- Email: [email protected]
Effectiveness of biopesticides in enhancing paddy growth for yield improvement
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
Contents lists available at ScienceDirect
Sustainable Chemistry and Pharmacy journal homepage: www.elsevier.com/locate/scp
Effectiveness of biopesticides in enhancing paddy growth for yield improvement
T
⁎
Noor Hafizah Ramli@Yusofa, Suzana Yusupa, , Benjamin Wei Bin Kueha, Puteri Sarah Diba Kamarulzamana, Noridah Osmana, Mardyahwati Abd. Rahimb, Ramlan Azizb, Sulaiman Mokhtarc, Abu Bakar Ahmadc a Biomass Processing Lab, Centre of Biofuel and Biochemical Research, Mission Oriented Research (Green Technology), Chemical Engineering Department, Universiti Teknologi PETRONAS, 32610 Seri Iskandar, Perak Darul Ridzuan, Malaysia b Plant Biosecurity Unit, Department of Agriculture of Perak Tengah, Simpang 3, 34200 Parit Buntar, Perak Darul Ridzuan, Malaysia c Bio-X Techno Sdn. Bhd., LK1-1-3A, Blok LK1, Laman Komersil ARECA, Putra Nilai, 71800 Nilai, Negeri Sembilan Darul Khusus, Malaysia
A R T I C L E I N F O
A B S T R A C T
Keywords: Biopesticides Paddy growth Yield Paddy cultivation
There is an urgent need to enhance the productivity of irrigated paddy fields to increase the rice production that meets population growth. The focus of this study is to investigate the effectiveness of biopesticides in enhancing the paddy growth for yield improvement after controlling the pest and rice diseases. This study aims to determine either one or two types of biopesticides should be used in paddy cultivation. Thus, three conditions were adopted in paddy cultivation at the insect house as follows; T1 where the paddy was sprayed using BV500WS and BV612EC to study the effectiveness of the biopesticides, T2 where the paddy was sprayed using BV500WS only and C1 where the paddy was cultivated following the conventional technique as a control. BV500WS was sprayed into T1 and T2 on the first day of paddy growth while BV612EC was sprayed later into T1, at 55 and 67 days of paddy cultivation a week after releasing the brown planthopper (BPH). Based on the analysis of correlation obtained for each growth, application of BV500WS and BV612EC of T1 resulted in better performance in terms of height of plant, number of tillers, width of leaves, and number of leaves compared to T2 and C1. The excellence performance of plant growth at T1 produced the highest paddy yield of 2304.20 ± 491.42 g−2, while T2 produced comparable yield as C1 which are 663.88 ± 128.41 g−2 and 665.42 ± 183.30 g−2 respectively. Hence, organic plants based biopesticides are successful in enhancing the plant growth of paddy and ultimately increase the paddy yield.
1. Introduction Malaysia warm and humid climate attracts a number of pests and rice diseases that severely reduce the productivity and yield of paddy fields. Approximately over 800 herbivore insect species inhabit the ecosystem. Thus, there is an urgent need to enhance the productivity of irrigated paddy fields in order to increase the production of rice to meet the population growth (Prabhu, 2010). In addition, the brown planthopper (BPH) also causes serious damage to the rice by sucking rice sap, ovipositing in the rice tissues and transmitting rice diseases, especially during tillering stage thus causing less production (Zakaria et al., 2016; Xiaolei Zhang et al., 2016). Rice diseases like bacterial blight (BB), leaf blast, brown spot and sheath blight frequently affected the paddy growth and yield. BB, caused by Xanthomonas oryzaepv. oryzae (Xoo), caused yield reductions of 20–50% at the tiller stage in
⁎
severe cases; while rice blast, caused by the fungal pathogen Magnaporthe oryzae, is responsible for large yield losses (Ni et al., 2015; Kim et al., 2015; Ji Zhi-juan et al., 2016). Besides, there is a growing interest worldwide in alternative agricultural practices that can help farmers reduce their dependence on synthetic fertilizers and pesticides. The excessive use and misuse of the synthetic chemicals have raised serious questions regarding their longterm effects on human health, soil quality, and the environment (Ephraums, 1990; Samy et al., 1995). For the improvement of grain yield, the knowledge of the association between grain yield and its component characters such as number of effective tillers per plant, plant height, and number of grains per panicle will be helpful (Lakshmi et al., 2014). The relative contribution of different traits on yield determination can be explained by correlation studies. The existence of correlation is attributed to the
Corresponding author. E-mail address: [email protected] (S. Yusup).
https://doi.org/10.1016/j.scp.2017.11.002 Received 19 August 2017; Received in revised form 15 November 2017; Accepted 16 November 2017 2352-5541/ © 2017 Elsevier B.V. All rights reserved.
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Only the growing seeds were selected to be planted in the pots. The seeds were sowed in the pots by adopting the paddy transplantation cultivation. The BV500WS solution was diluted in the remaining water with ratio of 1:2500 and was sprayed onto the T1 and T2. While the C1 were sprayed with the balance purified water from the curing process. The BV612EC was applied onto T1 only during 55 and 67 days of cultivation which is a week after the BPH population was released onto the paddy plants of T1 and T2. The BPH were not released to C1 as to maintain the standard cultivation and to study how the BPH infest the paddy naturally.
environmental effect or genetic effect of genes such as grain yield in rice, is significantly correlated with total tillers, flag leaf length, flag leaf width, number and grain weight (Ranawake et al., 2014). Biopesticides are employed in agricultural uses to control the insect, disease, nematode and weed. They also can improve plant physiology and productivity. As biopesticides commonly are biodegradable, they are able to decompose quickly and do not negatively impact surface water and groundwater (Alliance, 2015). The BV500WS, which has high azadirachtin (A + B) content with proprietary water soluble concentrates is formulated for root feeding, drip and foliar for crop protection and growth enhancement (Okada Ecotech, 2014). Santos, M.S. also reported that azadiracthin which can be found abundantly in neem has high insecticide and rapid degradation in the environment (Santos et al., 2015). Besides that, BV612EC is a series of environmental friendly insecticidal formulations based on a unique and extremely safe active ingredient known as Etofenprox. It has high insecticidal activities which provide a very quick killing action against a broad spectrum of crop insect pests including lepidoptera, hemiptera, coleopteran, diptera and orthoptera, and yet has remarkably weak toxicity to mammals (Okada Ecotech, 2014). Moreover, Etofenprox was reported to be highly toxic to the insect including BPH but relatively safe to non-target organisms (Xiaolei Zhang et al., 2016; Trisyono et al., 2017). The main focus of the study is to determine either one or two types of biopesticides should be used in paddy cultivation to control the pest and reduce the rice diseases for enhancing the paddy growth for yield improvement. This research also aimed to study the efficacy of new biopesticides in playing their roles as crop protection and growth enhancement for BV500WS and as insecticide for BV612EC in paddy cultivation. The correlation study on paddy growth and development were carried out at every stage of cultivation by focusing on plant height, number of tillers, number of leaves, and width of leaves. The efficacy of biopesticides was evaluated by analyzing the yield components using analysis of variance (ANOVA) during the harvesting stage.
2.2.2. Paddy growth development The plant growths were observed in every cultivation stage. The observation was focused on plant diseases, color of leaves, weeds and pest problems. In addition, the correlation study of paddy growth was also conducted by taking record on the plant height, number of tillers, number of leaves, and width of leaves of all samples. The stages of paddy cultivation are as per stated below (Malaysia and Padi, 2008):
• Stage 1: Plantation Day 1–5 days) • Stage 2: Tillering Stage 15–40 days) • Stage 3: Panicle Initiation Stage 40–69 days) • Stage 4: Heading Stage 70–89 days) • Stage 5: Ripening Stage 90–104 days) • Stage 6: Harvest Stage 105–112 days) 2.3. One way ANOVA analysis One way ANOVA is used to determine whether there are any statistically significant differences between T1, T2 and C1. The data were transformed using various algorithms so that the frequency distributions are normally distributed (Ltd, 2013). The mean with standard error, the minimum and maximum values, and the standard deviation of all recorded data were calculated and treated using ANOVA. The degree of freedom, the F-value and P-value were obtained from the analysis.
