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Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
Effects of Acalypha wilkesiana Leaf Extract, Hot and Boiling 
Water on Plantain Growth Trend and Soil Nematode Densities 
M. Omolara Olaniyi1 
Department of Crop, Soil and Pest Management, School of Agriculture and Agricultural Technology, 
Federal University of Technology, Akure, Ondo State, Nigeria 
The study was funded in part by the International Foundation for Science through research grant number 
C/3859-1. Olumide Alabi gave technical assistance with data collection. 
Abstract 
An experiment was laid out at the Teaching and Research Farm of the Federal University of Technology, Akure 
to investigate the trend in plantain growth response and nematode population density build up in response to 
paring, red acalypha leaf extract, hot water and boiling water dip. Above ground plant growth parameters were 
assessed 55, 84 and 111 days after planting, approximately 8, 12 and 16 weeks respectively. Population densities 
of plant parasitic nematodes in root rhizosphere at these dates were also investigated. The results indicated that 
paring enhanced plantain resistance to plant parasitic and edaphic stressors. In a holistic approach, paring and 
further treating pared suckers in red acalypha leaf extract for 5-15 mins conferred protection against plant 
parasitic nematodes and liming effect on the soil. This supposed liming effect merits further investigations. 
However, pre-plant dip of pared suckers in red acalypha leaf extract for 20 mins resulted in stress on the plant, 
which encouraged high nematode density build up and concomitant parasitism. 
Keywords: Helicotylenchus multicinctus, Pratylenchus coffeae, Radopholus similis, plantain, red acalypha, hot 
water treatment, plant extract 
1. Introduction 
Plant parasitic nematodes are major constraints to the production of plantain (Musa spp., AAB-group) in Nigeria. 
With reference to the, black sigatoka leaf disease and the banana weevils, the nematode species, Pratylenchus 
coffeae and Radopholus similis are considered the key pathogens causing great damage to plantain in Nigeria 
(Speijer et al., 2001). P. coffeae is strongly associated with root damage and plant toppling over, which are 
strong indices of plant parasitic nematode damage. The species has been ranked the number one constraint to 
plantain production in Nigeria (Olaniyi, 2011). The plant parasitic nematodes species most frequently associated 
with plantain in Nigeria are Helicotylenchus multicintus, Hoplolaimus pararobustus, Meloidogyne spp., 
Pratylenchus spp. and Radopholus similis (Rotimi et al., 1999; Speijer et al., 2001). 
A complex of Radopholus similis, Helicotylenchus dihystera, H. multicinctus, Hoplolaimus pararobustus and 
Meloidogyne species in South eastern Nigeria cause on average, 50% loss in plantain production in Nigeria 
depending on the cultivar type and cultural practices employed (Olaniyi, 2011). Pratylenchus coffeae was 
reported to be important in southwestern Nigeria, yet in spite of its seriousness, it damage potential in 
Southwestern Nigeria is yet to be quantified and described. 
These damaging nematodes are introduced to new and otherwise clean fields through infested soil adhering to 
planting materials and also infected roots of sucker planting materials (Speijer et al., 2001, Rotimi and Opadare, 
2006). This makes sanitation an important nematode management principle in plantain. To improve root and 
rhizome health, sucker planting materials are often pared (i.e. removal of adhering soil and peeling of the skin of 
the rhizome in order to expose infected tissues, remove where possible before planting or discard materials with 
serious infection (Blake, 1961). However, farmers do not feel comfortable with this procedure and are slow to 
adopt it because they often believe that such pared suckers would not survive and if they do would have low 
productivity. In order for farmers to adopt this cultural practice, the benefits of paring need to be demonstrated. 
Also in developing effective environmentally friendly interventions, several options are considered. Acalypha 
wilkesiana (red Acalypha) leaf extract has been demonstrated to be effective against plant parasitic nematodes 
(Rotimi and Moens, 2005) and documented as effective plantain root health enhancer. In order to develop an 
effective control tactic for plant parasitic nematodes on plantain an integrated management package should be 
developed. Therefore, this study investigated the efficacy of combining paring with red Acalypha leaf extract at 
different exposure duration, hot water or boiling water treatments in the control of plant parasitic nematodes on 
plantain, cultivar Agbagba, in P. coffeae endemic Southwestern Nigeria. 
105 
2. Materials and Methods 
2.1 Site Description and Field Layout 
1 Current Address: Biology Unit, National Open University of Nigeria, 14/16 Ahmadu Bello Way, Victoria Island, Lagos, 
Lagos State. E-mail: molaniyi@noun.edu.ng
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
The trial was conducted at the Teaching and Research Farm (Crop section) of the Federal university of 
Technology, Akure. Akure lies within the Tropical rainforest belt between latitude 5oN and longitude 150E of the 
equator, with an annual mean temperature of about 27oC.the dry season is usually witnessed in Akure between 
November and March, while the rainy season ranged from April to October. 
The experimental site covered a total area of 960m2. Previously, the site was used for a mulch trial to study the 
vegetative response of plantain to two organic mulch types. The experiment had been terminated five months 
earlier and left to re-vegetate naturally before the site was opened for the present study. The trial was arranged in 
a completely randomized design (CRD) of eight treatments in all and ten replicates per treatment. The site was 
slashed and burned before marking out and establishing the field. The spacing used was 3 metres between the 
rows and 2 metres within the rows, there were 10 suckers per row and eighty suckers in all for the trial. 
2.2 Preparation of Red Acalypha Leaf Extracts and Sucker Treatment 
Plantain (cultivar Agbagba) suckers were sourced from within the vicinity of the Teaching and Research Farm of 
the University (Crop section), while the red Acalypha was sourced from Owena, a suburb of Akure. There were 
eight treatments in the experiment including the controls. The treatments were: pared control (T1), pared suckers 
with 5 mins dip in Acalypha extract (T2), pared suckers with 10 mins dip in Acalypha extract (T3), pared suckers 
with 15 mins dip in Acalypha extract (T4), pared suckers with 20 mins dip in Acalypha extract (T5), Hot water 
treatment at 52oC for 20 mins (T6), Boiling water treatment at 100oC for 30 sec. (T7) and Non-pared control 
(T8). 
Air-dried leaves of red Acalypha plant were pulverized and 100g of the powder was dissolved in 9 litres of cold 
water in four different buckets each. A wooden rod was used to stir the mixture to ensure homogenization after 
which they were left to stand for 30 mins. The buckets were respectively labelled as 5, 10, 15 and 20 minutes 
with a marker to avoid mix-up. Thereafter, 10 suckers were dipped into each of the buckets and they were left to 
stand for the duration on each bucket’s label. The suckers were removed and left to air dry under shade 
afterwards. 
For the hot water treatment, 10 suckers were dipped in water at 52oC ±2oC for 20 mins and they were removed 
and left to cool for about 24 hours before planting, while for the boiling water treatment, 10 suckers were dipped 
in water at 100oC for 30 sec. Suckers were planted on 21st December, 2006. Due to cessation of rainfall during 
this period of the year, manual irrigation of the plant once in two days was adopted until the resumption of 
rainfall in late March, 2007. 
2.3 Above Ground Plant Growth Parameters 
Growth parameters were taken on the above ground parts of the sprouted plants at 55, 84 and 111 days after 
planting (DAP), giving approximately 8, 12 and 16 weeks after planting (WAP) respectively. Data taken 
included the height of the pseudostem of the mother plant from the surface to the point of emergence (axils) of 
the youngest leaf, the girth of the pseudostem at soil level, number of functional (green) and non-functional (dry) 
leaves. A leaf was considered functional when at least 75% of the leaf area is green and non-functional if 
otherwise (Rotimi et al., 2004b). The length and the width of the youngest leaf were measured: the length of the 
youngest leaf opened was taken from the stalk to the leaf apex while the width was taken from the widest portion 
of the leaf. The leaf area was calculated as length x width x 0.83 (a constant) according to Obiefuna and Ndubizu 
(1979). 
2.4 Nematode Extraction and Identification 
Soil samples were taken from the rhizosphere of each plant at 8, 12 and 16 WAP. One hundred ml of each 
sample was measured unto a serviette placed in a plastic sieve. The sieve was then placed in a plastic plate and 
water was carefully added to the plate until the soil on the serviette-covered sieve appeared moist on top 
(modified Baermann tray). The setup was left to stand for 24 hours after which the resulting suspension was 
decanted, left to stand for 4 hours afterwards and then reduced to 30ml with a syringe. 
