Introduction

Groundnut (Arachis hypogaea L.) is a vital oil kernel crop in the tropics and subtropics countries (Pasupuleti et al. 2013). The groundnut is cultivated in many countries between latitudes 40˚N and 40˚S in semiarid tropics and subtropics (Kayondo et al. 2014). Groundnut seeds contain about 12–15% carbohydrates, 25–30% protein and 40–50% fats (Saeed and Hassan 2009). Additionally, groundnuts contain a good source of vitamin, dietary fibres, and minerals such as niacin, magnesium, iron, phosphorus, calcium, zinc, and riboflavin (Tshilenge-Lukanda et al. 2013). In addition, groundnut as a legume crop improves soil fertility by converting the atmospheric nitrogen to nitrates, ammonia, and organic nitrogen (Pasupuleti et al. 2013). Thus, groundnut production plays a great role in developing and developed nations in improving the economic status (Tsigbey et al. 2003).

In Tanzania, the crop is cultivated mainly by smallholder farmers in Tabora, Mtwara, Dodoma, Singida, Shinyanga and Mwanza regions. The crop acts as a good source of food, cash crop, and animal feed (Osei et al. 2013). Despite the importance of groundnut in Tanzania, its average yield is still low amounting to 960 kg ha^-1 as opposed to the predictable yield potential of 2500 kg ha^-1 in developing countries (Philipo and Nchimbi-Msolla 2019). The crop is mainly constrained by drought stress, low level of inputs, foliar fungal diseases, and insect pest attacks.

The foliar fungal diseases namely, early leaf spot, late leaf spot, and rust are among the most destructive diseases, which account for huge economic yield losses (Naidu et al. 1999). The late leaf spot disease is one among the three diseases, which has been identified as a major constrain wherever groundnut is cultivated in Tanzania. The late leaf spot disease infection reduces the photosynthetic area by causing intense lesions on stems, leaves, and petioles consequently leading to defoliation and hence high yield losses (Monfort et al. 2004; Khedikar et al. 2010). According to Ghewande (1989), when the late leaf spot disease attacks groundnuts, it reduces about 80 and 93% canopy carbon exchange rate and carbon uptake respectively. Late leaf spot disease also causes adverse outcomes on seed and folder quality becoming unsuitable for animal feed (Monfort et al. 2004).

Efforts have been made in controlling late leaf spot disease through the development and the use of synthetic fungicides, which have been proven effective. However, the effectiveness of synthetic fungicides depends upon multiple fungicide applications; hence, smallholder farmers cannot afford the cost. Moreover, there are issues of environmental and health concerns (Jordan et al. 2012). The plants possess phytochemical compounds, which are effective against different pests including insects, fungi, virus, nematode, bacteria etc. (Khan et al. 2020; Javaid et al. 2020, 2021). Thus, plant based materials can be used as a substitute for synthetic pesticides in order to ensure the safety of human and the environment (Engindeniz and Engindeniz 2013; Khan and Javaid 2020; Banaras et al. 2021).

The Jatropha curcas belonging to Euphorbiaceae family is a multipurpose plant that survives in both tropics and arid regions. Almost all the plant parts of Jatropha are reported to possess antimicrobial potential against disease causing pathogens including fungi (Prasad et al. 2012; Dada et al. 2014). Moreover, J. curcas leaf possesses important compounds such as sterols, terpenes, flavonoids, saponin and steroids steroids, which play a great antifungal role (Nwosu and Okafor 1995; Campa et al. 2008; Saetae and Worapot 2010). Many findings confirm the antifungal potential of J. curcas in managing fungal diseases. A study by Thangavelu et al. (2004) found that J. curcas was effective in managing banana anthracnose disease. In addition, J. curcas leaf extracts were found effective in managing Azolla disease caused by Sclerotium spp. (Garcia and Lawas 1990). However, little is known about the effectiveness of J. curcas against P. personata pathogen causing the late leaf spot disease on groundnuts. Thus, the present study evaluated the efficacy of J. curcas against the groundnut late leaf spot disease and analysed the possible compounds exhibiting antifungal properties.