2. Material and method 2.1. Experimental site
2.3.1. Yield component calculation The experimental site was located at Plant Biosecurity Unit of Agriculture Department, Titi Serong, Parit Buntar, Perak, Malaysia. 2.2. Insect house experiment BV500WS and BV612EC were obtained from Bio-X Techno Sdn. Bhd. as a part of collaborative research. The application of BV 500WS is for crop protection and growth enhancement at the early stage of cultivation while BV612EC as insecticide to protect the paddy during pest infestation (Okada Ecotech, 2014). Three conditions were adopted in paddy cultivation at the insect house as follows which are T1 where the paddy was sprayed using BV500WS and BV612EC to study the effectiveness of both types of biopesticides, T2 where the paddy was sprayed using BV500WS only and C1 where the paddy was cultivated following the conventional technique as a control. Three different conditions with five replications each were performed in the Insect House. All pots (9 cm diameter) were filled with 15 cm depth of soil collected from paddy field at the Rice of Excellence Centre of Agriculture Department, Titi Serong, Parit Buntar, Perak, Malaysia.
mass of 1000 productive spikelets (g ) mass of 800 productive spikelets = × 1000 800
(1)
acurate mass of 14% moisture content, MC (g) 100−actualMC = × [1] 86
(2)
mass of 1 productive spikelet (g ) =
% Productives spikelets =
(2) 1000
Total productive spikelets × 100% Total spikelets
(3)
(4)
Number of panicle per m2 = Total panicles × area of sampling (5)
Gain yield
2.2.1. Cultivation of paddy The rice variety of MR 219 has been used in this experiment. The seeds for T1 and T2 were immersed in the BV500WS solution with ratio of 1:1000 to the purified water while the seeds for C1 were immersed in purified water only. All the seeds were immersed for 24 h before filtered. All the floated seeds were eliminated. Then, the seeds were dried under room temperature for another 24 h before seedling in the pot. The remaining water was used to spray in the pot during cultivation.
(gm−2) =
Total spikelets × (4) × (3) × (5) Total panicles
(6)
All calculations were performed according to standard protocol from Agriculture Department (Malaysia and Padi, 2008). 3. Results and discussion 3.1. Paddy growth observation The observation of paddy growth is to study the effectiveness of 2
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Paddy plant is in good condition
Paddy plant is in good condition
Paddy plant is in good condition
Brown Leaf Blight, and neck rot and detected
Brown Leaf Blight, neck rot and hole leaves detected
Brown Leaf Blight, neck rot and hole leaves detected
-The affected leaves and tillers increased -The brown planthopper has been released
-The paddy plants become more healthier and the panicles start to develop after spray 612EC. -The population of brown planthopper disappeared.
The paddy plant continuous to growth healthy. The panicles has fully develop and visible but few of the plants has weedy rice .
-The affected leaves and tillers increased -The brown planthopper has been released
The plant attacked by pest and diseases rapidly due to increasing of brown planthopper population.
The plant still suffering the pest attacked and diseases due to the present of brown planthopper. The number of weedy rice higher than T1
The condition of the paddy plants are worst compared to T1 & T2. Pest and diseased attacked the plants
Pest and diseases attacked continue to increasing and the present of weedy rice is the highest compare to T1 and T2
The affected leaves and tillers increased
Fig. 1. Condition of paddy plants at every stage on selected days of cultivation.
3.1.2. Color of leaves Fig. 2 showed that the color of the leaves started to change to yellowish after 42 days of paddy cultivation. From the tillering stage till the end of cultivation, the pest and bacteria began to infest the leaves especially at C1, resulted in increasing of yellowish leaves and directly increased the dead leaves as C1 was not applied with any treatment. The application of BV612EC on 55 and 67 days of cultivation at T1, resulted in yellowish leaves turned back to green and accelerated plant growth as per shown on Day 71 and Day 101.
BV500WS and BV 612EC in terms of physical appearance, weeds and pest problem. There are five major rice diseases which are brown spot, leaf blast, bacterial leaf blight, sheath blight, and tungro that frequently occur in tropical Asia. The diseases are caused by fungal pathogens and bacterial pathogen led in reducing both grain yield and quality (Ji Zhijuan et al., 2016; Kim et al., 2015). During early stage of cultivation, the paddy at T1 and T2 were healthier than C1 as the seeds were immersed and also sprayed using BV500WS to improve photosynthesis and assimilated translocation (Okada Ecotech, 2014). However, after BPH was released at 46 and 59 days of cultivation, at T1 and T2 to create a worst case scenario of infestation, the paddy started to show the impact of the pests. Even though the BPH at C1 was not released to let the pest infests naturally, C1 had a severe infestation by the BPH. Due to BPH infestation, the yellowish and dead leaves, rice diseases, and the affected tillers increased after 71 days of cultivation. Only T1 showed positive growth since it was sprayed with BV612EC during 55 and 67 days of cultivation for crop protection. The observation in Figs. 1–4 described the physical appearance of paddy and the weed and pest problems on the selected days of paddy cultivation in details.
3.1.3. Weed problems The weed population like yellow bur-head distracted the paddy growth as they competed with paddy for nutrients from water and soil while duckweeds covered the surface water to get enough oxygen as per described in Fig. 3 aon selected days of cultivation. Increasing the weed population caused reduced of nutrients availability for paddy, which subsequently reduced its yield (Awan et al., 2011). However, the biopesticides application showed a positive effect in reducing the weed population. T1 showed decreasing of weed population on 101 days of cultivation while T2, the population of weed remained the same started from 71 days of paddy cultivation as per shown in Fig. 3. Only C1 showed the population of weed increased till the harvesting stage.
3.1.1. Paddy condition The plant diseases such as bacterial leaf blight and leaf blast started to appear after 42 days of paddy cultivation. The grasshopper also ate the leaves. Fig. 1 showed that the pest and bacteria infestation increased the number of affected tillers resulted in reduction of panicles during heading stages. Therefore, T1 produced the highest panicles compared to T2 and C1, due to the application of BV500WS and BV612EC which reduced the rice diseases and pest infestation as per illustrated in the Fig. 1 on 101 days of paddy cultivation.
3.1.4. Pest problems There were three dead paddy plants at C1 during observation during harvesting stage due to bad infestation from pest especially BPH resulted in higher unfilled spikelets and lighter grains compared to T1 and T2. Fig. 4 described that the pest population at T1 started to decrease from 71 days of paddy cultivation due to repeated application of BV612EC contrary with T2 and C1, in which the pest population 3
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Remain green
Remain green
Remain green
The leaves turned yellowish
The yellowish leaves increased due to release brown plant hopper
The leaves turned back to green after sprayed with 612EC
Remain green
Remain green
The yellowish leaves increased due to release brown plant hopper
The yellowish leaves increased due to not sprayed with 612EC
The yellowish leaves increased
The leaves turned yellowish
The yellowish leaves increased due to pest infestation
The yellowish leaves increased
Dead leaves increased
Fig. 2. Color of leaves at every stage on selected days of cultivation. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article).
to only one type of biopesticide. Ji Zhi-juan et al. (2016) studied that the damage caused by BPH feeding has the greatest effect on the growth and crop yield of the susceptible rice plant through the removal of assimilates and the reduction in photosynthetic rate of leaves, ultimately causing plant death in its severe form. BV612EC which has Etofenprox as active ingredient
increased during observation on 71 and 101 days of paddy cultivation. The BPH and golden apple snail affected the tillers, while the insect such as grasshopper and bugs; and bacteria affected the leaves which led to increasing of dead leaves and tillers (Alexander et al., 2014; Ji Zhi-juan et al., 2016). Thus, the combination of the biopesticides application were more effective in reducing the pest infestation compared
Presence of Yellow bur-head (paku rawan)
-Yellow bur-head increased -presence of duckweeds
Weeds increased
Weeds population remain the same
-Duckweeds decreased -Yellow bur-head remains the same
Presence of duckweeds
-Presence of Yellow burhead -duckweeds increased
Weeds increased
Weeds population remain the same
Weeds population remain the same
Presence of duckweeds
-Presence of Yellow burhead -duckweeds increased
Weeds population remain the same
Weeds population remain the same
Fig. 3. Weed problems at every stage on selected days of cultivation.
4
Weeds increased
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
No pest
No pest
No pest
Presence of pest
No pest
Presence of grasshopper
Pest and rice diseases increased
Pest population increased.
Pest population increased.
Pest population decreased rapidly due to sprayed with 612EC
Pest population decreased
Pest population increased rapidly due to not sprayed with 612EC
Pest population increased
Pest population increased rapidly due to not sprayed with 612EC
Pest population increased
Fig. 4. Pest problems at every stage on selected days of cultivation.
infestation resulted in lower performance of plant growth development and reduction in crop yield as pest infestation commonly transmitted virus and diseases to the plants (Xiaolei Zhang et al., 2016). Unlike T1, the positive paddy growth led the paddy to produce healthier panicles compared to T2 and C1 due to the application of two types of biopesticides during cultivation. Based on the observation, it showed that the application of biopesticide, at the early stage of cultivation is not sufficient to prevent the plants from pest infestation. Application of biopesticide needs to also be applied upon infestation during tillering and panicle initiation stages.