Plant parasitic nematodes were identified to species level with the light microscope and all developmental stages 
of the nematode species were counted, except for the root knot nematode which was identified only to genus 
level and only vermiform juveniles and males that could be extracted with the extraction technique were 
counted. The total of all the developmental stages counted was presented in each case. Densities of the 
nematodes were estimated and presented per litre of soil. 
2.5 Soil Chemical Analysis 
At time of field establishment, soil samples were taken from each replicate hole and bulked per treatment for 
physico-chemical analysis. Soil samples were also taken at termination of the experiment, 16 WAP. Soil 
chemical analysis was carried out in the soil Analytical laboratory of the Department of Crop, Soil and Pest 
106
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
Management of the Federal University of Technology, Akure. Soil properties determined included pH, Nitrogen, 
cations such as Ca2+, Mg2+ and K+ content. Others were Phosphorus, Sodium, organic matter, and organic carbon. 
3. Results 
3.1 Effects of Treatments on Plantain Growth Pattern across the Sampling Dates 
The effect of the treatments on plant height relative to time is presented in Figure 1(a - h) the R2 value presented 
on each graph is the coefficient of determination of each regression equation. The 5 mins dip in red acalypha leaf 
extract (Figure 1b), 10 mins dip in red acalypha leaf extract (Figure 1c) and 30 sec dip in boiling water (Figure 
1g) treatments exhibited linear trend in growth response over time. A quadratic polynomial functional trend in 
height increase was observed with the 20 minutes dip in Acalypha extract treatment over time (T5), hot water 
treatment and non-pared control which is the farmer’s method followed (Figure 1e, f, h). Only the pared (Figure 
1a) and the 15 mins dip in Acalypha extract (Figure 1d) displayed a cubic polynomial trend in height increase 
over time. 
Pseudostem thickness increased at a linear rate over time only for the pared treatment (Figure 2a) while for the 5 
mins dip in red acalypha leaf extract, pseudostem thickness over time responded with a quadratic polynomial 
function (Figure 2b). All other treatments (Figure 2c - h) displayed a cubic polynomial trend in increase in 
thickness over time. 
In the number of functional leaves produced, the pared treatment followed a cubic polynomial function over time 
(Figure 3a) while boiling water dip (Figure 3g) and the farmer’s material (Figure 3h) displayed a quadratic 
polynomial trend from 55 to 111 days after planting. The number of green (functional) leaves produced in the 5, 
10, 15 and 20 minutes dip in Acalypha extract treatments along side the hot water treatment exhibited a linear 
response over the sampling dates (Figure 3b –f). 
Ten, 15 and 20 min dip in red acalypha leaf extract as well as 30 sec dip in boiling water resulted in linear 
response in leaf area expansion from 55 to 111 days after planting (Figure 4 c, d, e and g) while expansion trend 
in leaf area in all the other treatments can be explained by the quadratic polynomial function. 
107
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
25 
20 
15 
10 
5 
a b 
30 
25 
20 
15 
10 
5 
c d 
25 
20 
15 
10 
5 
e f 
35 
30 
25 
20 
15 
10 
5 
g h 
18 
16 
14 
12 
10 
8 
6 
4 
2 
30 
25 
20 
15 
10 
5 
30 
25 
20 
15 
10 
5 
20 
15 
10 
5 
Figure 1. Effects of red Acalypha extract, hot and boiling water treatments on height response of plantain from 
55 to 111 days after planting. 
108 
Pared control 
y = 7E-05x3 - 0.0155x2 + 1.0978x - 7.8614 
R2 = 0.7274 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
5 minutes dip in Acalypha 
extract 
y = 0.0747x + 12.09 
R2 = 0.9329 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Height (cm) 
10 minutes dip in Acalypha 
extract 
y = 0.1942x + 3.9017 
R2 = 0.9773 
0 
0 20 40 60 80 100 120 
days after planting (days) 
15 minutes dip in Acalypha 
extract 
y = -8E-05x3 + 0.0208x2 - 1.6214x + 
64.412 
R2 = 0.2923 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Height (cm) 
20 minutes dip in Acalypha 
extract 
y = 0.003x2 - 0.3532x + 27.632 
R2 = 0.7396 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Height (cm) 
Hot-water treatment 
y = -0.0024x2 + 0.4299x + 1.4399 
R2 = 0.6771 0 
0 20 40 60 80 100 120 
days after planting (days) 
Height (cm) Boiling-water treatment 
y = 0.0632x + 11.097 
R2 = 0.7666 
0 
0 20 40 60 80 100 120 
days af t er plant ing (days) 
Non-pared control 
y = -0.0042x2 + 0.5441x + 10.251 
R2 = 0.6654 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days)
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
109 
10 
8 
6 
4 
2 
a b 
12 
10 
8 
6 
4 
2 
c d 
12 
10 
8 
6 
4 
2 
e f 
10 
8 
6 
4 
2 
g h. 
10 
8 
6 
4 
2 
14 
12 
10 
8 
6 
4 
2 
10 
9.5 
9 
8.5 
15 
10 
5 
Figure 2. Effects of red Acalypha extract, hot and boiling water treatments on pseudostem girth of plantain 
response from 55 to 111 days after planting. 
Pared control 
y = -0.0442x + 10.391 
R2 = 0.945 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
5 minutes dip in Acalypha 
extract 
y = 0.0007x2 - 0.1269x + 13.265 
R2 = 0.6359 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
10 minutes dip in Acalypha 
extract 
y = 2E-05x3 - 0.0052x2 + 0.3831x - 
0.9432 
R2 = 0.4287 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Girth (cm) 
15 minutes dip in Acalypha 
extract 
y = -9E-05x3 + 0.0233x2 - 1.8922x + 
58.767 
R2 = 0.6689 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
20 minutes dip in Acalypha 
extract 
y = -9E-05x3 + 0.0229x2 - 1.7801x + 51.469 
R2 = 0.3556 
0 
0 20 40 60 80 100 120 
days af t er pl ant i ng ( days) 
Hot-water treatment 
y = 3E-05x3 - 0.006x2 + 0.4357x - 
0.9446 
R2 = 0.4061 
8 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
Boiling-water treatment 
y = -4E-05x3 + 0.0108x2 - 0.8369x + 
28.418 
R2 = 0.8299 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Girth (cm) 
Non-pared control 
y = -0.0016x2 + 0.1399x + 10.737 
R2 = 0.9049 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Girth (cm)
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
110 
4.5 
4 
3.5 
3 
2.5 
2 
1.5 
1 
0.5 
a b 
8 
7 
6 
5 
4 
3 
2 
1 
c d 
6 
5 
4 
3 
2 
1 
e f 
6 
5 
4 
3 
2 
1 
g h 
7 
6 
5 
4 
3 
2 
1 
8 
7 
6 
5 
4 
3 
2 
1 
6 
5 
4 
3 
2 
1 
4.5 
4 
3.5 
3 
2.5 
2 
1.5 
1 
0.5 
Figure 3. Effects of red Acalypha extract, hot and boiling water treatments on number of green leaves produced 
by the plant from 55 to 111 days after planting. 
