Maximizing potato yields and quality while minimizing nitrogen loss is a major challenge for potato producers. It is crucial to use nitrogen fertilizer for the plant growth, both economically and environmentally. A similar experiment was conducted in India for two years to investigate the response of potato varieties to nitrogen fertilization, and for this study two potato varieties, i.e., Kufri Surya and Kufri Sadabahar and five nitrogen levels (0, 75, 150, 225 and 300 kg/ha) were carried out in a randomized block design (factorial) with three replications. The results reveal that most of the growth parameters and NPK uptake by haulms showed maximum value at nitrogen 300 kg/ha closely followed by the nitrogen dose 225 kg/ha, while for yield parameters and NPK uptake by tubers, nitrogen level 225 kg/ha was the best with the highest value. It depicts that crop yield parameters are negatively affected by the nitrogen dose of above 225 kg/ha. So far the variety is concerned, Kufri Sadabahar was significantly better for all the noted growth and yield parameters as well as NPK uptake by tubers and haulms than the variety Kufri Surya. As far concern about the soil fertility behavior after harvest of the crop, higher left over nitrogen in soil was recorded for the highest applied nitrogen dose, whereas, phosphorus and potassium left over in soil was recorded more where no nitrogen was applied, which was closely followed by 75 kg/ha nitrogen level. The results provide useful recommendations for the improvement of nitrogen fertilization rate for these two varieties in sub-tropical region of north western India in Haryana state.
Research Article
Response of Potato Varieties to Nitrogen Fertilization for Growth, Yield and Nutrient Uptake
https://doi.org/10.21203/rs.3.rs-2890456/v1
This work is licensed under a CC BY 4.0 License
Journal Publication
published 13 Apr, 2024
You are reading this latest preprint version
Potato varieties
Tuber size
NPK uptake
Agronomic Nitrogen Use Efficiency
Soil fertility behavior
Potato is a short duration and nutrient exhaustive high yielding crop. To ensure better tuber yield, balanced fertilizers, along with optimum management practices, must be applied as a prerequisite. Beside other agronomic strategies, adequate supply of nutrients is of main importance for achieving desired potato yield (Koch et al., 2020). The selection of a high-yielding variety and the application of nitrogen are two of the most important management practices in potato cropping systems, which can affect the quantity, yield, chemical composition, and quality of the tuber (Kolodziejczyk, 2014). Potato is very responsive to nutrients application. Nitrogen in all kind of soils, affects tuber yield directly, which is also the main limiting nutrient for potato production. Its requirement, however, depends on soil type and nutrient status.
In the early growth stages of the crop, excessive use of nitrogenous fertilizers can result in increased vegetative growth at the cost of tuber formation which lead to higher production costs as well as environmental pollution (Yadav et al., 2022). However, nitrogen management in the crop is very challenging for obtaining higher yield and quality tubers. The symptoms of nitrogen deficiency are stunted plant growth with few thin stems and small pale-leaves resulting into low yield with few tubers only (Hazra and Som, 2015). Therefore, the crop nitrogen requirement understanding is very essential to ensure economical crop production and to mitigate the adverse environmental effects caused by leaching and runoff.
The fertility status of the soil and the genetic potential of the variety both are critical for crop growth and yield. As response of different varieties vary with nutrient levels, selection of variety with promising yield is very crucial (Kumar et al., 2015). Two high yielding potato varieties, viz., Kufri Surya and Kufri Sadabahar, released by Central Potato Research Institute, Shimla resulted into low yields on cultivation with the recommended dose of fertilizers for the region which obliged the standardization of nutritional requirements for these two varieties. In order to maximize the yield and quality of these potato varieties in Haryana, it is necessary to optimize the nitrogen level. Keeping the above points in mind, the present investigation was designed with the aim of examining the productivity and nutrient efficiency of various potato varieties in response to various nitrogen levels, while, also minimizing environmental pollution.
Description of the Study Area
The experiment was conducted at Research Farm and laboratory of the Department of Vegetable Science, CCS Haryana Agricultural University Hisar during Rabi season of 2015-16 and 2016-17. Hisar is situated in the semi-arid, sub-tropical region of north western India in Haryana state. The elevation of this site is 215.2 meters above sea level and lies at 29°10’ N latitude and longitude 75°46’ E.