Materials and Methods

Isolation of pathogen and culture preparation

Groundnut leaves showing symptoms of P. personata (circular spots underneath the leaflets) were obtained from the farmer’s fields in Singida and Dodoma regions, Tanzania. Fungal isolation was done by adopting the method described by Kishore et al. (2007) with some modification. The diseased portions of leaves were cut into pieces (0.5–1 cm) and sterilized with 5% NaOCl. Thereafter, the pieces were rinsed three times in the sterilized distilled water and dried on a blotter paper in Petri dishes. Thereafter, the pieces were plated onto potato dextrose agar (PDA) medium in a laminar hood then incubated at 28 ± 2°C for 7 days. After the emergence of mycelial growth, each fungal colony was sub-cultured into fresh PDA plates and incubated at 28 ± 2°C for 7 days to obtain the pure culture of P. personata. The P. personata pathogen was identified by a single spore method under compound microscope (magnification 40X).

Preparation of J. curcas leaf extracts

J. curcas leaves were collected from different parts in Arusha, Tanzania. The leaves were washed thoroughly and then air-dried at room temperature. Thereafter, these were pounded into powder. The fine powdered leaf sample (1 kg) was successively extracted through chloroform, ethyl acetate, and lastly methanol (48 h in each) at standard room temperature. Then, the leaf extracts were filtered with a Whatman No.1 filter paper and concentrated by a rotary evaporator. The product, which was a dark sticky semisolid extracts, was then stored under cold condition (4°C) for further experiment.

Greenhouse experiment on antifungal assay

The study was carried out to test the effectiveness of methanol, chloroform and ethyl acetate leaf extracts of J. curcas against late leaf spot disease under greenhouse condition. Three groundnut seeds (Upendo variety) were grown in a plastic pots filled with a mixture of black soil (later thinned to one). The plants were artificially inoculated by J. curcas conidial suspension 30 days after sowing (DAS). After inoculation, the plants were shielded with plastic sheets for 48 h to maintain leaf wetness during the nights. Four foliar sprays were applied onto the plants each J. curcas leaf extract at 0.1, 0.25 and 0.5 mg mL^-1 concentration, chlorothalonil (2 mL L^-1) (positive control) and sterile distilled water (negative control) using a hand sprayer at 14 days interval. The plants were sprayed starting from 48 DAS and completed two weeks before the harvest. The experiment was inspected often and the data on disease, growth, and yield were recorded. The trial was set in a completely randomized design replicated three times. The experiment was repeated twice. The disease incidence was assessed on each plant by evaluating the percent of the infected leaves per plant by adopting the formula by Subrahmanyam et al. (1995).

The late leaf spot disease severity under different treatments was scored using (1–9) disease rating scale (Chiteka et al. 1988).

Gas chromatography mass spectroscopy analysis

The phytochemical analysis of J. curcas leaf extracts was done using gas chromatography-mass spectroscopy (GC-MS) on Agilent technologies 7890A GC connected to Agilent 5975 MSD (Agilent technology, USA), comprising a 30 m length and film 0.25 µm and internal diameter of 0.250 mm and temperature limit of 50°C to 340°C (360°C). The inert gas helium was used as a carrier gas with 1.2 mL min^-1 flow rate. The inlet temperature was 250°C and the total running time was 35 min. The obtained peaks were compared with the known compounds spectra stored in the National Institute Standard and Technology library.

Data analysis

Data were subjected to 3-way ANOVA (analysis of variance) in factorial arrangement, using STATISTICA program. The treatment means were compared by applying Fischer’s least significant difference (LSD) at 5% level of significance.