Pared control 
y = -0.001x2 + 0.1532x - 1.972 
R2 = 0.887 0 
0 20 40 60 80 100 120 
days af t er plant ing ( d ays) 
5 minutes dip in Acalypha 
extract 
y = 0.0511x + 0.04 
R2 = 0.8296 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
10 minutes dip in Acalypha 
extract 
y = 0.0704x - 1.0638 
R2 = 0.8859 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
15 minutes dip in Acalypha 
extract 
y = 0.069x - 0.9813 
R2 = 0.9529 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
20 minutes dip in Acalypha 
extract 
y = 0.0597x - 1.0026 
R2 = 0.9944 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
Hot-water treatment 
y = 0.0545x - 0.4345 
R2 = 0.9548 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
Boiling-water treatment 
y = 0.0006x2 - 0.0677x + 5.6733 
R2 = 0.5818 
0 
0 20 40 60 80 100 120 
days after planting (days) 
no. of green leaves 
Non-pared control 
y = -0.0013x2 + 0.2194x - 5.3017 
R2 = 0.9004 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days)
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
leaf area (cm2) 
leaf area (cm2) 
111 
Pared control 
y = -0.0835x2 + 12.06x - 190.27 
R2 = 0.8809 
300 
250 
200 
150 
100 
50 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
a b 
10 minutes dip in Acalypha 
extract 
y = 4.6715x - 80.986 
R2 = 0.8613 
500 
400 
300 
200 
100 
0 
0 20 40 60 80 100 120 
days after planting (days) 
leaf area (cm2) 
c d 
20 minutes dip in Acalypha 
extract 
y = 3.5363x + 16.008 
R2 = 0.74 
500 
400 
300 
200 
100 
0 
0 20 40 60 80 100 120 
days after planting (days) 
leaf area (cm2) 
e f 
Boiling-water treatment 
y = 1.6598x + 143.12 
R2 = 0.633 
400 
300 
200 
100 
0 
0 20 40 60 80 100 120 
days after planting (days) 
leaf area (cm2) 
g h 
5 minutes dip in Acalypha 
extract 
y = -0.0955x2 + 15.076x - 276.73 
R2 = 0.7995 
350 
300 
250 
200 
150 
100 
50 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
15 minutes dip in Acalypha 
extract 
y = 1.697x + 333.63 
R2 = 0.5318 
600 
500 
400 
300 
200 
100 
0 
0 20 40 60 80 100 120 
days af t er plant ing ( days) 
Hot-water treatment 
y = -0.1562x2 + 27.117x - 
768.53 
R2 = 0.7773 
500 
400 
300 
200 
100 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Non-pared control 
y = -0.3791x2 + 57.51x - 
1560.6 
R2 = 0.7745 
800 
600 
400 
200 
0 
0 20 40 60 80 100 120 
days after planting (days) 
Figure 4. Effects of Acalypha wilkesiana leaf extract, hot and boiling water treatments on the leaf area growth 
trend of plantain from 55 to 111 days after planting. 
3.2 Effects of Treatments on Plant Parasitic Nematode Species and Densities Recovered 
Three species of plant parasitic nematodes namely: Helicotylenchus multicinctus, Pratylenchus coffeae and 
Radopholus similis were recovered from plantain rhizosphere in this study. At 55 days (8 weeks) after planting, 
only the 5 minutes dip in extract (T2) had a combination of two species of nematodes namely Radopholus similis
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Vol.4, No.18, 2014 
and Pratylenchus coffeae while P. coffeae was recovered under plantain in the other treatments except 20 mins 
dip in extract and the not pared control (Figure 5a), at 84 days (12 weeks) after planting, the highest population 
density and species combination was observed in the 20 minutes dip in extract (T5) (Figure 5b) while at 111 
days (16 weeks), the nematode population recovered from the roots had reduced relatively to the earlier 
population densities observed in preceding samplings (Figure 5c). 
density/litre soil 
112 
nematode from soil 55 DAP 
700 
600 
500 
400 
300 
200 
100 
0 
A 
A 
a 
A 
a 
a 
A 
a 
A 
A a 
a b 
a 
T1 T2 T3 T4 T5 T6 T7 T8 
Treatments 
density/litre soil 
R.s 
Praty 
a 
a b 
c 
nematode from soil 84 DAP 
700 
600 
500 
400 
300 
200 
100 
0 
A 
a a 
a1 
a1 
a 
A 
T1 T2 T3 T4 T5 T6 T7 T8 
Treatments 
R.s 
Praty 
H.m 
nematode from soil 111 DAP 
1000 
800 
600 
400 
200 
0 
A 
a a H.m 
A 
T1 T2 T3 T4 T5 T6 T7 T8 
Treatments 
density/litre soil 
R.s 
Praty 
Figure 5. Effects of treatments on rhizosphere nematode densities at 55 (a), 84 (b) and 111 (c) days after planting 
(DAP). 
T1 = Pared control (no dip treatment), T2 = 5 mins dip in Acalypha extract, T3: 10 mins dip in Acalypha 
extract, T4 = 15 mins dip in Acalypha extract, T5 = 20 mins dip in Acalypha extract, T6 = Hot-water treatment 
for 20 mins, T7 = Boiling-water treatment for 30 sec, T8 = Non-pared control. Column fractions followed by 
same letters (and numbers) are not significantly different at P≤0.05. 
3.3 Effects of Treatments on Soil Chemical Properties 
Chemical properties of the soil at pre-plant are presented on table 1 while table 2 presents the soil chemical 
properties 111 days after planting. Ten and 15 mins dip in Acalypha leaf extract, and 30 sec dip in boiling water 
depressed soil pH, while 5 and 20 mins dip in red Acalypha extract as well as 20 mins dip in hot water resulted 
in slightly raised pH. The non-pared control, which represented the farmer’s method, also resulted in depressed 
soil pH, while the pared suckers gave relatively constant pH. Generally, soil potassium content declined between 
planting and termination of the experiment 111 days afterwards. Except for the 15 mins dip in Acalypha extract 
treatments, soil phosphorus also declined from planting till end of experiment. Nitrogen content also declined 
marginally from planting to termination of the experiment; except for the 20 mins hot water dip and 30 mins 
boiling water dip treatments.
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Vol.4, No.18, 2014 
Table 1. Pre-plant soil chemical properties in plots where the treatments shown were laid out 
T1 = Pared control (no dip treatment), T2 = 5 mins dip in Acalypha extract, T3: 10 mins dip in 
Acalypha extract, T4 = 15 mins dip in Acalypha extract, T5 = 20 mins dip in Acalypha extract, T6 
= Hot-water treatment for 20 mins, T7 = Boiling-water treatment for 30 sec, T8 = Non-pared 
control. 
Treatment pH Mg Ca %OM %OC P Na K N 
T1 5.87 2.30 2.90 11.31 6.55 1.42 7.13 1.13 0.31 
T2 5.59 1.10 3.50 6.49 3.75 1.41 12.3 0.41 0.38 
T3 5.73 0.70 3.40 10.02 5.72 1.10 13.3 0.56 0.21 
T4 5.79 1.30 3.10 8.62 4.99 0.96 11.39 1.24 0.20 
T5 5.45 1.10 2.00 9.93 5.75 0.88 10.52 0.87 0.32 
T6 5.46 1.10 2.40 13.25 7.66 1.43 12.35 0.46 0.17 
T7 6.08 0.80 2.90 9.87 5.71 2.41 10.96 0.36 0.27 
T8 5.68 1.80 2.90 10.97 6.34 1.29 13.04 0.36 0.18 
Table 2. Effects of paring, red acalypha leaf extract , hot and boiling water treatments of plantain 
sucker planting materials on soil chemical properties 16 weeks after planting 
T1 = Pared control (no dip treatment), T2 = 5 mins dip in Acalypha extract, T3: 10 mins 
dip in Acalypha extract, T4 = 15 mins dip in Acalypha extract, T5 = 20 mins dip in 
Acalypha extract, T6 = Hot-water treatment for 20 mins, T7 = Boiling-water treatment for 
30 sec, T8 = Non-pared control. 
Treatment pH Mg Ca %OM %OC P Na K N 
T1 5.86 1.10 3.80 11.18 6.46 0.94 11.39 0.56 0.28 
T2 5.94 0.10 3.80 11.31 6.54 0.83 14.43 0.87 0.31 
T3 5.24 0.50 3.80 9.73 5.63 0.59 11.83 0.41 0.15 
T4 5.60 1.70 2.70 10.69 6.19 0.90 15.13 0.36 0.18 
T5 5.84 2.00 2.40 11.24 6.50 1.05 11.74 0.42 0.18 
T6 5.52 0.70 2.50 9.38 5.43 1.24 12.17 0.29 0.31 
T7 5.56 1.60 2.80 9.31 5.39 0.88 11.83 0.25 0.27 
T8 5.55 0.50 2.80 9.31 5.31 1.04 10.26 0.27 0.13 
4. Discussion 
The linear response obtained for some of the growth parameters for 5, 10, 15 and 20 minutes dip in acalypha 
extract, hot and boiling water treatments in this study connotes that the rate of change in the growth of the plant 
in the treatments increased over time at a constant rate in spite of the prevailing harsh dry condition. Polynomial 
functional relationships obtained in the non-pared control (Farmer’s method) might be an indication of the high 
susceptibility of unpared planting materials to dry weather. This then suggests that paring the sucker before 
planting enhanced the plant’s ability to withstand biotic and edaphic stress. Further treating the planting 
materials with plant extract or hot water (52 ±2oC) also conferred some level of resistance to these stress factors. 