3.2. Correlation analysis of paddy growth The correlation analysis of paddy growth in terms of plant height, number of tillers, number of leaves and width of leaves has been recorded during 19, 38, 66, 81 and 101 days on selected days of cultivation to study the effectiveness of one type and both types of biopesticides.
Fig. 5. Plant height (cm) at T1, T2 and C1 on selected days of cultivation.
3.2.1. Plant height Fig. 5 showed that all the three types of conditions have positive increment on plant height. T2 showed higher plant height at 38 days of cultivation with 86.21 ± 3.04 cm followed by T1 with 73.59 ± 2.36 cm and C1 with 45.52 ± 1.67 cm respectively. It showed that BV500WS was successful in accelerated the paddy growth of T1 and T2 at early stage of cultivation. However, releasing the BPH at 46 and 59 days of cultivation reduced the increment rate of plant height at T2 and C1 due to severity of pest infestation. Only the plant height at T1 increased rapidly achieved the highest plant height with 113.55 ± 10.05 cm on 101 day of cultivation as were sprayed with BV612EC during 55 and 67 days of cultivation to prevent the plant from infestation. Ranawake et al. (2014) and Kamarulzaman et al. (2016) reported that plant height had a positive relationship with the grain yield. Thus, higher plant height will lead to have a higher grain yield. Hence, the application of two types of biopesticides led to better performance in plant height compared to only one type of biopesticide.
Fig. 6. Number of tillers at T1, T2 and C1 on selected days of cultivation.
was successfully reduced the BPH population and rice diseases. Xiaolei Zhang et al. (2016) and Trisyono et al. (2017) also reported similar observation using Etofenprox to control the BPH in paddy. The results showed it was effective in reducing the BPH population and relatively safe to its natural enemies. C1 and T2 which had the severity of pest 5
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
of cultivation after BPH was released. The pest affected the tillers and directly increased the number of dead tillers towards the end of the cultivation. However, decreasing in the number of tillers can increase the paddy yield as the unproductive tillers will compete for resources. Thus, eliminating the unproductive tillers will allow more solar energy and mineral nutrients for the growth of productive tillers (Peng et al., 2014). T1 had highest remaining tillers with average 18 tillers per pot during observation on 101 days of cultivation compared with C1 with average 14 remaining tillers per pot and T2 with average 6 remaining tillers per pot. Even though the number of tillers started to decrease on 81 days of cultivation due to affected by pest infestation at T1, T2, and C1, by applying BV612EC twice at T1, it accelerated the growth of productive tillers which resulted in healthier panicles and higher yield. Fig. 7. Number of leaves at T1, T2 and C1on selected days of cultivation.
3.2.3. Number of leaves Plant leaves are important organs which have an active role in photosynthesis and yield contribution (Morteza Siavoshi et al., 2013). Fig. 7 showed that T1 has the highest number of leaves with average 94 leaves per pot as T1 was less affected by pest infestation and rice diseases compared to T2 with average 48 leaves per pot and C1 with average 46 leaves per pot from day 66 to day 101 of paddy cultivation. Thus, the application of two types of biopesticide was more effective in increasing the number of leaves and directly led to higher yield. 3.2.4. Width of leaves The available nutrients and crop protection might have helped in enhancing the leaf area, which thereby resulted in higher photo-assimilates and more dry matter accumulation (Morteza Siavoshi et al., 2013). The application of BV500WS and BV612EC enhanced the plant growth and led T1 with highest width of leaves with 2.15 ± 0.12 cm followed by T2 with 1.81 ± 0.10 cm and C1 with 1.21 ± 0.64 cm respectively as shown in Fig. 8.
Fig. 8. Width of leaves at T1, T2 and C1on selected days of cultivation.
3.2.2. Number of tillers Tillering is an important trait for grain production and is thereby an important aspect in rice yield (Morteza Siavoshi et al., 2013). BV500WS which is neem based biopesticide influenced the paddy in increasing the number of tillers at early stage of cultivation as a part of growth enhancement (Okada Ecotech, 2014). This is because organic sources offer more balanced nutrition to the plants especially micronutrients which, positively affects the number of tillers in plant (Morteza Siavoshi et al., 2013). Fig. 6 showed the reduction of tillers starting from 66 days
3.3. One way ANOVA analysis of T1, T2 and C1 During the harvesting stage, the data of plant height, panicle length, total panicle/m2, total spikelet per panicle, total productive spikelet per panicle, percentage of productive spikelet, the 1000 of grains weight and the grain yield were calculated and analyzed using one way ANOVA. Better performance in yield components at T1 as per shown in
Table 1 Descriptive statistics for T1, T2 and C1 yield components. Components
Type of condition
Plant Height (cm)
T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1
Panicle Length (cm)
Total panicle/m2
Total spikelet/panicle
Total productive spikelet/ panicle
% of productive spikelet
1000 grain weight (g)
Grain yield (g/m2)
(sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control)
Mean ± SE 500WS & 612EC) 500WS)
108.03 ± 1.99 98.32 ± 1.04 86.87 ± 3.71 27.00 ± 0.00 22.60 ± 0.60 20.20 ± 1.43 985.00 ± 57.78 318.40 ± 22.91 785.40 ± 156.97 123.40 ± 13.94 100.00 ± 9.20 56.40 ± 7.44 91.00 ± 17.92 78.43 ± 11.65 40.20 ± 8.68 71.80 ± 7.98 77.00 ± 6.58 69.20 ± 6.97 25.50 ± 0.00 26.00 ± 0.00 20.20 ± 0.00 2304.20 ± 491.42 663.88 ± 128.41 665.42 ± 183.30
500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS)
6
Min–Max
Standard Deviation
102.70–112.70 94.93–101.27 74.75–94.67 27.00 22.00–25.00 18.00–25.00 815.00–1111.00 278.00–407.00 278.00–407.00 98.00–164.00 75.00 –131.00 33.00–80.00 49.00–146.00 48.00–108.00 21.00–71.00 49.00–89.00 59.00–93.00 49.00–89.00 25.55 26.00 20.20 1133.81–3644.04 373.63–1050.15 117.83–1221.73
4.46 2.32 8.29 0.00 0.60 3.19 129.18 51.23 351.00 40.08 20.58 16.64 40.08 26.06 19.41 17.85 14.71 15.58 0.00 0.00 0.00 1098.86 287.13 409.89
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Fig. 9. Yield components of T1, T2 and C1.
Table 2 ANOVA table of yield components.
Plant Height (cm)
Panicle Length (cm)
Total panicle/m2
Number spikelet/panicle
Total productive spikelet/panicle
% of productive spikelet
1000 grain weight (g)
Grain Yield (g/m2)
Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total
Sum of Squares
df
Mean Square
F-value
P-value
1122.128 376.264 1498.391 118.933 48.000 166.933 1,170,474.533 570,056.400 1,740,530.933 11,562.533 6688.400 18,250.933 7000.296 10,647.209 17,647.506 0.016 0.311 0.327 103.30 0.000 0.000 8,960,824.318 5,831,845.090 1.479E7
2 12 14 2 12 14 2 12 14 2 12 14 2 12 14 2 12 14 2 12 14 2 12 14
561.064 31.355
17.894
*0.000
59.467 4.000
14.867
*0.001
585,237.267 47,504.700
12.320
*0.001
5781.267 557.367
10.372
*0.002
3500.148 887.267
3.945
*0.048
0.008 0.026
0.304
0.743
0.000 0.000
2.058E32
*0.000
4,480,412.159 485,987.091
9.219
*0.004
end of cultivation. In addition, improvement in growth characteristics as the result of organic pesticides application might be due to enhanced metabolic activities that led to an increase in various plant metabolites which responsible for cell division and elongation (Morteza Siavoshi et al., 2013). Based on the result in Table 1 and Fig. 9, T1 produced higher yield of 23.04 t/ha followed by C1 of 6.65 t/ha and T2 of 6.64 t/ ha respectively. Muazu et al. (2015) reported that paddy cultivation in Malaysia which normally use chemical pesticides had 7.625 t/ha. The excellent production of T1 is due effective role of both BV500WS and BV612EC. The production might be less if applied in actual paddy field. Besides, T1 had 0.0255 g of grain weight and T2 had 0.026 g of grain weight while Kamarulzaman et al. (2016) reported had 0.024 g of grain yield for the paddy field that was applied with chemical pesticides. Thus, it showed that the application of biopesticides was able to have comparable result with used of chemical pesticides in paddy cultivation. This preliminary test proved that the application of biopesticides were successful in enhancing the plant growth and increased the grain yield. Ning et al. (2017) reported that effective method in controlling pest and rice diseases lead to higher paddy yield. Even though T2 was
Table 1 showed that the application of biopesticides were successful in increasing the productivity of paddy. This preliminary test proved that biopesticides were able to act as plant growth regulator which, can promote, inhibit and modify the physiology of plants to have a higher yield (Alliance, 2015). Transplanting cultivation provided proper space between plant to plant led the paddy to produce more tillers and increase the number of panicle per m2 as the plants utilized the soil nutrients more effectively (Awan et al., 2011). Hence, even though the paddy plants for T1, T2, and C1 were infected from pest and bacteria, they still produced positive grain yield due to adoption of the transplanting cultivation. Besides, higher results of plant height, number of tillers, number of leaves, and width of leaves for T1 compared to T2 and C1 as a result of the application of BV500WS and BV612EC that increased the number of panicle per m2, total spikelet per panicle, total productive spikelet per panicle and eventually grain yield. Ranawake et al. (2014) and Kamarulzaman et al. (2016) also reported similar that the paddy growth characteristics had a positive relationship with yield components. Better performance in paddy growth led higher grain yield at the 7
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
severely infected by the pest after BPH was released, it still managed to produce comparable yield with C1 due to application of BV 500WS at earlier stage of cultivation. Furthermore, Table 2 described that all yield components which are plant height, panicle length, total panicle per m2, number of spikelet per panicle, total productive spikelet per panicle, 1000 of grain weight, and grain yield except the percentage of productive spikelet were significantly affected by the application of biopesticides during paddy cultivation as the P-value is lower than 0.05 (Ltd, 2013). This research showed that the application of this biopesticides was effective in controlling the pest and reducing the rice diseases in order to enhance the paddy growth for yield improvement. Kamarulzaman et al. (2016) also promoted that effective biopesticide in increasing the grain yield will lead to higher preference to farmers to apply it to paddy cultivation.