Results

In this study, the effectiveness of J. curcas leaf extracts against the late leaf spot disease was determined by observing their effect on reducing the disease incidence and severity. Moreover, growth and yield attributes were also assessed. Three leaf extract of J. curcas (methanolic, chloroform, and ethyl acetate), one standard fungicide chlorothalonil (positive control) and one negative control (distilled water) as foliar spray were used as treatments against late leaf spot disease. The effects of treatments, solvents and concentrations on late leaf spot disease incidence, severity, growth and yield attributes are presented in Table 1, 2 and 3.

The late leaf spot disease varied significantly (P ≤ 0.001) with the effect to treatments. The plants treated by Jatropha leaf extracts had lower late leaf spot disease incidence (13.33%) similar to the standard fungicide chlorothalonil (5.41%). Moreover, the late leaf spot disease incidence differed significantly (P ≤ 0.01) with the type of solvents used for extraction. Methanolic leaf extract of J. curcas had lower late leaf spot disease incidence (24.7%) as compared to chloroform (36.89%) and ethyl acetate (36.59%,) extracts (Table 1). Moreover, the results showed that the late leaf spot disease incidence and severity differed significantly (P ≤ 0.001) from the effect of J. curcas leaf extracts concentration. J. curcas leaf extracts at the highest concentration (0.5 mg mL^-1) significantly reduced the late leaf spot disease incidence as compared to the lowest concentration (0.1 mg mL^-1) (Table 1).

Growth parameters varied significantly (P ≤ 0.001) with the effect of treatments, solvents and concentrations of J. curcas leaf extracts (Table 2). The plants treated with chlorothalonil and J. curcas had taller shoots and big number of leaves per plant both at flowering and maturity (38.59 cm, 8.93) (47.26 cm, 12.89), (27.78 cm, 6.74) and (36.07 cm, 9.48) respectively compared with the control. Similarly, growth parameter differed significantly (P ≤ 0.05) from the type of solvents used for extraction. The methanolic and chloroform leaf extracts of J. curcas had taller shoots (30.48 cm, 29.56 cm), (38.63 cm, and 36.96 cm) at both flowering and maturity respectively compared with ethyl acetate extracts (27.52 cm, 34.93 cm). Additionally, shoot length and big number of leaves per plant differed significantly (P ≤ 0.001) at different J. curcas leaf extracts concentrations. The plants treated with plant extracts at the highest concentration (0.5 mg mL^-1) had taller shoots and bigger number of leaves per plant at flowering and maturity (31.89 cm, 7.88), (39.9 3 cm, 9.93), respectively compared with the control (25.91 cm, 5.82), (33.48 cm, 7.81) at both flowering and maturity (Table 2).

Likewise, yield attributes components varied significantly (P ≤ 0.001) with the treatments where J. curcas leaf extracts had bigger number of pods per plant, seeds per plant and seed yield (ton ha^-1). J. curcas was less similar to the standard fungicides (chlorothalonil) that is, the number of pods per plant (32.9), the number of seeds per plant (61.2) and seed yield (1.6 ton ha^-1) (Table 3). Yield data did not differ significantly with the effect to solvents, similar results were observed. The yield attributes differed significantly (P ≤ 0.01) with the effect to J. curcas leaf extracts concentration. J. curcas leaf extracts at the highest concentration had bigger number of pods plant^-1 (34.59), the number of seeds per plant (63.19), 100 seed weight (53.09 g) and seed yield (1.85 ton ha^-1) (Table 3).