Olaniyi (2014b, In Press) also reported functional growth trend in time for plantain in reaction to organic mulch. 
The author noted that organic mulch enhanced the plants ability to withstand biotic stressors like plant parasitic 
nematodes in the environment. 
The effects of treatments in the present study on growth parameters such as the leaf area, number of functional 
leaves, plant height and girth revealed that 15 minutes dip of planting materials in the extract showed some 
promise in enhancing plant growth performance. However, it is not clear what this additional benefit of dipping 
113
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
pared suckers translates to in maintaining low nematode populations since the suckers that were only pared had 
very low densities of P. coffeae (about 20/litre soil) and maintained no detectable level of nematodes 
subsequently. Where as the highest density of P. coffeae recorded in this study was from the 15 mins dip in 
extract, earlier report (Olaniyi, 2014a, In Press) showed that 15 min dip reduced nematode damage to root and 
rhizome of plantain and also reduced nematode densities. In the present study 5 and 10 mins dip treatment 
seemed to better reduce nematode population densities on plantain compared to the 15 mins dip. Generally, 
Helicotylenchus multicinctus, Pratylenchus coffeae and Radopholus similis were the nematode species observed 
from plantain roots and the rhizosphere in this study. There is need to monitor plant response to these treatments 
and nematode development on plantain under these treatments till yield stage. 
Although densities of the species in the rhizosphere were low with maximum of 100 H. multicinctus / litre, 600 
P. coffeae / litre and 200 R. similis / litre, it is clear that densities had increased from the initial pre-plant 
densities of 1 P. coffeae / litre soil and 2 H. multicinctus / litre soil. This shows that planting a host plant in a soil 
would quickly increase the densities of the nematode species to a damaging level. It would be informative to 
investigate the effect of these densities on root and rhizome damage of plantain. Before the choice of a plant to 
put in a soil is made, it would be helpful to first do a diagnostic nematode assessment in order not to put in a host 
crop in a field where a damaging species is prevalent. For instance, Speijer et al. (2001) established Pc as the 
most damaging on plantain in Nigeria and the fact that it could persist in a field without the plantain host makes 
its control a big challenge as this might signify that it has a wide host range. The trend in population densities 
recovered from plantain rhizosphere is at variance with earlier record of H. multicinctus often in higher densities 
than other species on plantain in Nigeria (Speijer et al., 2001; Rotimi et al., 2004b, c; Coyne et al., 2005). 
Radopholus similis was not recovered from the soil before the establishment of this trial but was recovered from 
the rhizosphere of plantain 55 days (8 weeks) after planting in 5 min dip in red acalypha leaf extract treatment 
but subsequently, it was undetected in soil samples. This shows that first R. similis may not have as wide host 
range as P. coffeae or probably it could not persist as long as P. coffeae in the absence of a preferred host hence 
the absence of the species in the soil pre plant. The land had been fallowed for about 5 months before it was 
opened up for this study. However, the presence and disappearance of the species at subsequent sampling date 
supported Olaniyi’s (2011) submission that for accurate diagnosis, nematode sampling should be done across 
seasons. The study reported here was done in the dry season and Rotimi et al. (2004a) similarly observed that the 
densities of R. similis could decline to an undetected level in dry soil. 
No specific trend emerged in plant parasitic nematode community structure under plantain in this study. This 
suggests that several factors confound field studies and it would be useful to investigate nematode reactions to 
these treatments in controlled environment. For instance in the unpared control treatment, no nematode was 
detected in plantain rhizosphere 8 and 16 weeks after planting whereas at 12 weeks after planting P. coffeae was 
detected at an average of 400 nematodes / litre soil. However under the pared treatment, only very low density of 
Pc was recovered 8 weeks after planting. Subsequently no nematode was detected implying that it may suffice 
for nematode control to simply pare plantain sucker planting materials before planting without any further 
treatment. Olaniyi (2014a, In Press) also did not recover any plant parasitic nematode species from the roots of 
pared plantain at those dates. 
A number of factors could be responsible for the variations observed in the nematode incidence across the 
sampling dates. The variations obtained in the result across the three samplings established the rationale and the 
importance of sampling more than once in the diagnostic study of plant parasitic nematodes so that correct 
assertion can finally be made (Olaniyi, 2011, 2014a, In Press). It is a common phenomenon in nematode 
population dynamics, and that is why it is advisable to sample several times in time and season. Also, sampling 
only the rhizosphere could give misleading information as the species recorded in this study are migratory 
endoparasites and several factors could be responsible for their density fluctuation in the rhizosphere. To confirm 
this, the study reported by Olaniyi (2014a, In Press) investigated root densities and a different pattern emerged. 
Therefore, for a more holistic overview, it is important to sample both root and soil complementarily as this 
would give more reliable information and better guide management decision. Other factors that may be 
responsible for variations include the soil condition, environmental condition and availability of moisture in the 
soil. 
As expected, N, P & K values declined over time in this study. Results of this study revealed that introducing 
soil from other sources with planting materials as with the farmer’s method of not paring may result in increased 
soil acidity. But this would be dependent on the chemical status of the introduced soil. This is revealed by the 
stable pH of soils from pared suckers, which had all adhering soils removed before planting. Dipping rhizomes 
of planting materials in red Acalypha leaf extract for 5 or 20 minutes tended to reduce soil acidity. The liming 
effect of red Acalypha should be further explored. Considering the overall effect of treatments on plant response 
and nematode density, it would be safe to recommend paring as a useful cultural practice that farmers should be 
114
Journal of Biology, Agriculture and Healthcare www.iiste.org 
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) 
Vol.4, No.18, 2014 
encouraged to engage in. Further treating pared suckers in red acalypha extract for 5-15 mins might confer 
protection against plant parasitic nematode damage (Olaniyi, 2014a, In Press) and improve soil quality. 
References 
Blake, C.D. (1961), “Root rot of bananas caused by Radopholus similis (Cobb) and its control in New South 
115 
Wales”. Nematologica 6, 295-310. 
Coyne, D.L., Rotimi, M.O., Speijer, P., De Schutter, B., Dubois, T., Auwerkerken, A., Tenkouano, A. and De 
Waele, D. (2005), “Effects of nematode infection and mulching on the yield of plantain (Musa spp., 
AAB-group) ratoon crops and plantation longevity in southeastern Nigeria”. Nematology 7(4), 531-541. 
Obiefuna, J.C. and Ndubizu., T.O.C. (1979), “Estimating leaf area in plantain”. Scientia Horticulturae 11, 31-36. 
Olaniyi, M.O. (2006), “Effect of organic mulch and root dips on the establishment and root health of plantain”. 
Xvill Reunia International Acorbat Anais Proceedings Memorias, 321. 
Olaniyi, M.O. (2011), “Plant Parasitic Nematode Constraint to Plantain production in Nigeria”. LAP Lambert 
Publishing, Germany. September 2011. ISBN 978-3-8454-2312-8, 240pp. 
Olaniyi, M.O. (2014a), “Effects of Acalypha wilkesiana leaf extract, hot and boiling water on plantain growth 
response and nematode damage”. Nigerian Journal of Nematology, In Press. 
Olaniyi, M.O. (2014b), “Functional trend in leaf and sucker production of plantain in response to paring and 
organic mulch”. The International Journal of Science and Technoledge, In Press. 
Olaniyi, M.O. and Opadare, S.O. (2006), “Effect of paring on the establishment and root damage of French 
plantain suckers”. In: E. Soprano, F.A. Tcacenco, L.A. Litchemberg & M.C. Silva (Eds.). Banana: A 
Sustainable Business. XVII ACORBAT International Meeting, Joinville, Brazil. 15-20 October 2006, 
345. 
Rotimi, M.O. and Moens, M. (2005), “Effect of leaf extracts of some herbs on the juveniles of Meloidogyne 
incognita (Kofoid & White) Chitwood”. Nigerian Journal of Plant Protection 22, 95- 103. 
Rotimi, M.O., Speijer, P.R. and De Waele D. (2004a), “Effect of soil moisture and organic matter levels on 
plantain root reaction to Radopholus similis”. International Journal of Nematology 14(2), 122-128. 