Ephraums, G.L.J.A.J.J., 1990. IPCC. 1990. Climate Change. The IPCC Scientific Assessment in J.T. Houghton. In: Paper Presented at the Conference Proceedings on WHO/United Nations Programme, Cambridge University Press, Cambridge, England. Ji Zhi-juan, Y.S.-D., Yu-Xiang, Z.E.N.G., Yan, L.I.A.N.G., Chang-deng, Y.A.N.G., Qian, Q.I.A.N., 2016. Pyramiding blast, bacterial blight and brown planthopper resistance genes in rice restore line. J. Intregrative Agric. 15 (7), 1432–1440. Kamarulzaman, P.S.D., Dailin, D.J., Yusup, S., Osman, N.B., Chuah, L.F., Bokhari, A., 2016. Trait associations of rice (Oryza sativa) productivity upon neem-based biopesticide treatment by SPSS. Am. J. Biochem. 6 (6), 137–144. Kim, K.H., Cho, J., Lee, Y.H., Lee, W.S., 2015. Predicting potential epidemics of rice leaf blast and sheath blight in South Korea under the RCP 4.5 and RCP 8.5 climate change scenarios using a rice disease epidemiology model, EPIRICE. Agric. For. Meteorol. 203, 191–207. Lakshmi, M.V., Suneetha, Y., Yugandhar, G., Lakshmi, N.V., 2014. Correlation studies in rice (Oryza sativa L.). Int. J. Genet. Eng. Biotechnol. 5 (2), 121–126. Ltd, L.R., 2013. One-way ANOVA. from 〈https://statistics.laerd.com/statistical-guides/ one-way-anova-statistical-guide.php〉. Malaysia, Jabatan Pertanian, K.P.D.I.A.T., 2008. Pakej Teknologi Padi, 1st edition 2008 ed. Perpustakaan Negara Malaysia, Malaysia. Morteza Siavoshi, S.D., Yassari, Esmail, Laware, Shankar L., 2013. Role of organic fertilizers on morphological and yield parameters in rice (Oryza sativa L.). Int. J. Agron. Plant Prod. 4 (6), 1220–1225. Muazu, A., Yahya, A., Ishak, W.I.W., Khairunniza-Bejo, S., 2015. Energy audit for sustainable wetland paddy cultivation in Malaysia. Energy 87, 182–191. Ni, D., Song, F., Ni, J., Zhang, A., Wang, C., Zhao, K., Li, L., 2015. Marker-assisted selection of two-line hybrid rice for disease resistance to rice blast and bacterial blight. Field Crops Res. 184, 1–8. Ning, C., Qu, J., He, L., Yang, R., Chen, Q., Luo, S., Cai, K., 2017. Improvement of yield, pest control and Si nutrition of rice by rice-water spinach intercropping. Field Crops Res. 208, 34–43. Okada Ecotech Pte. Ltd., 500WS, 2014. 〈http://www.okada-ecotech.com/EN/products_ 500WS.html〉. Okada Ecotech Pte. Ltd., 612EC, 2014. 〈http://www.okada-ecotech.com/EN/products_ 612EC.html〉. Peng, S., Khush, G.S., Cassman, K.G., November 1994. Evolution of the new plant ideotype for increased yield potential. In: Breaking the Yield Barrier. Proceedings of a Workshop on Rice Yield Potential in Favourable Environments. International Rice Research Institute, Philippines. pp. 5–20. Prabhu, S.T., 2010. Status of Paddy Insect Pests and Their Natural Enemies in Rainfed Ecosystem of Uttara Kannada District and Management of Rice Leaf Folder (PhD dissertation). UAS, Dharwad. Ranawake, A.L., Amarasinghe, U.G.S., Senanayake, S.G.J.N., 2014. Ranking traditional rice cultivars based on yield and Some morphological traits using path analysis, multi-criteria decision making model and compromise programming. Am. J. Exp. Agric. 4 (12), 1579. Samy, S.J., Xaviar, A., Rahman, A.B., Sharifuddin, H.A.H., June 1995. Effect of EM on rice production and methane emission from paddy fields in Malaysia. In: Proceedings of the Fourth International Conference on Kyusei Nature Farming. Santos, M.S., Zanardi, O.Z., Pauli, K.S., Forim, M.R., Yamamoto, P.T., Vendramim, J.D., 2015. Toxicity of an azadirachtin-based biopesticide on Diaphorina citri Kuwayama (Hemiptera: liviidae) and its ectoparasitoid Tamarixia radiata (Waterston) (Hymenoptera: eulophidae). Crop Prot. 74, 116–123. Trisyono, Y.A., Aryuwandari, V.E., Rahayu, T., Martono, E., 2017. Effects of etofenprox applied at the sublethal concentration on the fecundity of rice brown planthopper, Nilaparvata lugens. J. Asia-Pac. Entomol. 20 (2), 547–551. Xiaolei Zhang, X.L., Mao, Kaikai, Zhang, Kaixiong, Wan, Hu, Li, Jianhong, 2016. Insecticide resistance monitoring and correlation analysis of insecticides in field populations of the brown planthopper Nilaparvata lugens (stal) in China 2012–2014. Pestic. Biochem. Physiol. 132, 13–20. Zakaria, Ismadi, Malaysia, Jabatan Pertanian, K.P.D.I.A.T, 2016. Perosak Tanaman Padi, 1st edition 2016 ed. Perpustakaan Negara Malaysia, Malaysia.
4. Conclusions According to correlation on the plant growth development and oneway ANOVA analysis, the grain yield is influenced by the plant height, number of tillers, number of leaves, width of leaves, panicle length, number panicle per m2, number of spikelet per panicle, total productive spikelet per panicle, and 1000 of grain weight. In conclusion, combination of BV500WS and BV612EC are more effective in enhancing the paddy growth compared to if only BV500WS is used. The application of biopesticide at the early stage of cultivation is not sufficient to prevent the plants from pest infestation. Application of biopesticide needs to also be applied upon infestation. Acknowledgements This is a collaboration research between Universiti Teknologi PETRONAS, Bio-X Techno Sdn Bhd and Agriculture Department of Perak Tengah. The authors would like to thank all those who have contributed for their joint efforts in conducting this research and also to Universiti Teknologi PETRONAS (0153AB-F02) for supporting this work and Elsevier Foundation Green and Sustainable Chemistry Challenge for the grant to undertake the research. References Alexander, M., Stuart, A.N.P., Bernal, Carmencita C., Ramal, Angelee Fame, Horgan, Finbarr G., 2014. Effects of fertiliser applications on survival and recruitment of the apple snail, Pomacea canaliculata (Lamarck). Crop Prot. 64, 78–87. Alliance, B.I., 2015. Biopesticides Offer Multiple Benefits for Agricultural Dealers and Consultants. From 〈http://www.bpia.org/wp-content/uploads/2014/01/dealer_ consultant-final.pdf〉. Awan, T.H., Ahmad, M., Ashraf, M.M., Ali, I., 2011. Effect of different transplanting methods on paddy yield and its components at farmer's field in rice zone of Punjab. J. Anim. Plant Sci. 21 (3), 498–502.
