essential nutrient elements due to mineral stress deficiency, lack of
availability or toxicity (Pathak et al., 2012; Moller, 2009). The legumes
are important to boost soil fertility and to increase crop productivity in
developing countries’ cultivation systems of arid and semi-arid areas.
Furthermore, Bio fertilizers are products made of living microorganisms that
can increase crop production in an environmentally friendly, sustainable way,
through a variety of direct and indirect mechanisms (Amjad,
2002). Chickpea constitutes a significant source of human dietary protein,
particularly for a large vegetarian population. It produces an average of 126
kg of protein per hectare and is potentially the top protein producing
vegetables alongside soybean. Peas have been
cultivated for many centuries as a significant source of animal feed and human
food Cousin, 1997). Field pea is a cool legume crop cultivated worldwide in the
cool season. 20% of the available protein is provided by beans in many
developing countries. Beans are also a key component of dietary protein in 50%
of the world’s population (Deshpande et al., 1984; Sai
et al., 2017).
Stress is any external elements that affect physiology,
development, productivity, metabolism and plant survival. Stress has been
divided into two categories: abiotic and biotic (Mahajan et al., 2018). Abiotic stress is a stress mainly caused by environmental
changes such as drought , salinity, heat, colds, water recesses, extreme soil
pH changes, mechanical stress (for example wind, hail, wounds, etc.) and
effects of herbicides and weedicides and exposure to
other heavy metals. Although, the biotic stress is often pathogenic stress which
is caused by living organisms like bacteria, fungi, viruses, nematodes, etc.,
insects and weeds, which caused higher loss of crop productivity (Chen, 2006; Sangolli et al., 2018). Across
different terrestrial habitats around the world, heavy metal toxicity has
become a key concern. The damage to soil texture i.e. pH of soil, the presence
of various elements, heavy metal causes a direct and indirect decrease in plant
growth by adversely affecting diverse physiological and molecular activity of
plants. Heavy metals, including Hg, Co, Cd, Fe, Ni,
Al, Cr, Ar, Zn, Cu, Mo and Mn,
etc., today extensively used in industries and imparts damage to soil as well
as to crop productivity (Mahanty et al., 2017; Ranpariya
et al., 2017; Zubair
et al., 2016).
Materials and methods
Chickpea
(Cicer arietinum L.),
pea (Pisum sativum L.),
green mung bean (Vigna radiate L.) and black mung bean (Vigna mungo L.) seeds
were collected from local market of Lahore. The present experiment was
conducted at the greenhouse of Institute of Molecular Biology and
Biotechnology, The University of Lahore, Lahore during the season of July to
August of 2020 to study the effects of different treatments of ZnSO4 and
biogas wastewater on the growth of chickpea, pea, green bean and black bean.
The experiment consisted of two treatment combinations comprising of two levels
of ZnSO4 at 0.5M and 0.25M along with the two levels of biogas
wastewater at 500ml and 250ml along with including control group. Seeds were sown and after complete germination five plants
from each pot of chickpea, pea, green mung bean and
black mung bean were taken to collect data and after
removing the plants from each pot, treatments of biogas wastewater and ZnSO4
was applied. Five groups were made; the first group was given no treatment
because it was kept as control group. The next four groups were included; second
group was treated with 0.5M ZnSO4, third group with 0.25M ZnSO4,
fourth group with 500ml of biogas wastewater and fifth group was 250 ml of
biogas wastewater. After a period of one week of first treatment, five plants
were again removed from each pot to record data. After that second treatment
was given to each pot and after period of one week again plants were removed to
collect the data. The application of treatments and data recording was carried
out five times. After collection of data, pooled analysis of variance,
correlation and regression analysis was carried out for traits including root length,
roots per plant, shoot length and leaf length.