Rotimi, M.O., Speijer, P., De Waele, D. and Swennen, R. (2004b), “Effect of mulching on the response of 
plantain (cv. Agbagba, Musa spp. AAB-group) to plant parasitic nematodes in Southeastern Nigeria. I: 
Vegetative growth”. Nigerian Journal of Forestry. 34 (1), 61-71. 
Rotimi, M.O., Speijer, P., De Waele, D. and Swennen, R. (2004c). “Effect of mulching on the response of 
plantain (cv. Agbagba, Musa spp. AAB-group) to plant parasitic nematodes in southern Nigeria II: 
Reproductive growth and yield” Nigerian Journal of Forestry. 34(2), 61-71. 
Speijer, P. R., Rotimi, M.O. and De Waele, D. (2001), “Plant parasitic nematodes associated with plantain in 
southern Nigeria and their relative importance compared with other abiotic constraints”, Nematologie 3, 
423-436.
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  • 1. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 Effects of Acalypha wilkesiana Leaf Extract, Hot and Boiling Water on Plantain Growth Trend and Soil Nematode Densities M. Omolara Olaniyi1 Department of Crop, Soil and Pest Management, School of Agriculture and Agricultural Technology, Federal University of Technology, Akure, Ondo State, Nigeria The study was funded in part by the International Foundation for Science through research grant number C/3859-1. Olumide Alabi gave technical assistance with data collection. Abstract An experiment was laid out at the Teaching and Research Farm of the Federal University of Technology, Akure to investigate the trend in plantain growth response and nematode population density build up in response to paring, red acalypha leaf extract, hot water and boiling water dip. Above ground plant growth parameters were assessed 55, 84 and 111 days after planting, approximately 8, 12 and 16 weeks respectively. Population densities of plant parasitic nematodes in root rhizosphere at these dates were also investigated. The results indicated that paring enhanced plantain resistance to plant parasitic and edaphic stressors. In a holistic approach, paring and further treating pared suckers in red acalypha leaf extract for 5-15 mins conferred protection against plant parasitic nematodes and liming effect on the soil. This supposed liming effect merits further investigations. However, pre-plant dip of pared suckers in red acalypha leaf extract for 20 mins resulted in stress on the plant, which encouraged high nematode density build up and concomitant parasitism. Keywords: Helicotylenchus multicinctus, Pratylenchus coffeae, Radopholus similis, plantain, red acalypha, hot water treatment, plant extract 1. Introduction Plant parasitic nematodes are major constraints to the production of plantain (Musa spp., AAB-group) in Nigeria. With reference to the, black sigatoka leaf disease and the banana weevils, the nematode species, Pratylenchus coffeae and Radopholus similis are considered the key pathogens causing great damage to plantain in Nigeria (Speijer et al., 2001). P. coffeae is strongly associated with root damage and plant toppling over, which are strong indices of plant parasitic nematode damage. The species has been ranked the number one constraint to plantain production in Nigeria (Olaniyi, 2011). The plant parasitic nematodes species most frequently associated with plantain in Nigeria are Helicotylenchus multicintus, Hoplolaimus pararobustus, Meloidogyne spp., Pratylenchus spp. and Radopholus similis (Rotimi et al., 1999; Speijer et al., 2001). A complex of Radopholus similis, Helicotylenchus dihystera, H. multicinctus, Hoplolaimus pararobustus and Meloidogyne species in South eastern Nigeria cause on average, 50% loss in plantain production in Nigeria depending on the cultivar type and cultural practices employed (Olaniyi, 2011). Pratylenchus coffeae was reported to be important in southwestern Nigeria, yet in spite of its seriousness, it damage potential in Southwestern Nigeria is yet to be quantified and described. These damaging nematodes are introduced to new and otherwise clean fields through infested soil adhering to planting materials and also infected roots of sucker planting materials (Speijer et al., 2001, Rotimi and Opadare, 2006). This makes sanitation an important nematode management principle in plantain. To improve root and rhizome health, sucker planting materials are often pared (i.e. removal of adhering soil and peeling of the skin of the rhizome in order to expose infected tissues, remove where possible before planting or discard materials with serious infection (Blake, 1961). However, farmers do not feel comfortable with this procedure and are slow to adopt it because they often believe that such pared suckers would not survive and if they do would have low productivity. In order for farmers to adopt this cultural practice, the benefits of paring need to be demonstrated. Also in developing effective environmentally friendly interventions, several options are considered. Acalypha wilkesiana (red Acalypha) leaf extract has been demonstrated to be effective against plant parasitic nematodes (Rotimi and Moens, 2005) and documented as effective plantain root health enhancer. In order to develop an effective control tactic for plant parasitic nematodes on plantain an integrated management package should be developed. Therefore, this study investigated the efficacy of combining paring with red Acalypha leaf extract at different exposure duration, hot water or boiling water treatments in the control of plant parasitic nematodes on plantain, cultivar Agbagba, in P. coffeae endemic Southwestern Nigeria. 105 2. Materials and Methods 2.1 Site Description and Field Layout 1 Current Address: Biology Unit, National Open University of Nigeria, 14/16 Ahmadu Bello Way, Victoria Island, Lagos, Lagos State. E-mail: molaniyi@noun.edu.ng
  • 2. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 The trial was conducted at the Teaching and Research Farm (Crop section) of the Federal university of Technology, Akure. Akure lies within the Tropical rainforest belt between latitude 5oN and longitude 150E of the equator, with an annual mean temperature of about 27oC.the dry season is usually witnessed in Akure between November and March, while the rainy season ranged from April to October. The experimental site covered a total area of 960m2. Previously, the site was used for a mulch trial to study the vegetative response of plantain to two organic mulch types. The experiment had been terminated five months earlier and left to re-vegetate naturally before the site was opened for the present study. The trial was arranged in a completely randomized design (CRD) of eight treatments in all and ten replicates per treatment. The site was slashed and burned before marking out and establishing the field. The spacing used was 3 metres between the rows and 2 metres within the rows, there were 10 suckers per row and eighty suckers in all for the trial. 2.2 Preparation of Red Acalypha Leaf Extracts and Sucker Treatment Plantain (cultivar Agbagba) suckers were sourced from within the vicinity of the Teaching and Research Farm of the University (Crop section), while the red Acalypha was sourced from Owena, a suburb of Akure. There were eight treatments in the experiment including the controls. The treatments were: pared control (T1), pared suckers with 5 mins dip in Acalypha extract (T2), pared suckers with 10 mins dip in Acalypha extract (T3), pared suckers with 15 mins dip in Acalypha extract (T4), pared suckers with 20 mins dip in Acalypha extract (T5), Hot water treatment at 52oC for 20 mins (T6), Boiling water treatment at 100oC for 30 sec. (T7) and Non-pared control (T8). Air-dried leaves of red Acalypha plant were pulverized and 100g of the powder was dissolved in 9 litres of cold water in four different buckets each. A wooden rod was used to stir the mixture to ensure homogenization after which they were left to stand for 30 mins. The buckets were respectively labelled as 5, 10, 15 and 20 minutes with a marker to avoid mix-up. Thereafter, 10 suckers were dipped into each of the buckets and they were left to stand for the duration on each bucket’s label. The suckers were removed and left to air dry under shade afterwards. For the hot water treatment, 10 suckers were dipped in water at 52oC ±2oC for 20 mins and they were removed and left to cool for about 24 hours before planting, while for the boiling water treatment, 10 suckers were dipped in water at 100oC for 30 sec. Suckers were planted on 21st December, 2006. Due to cessation of rainfall during this period of the year, manual irrigation of the plant once in two days was adopted until the resumption of rainfall in late March, 2007. 