8
Contents lists available at ScienceDirect
Sustainable Chemistry and Pharmacy journal homepage: www.elsevier.com/locate/scp
Effectiveness of biopesticides in enhancing paddy growth for yield improvement
T
⁎
Noor Hafizah Ramli@Yusofa, Suzana Yusupa, , Benjamin Wei Bin Kueha, Puteri Sarah Diba Kamarulzamana, Noridah Osmana, Mardyahwati Abd. Rahimb, Ramlan Azizb, Sulaiman Mokhtarc, Abu Bakar Ahmadc a Biomass Processing Lab, Centre of Biofuel and Biochemical Research, Mission Oriented Research (Green Technology), Chemical Engineering Department, Universiti Teknologi PETRONAS, 32610 Seri Iskandar, Perak Darul Ridzuan, Malaysia b Plant Biosecurity Unit, Department of Agriculture of Perak Tengah, Simpang 3, 34200 Parit Buntar, Perak Darul Ridzuan, Malaysia c Bio-X Techno Sdn. Bhd., LK1-1-3A, Blok LK1, Laman Komersil ARECA, Putra Nilai, 71800 Nilai, Negeri Sembilan Darul Khusus, Malaysia
A R T I C L E I N F O
A B S T R A C T
Keywords: Biopesticides Paddy growth Yield Paddy cultivation
There is an urgent need to enhance the productivity of irrigated paddy fields to increase the rice production that meets population growth. The focus of this study is to investigate the effectiveness of biopesticides in enhancing the paddy growth for yield improvement after controlling the pest and rice diseases. This study aims to determine either one or two types of biopesticides should be used in paddy cultivation. Thus, three conditions were adopted in paddy cultivation at the insect house as follows; T1 where the paddy was sprayed using BV500WS and BV612EC to study the effectiveness of the biopesticides, T2 where the paddy was sprayed using BV500WS only and C1 where the paddy was cultivated following the conventional technique as a control. BV500WS was sprayed into T1 and T2 on the first day of paddy growth while BV612EC was sprayed later into T1, at 55 and 67 days of paddy cultivation a week after releasing the brown planthopper (BPH). Based on the analysis of correlation obtained for each growth, application of BV500WS and BV612EC of T1 resulted in better performance in terms of height of plant, number of tillers, width of leaves, and number of leaves compared to T2 and C1. The excellence performance of plant growth at T1 produced the highest paddy yield of 2304.20 ± 491.42 g−2, while T2 produced comparable yield as C1 which are 663.88 ± 128.41 g−2 and 665.42 ± 183.30 g−2 respectively. Hence, organic plants based biopesticides are successful in enhancing the plant growth of paddy and ultimately increase the paddy yield.
1. Introduction Malaysia warm and humid climate attracts a number of pests and rice diseases that severely reduce the productivity and yield of paddy fields. Approximately over 800 herbivore insect species inhabit the ecosystem. Thus, there is an urgent need to enhance the productivity of irrigated paddy fields in order to increase the production of rice to meet the population growth (Prabhu, 2010). In addition, the brown planthopper (BPH) also causes serious damage to the rice by sucking rice sap, ovipositing in the rice tissues and transmitting rice diseases, especially during tillering stage thus causing less production (Zakaria et al., 2016; Xiaolei Zhang et al., 2016). Rice diseases like bacterial blight (BB), leaf blast, brown spot and sheath blight frequently affected the paddy growth and yield. BB, caused by Xanthomonas oryzaepv. oryzae (Xoo), caused yield reductions of 20–50% at the tiller stage in
⁎
severe cases; while rice blast, caused by the fungal pathogen Magnaporthe oryzae, is responsible for large yield losses (Ni et al., 2015; Kim et al., 2015; Ji Zhi-juan et al., 2016). Besides, there is a growing interest worldwide in alternative agricultural practices that can help farmers reduce their dependence on synthetic fertilizers and pesticides. The excessive use and misuse of the synthetic chemicals have raised serious questions regarding their longterm effects on human health, soil quality, and the environment (Ephraums, 1990; Samy et al., 1995). For the improvement of grain yield, the knowledge of the association between grain yield and its component characters such as number of effective tillers per plant, plant height, and number of grains per panicle will be helpful (Lakshmi et al., 2014). The relative contribution of different traits on yield determination can be explained by correlation studies. The existence of correlation is attributed to the
Corresponding author. E-mail address: [email protected] (S. Yusup).
https://doi.org/10.1016/j.scp.2017.11.002 Received 19 August 2017; Received in revised form 15 November 2017; Accepted 16 November 2017 2352-5541/ © 2017 Elsevier B.V. All rights reserved.
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Only the growing seeds were selected to be planted in the pots. The seeds were sowed in the pots by adopting the paddy transplantation cultivation. The BV500WS solution was diluted in the remaining water with ratio of 1:2500 and was sprayed onto the T1 and T2. While the C1 were sprayed with the balance purified water from the curing process. The BV612EC was applied onto T1 only during 55 and 67 days of cultivation which is a week after the BPH population was released onto the paddy plants of T1 and T2. The BPH were not released to C1 as to maintain the standard cultivation and to study how the BPH infest the paddy naturally.
environmental effect or genetic effect of genes such as grain yield in rice, is significantly correlated with total tillers, flag leaf length, flag leaf width, number and grain weight (Ranawake et al., 2014). Biopesticides are employed in agricultural uses to control the insect, disease, nematode and weed. They also can improve plant physiology and productivity. As biopesticides commonly are biodegradable, they are able to decompose quickly and do not negatively impact surface water and groundwater (Alliance, 2015). The BV500WS, which has high azadirachtin (A + B) content with proprietary water soluble concentrates is formulated for root feeding, drip and foliar for crop protection and growth enhancement (Okada Ecotech, 2014). Santos, M.S. also reported that azadiracthin which can be found abundantly in neem has high insecticide and rapid degradation in the environment (Santos et al., 2015). Besides that, BV612EC is a series of environmental friendly insecticidal formulations based on a unique and extremely safe active ingredient known as Etofenprox. It has high insecticidal activities which provide a very quick killing action against a broad spectrum of crop insect pests including lepidoptera, hemiptera, coleopteran, diptera and orthoptera, and yet has remarkably weak toxicity to mammals (Okada Ecotech, 2014). Moreover, Etofenprox was reported to be highly toxic to the insect including BPH but relatively safe to non-target organisms (Xiaolei Zhang et al., 2016; Trisyono et al., 2017). The main focus of the study is to determine either one or two types of biopesticides should be used in paddy cultivation to control the pest and reduce the rice diseases for enhancing the paddy growth for yield improvement. This research also aimed to study the efficacy of new biopesticides in playing their roles as crop protection and growth enhancement for BV500WS and as insecticide for BV612EC in paddy cultivation. The correlation study on paddy growth and development were carried out at every stage of cultivation by focusing on plant height, number of tillers, number of leaves, and width of leaves. The efficacy of biopesticides was evaluated by analyzing the yield components using analysis of variance (ANOVA) during the harvesting stage.
2.2.2. Paddy growth development The plant growths were observed in every cultivation stage. The observation was focused on plant diseases, color of leaves, weeds and pest problems. In addition, the correlation study of paddy growth was also conducted by taking record on the plant height, number of tillers, number of leaves, and width of leaves of all samples. The stages of paddy cultivation are as per stated below (Malaysia and Padi, 2008):
• Stage 1: Plantation Day 1–5 days) • Stage 2: Tillering Stage 15–40 days) • Stage 3: Panicle Initiation Stage 40–69 days) • Stage 4: Heading Stage 70–89 days) • Stage 5: Ripening Stage 90–104 days) • Stage 6: Harvest Stage 105–112 days) 2.3. One way ANOVA analysis One way ANOVA is used to determine whether there are any statistically significant differences between T1, T2 and C1. The data were transformed using various algorithms so that the frequency distributions are normally distributed (Ltd, 2013). The mean with standard error, the minimum and maximum values, and the standard deviation of all recorded data were calculated and treated using ANOVA. The degree of freedom, the F-value and P-value were obtained from the analysis.