Description of Climatic Conditions
This region experiences hot and dry winds during the summer and dry and severe cold in the winter. The annual rainfall for this area is approximately 400 mm, though it varies greatly in distribution. Eighty to ninety percent of all rainfall is seen from July to September months during the Southwest monsoon, with only a few showers during the winter and spring. Both years of study showed wide fluctuations in the maximum and minimum temperatures during the growing season. The monthly mean air temperature, bright sunshine hours as well as morning and evening humidity during the crop season of both the years are given in Table 1.
|
Month |
2015-16 |
2016-17 |
|||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
|
Monthly mean air temperature (oC) |
BSS* (h) |
Humidity (%) |
Monthly mean air temperature (oC) |
BSS* (h) |
Humidity (%) |
||||||||
|
Max. |
Mini. |
Mean |
Morn. |
Even. |
Max. |
Mini. |
Mean |
Morn. |
Even. |
||||
|
October |
34.1 |
18.4 |
26.3 |
8.5 |
81.0 |
33.6 |
34.0 |
17.8 |
25.9 |
7.0 |
85.8 |
43.4 |
|
|
November |
27.6 |
11.7 |
19.6 |
5.6 |
91.5 |
40.8 |
29.1 |
10.2 |
19.6 |
6.4 |
91.3 |
46.5 |
|
|
December |
22.2 |
5.9 |
14.0 |
5.8 |
95.8 |
45.5 |
24.1 |
7.4 |
15.8 |
6.3 |
97.5 |
56.5 |
|
|
January |
19.2 |
6.6 |
12.9 |
4.1 |
94.8 |
64.5 |
19.0 |
7.0 |
13.0 |
4.4 |
98.4 |
70.2 |
|
|
February |
24.4 |
8.3 |
16.4 |
7.0 |
92.6 |
49.6 |
25.0 |
8.0 |
16.5 |
7.9 |
91.0 |
43.0 |
|
| Source: Experimental meteorological station near the research farm, Department of Agriculture Meteorology, CCS Haryana Agricultural University, Hisar | |||||||||||||
| *BSS: Bright sunshine hours | |||||||||||||
Treatments and Experimental Design
The treatments comprised of two potato varieties, i.e. Kufri Surya and Kufri Sadabahar with five doses of nitrogen, i.e. nitrogen 0 kg/ha (control), 75, 150, 225 and 300 kg/ha, laid out in a randomized block design (factorial) with three replications. The mean values of different parameters presented in the Tables were obtained from a pool of data and analyzed by the statistical method demarcated by Panse and Sukhatme (1978). The OPSTAT statistical software (http://14.139.232.166/opstat/index.asp) developed by Chaudhry Charan Singh Haryana Agricultural University, Hisar, Haryana, India was used for this purpose. To evaluate the significance of difference between the means of two treatments, the Critical Difference was computed and all tests of significance were performed at the 5% level of significance.
Crop Management Practices
Well sprouted seed tubers (Approx. 50 to 60 g weight) were planted at a spacing of 60 x 20 cm (83,333 plants per hectare) in plot size of 4.8 x 4 m on 25 October in 2015 and on 20 October in 2016. Recommended doses of phosphorus (50 kg/ha) and potassium (100 kg/ha) for crops and half of the nitrogen dose calculated according to the treatment were applied as bend placement during planting of potato. The remaining half dose of the nitrogen was applied at earthing up time (35 DAP). Nitrogen, phosphorus and potassium were applied in the form of urea, single super phosphate and muriate of potash, respectively. Recommended package of practices was followed for successful crop raising. Haulm killing was performed at 100 days after planting. Two weeks after the skin set, harvesting was carried out in both the years and observations were recorded on various growth and yield parameters as well as uptake of NPK by the tubers and haulms.