2.3 Above Ground Plant Growth Parameters Growth parameters were taken on the above ground parts of the sprouted plants at 55, 84 and 111 days after planting (DAP), giving approximately 8, 12 and 16 weeks after planting (WAP) respectively. Data taken included the height of the pseudostem of the mother plant from the surface to the point of emergence (axils) of the youngest leaf, the girth of the pseudostem at soil level, number of functional (green) and non-functional (dry) leaves. A leaf was considered functional when at least 75% of the leaf area is green and non-functional if otherwise (Rotimi et al., 2004b). The length and the width of the youngest leaf were measured: the length of the youngest leaf opened was taken from the stalk to the leaf apex while the width was taken from the widest portion of the leaf. The leaf area was calculated as length x width x 0.83 (a constant) according to Obiefuna and Ndubizu (1979). 2.4 Nematode Extraction and Identification Soil samples were taken from the rhizosphere of each plant at 8, 12 and 16 WAP. One hundred ml of each sample was measured unto a serviette placed in a plastic sieve. The sieve was then placed in a plastic plate and water was carefully added to the plate until the soil on the serviette-covered sieve appeared moist on top (modified Baermann tray). The setup was left to stand for 24 hours after which the resulting suspension was decanted, left to stand for 4 hours afterwards and then reduced to 30ml with a syringe. Plant parasitic nematodes were identified to species level with the light microscope and all developmental stages of the nematode species were counted, except for the root knot nematode which was identified only to genus level and only vermiform juveniles and males that could be extracted with the extraction technique were counted. The total of all the developmental stages counted was presented in each case. Densities of the nematodes were estimated and presented per litre of soil. 2.5 Soil Chemical Analysis At time of field establishment, soil samples were taken from each replicate hole and bulked per treatment for physico-chemical analysis. Soil samples were also taken at termination of the experiment, 16 WAP. Soil chemical analysis was carried out in the soil Analytical laboratory of the Department of Crop, Soil and Pest 106
  • 3. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 Management of the Federal University of Technology, Akure. Soil properties determined included pH, Nitrogen, cations such as Ca2+, Mg2+ and K+ content. Others were Phosphorus, Sodium, organic matter, and organic carbon. 3. Results 3.1 Effects of Treatments on Plantain Growth Pattern across the Sampling Dates The effect of the treatments on plant height relative to time is presented in Figure 1(a - h) the R2 value presented on each graph is the coefficient of determination of each regression equation. The 5 mins dip in red acalypha leaf extract (Figure 1b), 10 mins dip in red acalypha leaf extract (Figure 1c) and 30 sec dip in boiling water (Figure 1g) treatments exhibited linear trend in growth response over time. A quadratic polynomial functional trend in height increase was observed with the 20 minutes dip in Acalypha extract treatment over time (T5), hot water treatment and non-pared control which is the farmer’s method followed (Figure 1e, f, h). Only the pared (Figure 1a) and the 15 mins dip in Acalypha extract (Figure 1d) displayed a cubic polynomial trend in height increase over time. Pseudostem thickness increased at a linear rate over time only for the pared treatment (Figure 2a) while for the 5 mins dip in red acalypha leaf extract, pseudostem thickness over time responded with a quadratic polynomial function (Figure 2b). All other treatments (Figure 2c - h) displayed a cubic polynomial trend in increase in thickness over time. In the number of functional leaves produced, the pared treatment followed a cubic polynomial function over time (Figure 3a) while boiling water dip (Figure 3g) and the farmer’s material (Figure 3h) displayed a quadratic polynomial trend from 55 to 111 days after planting. The number of green (functional) leaves produced in the 5, 10, 15 and 20 minutes dip in Acalypha extract treatments along side the hot water treatment exhibited a linear response over the sampling dates (Figure 3b –f). Ten, 15 and 20 min dip in red acalypha leaf extract as well as 30 sec dip in boiling water resulted in linear response in leaf area expansion from 55 to 111 days after planting (Figure 4 c, d, e and g) while expansion trend in leaf area in all the other treatments can be explained by the quadratic polynomial function. 107
  • 4. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 25 20 15 10 5 a b 30 25 20 15 10 5 c d 25 20 15 10 5 e f 35 30 25 20 15 10 5 g h 18 16 14 12 10 8 6 4 2 30 25 20 15 10 5 30 25 20 15 10 5 20 15 10 5 Figure 1. Effects of red Acalypha extract, hot and boiling water treatments on height response of plantain from 55 to 111 days after planting. 108 Pared control y = 7E-05x3 - 0.0155x2 + 1.0978x - 7.8614 R2 = 0.7274 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 5 minutes dip in Acalypha extract y = 0.0747x + 12.09 R2 = 0.9329 0 0 20 40 60 80 100 120 days after planting (days) Height (cm) 10 minutes dip in Acalypha extract y = 0.1942x + 3.9017 R2 = 0.9773 0 0 20 40 60 80 100 120 days after planting (days) 15 minutes dip in Acalypha extract y = -8E-05x3 + 0.0208x2 - 1.6214x + 64.412 R2 = 0.2923 0 0 20 40 60 80 100 120 days after planting (days) Height (cm) 20 minutes dip in Acalypha extract y = 0.003x2 - 0.3532x + 27.632 R2 = 0.7396 0 0 20 40 60 80 100 120 days after planting (days) Height (cm) Hot-water treatment y = -0.0024x2 + 0.4299x + 1.4399 R2 = 0.6771 0 0 20 40 60 80 100 120 days after planting (days) Height (cm) Boiling-water treatment y = 0.0632x + 11.097 R2 = 0.7666 0 0 20 40 60 80 100 120 days af t er plant ing (days) Non-pared control y = -0.0042x2 + 0.5441x + 10.251 R2 = 0.6654 0 0 20 40 60 80 100 120 days af t er plant ing ( days)
  • 5. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 109 10 8 6 4 2 a b 12 10 8 6 4 2 c d 12 10 8 6 4 2 e f 10 8 6 4 2 g h. 10 8 6 4 2 14 12 10 8 6 4 2 10 9.5 9 8.5 15 10 5 Figure 2. Effects of red Acalypha extract, hot and boiling water treatments on pseudostem girth of plantain response from 55 to 111 days after planting. Pared control y = -0.0442x + 10.391 R2 = 0.945 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 5 minutes dip in Acalypha extract y = 0.0007x2 - 0.1269x + 13.265 R2 = 0.6359 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 10 minutes dip in Acalypha extract y = 2E-05x3 - 0.0052x2 + 0.3831x - 0.9432 R2 = 0.4287 0 0 20 40 60 80 100 120 days after planting (days) Girth (cm) 15 minutes dip in Acalypha extract y = -9E-05x3 + 0.0233x2 - 1.8922x + 58.767 R2 = 0.6689 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 20 minutes dip in Acalypha extract y = -9E-05x3 + 0.0229x2 - 1.7801x + 51.469 R2 = 0.3556 0 0 20 40 60 80 100 120 days af t er pl ant i ng ( days) Hot-water treatment y = 3E-05x3 - 0.006x2 + 0.4357x - 0.9446 R2 = 0.4061 8 0 20 40 60 80 100 120 days af t er plant ing ( days) Boiling-water treatment y = -4E-05x3 + 0.0108x2 - 0.8369x + 28.418 R2 = 0.8299 0 0 20 40 60 80 100 120 days after planting (days) Girth (cm) Non-pared control y = -0.0016x2 + 0.1399x + 10.737 R2 = 0.9049 0 0 20 40 60 80 100 120 days after planting (days) Girth (cm)
  • 6. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 110 4.5 4 3.5 3 2.5 2 1.5 1 0.5 a b 8 7 6 5 4 3 2 1 c d 6 5 4 3 2 1 e f 6 5 4 3 2 1 g h 7 6 5 4 3 2 1 8 7 6 5 4 3 2 1 6 5 4 3 2 1 4.5 4 3.5 3 2.5 2 1.5 1 0.5 Figure 3. Effects of red Acalypha extract, hot and boiling water treatments on number of green leaves produced by the plant from 55 to 111 days after planting. Pared control y = -0.001x2 + 0.1532x - 1.972 R2 = 0.887 0 0 20 40 60 80 100 120 days af t er plant ing ( d ays) 5 minutes dip in Acalypha extract y = 0.0511x + 0.04 R2 = 0.8296 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 10 minutes dip in Acalypha extract y = 0.0704x - 1.0638 R2 = 0.8859 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 15 minutes dip in Acalypha extract y = 0.069x - 0.9813 R2 = 0.9529 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 20 minutes dip in Acalypha extract y = 0.0597x - 1.0026 R2 = 0.9944 0 0 20 40 60 80 100 120 days af t er plant ing ( days) Hot-water treatment y = 0.0545x - 0.4345 R2 = 0.9548 0 0 20 40 60 80 100 120 days af t er plant ing ( days) Boiling-water treatment y = 0.0006x2 - 0.0677x + 5.6733 R2 = 0.5818 0 0 20 40 60 80 100 120 days after planting (days) no. of green leaves Non-pared control y = -0.0013x2 + 0.2194x - 5.3017 R2 = 0.9004 0 0 20 40 60 80 100 120 days af t er plant ing ( days)