2. Material and method 2.1. Experimental site
2.3.1. Yield component calculation The experimental site was located at Plant Biosecurity Unit of Agriculture Department, Titi Serong, Parit Buntar, Perak, Malaysia. 2.2. Insect house experiment BV500WS and BV612EC were obtained from Bio-X Techno Sdn. Bhd. as a part of collaborative research. The application of BV 500WS is for crop protection and growth enhancement at the early stage of cultivation while BV612EC as insecticide to protect the paddy during pest infestation (Okada Ecotech, 2014). Three conditions were adopted in paddy cultivation at the insect house as follows which are T1 where the paddy was sprayed using BV500WS and BV612EC to study the effectiveness of both types of biopesticides, T2 where the paddy was sprayed using BV500WS only and C1 where the paddy was cultivated following the conventional technique as a control. Three different conditions with five replications each were performed in the Insect House. All pots (9 cm diameter) were filled with 15 cm depth of soil collected from paddy field at the Rice of Excellence Centre of Agriculture Department, Titi Serong, Parit Buntar, Perak, Malaysia.
mass of 1000 productive spikelets (g ) mass of 800 productive spikelets = × 1000 800
(1)
acurate mass of 14% moisture content, MC (g) 100−actualMC = × [1] 86
(2)
mass of 1 productive spikelet (g ) =
% Productives spikelets =
(2) 1000
Total productive spikelets × 100% Total spikelets
(3)
(4)
Number of panicle per m2 = Total panicles × area of sampling (5)
Gain yield
2.2.1. Cultivation of paddy The rice variety of MR 219 has been used in this experiment. The seeds for T1 and T2 were immersed in the BV500WS solution with ratio of 1:1000 to the purified water while the seeds for C1 were immersed in purified water only. All the seeds were immersed for 24 h before filtered. All the floated seeds were eliminated. Then, the seeds were dried under room temperature for another 24 h before seedling in the pot. The remaining water was used to spray in the pot during cultivation.
(gm−2) =
Total spikelets × (4) × (3) × (5) Total panicles
(6)
All calculations were performed according to standard protocol from Agriculture Department (Malaysia and Padi, 2008). 3. Results and discussion 3.1. Paddy growth observation The observation of paddy growth is to study the effectiveness of 2
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Paddy plant is in good condition
Paddy plant is in good condition
Paddy plant is in good condition
Brown Leaf Blight, and neck rot and detected
Brown Leaf Blight, neck rot and hole leaves detected
Brown Leaf Blight, neck rot and hole leaves detected
-The affected leaves and tillers increased -The brown planthopper has been released
-The paddy plants become more healthier and the panicles start to develop after spray 612EC. -The population of brown planthopper disappeared.
The paddy plant continuous to growth healthy. The panicles has fully develop and visible but few of the plants has weedy rice .
-The affected leaves and tillers increased -The brown planthopper has been released
The plant attacked by pest and diseases rapidly due to increasing of brown planthopper population.
The plant still suffering the pest attacked and diseases due to the present of brown planthopper. The number of weedy rice higher than T1
The condition of the paddy plants are worst compared to T1 & T2. Pest and diseased attacked the plants
Pest and diseases attacked continue to increasing and the present of weedy rice is the highest compare to T1 and T2
The affected leaves and tillers increased
Fig. 1. Condition of paddy plants at every stage on selected days of cultivation.
3.1.2. Color of leaves Fig. 2 showed that the color of the leaves started to change to yellowish after 42 days of paddy cultivation. From the tillering stage till the end of cultivation, the pest and bacteria began to infest the leaves especially at C1, resulted in increasing of yellowish leaves and directly increased the dead leaves as C1 was not applied with any treatment. The application of BV612EC on 55 and 67 days of cultivation at T1, resulted in yellowish leaves turned back to green and accelerated plant growth as per shown on Day 71 and Day 101.
BV500WS and BV 612EC in terms of physical appearance, weeds and pest problem. There are five major rice diseases which are brown spot, leaf blast, bacterial leaf blight, sheath blight, and tungro that frequently occur in tropical Asia. The diseases are caused by fungal pathogens and bacterial pathogen led in reducing both grain yield and quality (Ji Zhijuan et al., 2016; Kim et al., 2015). During early stage of cultivation, the paddy at T1 and T2 were healthier than C1 as the seeds were immersed and also sprayed using BV500WS to improve photosynthesis and assimilated translocation (Okada Ecotech, 2014). However, after BPH was released at 46 and 59 days of cultivation, at T1 and T2 to create a worst case scenario of infestation, the paddy started to show the impact of the pests. Even though the BPH at C1 was not released to let the pest infests naturally, C1 had a severe infestation by the BPH. Due to BPH infestation, the yellowish and dead leaves, rice diseases, and the affected tillers increased after 71 days of cultivation. Only T1 showed positive growth since it was sprayed with BV612EC during 55 and 67 days of cultivation for crop protection. The observation in Figs. 1–4 described the physical appearance of paddy and the weed and pest problems on the selected days of paddy cultivation in details.
3.1.3. Weed problems The weed population like yellow bur-head distracted the paddy growth as they competed with paddy for nutrients from water and soil while duckweeds covered the surface water to get enough oxygen as per described in Fig. 3 aon selected days of cultivation. Increasing the weed population caused reduced of nutrients availability for paddy, which subsequently reduced its yield (Awan et al., 2011). However, the biopesticides application showed a positive effect in reducing the weed population. T1 showed decreasing of weed population on 101 days of cultivation while T2, the population of weed remained the same started from 71 days of paddy cultivation as per shown in Fig. 3. Only C1 showed the population of weed increased till the harvesting stage.
3.1.1. Paddy condition The plant diseases such as bacterial leaf blight and leaf blast started to appear after 42 days of paddy cultivation. The grasshopper also ate the leaves. Fig. 1 showed that the pest and bacteria infestation increased the number of affected tillers resulted in reduction of panicles during heading stages. Therefore, T1 produced the highest panicles compared to T2 and C1, due to the application of BV500WS and BV612EC which reduced the rice diseases and pest infestation as per illustrated in the Fig. 1 on 101 days of paddy cultivation.
3.1.4. Pest problems There were three dead paddy plants at C1 during observation during harvesting stage due to bad infestation from pest especially BPH resulted in higher unfilled spikelets and lighter grains compared to T1 and T2. Fig. 4 described that the pest population at T1 started to decrease from 71 days of paddy cultivation due to repeated application of BV612EC contrary with T2 and C1, in which the pest population 3
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Remain green
Remain green
Remain green
The leaves turned yellowish
The yellowish leaves increased due to release brown plant hopper
The leaves turned back to green after sprayed with 612EC
Remain green
Remain green
The yellowish leaves increased due to release brown plant hopper
The yellowish leaves increased due to not sprayed with 612EC
The yellowish leaves increased
The leaves turned yellowish
The yellowish leaves increased due to pest infestation
The yellowish leaves increased
Dead leaves increased
Fig. 2. Color of leaves at every stage on selected days of cultivation. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article).
to only one type of biopesticide. Ji Zhi-juan et al. (2016) studied that the damage caused by BPH feeding has the greatest effect on the growth and crop yield of the susceptible rice plant through the removal of assimilates and the reduction in photosynthetic rate of leaves, ultimately causing plant death in its severe form. BV612EC which has Etofenprox as active ingredient
increased during observation on 71 and 101 days of paddy cultivation. The BPH and golden apple snail affected the tillers, while the insect such as grasshopper and bugs; and bacteria affected the leaves which led to increasing of dead leaves and tillers (Alexander et al., 2014; Ji Zhi-juan et al., 2016). Thus, the combination of the biopesticides application were more effective in reducing the pest infestation compared
Presence of Yellow bur-head (paku rawan)
-Yellow bur-head increased -presence of duckweeds
Weeds increased
Weeds population remain the same
-Duckweeds decreased -Yellow bur-head remains the same
Presence of duckweeds
-Presence of Yellow burhead -duckweeds increased
Weeds increased
Weeds population remain the same
Weeds population remain the same
Presence of duckweeds
-Presence of Yellow burhead -duckweeds increased
Weeds population remain the same
Weeds population remain the same
Fig. 3. Weed problems at every stage on selected days of cultivation.
4
Weeds increased
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
No pest
No pest
No pest
Presence of pest
No pest
Presence of grasshopper
Pest and rice diseases increased
Pest population increased.
Pest population increased.
Pest population decreased rapidly due to sprayed with 612EC
Pest population decreased
Pest population increased rapidly due to not sprayed with 612EC
Pest population increased
Pest population increased rapidly due to not sprayed with 612EC
Pest population increased
Fig. 4. Pest problems at every stage on selected days of cultivation.
infestation resulted in lower performance of plant growth development and reduction in crop yield as pest infestation commonly transmitted virus and diseases to the plants (Xiaolei Zhang et al., 2016). Unlike T1, the positive paddy growth led the paddy to produce healthier panicles compared to T2 and C1 due to the application of two types of biopesticides during cultivation. Based on the observation, it showed that the application of biopesticide, at the early stage of cultivation is not sufficient to prevent the plants from pest infestation. Application of biopesticide needs to also be applied upon infestation during tillering and panicle initiation stages.
3.2. Correlation analysis of paddy growth The correlation analysis of paddy growth in terms of plant height, number of tillers, number of leaves and width of leaves has been recorded during 19, 38, 66, 81 and 101 days on selected days of cultivation to study the effectiveness of one type and both types of biopesticides.