The GC-MS results led to the identification of different phytochemical constituents from fractions of ethyl acetate, methanolic, and chloroform leaf extracts of J. curcas. The mass spectra of the detected compounds from methanolic, chloroform and ethyl acetate leaf extracts of J. curcas were compared with the spectra of the recognized compounds in the NIST library. The name of compound, molecular weight, retention time, and molecular formula of the compounds contained in these leaf extracts are presented in Tables 4, 5 and 6. The following phytoconstituents with antifungal properties were recognized by GC-MS from the chloroform leaf extract of J. curcas; dodecane; 2,6,11-trimethyl-2-tetradecene; tetradecane; pentadecane; octacosane; sulfurous acid butyl decyl ester; 2-bromo heneicosane; phenol 2,4-bis (1, 1-dimethylethyl); hexadecane; heptadecane; heptacosane; 2,4-dimethyldodecane; n-hexadecanoic acid; ethanol 2-(octadecyloxy)-; hentriacontane; geranylgeraniol; octadecane; 12-methyl-E-E-2 13-octadecadien-1-ol; tetradecanal and cyclotetracosane (Table 4). Among them n-hexadecanoic acid (7.89%); phenol 2,4-bis (1,1-dimethylethyl) (4.04%); cyclotetracosane (1.23%); hexadecane (1.20%) and octacosane (1.02%) were the major identified phytoconstituents compounds (Table 4).

Table 1: Late leaf spot disease incidence and severity as affected with treatments, solvent and concentration

Factors	Incidence	Severity
Treatments
Jatropha curcas	13.33 ± 2.02^b	2.26 ± 0.31^b
Chlorothalonil	5.41 ± 1.07^a	1.33 ± 0.22^a
Control	89.41 ± 0.92^c	8.96 ± 0.04^c
Solvents
Chloroform	36.89 ± 7.49^b	3.89 ± 0.63^b
Ethyl acetate	36.59 ± 7.59^b	3.00 ± 0.59^b
Methanol	24.67 ± 7.60^a	2.67 ± 0.63^a
Concentrations
0.1 mg mL^-1	41.89 ± 7.36^c	5.42 ± 0.53^c
0.25 mg mL^-1	35.93 ± 7.39^b	4.00 ± 0.59^b
0.5 mg mL^-1	20.33 ± 7.76^a	2.07 ± 0.69^a
3-WAY ANOVA (F-value)
Treatments	4332.52***	512.564***
Solvents	2390.38**	1.14**
Concentrations	1.62***	20.34***
Treatments*Solvents	37.11ns	4.555**
Treatments*Concentrations	1.57ns	8.04***
Solvents*Concentrations	7.751***	0.38ns
TreatmentsSolventsConcentrations	0.381ns	0.56ns

Means with the same letter(s) were considered statistically not significant at (P = 0.05), Fischer’s least significant difference (LSD) test

Table 2: Growth attributes of groundnut Upendo genotype as affected by treatments, solvents and concentrations

Factors	Shoot length (cm) flowering	Number of branches at flowering	Shoot length at maturity	Number of branches at maturity
Treatments
Jatropha curcas	27.78 ± 1.16^b	6.74 ± 0.27^b	36.07 ± 1.22^b	9.48 ± 0.43^b
Chlorothalonil	38.59 ± 1.10^c	8.93 ± 0.35^c	47.26 ± 1.19^c	12.89 ± 0.47^c
Control	21.19 ± 0.72^a	4.85 ± 0.17^a	27.19 ± 0.71^a	4.48 ± 0.14^a
Solvents
Chloroform	29.56 ± 1.76^b	6.96 ± 0.42^a	36.96 ± 2.00^b	9.33 ± 0.85^b
Ethyl acetate	27.52 ± 1.63^a	6.59 ± 0.40^a	34.93 ± 1.79^a	8.15 ± 0.71^a
Methanol	30.48 ± 1.75^b	6.69 ± 0.45^a	38.63 ± 1.93^c	9.37 ± 0.73^b
Concentrations
0.1 mg mL^-1	25.91 ± 0.95^a	5.82 ± 0.31^a	33.48 ± 1.53a	7.81 ± 0.62^a
0.25 mg mL^-1	29.58 ± 1.10^b	6.81 ± 0.39^b	37. 11 ± 1.95^b	9.11 ± 0.72^b
0.5 mg mL^-1	31.89 ± 1.18^c	7.88 ± 0.47^c	39.93 ± 2.07^c	9.93 ± 0.89^c
3-Way ANOVA (F-value)
Treatments	153.15***	108.23***	179.32***	252.32***
Solvents	4.56*	1.190ns	6.098**	6.819**
Concentrations	17.54***	28.012***	18.49***	15.994***
Treatments*Solvents	10.29***	4.226**	8.58***	7.72***
Treatments*Concentrations	2.251ns	3.51*	3.15*	4.74**
Solvents*Concentrations	0.15ns	1.048ns	0.09ns	0.18ns
TreatmentsSolventsConcentrations	1.20ns	0.896ns	0.923ns	0.96ns