  • 7. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 leaf area (cm2) leaf area (cm2) 111 Pared control y = -0.0835x2 + 12.06x - 190.27 R2 = 0.8809 300 250 200 150 100 50 0 0 20 40 60 80 100 120 days af t er plant ing ( days) a b 10 minutes dip in Acalypha extract y = 4.6715x - 80.986 R2 = 0.8613 500 400 300 200 100 0 0 20 40 60 80 100 120 days after planting (days) leaf area (cm2) c d 20 minutes dip in Acalypha extract y = 3.5363x + 16.008 R2 = 0.74 500 400 300 200 100 0 0 20 40 60 80 100 120 days after planting (days) leaf area (cm2) e f Boiling-water treatment y = 1.6598x + 143.12 R2 = 0.633 400 300 200 100 0 0 20 40 60 80 100 120 days after planting (days) leaf area (cm2) g h 5 minutes dip in Acalypha extract y = -0.0955x2 + 15.076x - 276.73 R2 = 0.7995 350 300 250 200 150 100 50 0 0 20 40 60 80 100 120 days af t er plant ing ( days) 15 minutes dip in Acalypha extract y = 1.697x + 333.63 R2 = 0.5318 600 500 400 300 200 100 0 0 20 40 60 80 100 120 days af t er plant ing ( days) Hot-water treatment y = -0.1562x2 + 27.117x - 768.53 R2 = 0.7773 500 400 300 200 100 0 0 20 40 60 80 100 120 days after planting (days) Non-pared control y = -0.3791x2 + 57.51x - 1560.6 R2 = 0.7745 800 600 400 200 0 0 20 40 60 80 100 120 days after planting (days) Figure 4. Effects of Acalypha wilkesiana leaf extract, hot and boiling water treatments on the leaf area growth trend of plantain from 55 to 111 days after planting. 3.2 Effects of Treatments on Plant Parasitic Nematode Species and Densities Recovered Three species of plant parasitic nematodes namely: Helicotylenchus multicinctus, Pratylenchus coffeae and Radopholus similis were recovered from plantain rhizosphere in this study. At 55 days (8 weeks) after planting, only the 5 minutes dip in extract (T2) had a combination of two species of nematodes namely Radopholus similis
  • 8. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 and Pratylenchus coffeae while P. coffeae was recovered under plantain in the other treatments except 20 mins dip in extract and the not pared control (Figure 5a), at 84 days (12 weeks) after planting, the highest population density and species combination was observed in the 20 minutes dip in extract (T5) (Figure 5b) while at 111 days (16 weeks), the nematode population recovered from the roots had reduced relatively to the earlier population densities observed in preceding samplings (Figure 5c). density/litre soil 112 nematode from soil 55 DAP 700 600 500 400 300 200 100 0 A A a A a a A a A A a a b a T1 T2 T3 T4 T5 T6 T7 T8 Treatments density/litre soil R.s Praty a a b c nematode from soil 84 DAP 700 600 500 400 300 200 100 0 A a a a1 a1 a A T1 T2 T3 T4 T5 T6 T7 T8 Treatments R.s Praty H.m nematode from soil 111 DAP 1000 800 600 400 200 0 A a a H.m A T1 T2 T3 T4 T5 T6 T7 T8 Treatments density/litre soil R.s Praty Figure 5. Effects of treatments on rhizosphere nematode densities at 55 (a), 84 (b) and 111 (c) days after planting (DAP). T1 = Pared control (no dip treatment), T2 = 5 mins dip in Acalypha extract, T3: 10 mins dip in Acalypha extract, T4 = 15 mins dip in Acalypha extract, T5 = 20 mins dip in Acalypha extract, T6 = Hot-water treatment for 20 mins, T7 = Boiling-water treatment for 30 sec, T8 = Non-pared control. Column fractions followed by same letters (and numbers) are not significantly different at P≤0.05. 3.3 Effects of Treatments on Soil Chemical Properties Chemical properties of the soil at pre-plant are presented on table 1 while table 2 presents the soil chemical properties 111 days after planting. Ten and 15 mins dip in Acalypha leaf extract, and 30 sec dip in boiling water depressed soil pH, while 5 and 20 mins dip in red Acalypha extract as well as 20 mins dip in hot water resulted in slightly raised pH. The non-pared control, which represented the farmer’s method, also resulted in depressed soil pH, while the pared suckers gave relatively constant pH. Generally, soil potassium content declined between planting and termination of the experiment 111 days afterwards. Except for the 15 mins dip in Acalypha extract treatments, soil phosphorus also declined from planting till end of experiment. Nitrogen content also declined marginally from planting to termination of the experiment; except for the 20 mins hot water dip and 30 mins boiling water dip treatments.
  • 9. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 Table 1. Pre-plant soil chemical properties in plots where the treatments shown were laid out T1 = Pared control (no dip treatment), T2 = 5 mins dip in Acalypha extract, T3: 10 mins dip in Acalypha extract, T4 = 15 mins dip in Acalypha extract, T5 = 20 mins dip in Acalypha extract, T6 = Hot-water treatment for 20 mins, T7 = Boiling-water treatment for 30 sec, T8 = Non-pared control. Treatment pH Mg Ca %OM %OC P Na K N T1 5.87 2.30 2.90 11.31 6.55 1.42 7.13 1.13 0.31 T2 5.59 1.10 3.50 6.49 3.75 1.41 12.3 0.41 0.38 T3 5.73 0.70 3.40 10.02 5.72 1.10 13.3 0.56 0.21 T4 5.79 1.30 3.10 8.62 4.99 0.96 11.39 1.24 0.20 T5 5.45 1.10 2.00 9.93 5.75 0.88 10.52 0.87 0.32 T6 5.46 1.10 2.40 13.25 7.66 1.43 12.35 0.46 0.17 T7 6.08 0.80 2.90 9.87 5.71 2.41 10.96 0.36 0.27 T8 5.68 1.80 2.90 10.97 6.34 1.29 13.04 0.36 0.18 Table 2. Effects of paring, red acalypha leaf extract , hot and boiling water treatments of plantain sucker planting materials on soil chemical properties 16 weeks after planting T1 = Pared control (no dip treatment), T2 = 5 mins dip in Acalypha extract, T3: 10 mins dip in Acalypha extract, T4 = 15 mins dip in Acalypha extract, T5 = 20 mins dip in Acalypha extract, T6 = Hot-water treatment for 20 mins, T7 = Boiling-water treatment for 30 sec, T8 = Non-pared control. Treatment pH Mg Ca %OM %OC P Na K N T1 5.86 1.10 3.80 11.18 6.46 0.94 11.39 0.56 0.28 T2 5.94 0.10 3.80 11.31 6.54 0.83 14.43 0.87 0.31 T3 5.24 0.50 3.80 9.73 5.63 0.59 11.83 0.41 0.15 T4 5.60 1.70 2.70 10.69 6.19 0.90 15.13 0.36 0.18 T5 5.84 2.00 2.40 11.24 6.50 1.05 11.74 0.42 0.18 T6 5.52 0.70 2.50 9.38 5.43 1.24 12.17 0.29 0.31 T7 5.56 1.60 2.80 9.31 5.39 0.88 11.83 0.25 0.27 T8 5.55 0.50 2.80 9.31 5.31 1.04 10.26 0.27 0.13 4. Discussion The linear response obtained for some of the growth parameters for 5, 10, 15 and 20 minutes dip in acalypha extract, hot and boiling water treatments in this study connotes that the rate of change in the growth of the plant in the treatments increased over time at a constant rate in spite of the prevailing harsh dry condition. Polynomial functional relationships obtained in the non-pared control (Farmer’s method) might be an indication of the high susceptibility of unpared planting materials to dry weather. This then suggests that paring the sucker before planting enhanced the plant’s ability to withstand biotic and edaphic stress. Further treating the planting materials with plant extract or hot water (52 ±2oC) also conferred some level of resistance to these stress factors. Olaniyi (2014b, In Press) also reported functional growth trend in time for plantain in reaction to organic mulch. The author noted that organic mulch enhanced the plants ability to withstand biotic stressors like plant parasitic nematodes in the environment. The effects of treatments in the present study on growth parameters such as the leaf area, number of functional leaves, plant height and girth revealed that 15 minutes dip of planting materials in the extract showed some promise in enhancing plant growth performance. However, it is not clear what this additional benefit of dipping 113