Fig. 5. Plant height (cm) at T1, T2 and C1 on selected days of cultivation.
3.2.1. Plant height Fig. 5 showed that all the three types of conditions have positive increment on plant height. T2 showed higher plant height at 38 days of cultivation with 86.21 ± 3.04 cm followed by T1 with 73.59 ± 2.36 cm and C1 with 45.52 ± 1.67 cm respectively. It showed that BV500WS was successful in accelerated the paddy growth of T1 and T2 at early stage of cultivation. However, releasing the BPH at 46 and 59 days of cultivation reduced the increment rate of plant height at T2 and C1 due to severity of pest infestation. Only the plant height at T1 increased rapidly achieved the highest plant height with 113.55 ± 10.05 cm on 101 day of cultivation as were sprayed with BV612EC during 55 and 67 days of cultivation to prevent the plant from infestation. Ranawake et al. (2014) and Kamarulzaman et al. (2016) reported that plant height had a positive relationship with the grain yield. Thus, higher plant height will lead to have a higher grain yield. Hence, the application of two types of biopesticides led to better performance in plant height compared to only one type of biopesticide.
Fig. 6. Number of tillers at T1, T2 and C1 on selected days of cultivation.
was successfully reduced the BPH population and rice diseases. Xiaolei Zhang et al. (2016) and Trisyono et al. (2017) also reported similar observation using Etofenprox to control the BPH in paddy. The results showed it was effective in reducing the BPH population and relatively safe to its natural enemies. C1 and T2 which had the severity of pest 5
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
of cultivation after BPH was released. The pest affected the tillers and directly increased the number of dead tillers towards the end of the cultivation. However, decreasing in the number of tillers can increase the paddy yield as the unproductive tillers will compete for resources. Thus, eliminating the unproductive tillers will allow more solar energy and mineral nutrients for the growth of productive tillers (Peng et al., 2014). T1 had highest remaining tillers with average 18 tillers per pot during observation on 101 days of cultivation compared with C1 with average 14 remaining tillers per pot and T2 with average 6 remaining tillers per pot. Even though the number of tillers started to decrease on 81 days of cultivation due to affected by pest infestation at T1, T2, and C1, by applying BV612EC twice at T1, it accelerated the growth of productive tillers which resulted in healthier panicles and higher yield. Fig. 7. Number of leaves at T1, T2 and C1on selected days of cultivation.
3.2.3. Number of leaves Plant leaves are important organs which have an active role in photosynthesis and yield contribution (Morteza Siavoshi et al., 2013). Fig. 7 showed that T1 has the highest number of leaves with average 94 leaves per pot as T1 was less affected by pest infestation and rice diseases compared to T2 with average 48 leaves per pot and C1 with average 46 leaves per pot from day 66 to day 101 of paddy cultivation. Thus, the application of two types of biopesticide was more effective in increasing the number of leaves and directly led to higher yield. 3.2.4. Width of leaves The available nutrients and crop protection might have helped in enhancing the leaf area, which thereby resulted in higher photo-assimilates and more dry matter accumulation (Morteza Siavoshi et al., 2013). The application of BV500WS and BV612EC enhanced the plant growth and led T1 with highest width of leaves with 2.15 ± 0.12 cm followed by T2 with 1.81 ± 0.10 cm and C1 with 1.21 ± 0.64 cm respectively as shown in Fig. 8.
Fig. 8. Width of leaves at T1, T2 and C1on selected days of cultivation.
3.2.2. Number of tillers Tillering is an important trait for grain production and is thereby an important aspect in rice yield (Morteza Siavoshi et al., 2013). BV500WS which is neem based biopesticide influenced the paddy in increasing the number of tillers at early stage of cultivation as a part of growth enhancement (Okada Ecotech, 2014). This is because organic sources offer more balanced nutrition to the plants especially micronutrients which, positively affects the number of tillers in plant (Morteza Siavoshi et al., 2013). Fig. 6 showed the reduction of tillers starting from 66 days
3.3. One way ANOVA analysis of T1, T2 and C1 During the harvesting stage, the data of plant height, panicle length, total panicle/m2, total spikelet per panicle, total productive spikelet per panicle, percentage of productive spikelet, the 1000 of grains weight and the grain yield were calculated and analyzed using one way ANOVA. Better performance in yield components at T1 as per shown in
Table 1 Descriptive statistics for T1, T2 and C1 yield components. Components
Type of condition
Plant Height (cm)
T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1 T1 T2 C1
Panicle Length (cm)
Total panicle/m2
Total spikelet/panicle
Total productive spikelet/ panicle
% of productive spikelet
1000 grain weight (g)
Grain yield (g/m2)
(sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control) (sprayed with (sprayed with (control)
Mean ± SE 500WS & 612EC) 500WS)
108.03 ± 1.99 98.32 ± 1.04 86.87 ± 3.71 27.00 ± 0.00 22.60 ± 0.60 20.20 ± 1.43 985.00 ± 57.78 318.40 ± 22.91 785.40 ± 156.97 123.40 ± 13.94 100.00 ± 9.20 56.40 ± 7.44 91.00 ± 17.92 78.43 ± 11.65 40.20 ± 8.68 71.80 ± 7.98 77.00 ± 6.58 69.20 ± 6.97 25.50 ± 0.00 26.00 ± 0.00 20.20 ± 0.00 2304.20 ± 491.42 663.88 ± 128.41 665.42 ± 183.30
500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS) 500WS & 612EC) 500WS)
6
Min–Max
Standard Deviation
102.70–112.70 94.93–101.27 74.75–94.67 27.00 22.00–25.00 18.00–25.00 815.00–1111.00 278.00–407.00 278.00–407.00 98.00–164.00 75.00 –131.00 33.00–80.00 49.00–146.00 48.00–108.00 21.00–71.00 49.00–89.00 59.00–93.00 49.00–89.00 25.55 26.00 20.20 1133.81–3644.04 373.63–1050.15 117.83–1221.73
4.46 2.32 8.29 0.00 0.60 3.19 129.18 51.23 351.00 40.08 20.58 16.64 40.08 26.06 19.41 17.85 14.71 15.58 0.00 0.00 0.00 1098.86 287.13 409.89
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
Fig. 9. Yield components of T1, T2 and C1.
Table 2 ANOVA table of yield components.
Plant Height (cm)
Panicle Length (cm)
Total panicle/m2
Number spikelet/panicle
Total productive spikelet/panicle
% of productive spikelet
1000 grain weight (g)
Grain Yield (g/m2)
Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total Between Groups Within Groups Total
Sum of Squares
df
Mean Square
F-value
P-value
1122.128 376.264 1498.391 118.933 48.000 166.933 1,170,474.533 570,056.400 1,740,530.933 11,562.533 6688.400 18,250.933 7000.296 10,647.209 17,647.506 0.016 0.311 0.327 103.30 0.000 0.000 8,960,824.318 5,831,845.090 1.479E7
2 12 14 2 12 14 2 12 14 2 12 14 2 12 14 2 12 14 2 12 14 2 12 14
561.064 31.355
17.894
*0.000
59.467 4.000
14.867
*0.001
585,237.267 47,504.700
12.320
*0.001
5781.267 557.367
10.372
*0.002
3500.148 887.267
3.945
*0.048
0.008 0.026
0.304
0.743
0.000 0.000
2.058E32
*0.000
4,480,412.159 485,987.091
9.219
*0.004
end of cultivation. In addition, improvement in growth characteristics as the result of organic pesticides application might be due to enhanced metabolic activities that led to an increase in various plant metabolites which responsible for cell division and elongation (Morteza Siavoshi et al., 2013). Based on the result in Table 1 and Fig. 9, T1 produced higher yield of 23.04 t/ha followed by C1 of 6.65 t/ha and T2 of 6.64 t/ ha respectively. Muazu et al. (2015) reported that paddy cultivation in Malaysia which normally use chemical pesticides had 7.625 t/ha. The excellent production of T1 is due effective role of both BV500WS and BV612EC. The production might be less if applied in actual paddy field. Besides, T1 had 0.0255 g of grain weight and T2 had 0.026 g of grain weight while Kamarulzaman et al. (2016) reported had 0.024 g of grain yield for the paddy field that was applied with chemical pesticides. Thus, it showed that the application of biopesticides was able to have comparable result with used of chemical pesticides in paddy cultivation. This preliminary test proved that the application of biopesticides were successful in enhancing the plant growth and increased the grain yield. Ning et al. (2017) reported that effective method in controlling pest and rice diseases lead to higher paddy yield. Even though T2 was
Table 1 showed that the application of biopesticides were successful in increasing the productivity of paddy. This preliminary test proved that biopesticides were able to act as plant growth regulator which, can promote, inhibit and modify the physiology of plants to have a higher yield (Alliance, 2015). Transplanting cultivation provided proper space between plant to plant led the paddy to produce more tillers and increase the number of panicle per m2 as the plants utilized the soil nutrients more effectively (Awan et al., 2011). Hence, even though the paddy plants for T1, T2, and C1 were infected from pest and bacteria, they still produced positive grain yield due to adoption of the transplanting cultivation. Besides, higher results of plant height, number of tillers, number of leaves, and width of leaves for T1 compared to T2 and C1 as a result of the application of BV500WS and BV612EC that increased the number of panicle per m2, total spikelet per panicle, total productive spikelet per panicle and eventually grain yield. Ranawake et al. (2014) and Kamarulzaman et al. (2016) also reported similar that the paddy growth characteristics had a positive relationship with yield components. Better performance in paddy growth led higher grain yield at the 7
Sustainable Chemistry and Pharmacy 7 (2018) 1–8
N.H. Ramli@Yusof et al.