Results and Discussions
It was persuaded from results given in
table 1 that significant differences were among treatments, genotypes and genotypes × treatment. It was found
that the average leaf length (5.524±0.0035cm), number of roots per plant
(5.125±0.0044cm), root length (5.1045±0.0011cm) and shoot length (3.5±0.0023cm)
were found under combined effects of all of the treatments. The genetic advance
was found higher for all of the studied traits of legume species. The
coefficient of variation was found lower for all of the studied traits which
indicated that the results for root length, shoot
length, leaf length and number of roots per plants were highly consistent and
reliable for making selection under heavy metal stress conditions. The results
from table 2 indicated that survival of legume genotypes was higher under
control conditions as compared with ZnSO4 and biogas wastewater treatment. The lowest
survival of black mung bean was found under the treatment
of 0.5M ZnSO4 followed by 0.25 M ZnSO4. The application of ZnSO4
showed adverse effects on black mung bean only to
reduce the survival. The lower survival percentage indicated that the genotype
was sensitive to application of ZnSO4. The genotypes which showed
higher survival percentage under applications of ZnSO4 and
biogas wastewater indicated that the legume genotype may be used as heavy metal
tolerance genotype (Akhtar et al., 2017; Bhardwaj et al., 2014; Garci-Gomez
et al., 2017).
Table 1 Pooled
analysis of variance
|
Source |
DF |
LL |
NRP |
RL |
SL |
|
Replication |
1 |
3.23761 |
2.025 |
0.03721 |
0.121 |
|
Genotypes |
3 |
0.04902* |
0.825* |
0.32551* |
0.15* |
|
Treatment |
4 |
0.67771* |
1* |
0.14857* |
0.00437* |
|
Genotypes × Treatment |
12 |
0.84646* |
1.53333* |
0.28637* |
0.10104* |
|
Error |
19 |
0.35954 |
0.28816 |
0.35686 |
0.03468 |
|
Grand Mean |
5.524 |
5.125 |
5.1045 |
3.5 |
|
|
Standard error |
0.0035 |
0.0044 |
0.0011 |
0.0023 |
|
|
Coefficient of variance |
10.85 |
10.47 |
11.7 |
5.32 |
|
|
Genetic Advance |
13.237 |
15.654 |
16.342 |
17.346 |
|
* = Significant at 5% probability level, LL = Leaf length, RL = Root
length, SL = Shoot length, NPR = Number of roots per plant
Table 2 Survival
percentage of seedlings under different treatments
|
Treatments |
Chickpea |
Pea |
Green mung bean |
Black mung bean |
|
Control |
94.94 |
97.14 |
94.94 |
92.50 |
|
0.5 M
ZnSO4 |
84.33 |
78.75 |
84.33 |
70.25 |
|
0.25 M
ZnSO4 |
89.59 |
80.13 |
84.61 |
72.67 |
|
500 ml
Biogas wastewater |
89.82 |
92.86 |
89.82 |
92.50 |
|
250 ml
Biogas wastewater |
89.41 |
87.14 |
89.41 |
85.34 |
The results from table 3 for chickpea indicated that the higher number of
roots per plant was found higher under control condition (9.10) while lowest
was found under 250ml biogas wastewater. The shoot length was also found under
control condition (18.19cm) while lowest for 0.5M ZnSO4
(13.18cm). The higher leaf length was found under control conditions (5.26cm)
while lowest for 0.5M ZnSO4 (4.15cm) while root length was found higher under
control condition (19.91cm) while lowest under 0.25M ZnSO4 (16.88cm). The higher shoot
length and root length under applications of ZnSO4 indicated that the genotype has
tolerance against heavy metals applications and selection may be helpful to
improve chickpea yield under stressful environment (Akhtar
et al., 2017; Bhardwaj
et al., 2014; Garci-Gomez
et al., 2017; Moller
and Stinner 2010; Verma et al., 2017). The results from table 3 for pea indicated that the
higher number of roots per plant was found higher under 250 ml Biogas wastewater condition (8.1) while lowest was found under 0.5M ZnSO4 (6.2). The shoot length was also found under control condition (15.28cm) while
lowest for 0.5M ZnSO4 (11.22cm). The higher leaf
length was found under 250 ml
Biogas wastewater
conditions (7.16cm) while lowest for 0.5M ZnSO4 (6.17cm) while root length was
found higher under 250 ml
Biogas wastewater
condition (17.91cm) while lowest under 0.5M ZnSO4 (15.89cm). The higher shoot
length and root length under applications of ZnSO4 indicated that the genotype has
tolerance against heavy metals applications and selection may be helpful to
improve pea yield under stressful environment (Akhtar
et al., 2017; Bhardwaj
et al., 2014; Garci-Gomez
et al., 2017; Usman et al., 2014).