From each plot, ten plants were randomly tagged and the heights and above ground biomass (leaves and stem weight) of these plants were measured. Afterwards, the average of the selected plants was calculated for each plot. Harvesting of each plot was done separately and potato tubers were graded in four grade, i.e. < 25 g, > 25–50 g, > 50–75 g and > 75 g and then the number of tubers in each grade were counted and weighted separately and then finally number per square meter and yield quintal per hectare was worked out. Agronomic nitrogen use efficiency (AEN) was computed by the following equation:
Soil Sampling and Analysis
Prior to conducting the study, composite soil samples were taken from the field to asoil depth of 15 cm depth of soil and subjected to analysis to assess the initial physico-chemical properties of the soil before application of treatments. To estimation of NPK leftover in soil after harvest of crop, soil samples collected from each plot up to a depth of 0 to 15 cm after harvesting of crop were dried under shade and then crushed into fine powder with wooden pestle and mortar and passed through 2 mm sieve. These prepared soil samples were used to analyse for N, P and K content available in soil as per methods described in Table 2.
|
Constituent |
Values |
Method |
|---|---|---|
|
Sand (%) |
73.4 |
International pipette method (Piper, 1966) |
|
Silt (%) |
15.6 |
International pipette method (Piper, 1966) |
|
Clay (%) |
11.0 |
International pipette method (Piper, 1966) |
|
pH (1:2) soil:water suspension |
8.0 |
pH meter having glass electrode (Walkley and Black, 1934) |
|
EC (dS/m at 25° C) |
0.3 |
Walkley and Black (1934) |
|
Organic carbon (%) |
0.4 |
Walkley and Black (1934) |
|
Available nitrogen (kg/ha) |
126.7 |
Alkaline Permanganate method (Subbiah and Asija, 1956) |
|
Available phosphorus (kg/ha) |
18.5 |
Sodium bicarbonate soluble P method (Olsen et al., 1954) |
|
Available potassium (kg/ha) |
210.0 |
Ammonium acetate method (Jackson, 1973) |
Plant Sampling and Analysis
To estimation of nitrogen, phosphorus and potassium uptake by the tubers and haulms, samples of tuber and haulms were collected just after harvesting from each plot. After washing, samples were dried in hot air oven at 65°C for three days. The dried samples were ground in stainless steel Wiley mill, passed through sieve of 40-mesh size. To analyse nitrogen, phosphorus and potassium (NPK) uptake by tubers and haulms, the prepared tuber samples were digested in diacid mixture of H2SO4 and HClO4 (4:1). The final volume of the digested samples was made 50 ml by using distilled water, followed by filtration and storage for further assessment. The NPK uptake was analysed by the methods as mentioned in Table 3.
|
Sr. No. |
Nutrient |
Method used |
Reference |
|---|---|---|---|
|
1. |
N |
Nessler’s reagent method |
Lindner (1944) |
|
2. |
P |
Ammonium Vanadomolybdo-Phosphoric acid yellow colour method |
Koenig and Johnson (1942) |
|
3. |
K |
Flame photometric method |
Jackson (1973) |
| N nitrogen, P phosphorus, K potassium | |||
Growth Characters
As the nitrogen levels increased from zero to 300 kg/ha, the plant height, leaves, and stem weight per hill, all increased continuously (Table 4). The nitrogen level 300 kg/ha recorded the maximum plant heights (71.3 and 80.0 cm at 60 and 90 days after planting), leaves and stem weights of 263.5 g and 145.1 g, respectively. The dose of 225 kg/ha was a close second, with plant heights of 68.3 and 76.4 cm, and leaves and stem weights of 262.9 g and 141.9 g, respectively. With increased levels of nitrogen fertilizer, potato plants grew taller and more vigorous which could be possibly due to better uptake of nitrogen by the plants and its positive effect on promoting synthesis and accumulation of proteins and carbohydrates, essentially needed for the multipication and elongation of cells, contributing to the plant growth. The outcomes of the present study are in agreement with those of Banjare et al. (2014), Mozumder et al. (2014), Sharma et al. (2015) and Sriom et al. (2017), who concluded that the consecutive increase in nitrogen levels had a significant effect on plant height, weight of stems and leaves per hill as compared to lower levels of nitrogen.
As compared to variety Kufri Surya, Kufri Sadabahar gave significant higher values for plant height at 60 and 90 days after planting (66.3 and 73.9 cm), weight of leaves (232.1 g) and weight of stems (122.1 g) per hill (Table 4). The effective nitrogen use by the plant foliage for carbohydrates synthesis, essential for growth could be the reason behind the vigorous growth of Kufri Sadabahar plants. The growth parameters of the variety may differ due to its genetic makeup (Kumar et al., 2004). The results of this study are in agreement with the findings of Dubey et al. (2012) and Mozumder et al. (2014) regarding the various growth parameters among various varieties. Additionally, there was no significant interaction effect between nitrogen levels and varieties for all the growth parameters under study.