Means with the same letter(s) were considered statistically not significant at (P = 0.05), Fischer’s least significant difference (LSD) test

From ethyl acetate leaf extract of J. curcas, the following phytoconstituents with antifungal activity were identified by GC-MS; 1,2,3-ropanetriol; monoacetate; 2,5-pyrrolidinedione; thiomorpholine; methyl salicylate; triacetin, 1-naphthalenol; 8-hexadecenal; 14-methyl-, (Z)-, undecane; phenol, 2,4-bis (1,1-dimethylethyl); hexadecane; heptadecane; 1H-indene 1-ethylideneoctahydro-7 a-methyl- cis-, E-14-hexadecenal; 1-tetradecene; tetramethyl-2-hexadecen-1-ol; 9,12-octadecadienoic acid (Z,Z); 5-eicosene, (E); hexadecanoic acid ethyl ester; 2-methyl-Z,Z-3,13-octadecadienol,; (Z)-; n-hexadecanoic acid; phytol; 9,12,15-octadecatrienoic acid ethyl ester; (Z,Z,Z)-; heptadecanoic acid ethyl ester and eicosane (Table 5). The major phytoconstituents were phytol (9.31%); thiomorpholine (4.83%); hexadecanoic acid ethyl ester (3.97%); phenol 2,4-bis (1,1-dimethylethyl) (3.37%); 9,12,15-octadecatrienoic acid ethyl ester, (Z,Z,Z)- (2.75%); 5-eicosene, (E)- (2. 11%) and 1-heneicosyl (1.92%) (Table 5).

The phytoconstituents with antifungal property identified by GC-MS in J. curcas methanolic leaf extract were; 1,2,3-propanetriol monoacetate; methyl salicylate; 2-undecanone; decanoic acid methyl ester; 2-methoxy-4-vinylphenol; tert-hexadecanethiol; phenol 2,6-dimethoxy; tetradecane; cyclotetradecane; pentanoic acid ethyl ester; 2-propenoic acid 3-phenyl- methyl ester; diphenyl ether; pentadecane; tridecane; hexadecane; heptadecane; 17-pentatriacontene, 1-nonadecene; E-15-heptadecenal; 8-hexadecenal 14-methyl; cyclopentadecane; hexadecanoic acid methyl ester; 1-octadecene; 2-methyl-Z, Z-3, Table 3: Yield attributes of groundnut Upendo genotype as affected by treatments, solvents and concentrations