Table 3 Mean
comparison among legumes for different traits of seedlings
|
Genotypes |
Treatment |
NRP |
SL |
LL |
RL |
|
Chickpea |
Control |
9.1a |
18.19a |
5.26a |
19.91a |
|
0.5 M
ZnSO4 |
7.3b |
13.18d |
4.15b |
16.89c |
|
|
0.25 M
ZnSO4 |
6.4c |
14.23c |
4.19b |
16.88c |
|
|
500 ml
Biogas wastewater |
6.5c |
13.27d |
5.11a |
18.87b |
|
|
250 ml
Biogas wastewater |
5.1d |
15.20 b |
5.19a |
18.92b |
|
|
Pea |
Control |
7.6b |
15.28a |
7.14a |
16.87b |
|
0.5 M
ZnSO4 |
6.2c |
11.22d |
6.17b |
15.89c |
|
|
0.25 M
ZnSO4 |
7.3b |
12.26c |
6.19b |
16.78b |
|
|
500 ml
Biogas wastewater |
7.7b |
14.19b |
7.14a |
16.78b |
|
|
250 ml
Biogas wastewater |
8.1a |
14.25b |
7.16a |
17.91a |
|
|
Green mung bean |
Control |
8.3a |
17.24a |
8.19a |
15.86 b |
|
0.5 M
ZnSO4 |
7.2b |
13.23c |
6.18c |
14.87 c |
|
|
0.25 M
ZnSO4 |
6.3c |
14.27b |
6.17c |
13.88d |
|
|
500 ml
Biogas wastewater |
8.1a |
13.21c |
7.19b |
16.89a |
|
|
250 ml
Biogas wastewater |
8.4a |
12.26d |
7.13b |
16.79a |
|
|
Black mung bean |
Control |
8.5a |
16.29a |
8.14a |
17.91a |
|
0.5 M
ZnSO4 |
5.9c |
13.21d |
6.17c |
15.85c |
|
|
0.25 M
ZnSO4 |
5.8c |
12.24e |
6.12c |
14.87d |
|
|
500 ml
Biogas wastewater |
7.1b |
15.19b |
7.18b |
16.84 b |
|
|
250 ml
Biogas wastewater |
7.4b |
14.27c |
7.16b |
17.82a |
LL = Leaf length, RL = Root length, SL
= Shoot length, NPR = Number of roots per plant
The results from table 3 for green mung bean
indicated that the higher number of roots per plant was found higher under 250 ml Biogas wastewater condition (8.4) while lowest was found under 0.25M ZnSO4 (6.3). The root length was also found under 250 ml Biogas wastewater condition (16.79cm) while lowest for
0.25M ZnSO4 (13.88cm). The higher shoot
length was found under control conditions (17.24cm) while
lowest for 250 ml
Biogas wastewater (12.26cm)
while leaf length was found higher under control condition (8.19cm) while lowest
under 0.25M ZnSO4 (6.17cm). The higher shoot length and root length
under applications of ZnSO4 indicated that the genotype has tolerance against
heavy metals applications and selection may be helpful to improve green mung bean yield under stressful environment. The results from table 3 for
black mung bean indicated that the higher number of
roots per plant was found higher under control condition (8.5) while lowest was found
under 0.25M ZnSO4 (5.8). The root length was also found under 250 ml Biogas wastewater condition (17.82cm) and control
conditions (17.91cm) while lowest for 0.25M ZnSO4 (14.87cm). The higher shoot
length was found under control conditions (16.29cm) while
lowest for 0.25M ZnSO4 (12.24cm) while leaf length was found higher under control
condition (8.14cm) while lowest under 0.25M ZnSO4 (6.12cm). The higher shoot
length and root length under applications of ZnSO4 indicated that the genotype has
tolerance against heavy metals applications and selection may be helpful to
improve black mung bean yield under stressful
environment (Ali et al., 2012; Akhtar et al., 2017; Bhardwaj
et al., 2014; Garci-Gomez
et al., 2017; Hussain et al., 2015; Usman et al., 2014).