Yield Characters
The data depicted in Table 4 portray that the different nitrogen levels and the varieties of potato, significantly increases the number and weight of tubers in grade > 25–50, > 50–75 and > 75 as well as total with increasing level of nitrogen up to 225 kg/ha which was significantly at par with higher dose, i.e. 300 kg/ha. It means that the increased dose of nitrogen more than 225 kg/ha has negative effect on tuber number and yield per unit area. Nitrogen dose of 225 kg/ha noted higher tuber number per square metre area of grade > 25–50 (11.1), > 50–75 (10.3), > 75 (13.4) and total (47.8), which increased to a tune of 32.1%, 41.0%, 42.5 and 19.2% as compared to control, respectively. Similarly, maximum weight of tubers of grade > 25–50 (54.5 q/ha), > 50–75 (85.8 q/ha), > 75 (228.1 q/ha) and total yield (388.7 q/ha) was recorded with nitrogen level 225 kg/ha followed by nitrogen level 300 kg/ha and increased to tune of 42.6%, 58.8%, 56.0% and 48.8% over control. The number and weight of tubers of grade < 25 g was recorded non-significant among different nitrogen levels and varieties.
The influence of nitrogen fertilization on different grade as well as total tuber number and yield could be explained that leaf chlorophyll content and plant growth are improved by the nitrogen fertilization, which might have stimulated ample metabolites to produce higher number of tubers and yield per unit area. The findings of present investigation are consistent with those reported by Sahu et al. (2016) and Banjare et al. (2014), who found the maximum number and tuber yield in 50–75 and > 75 g grade with the nitrogen level of 225 kg/ha, whereas, Etemad and Sarajuoghi (2012) noticed the highest tuber number per square metre by applying 200 kg/ha nitrogen.
The variety Kufri Sadabahar revealed significantly more number and weight of tuber of different grade as well as total per unit area over Kufri Surya (Table 4). The variations in these varieties might be attributable to their genetic composition. The higher yield of a variety could be attributed to its increased efficacy in transporting photosynthate from source to sink.
|
Treatments |
Plant height (cm) |
Leaf weight per hill (g) |
Stem weight per hill (g) |
Number of tubers per sq.m. |
Tubers yield (q/ha) |
|||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
|
60 DAP |
90 DAP |
< 25 g |
> 25–50 g |
> 50–75 g |
> 75 g |
Total |
< 25 g |
> 25–50 g |
> 50–75 g |
> 75 g |
Total |
|||
|
Nitrogen levels (kg/ha) |
||||||||||||||
|
0 |
54.7 |
59.6 |
138.0 |
61.8 |
15.0 |
8.4 |
7.3 |
9.4 |
40.1 |
22.8 |
38.2 |
54.0 |
146.2 |
261.2 |
|
75 |
63.4 |
69.3 |
204.1 |
95.1 |
14.9 |
9.7 |
7.6 |
11.0 |
43.2 |
18.5 |
46.2 |
69.2 |
183.3 |
317.2 |
|
150 |
65.7 |
72.3 |
257.0 |
136.9 |
13.0 |
10.7 |
9.4 |
12.5 |
45.6 |
15.8 |
48.7 |
74.5 |
212.0 |
351.0 |
|
225 |
68.3 |
76.4 |
262.9 |
141.6 |
13.0 |
11.1 |
10.3 |
13.4 |
47.8 |
20.3 |
54.5 |
85.8 |
228.1 |
388.7 |
|
300 |
71.3 |
80.0 |
263.5 |
145.1 |
13.6 |
11.0 |
9.8 |
13.1 |
47.5 |
21.2 |
49.2 |
80.4 |
220.1 |
370.9 |
|
SE± |
1.59 |
1.83 |
4.07 |
3.89 |
0.4 |
0.2 |
0.1 |
0.2 |
0.4 |
1.8 |
2.0 |
4.1 |
10.5 |
11.0 |
|
CD (p = 0.05) |
4.8 |
5.0 |
6.1 |
7.8 |
N.S. |
0.4 |
0.6 |
0.7 |
1.3 |
N.S. |
4.5 |
8.3 |
11.6 |
30.2 |
|
Potato varieties |
||||||||||||||
|
Kufri Surya |
63.0 |
69.2 |
218.1 |
110.1 |
14.5 |
10.0 |
8.4 |
11.2 |
44.1 |
20.6 |
45.1 |
66.0 |
179.0 |
310.7 |
|
Kufri Sadabahar |
66.3 |
73.9 |
232.1 |
122.1 |
13.5 |
10.6 |
9.2 |
12.5 |
45.8 |
18.7 |
49.6 |
79.5 |
217.0 |
364.8 |
|
SE± |
1.01 |
1.16 |
2.57 |
2.46 |
0.2 |
0.1 |
0.1 |
0.1 |
0.3 |
1.3 |
1.4 |
2.2 |
6.1 |
8.0 |
|
CD (p = 0.05) |
3.0 |
2.5 |
5.7 |
4.4 |