Factors	Number of pods/plant	Number of seeds/plant	100 kernel weight (g)	Seed yield (tonnes/ha)
Treatments
Jatropha curcas	32.96 ± 1.13^b	61.15 ± 2.25^b	49.00 ± 2.30^b	1.59 ± 0.09^b
Chlorothalonil	40.30 ± 1.27^c	75.81 ± 2.49^c	54.18 ± 1.50^c	2.49 ± 0.10^c
Control	16.0 ± 0.95^a	24.41 ± 1.67^a	33.85 ± 1.75^a	0.49 ± 0.03^a
Solvents
Chloroform	30.07 ± 2.40^b	53.96 ± 5.04^a	50.07 ± 1.79^a	1.54 ± 0.19^a
Ethyl acetate	28.59 ± 2.18^a	51.33 ± 4.56^a	50.82 ± 1.75^a	1.50 ± 0.18^a
Methanol	30.59 ± 2.25^b	56.07 ± 4.62^a	51.13 ± 1.57^b	1.54 ± 0.17^a
Concentration
0.1 mg mL^-1	24.59 ± 2.00^a	43.85 ± 4.15^a	49.16 ± 1.52^a	1.20 ± 0.14^a
0.25 mg mL^-1	30.07 ± 2.18^b	54.33 ± 4.55^b	49.78 ± 1.71^a	1.52 ± 0.17^b
0.5 mg mL^-1	34.59 ± 2.23^c	63.19 ± 4.76^c	53.09 ± 1.82^b	1.85 ± 0.20^c
3-Way ANOVA (F-value)
Treatments	282.57***	353.15***	38.74***	248.92***
Solvents	1.96ns	2.84ns	0.135*	0.15ns
Concentrations	45.63**	47.17**	0.47*	25.883***
Treatments*Solvents	1.707ns	1.92ns	0.44ns	0.63ns
Treatments*Concentrations	1.179ns	1.47ns	0.43ns	3.136*
Solvents*Concentrations	4.033*	3.669*	0.45ns	1.774ns
TreatmentsSolventsConcentrations	0.238ns	0.212ns	1.20ns	1.482ns

Means with the same letter(s) were considered statistically not significant at (P = 0.05), Fischer’s least significant difference (LSD) test

Table 4: Phytochemical compounds with antifungal activity obtained from chloroform leaf extract of J. curcas

Retention time (min)	Compound name	Molecular formula	Molecular weight (g/mol)	References
10.629	Dodecane, 2,6,11-trimethyl-	C₁₅H₃₂	212.41	(Zhang et al. 2015)
11.745	2-Tetradecene	C₁₄H₂₈	196.37	(Shirani et al. 2017)
11.905	Tetradecane	C₁₄H₃₀	198.39	(Begum et al.2016)
12.460	Pentadecane	C₁₈H₃₈	254.49	(Zhang et al. 2015)
12.958	Octacosane	C₂₈H₅₈	394.76	(Zhang et al.2018)
13.192	Sulfurous acid butyl decyl ester	C₁₆H₃₄O₃S	306.50	(Sharma et al. 2019)
13.267	Heneicosane	C₂₁H₄₄	296.57	(Ebrahimabadi et al. 2016)
13.461	Phenol 2,4-bis(1, 1-dimethylethyl)	C₁₄H₂₂O	206.32	(Manikandan et al. 2017)
14.011	2-Bromo dodecane	C₁₂H₂₅Br	249.23	(Manikandan et al. 2017)
14.503	Hexadecane	C₁₆H₃₄	226.44	(Zhang et al. 2015)
15.041	Heptadecane, 9-octyl-	C₂₅H₅₂	352.68	(Musa et al.2015)
15.401	Heptacosane	C₂₇H₅₆	380.73	(Bouzabata et al.2018)
16.002	2,4-Dimethyldodecane	C₁₄H₃₀	198.38	(Begum et al. 2016)
16.488	Pentadecane	C₁₅H₃₂	212.41	(Yuan et al.2012; Zhang et al. 2015)
17.009	Ethanol, 2-(octadecyloxy)-	C₂₀H₄₂O₂	314.50	(Mohy and Mohyeldin 2018)
18.067	Octacosane	C₂₈H₅₈	394.76	(Zhang et al.2018)
18.142	Hentriacontane	C₃₁H₆₄	436.84	(Ruban and Gajalakshmi 2012)
18.457	Geranylgeraniol	C₂₀H₃₄O	290.48	(Ashraf et al. 2017)
18.542	Octadecane	C₁₈H₃₈	254.49	(Zhang et al. 2018)
18.869	n-Hexadecanoic acid	C₁₆H₃₂O₂	256.42	(Omoruyi et al. 2014)
19.584	12-Methyl-E-E-2, 13-octadecadien-1-ol	C₁₉H₃₆O	280.00	(Vijayabaskar and Elango 2018).
20.013	Tetradecanal	C₁₄H₂₈O	212.37	(Passos et al. 2003)
29.037	Cyclotetracosane	C₂₄H₄₈	336.64	(Bughio et al. 2017)