The results from table 4 about
pooled correlation analysis of studied traits of legume crop species indicated
that there was significant and positive correlation among all of the studied
traits viz. root length, shoot length, number of roots per plant and leaf
length. The significant correlation indicated that the selection may be helpful
to improve legume seed yield under various stress conditions (Ali and Ahsan 2012; Ali et
al., 2010ab; Ali et al., 2013; Ali et al.,
2014; Ali et al., 216).
Table 4. Pooled correlation among morphological
traits of legumes under different treatments
|
Source |
LL |
NRP |
RL |
|
NRP |
0.4627* |
||
|
RL |
0.3832* |
0.6312* |
|
|
SL |
0.4972* |
0.5823* |
0.6759* |
* = Significant at 5% probability level, LL = Leaf length, RL = Root
length, SL = Shoot length, NPR = Number of roots per plant
Conflict of interest
The authors declared absence of any type
of conflict of interest.
Akhtar, M. F. u. Z., Jamil, M., Ahamd, M., & Abbasi, G. H. (2017). Evaluation of biofertilizer
in combination with organic amendments and rock phosphate enriched compost for
improving productivity of chickpea and maize. Soil and Environment, 36(1),
59–69.
Ali, Q., Ahsan, M., Ali, F., Aslam, M.,
Khan, N.H., Munzoor, M., Mustafa, H.S.B. and
Muhammad, S., (2013). Heritability, heterosis
and heterobeltiosis studies for morphological traits
of maize (Zea mays L.)
seedlings. Advancements in Life
sciences, 1(1): 53-62.
Ali,
Q., Ali, A., Ahsan, M., Nasir,
I.A., Abbas, H.G. and Ashraf,
M.A., (2014).
Line× Tester analysis for morpho-physiological traits
of Zea mays L seedlings. Advancements in Life sciences, 1(4): 242-253.
Ali,
Q., Ahsan, M., Kanwal, N.,
Ali, F., Ali, A., Ahmed, W., Ishfaq, M. and Saleem, M., (2016). Screening for drought tolerance: comparison of
maize hybrids under water deficit condition. Advancements in Life Sciences, 3(2), pp.51-58.
Ali,
Q., Ahsan, M. and Saleem,
M., (2010). Genetic variability and trait association in chickpea (Cicer arietinum L.). Electronic Journal of Plant Breeding, 1(3): 328-333.
Ali, Q., Muhammad,
A. and Farooq, J., (2010). Genetic
variability and trait association in chickpea (Cicer arietinum L.) genotypes at seedling stage. Electronic Journal of Plant Breeding,
1(3): 334-341.
Ali,
Q., Ahsan, M., Khaliq, I., Elahi, M., Shahbaz, M., Ahmed, W.
and Naees, M., (2011). Estimation of
genetic association of yield and quality traits in chickpea (Cicer arietinum L.).
International Research Journal of Plant
Sciences, 2(6): 166-169.
Ali,
Q., Ahsan, M., Khan, N.H., Ali, F., Elahi, M. and Elahi, F., (2012). Genetic
analysis for various quantitative traits of chickpea (Cicer
arietinum L.). International Journal for Agro Veterinary and Medical Sciences, 6(1): 51-57.
Ali,
Q. and Ahsan, M., (2012). Estimation of
genetic variability and correlation analysis for quantitative traits in
chickpea (Cicer arietinum
L.). International
Journal for Agro Veterinary and Medical Sciences, 6(4), pp.241-249.
Amjad, M., (2002). Performance of
nine pea cultivars under Faisalabad conditions. 39, 16–19.
Bhardwaj, D., Ansari, M. W., Sahoo, R. K.,
& Tuteja, N. (2014). Biofertilizers
function as key player in sustainable agriculture by improving soil fertility,
plant tolerance and crop productivity. Microbial
Cell Factories, 13(1), 1–10.
Chen,
J.-H. (2006).
The combined use of chemical and organic fertilizers
and/or biofertilizer for crop growth and soil
fertility Jen-Hshuan Chen ? Department of Soil and
Environmental Sciences, National Chung Hsing
University, Taiwan, R.O.C. International
Workshop on Sustained Management, October, 1–11.