N.S. |
0.3 |
0.4 |
0.6 |
0.9 |
N.S. |
3.0 |
6.4 |
9.3 |
19.5 |
| Interaction: nitrogen levels x variety was non-significant | ||||||||||||||
SE standard error, CD critical difference, DAP days after planting, N.S. not significant
Agronomic Nitrogen Use Efficiency
Agronomic nitrogen use efficiency (AEN) declined linearly with increase in nitrogen rate (Fig. 1) and it was 74.7 kg tuber kg− 1 N at 75 kg nitrogen dose per hectare, which reduced to 36.6 kg tuber kg− 1 N at the maximum nitrogen level (75 kg N/ha). The response of nitrogen rate was quadratic for agronomic nitrogen use efficiency (R2 = 0.935).
Nutrients Uptake by the Tubers and Haulms
The data depicted in Table 5 indicated that nitrogen, phosphorus and potash uptake by the tubers (74.7, 19.7and 173.0 kg/ha, respectively) was recorded maximum for the nitrogen rate 225 kg/ha (Fig. 2) closely followed by 300 kg/ha nitrogen dose (72.6, 19.0 and 168.4 kg/ha, respectively). The significantly highest nitrogen, phosphorus and potassium uptake by the haulms (41.9, 11.3 and 116.5 kg/ha, respectively) was recorded for the nitrogen rate 300 kg/ha closely followed by 225 kg/ha nitrogen dose (41.5, 10.6 and 114.5 kg/ha, respectively). The increased accumulation of nitrogen, phosphorus and potash in tubers and haulms might be due to more absorption of these elements from the soil caused by better growth and development of root and aerial portion with successive increase in fertility levels. In potato plants, Getie et al. (2015) reported a progressive increase in nitrogen, phosphorus and potassium uptake by the plants with the increase in their rates of application.
|
Treatments |
Nutrients uptake by tubers (kg/ha) |
Nutrients uptake by haulms (kg/ha) |
Nutrients available in soil after harvest (kg/ha) |
||||||
|---|---|---|---|---|---|---|---|---|---|
|
N |
P |
K |
N |
P |
K |
N |
P |
K |
|
|
Nitrogen levels (kg/ha) |
|||||||||
|
0 |
48.1 |
11.1 |
103.4 |
25.2 |
4.0 |
62.0 |
116.7 |
24.3 |
244.3 |
|
75 |
57.1 |
14.0 |
139.3 |
32.8 |
6.3 |
100.0 |
141.8 |
23.9 |
241.3 |
|
150 |
64.7 |
18.1 |
159.3 |
38.4 |
9.5 |
112.5 |
180.0 |
22.2 |
235.0 |
|
225 |
74.7 |
19.7 |
173.0 |
41.5 |
10.6 |
114.5 |
197.0 |
18.6 |
225.4 |
|
300 |
72.6 |
19.0 |
168.4 |
41.9 |
11.3 |
116.5 |
210.8 |
18.6 |
226.8 |
|
SE± |
1.48 |
0.9 |
1.20 |
1.11 |
0.43 |
0.99 |
1.98 |
1.31 |
1.04 |
|
CD (p = 0.05) |
4.4 |
2.7 |
3.6 |
3.3 |
1.3 |
3.0 |
5.9 |
3.9 |
3.1 |
|
Potato varieties |
|||||||||
|
Kufri Surya |
59.1 |
15.3 |
140.4 |
33.6 |
7.2 |
98.8 |
176.7 |
22.2 |
235.8 |
|
Kufri Sadabahar |
67.8 |
17.4 |
157.0 |
38.3 |
9.5 |
103.4 |
161.8 |
20.8 |
233.2 |
|
SE± |
0.94 |
0.57 |
0.76 |
0.71 |
0.27 |
0.63 |
1.25 |
0.83 |
0.65 |
|
CD (p = 0.05) |
2.8 |
1.7 |
2.3 |
2.1 |
0.8 |
1.9 |
3.7 |
N.S. |
2.0 |
N nitrogen, P phosphorus, K potassium, SE standard error, CD critical difference, N.S. not significant
The significantly highest value for available nitrogen in soil after harvesting of crop (210.8 kg/ha) was noticed with 300 kg/ha nitrogen level, while for phosphorus, the same was recorded for the treatment with zero nitrogen (control) application (24.3 kg/ha), closely followed by 75 and 150 kg/ha nitrogen levels, and for potassium, it was noticed significantly highest (244.3 kg/ha) for the control treatment, closely followed by the nitrogen rate 75 kg/ha (Table 5). The higher leftover values for nitrogen in soil where its highest rate was applied might be due to its application excess than the crop requirement, while the higher leftover values for phosphorus and potassium in soil where no nitrogen was given might be due to less utilization of these elements on account of poor plant growth and tuber yield.