  • 10. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 pared suckers translates to in maintaining low nematode populations since the suckers that were only pared had very low densities of P. coffeae (about 20/litre soil) and maintained no detectable level of nematodes subsequently. Where as the highest density of P. coffeae recorded in this study was from the 15 mins dip in extract, earlier report (Olaniyi, 2014a, In Press) showed that 15 min dip reduced nematode damage to root and rhizome of plantain and also reduced nematode densities. In the present study 5 and 10 mins dip treatment seemed to better reduce nematode population densities on plantain compared to the 15 mins dip. Generally, Helicotylenchus multicinctus, Pratylenchus coffeae and Radopholus similis were the nematode species observed from plantain roots and the rhizosphere in this study. There is need to monitor plant response to these treatments and nematode development on plantain under these treatments till yield stage. Although densities of the species in the rhizosphere were low with maximum of 100 H. multicinctus / litre, 600 P. coffeae / litre and 200 R. similis / litre, it is clear that densities had increased from the initial pre-plant densities of 1 P. coffeae / litre soil and 2 H. multicinctus / litre soil. This shows that planting a host plant in a soil would quickly increase the densities of the nematode species to a damaging level. It would be informative to investigate the effect of these densities on root and rhizome damage of plantain. Before the choice of a plant to put in a soil is made, it would be helpful to first do a diagnostic nematode assessment in order not to put in a host crop in a field where a damaging species is prevalent. For instance, Speijer et al. (2001) established Pc as the most damaging on plantain in Nigeria and the fact that it could persist in a field without the plantain host makes its control a big challenge as this might signify that it has a wide host range. The trend in population densities recovered from plantain rhizosphere is at variance with earlier record of H. multicinctus often in higher densities than other species on plantain in Nigeria (Speijer et al., 2001; Rotimi et al., 2004b, c; Coyne et al., 2005). Radopholus similis was not recovered from the soil before the establishment of this trial but was recovered from the rhizosphere of plantain 55 days (8 weeks) after planting in 5 min dip in red acalypha leaf extract treatment but subsequently, it was undetected in soil samples. This shows that first R. similis may not have as wide host range as P. coffeae or probably it could not persist as long as P. coffeae in the absence of a preferred host hence the absence of the species in the soil pre plant. The land had been fallowed for about 5 months before it was opened up for this study. However, the presence and disappearance of the species at subsequent sampling date supported Olaniyi’s (2011) submission that for accurate diagnosis, nematode sampling should be done across seasons. The study reported here was done in the dry season and Rotimi et al. (2004a) similarly observed that the densities of R. similis could decline to an undetected level in dry soil. No specific trend emerged in plant parasitic nematode community structure under plantain in this study. This suggests that several factors confound field studies and it would be useful to investigate nematode reactions to these treatments in controlled environment. For instance in the unpared control treatment, no nematode was detected in plantain rhizosphere 8 and 16 weeks after planting whereas at 12 weeks after planting P. coffeae was detected at an average of 400 nematodes / litre soil. However under the pared treatment, only very low density of Pc was recovered 8 weeks after planting. Subsequently no nematode was detected implying that it may suffice for nematode control to simply pare plantain sucker planting materials before planting without any further treatment. Olaniyi (2014a, In Press) also did not recover any plant parasitic nematode species from the roots of pared plantain at those dates. A number of factors could be responsible for the variations observed in the nematode incidence across the sampling dates. The variations obtained in the result across the three samplings established the rationale and the importance of sampling more than once in the diagnostic study of plant parasitic nematodes so that correct assertion can finally be made (Olaniyi, 2011, 2014a, In Press). It is a common phenomenon in nematode population dynamics, and that is why it is advisable to sample several times in time and season. Also, sampling only the rhizosphere could give misleading information as the species recorded in this study are migratory endoparasites and several factors could be responsible for their density fluctuation in the rhizosphere. To confirm this, the study reported by Olaniyi (2014a, In Press) investigated root densities and a different pattern emerged. Therefore, for a more holistic overview, it is important to sample both root and soil complementarily as this would give more reliable information and better guide management decision. Other factors that may be responsible for variations include the soil condition, environmental condition and availability of moisture in the soil. As expected, N, P & K values declined over time in this study. Results of this study revealed that introducing soil from other sources with planting materials as with the farmer’s method of not paring may result in increased soil acidity. But this would be dependent on the chemical status of the introduced soil. This is revealed by the stable pH of soils from pared suckers, which had all adhering soils removed before planting. Dipping rhizomes of planting materials in red Acalypha leaf extract for 5 or 20 minutes tended to reduce soil acidity. The liming effect of red Acalypha should be further explored. Considering the overall effect of treatments on plant response and nematode density, it would be safe to recommend paring as a useful cultural practice that farmers should be 114
  • 11. Journal of Biology, Agriculture and Healthcare www.iiste.org ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.4, No.18, 2014 encouraged to engage in. Further treating pared suckers in red acalypha extract for 5-15 mins might confer protection against plant parasitic nematode damage (Olaniyi, 2014a, In Press) and improve soil quality. References Blake, C.D. (1961), “Root rot of bananas caused by Radopholus similis (Cobb) and its control in New South 115 Wales”. Nematologica 6, 295-310. Coyne, D.L., Rotimi, M.O., Speijer, P., De Schutter, B., Dubois, T., Auwerkerken, A., Tenkouano, A. and De Waele, D. (2005), “Effects of nematode infection and mulching on the yield of plantain (Musa spp., AAB-group) ratoon crops and plantation longevity in southeastern Nigeria”. Nematology 7(4), 531-541. Obiefuna, J.C. and Ndubizu., T.O.C. (1979), “Estimating leaf area in plantain”. Scientia Horticulturae 11, 31-36. Olaniyi, M.O. (2006), “Effect of organic mulch and root dips on the establishment and root health of plantain”. Xvill Reunia International Acorbat Anais Proceedings Memorias, 321. Olaniyi, M.O. (2011), “Plant Parasitic Nematode Constraint to Plantain production in Nigeria”. LAP Lambert Publishing, Germany. September 2011. ISBN 978-3-8454-2312-8, 240pp. Olaniyi, M.O. (2014a), “Effects of Acalypha wilkesiana leaf extract, hot and boiling water on plantain growth response and nematode damage”. Nigerian Journal of Nematology, In Press. Olaniyi, M.O. (2014b), “Functional trend in leaf and sucker production of plantain in response to paring and organic mulch”. The International Journal of Science and Technoledge, In Press. Olaniyi, M.O. and Opadare, S.O. (2006), “Effect of paring on the establishment and root damage of French plantain suckers”. In: E. Soprano, F.A. Tcacenco, L.A. Litchemberg & M.C. Silva (Eds.). Banana: A Sustainable Business. XVII ACORBAT International Meeting, Joinville, Brazil. 15-20 October 2006, 345. Rotimi, M.O. and Moens, M. (2005), “Effect of leaf extracts of some herbs on the juveniles of Meloidogyne incognita (Kofoid & White) Chitwood”. Nigerian Journal of Plant Protection 22, 95- 103. Rotimi, M.O., Speijer, P.R. and De Waele D. (2004a), “Effect of soil moisture and organic matter levels on plantain root reaction to Radopholus similis”. International Journal of Nematology 14(2), 122-128. Rotimi, M.O., Speijer, P., De Waele, D. and Swennen, R. (2004b), “Effect of mulching on the response of plantain (cv. Agbagba, Musa spp. AAB-group) to plant parasitic nematodes in Southeastern Nigeria. I: Vegetative growth”. Nigerian Journal of Forestry. 34 (1), 61-71. Rotimi, M.O., Speijer, P., De Waele, D. and Swennen, R. (2004c). “Effect of mulching on the response of plantain (cv. Agbagba, Musa spp. AAB-group) to plant parasitic nematodes in southern Nigeria II: Reproductive growth and yield” Nigerian Journal of Forestry. 34(2), 61-71. Speijer, P. R., Rotimi, M.O. and De Waele, D. (2001), “Plant parasitic nematodes associated with plantain in southern Nigeria and their relative importance compared with other abiotic constraints”, Nematologie 3, 423-436.
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