Cousin,
R. (1997). Peas (Pisum sativum
L.). Field Crops Research, 53(1–3), 111–130.
Deshpande, S. S., Sathe, S. K., & Salunkhe, D.
K. (1984).
Interrelationships between certain physical and chemical properties of dry bean
(Phaseolus vulgaris L.). Qualitas Plantarum Plant
Foods for Human Nutrition, 34(1),
53–65.
García-Gómez, C., Obrador, A., González, D., Babín, M., & Fernández, M. D.
(2017).
Comparative effect of ZnO NPs, ZnO
bulk and ZnSO4 in the antioxidant defences of two
plant species growing in two agricultural soils under greenhouse conditions. Science of the Total Environment, 589, 11–24.
Hussain, N., Aslam, M., Ghaffar, A., & Irshad, M. (2015). Chickpea genotypes evaluation for morpho-yield traits under water stress conditions. JAPS:
Journal of Animal & Plant Sciences
25(1), 206–211.
Mahajan, R., Dar, A. A., Mukthar, S., & Zargar, S. M.
(2018). Pisum Improvement Against
Biotic Stress : Current Status and Future Prospects. In Pulse Improvement (pp.
109-136). Springer, Cham.
Mahanty, T., Bhattacharjee, S., Goswami, M.,
Bhattacharyya, P., Das, B., Ghosh, A., & Tribedi, P. (2017). Biofertilizers: a
potential approach for sustainable agriculture development. Environmental Science and Pollution Research,
24(4), 3315–3335.
Möller, K. (2009). Influence of
different manuring systems with and without biogas
digestion on soil organic matter and nitrogen inputs, flows and budgets in
organic cropping systems. Nutrient
Cycling in Agroecosystems, 84(2), 179–202.
Möller, K., & Stinner, W. (2010). Effects of organic wastes
digestion for biogas production on mineral nutrient availability of biogas
effluents. Nutrient Cycling in Agroecosystems, 87(3),
395–413.
Pathak, G. C., Gupta,
B., & Pandey, N. (2012). Improving
reproductive efficiency of chickpea by foliar application of zinc. Brazilian Journal of Plant Physiology, 24(3), 173–180.
Ranpariya, V. S., Polara, K. B., Hirpara, D. V,
& Bodar, K. H. (2017). Effect of potassium
, zinc and FYM on content and uptake of nutrients in seed of summer
green gram (Vigna radiata L
.) and post harvest soil fertility under medium black calcareous soil. International Journal of Chemical
Studies, 5(5), 1055–1058.
Sai, K., Rama, S., & George, P.
J. (2017). Effect of levels of phosphorus and zinc on growth and yield of
Kabuli chickpea (Cicer
kabulium L .). 6(4),
1013–1016.
Sangolli, V. O., Nawalagatti, C. M., Uppar, D. S.,
& Koti, R. V. (2018). Effect of zinc
nutrition on morphological characters in chickpea. 7(2), 902–904.
Serdjuk, M., Bodmer, U., & Hülsbergen, K.
J. (2018).
Integration of biogas production into organic arable farming systems: crop
yield response and economic effects. Organic Agriculture, 8(4), 301–314.
Usman, M., Tahir, M. and Majeed, M.A., 2014. Effect of zinc
sulphate as soil application and seed treatment on
green gram (Vigna radiata
L.). Pakistan
Journal of Life and Social Sciences, 12(2),
pp.87-91.
Verma, P. D., Swaroop,
N., Upadhyay, Y., Swamy,
A., & Dhruw, S. S. (2017). Role
of Phosphorus, Zinc and Rhizobium on Physico-Chemical Properties of Soil in Field Pea (Pisum sativum L.) cv. Rachna. International Journal of Current Microbiology and Applied
Sciences, 6(7), 4423–4428.
Zubair, M., Shakir, M., Ali, Q., Rani, N.,
Fatima, N., Farooq, S., Shafiq,
S., Kanwal, N., Ali, F. and Nasir,
I.A. (2016). Rhizobacteria
and phytoremediation of heavy metals. Environmental Technology Reviews, 5(1): 112-119.