Between the varieties significant maximum available nitrogen and potassium, in soil after harvest of crop, was observed in the variety Kufri Surya (176.7 and 235.8 kg/ha) as compared to Kufri Sadabahar (161.8 and 232.2 kg/ha, respectively), whereas, the effect of variety was found non-significant for available phosphorus in soil after harvesting of crop. No significant interaction was present between the nitrogen levels and the potato varieties for the available nitrogen, phosphorus and potassium in soil after harvesting of the crop.
The interaction effect was found significant between the nitrogen levels and the potato varieties for potassium uptake by the tubers, which was registered highest for the variety Kufri Sadabahar with 225 kg/ha nitrogen level, closely followed by 300 kg/ha nitrogen rate for the same variety (Table 6). These findings are in accordance with the results of Jamaati-e-Somarin et al. (2010), they recorded the significantly highest tuber potassium uptake with increasing nitrogen level up to 200 kg/ha. Patel et al. (2012) observed that 75% recommended dose of nitrogen and potassium (206 kg/ha each) supplied as fertigation in five splits resulted in increased total nitrogen, phosphorus and potash uptake by the potato tubers.
|
Nitrogen levels (kg/ha) |
Potassium uptake by the tubers (kg/ha) |
||
|---|---|---|---|
|
Varieties |
Mean |
|
|
|
Kufri Surya |
Kufri Sadabahar |
||
|
0 |
102.0 |
104.8 |
103.4 |
|
75 |
130.3 |
148.3 |
139.3 |
|
150 |
152.4 |
166.1 |
159.3 |
|
225 |
160.7 |
185.2 |
173.0 |
|
300 |
156.4 |
180.4 |
168.4 |
|
Mean |
140.4 |
157.0 |
|
|
CD (p = 0.05): Nitrogen: 3.6, Variety: 2.3, Nitrogen x Variety: 5.1 |
|||
| CD critical difference | |||
In general, this study indicates that yield and yield components of potato variety Kufri Sadabahar improved over Kufri Surya in Haryana state of India. The results have also revealed that the current recommendation rate of nitrogen for potato production (150 kg N/ha) is obsolete and should be revised for enhancing the yield of these newly developed variety. Thus, it could be concluded that the application of 225 kg/ha nitrogen along with variety Kufri Sadabahar leads to highest production of fresh tuber yields in Hisar district of Northern Plains of India and on soils with similar physico-chemical characteristics of the experimental soil.
Acknowledgements: The author thankfully acknowledges the Chaudhary Charan Singh Haryana Agricultural University for providing facilities and necessary help to carry out this research work.
Author Contribution: R.Y. contributed to the execution of experiment, data collection, analysis of study and preparing first draft of manuscript. V.P.S.P. contributed to the planning, designing, monitoring of the research work, data analysis and interpretation, drafting and revision of manuscript as well as final submission to journal. R and R.P. contributed to the study’s conception, monitoring of study, data analysis and interpretation.
All authors approved the final version of the manuscript and agree to be accountable for all aspects of the work.
Data Availability: Available with RY
Ethics approval: Not applicable
Consent to participate: Not applicable
Consent for publication: The authors provide their consent for publication.
Conflict of interest: The authors declare no competing interests.
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