severely infected by the pest after BPH was released, it still managed to produce comparable yield with C1 due to application of BV 500WS at earlier stage of cultivation. Furthermore, Table 2 described that all yield components which are plant height, panicle length, total panicle per m2, number of spikelet per panicle, total productive spikelet per panicle, 1000 of grain weight, and grain yield except the percentage of productive spikelet were significantly affected by the application of biopesticides during paddy cultivation as the P-value is lower than 0.05 (Ltd, 2013). This research showed that the application of this biopesticides was effective in controlling the pest and reducing the rice diseases in order to enhance the paddy growth for yield improvement. Kamarulzaman et al. (2016) also promoted that effective biopesticide in increasing the grain yield will lead to higher preference to farmers to apply it to paddy cultivation.
Ephraums, G.L.J.A.J.J., 1990. IPCC. 1990. Climate Change. The IPCC Scientific Assessment in J.T. Houghton. In: Paper Presented at the Conference Proceedings on WHO/United Nations Programme, Cambridge University Press, Cambridge, England. Ji Zhi-juan, Y.S.-D., Yu-Xiang, Z.E.N.G., Yan, L.I.A.N.G., Chang-deng, Y.A.N.G., Qian, Q.I.A.N., 2016. Pyramiding blast, bacterial blight and brown planthopper resistance genes in rice restore line. J. Intregrative Agric. 15 (7), 1432–1440. Kamarulzaman, P.S.D., Dailin, D.J., Yusup, S., Osman, N.B., Chuah, L.F., Bokhari, A., 2016. Trait associations of rice (Oryza sativa) productivity upon neem-based biopesticide treatment by SPSS. Am. J. Biochem. 6 (6), 137–144. Kim, K.H., Cho, J., Lee, Y.H., Lee, W.S., 2015. Predicting potential epidemics of rice leaf blast and sheath blight in South Korea under the RCP 4.5 and RCP 8.5 climate change scenarios using a rice disease epidemiology model, EPIRICE. Agric. For. Meteorol. 203, 191–207. Lakshmi, M.V., Suneetha, Y., Yugandhar, G., Lakshmi, N.V., 2014. Correlation studies in rice (Oryza sativa L.). Int. J. Genet. Eng. Biotechnol. 5 (2), 121–126. Ltd, L.R., 2013. One-way ANOVA. from 〈https://statistics.laerd.com/statistical-guides/ one-way-anova-statistical-guide.php〉. Malaysia, Jabatan Pertanian, K.P.D.I.A.T., 2008. Pakej Teknologi Padi, 1st edition 2008 ed. Perpustakaan Negara Malaysia, Malaysia. Morteza Siavoshi, S.D., Yassari, Esmail, Laware, Shankar L., 2013. Role of organic fertilizers on morphological and yield parameters in rice (Oryza sativa L.). Int. J. Agron. Plant Prod. 4 (6), 1220–1225. Muazu, A., Yahya, A., Ishak, W.I.W., Khairunniza-Bejo, S., 2015. Energy audit for sustainable wetland paddy cultivation in Malaysia. Energy 87, 182–191. Ni, D., Song, F., Ni, J., Zhang, A., Wang, C., Zhao, K., Li, L., 2015. Marker-assisted selection of two-line hybrid rice for disease resistance to rice blast and bacterial blight. Field Crops Res. 184, 1–8. Ning, C., Qu, J., He, L., Yang, R., Chen, Q., Luo, S., Cai, K., 2017. Improvement of yield, pest control and Si nutrition of rice by rice-water spinach intercropping. Field Crops Res. 208, 34–43. Okada Ecotech Pte. Ltd., 500WS, 2014. 〈http://www.okada-ecotech.com/EN/products_ 500WS.html〉. Okada Ecotech Pte. Ltd., 612EC, 2014. 〈http://www.okada-ecotech.com/EN/products_ 612EC.html〉. Peng, S., Khush, G.S., Cassman, K.G., November 1994. Evolution of the new plant ideotype for increased yield potential. In: Breaking the Yield Barrier. Proceedings of a Workshop on Rice Yield Potential in Favourable Environments. International Rice Research Institute, Philippines. pp. 5–20. Prabhu, S.T., 2010. Status of Paddy Insect Pests and Their Natural Enemies in Rainfed Ecosystem of Uttara Kannada District and Management of Rice Leaf Folder (PhD dissertation). UAS, Dharwad. Ranawake, A.L., Amarasinghe, U.G.S., Senanayake, S.G.J.N., 2014. Ranking traditional rice cultivars based on yield and Some morphological traits using path analysis, multi-criteria decision making model and compromise programming. Am. J. Exp. Agric. 4 (12), 1579. Samy, S.J., Xaviar, A., Rahman, A.B., Sharifuddin, H.A.H., June 1995. Effect of EM on rice production and methane emission from paddy fields in Malaysia. In: Proceedings of the Fourth International Conference on Kyusei Nature Farming. Santos, M.S., Zanardi, O.Z., Pauli, K.S., Forim, M.R., Yamamoto, P.T., Vendramim, J.D., 2015. Toxicity of an azadirachtin-based biopesticide on Diaphorina citri Kuwayama (Hemiptera: liviidae) and its ectoparasitoid Tamarixia radiata (Waterston) (Hymenoptera: eulophidae). Crop Prot. 74, 116–123. Trisyono, Y.A., Aryuwandari, V.E., Rahayu, T., Martono, E., 2017. Effects of etofenprox applied at the sublethal concentration on the fecundity of rice brown planthopper, Nilaparvata lugens. J. Asia-Pac. Entomol. 20 (2), 547–551. Xiaolei Zhang, X.L., Mao, Kaikai, Zhang, Kaixiong, Wan, Hu, Li, Jianhong, 2016. Insecticide resistance monitoring and correlation analysis of insecticides in field populations of the brown planthopper Nilaparvata lugens (stal) in China 2012–2014. Pestic. Biochem. Physiol. 132, 13–20. Zakaria, Ismadi, Malaysia, Jabatan Pertanian, K.P.D.I.A.T, 2016. Perosak Tanaman Padi, 1st edition 2016 ed. Perpustakaan Negara Malaysia, Malaysia.
4. Conclusions According to correlation on the plant growth development and oneway ANOVA analysis, the grain yield is influenced by the plant height, number of tillers, number of leaves, width of leaves, panicle length, number panicle per m2, number of spikelet per panicle, total productive spikelet per panicle, and 1000 of grain weight. In conclusion, combination of BV500WS and BV612EC are more effective in enhancing the paddy growth compared to if only BV500WS is used. The application of biopesticide at the early stage of cultivation is not sufficient to prevent the plants from pest infestation. Application of biopesticide needs to also be applied upon infestation. Acknowledgements This is a collaboration research between Universiti Teknologi PETRONAS, Bio-X Techno Sdn Bhd and Agriculture Department of Perak Tengah. The authors would like to thank all those who have contributed for their joint efforts in conducting this research and also to Universiti Teknologi PETRONAS (0153AB-F02) for supporting this work and Elsevier Foundation Green and Sustainable Chemistry Challenge for the grant to undertake the research. References Alexander, M., Stuart, A.N.P., Bernal, Carmencita C., Ramal, Angelee Fame, Horgan, Finbarr G., 2014. Effects of fertiliser applications on survival and recruitment of the apple snail, Pomacea canaliculata (Lamarck). Crop Prot. 64, 78–87. Alliance, B.I., 2015. Biopesticides Offer Multiple Benefits for Agricultural Dealers and Consultants. From 〈http://www.bpia.org/wp-content/uploads/2014/01/dealer_ consultant-final.pdf〉. Awan, T.H., Ahmad, M., Ashraf, M.M., Ali, I., 2011. Effect of different transplanting methods on paddy yield and its components at farmer's field in rice zone of Punjab. J. Anim. Plant Sci. 21 (3), 498–502.
8