13-octadecadienol; oleic acid, 9,17-octadecadienal, (Z); 2-methyl-Z,Z-3,13-octadecadienol; 9, 12-octadecadienoic acid (Z,Z)-methyl ester; phytol; octadecanoic acid methyl ester; behenic alcohol; octadecanoic acid ethyl ester; 3,7,11,15-tetramethyl-2-hexadecen-1-ol; 9,17-octadecadienal; eicosane and docosanoic acid methyl ester (Table 6). Phytol (26.75%); hexadecanoic acid methyl ester (14.32%); octadecanoic acid methyl ester (2.79%) and 9,12-octadecadienoic acid (Z,Z)- methyl ester (2.33%) were identified as major phytoconstituents (Table 6).

Discussion

The in vivo studies confirmed the efficacy of J. curcas by lowering the disease incidence and severity as the concentration increased. The lowest late leaf spot disease incidence and severity were achieved with both J. curcas leaf extracts, similar to the standard fungicide (chlorothalonil). This corresponds with the findings of Thangavelu et al. (2004), who revealed that the leaf extract of J. curcas effectively controlled Colletotrichum musae and Sclerotium spp. causal agents for anthracnose disease in banana and Azolla, respectively. Methanolic extracts showed the lowest late leaf spot disease incidence and severity compared to ethyl acetate and chloroform extracts. This suggests that more polar compounds Table 5: Phytochemical compounds with antifungal activity obtained from ethyl acetate leaf extract of J. curcas

Retention time (min)	Compound name	Molecular formula	Molecular weight (g/mol)	References
7.539	1,2,3-Ropanetriol, monoacetate	C₅H₁₀O₄	134.13	(Teoh and Mashitah 2012)
8.460	2,5-Pyrrolidinedione	C₈H₁₃NO₂	331.32	(Takayama et al. 1982)
8.826	Hexadecane	‎C₁₆H₃₄	‎226.44	(Adeleye et al. 2010)
9.273	Methyl salicylate	C₈H₈O₃	‎152.15	(Pawar and Thaker 2006)
11.321	Triacetin	C₉H₁₄O₆	218.21	(Osuntokun and Olajubu 2014)
11.813	Heptadecane	C₁₇H₃₆	240.5	(Zhang et al., 2015)
11.899	8-Hexadecenal, 14-methyl-, (Z)-	C₁₇H₃₂O	252.4	(Osuntokun and Olajubu 2014)
12.952	Undecane	C₁₁H₂₄	156.31	(Wanxi et al. 2013)
13.467	Phenol,2,4-bis(1,1-dimethylethyl)	C₁₇H₃₀OSi	278.50	(Jun et al. 2018)
13.993	1-Naphthalenol	C₁₀H₈O	144.17	(Kumar et al. 2012)
14.503	Hexadecane	C₁₆H₃₄	226.41	(Oliveira et al. 2014)
15.658	Heptadecane	C₁₇H₃₆	240.48	(Zhang et al. 2015)
16.889	E-14-Hexadecenal	C₁₆H₃₀O	238.41	(Devakumar et al. 2017)
17.106	1-Tetradecene	‎C₁₄H₂₈	196.37	(Tayung and Jha 2014)
17.896	Tetramethyl-2-hexadecen-1-ol	C₂₀H₄₀O	296.50	(Mohy and Mohyeldin 2018)
18.868	n-Hexadecanoic acid	‎C₁₆H₃₂O₂	256.42	(Tyagi and Agarwal 2017)
18.983	9,12-Octadecadienoic acid (Z,Z)-	C₁₉H₃₄O₂	280.40	(Mohy and Mohyeldin, 2018)
19.109	5-Eicosene, (E)-	C₂₀H₄₀	280.50	(Adibe et al. 2019)
19.172	Hexadecanoic acid ethyl ester	C₁₈H₃₆O₂	284.47	(Mohy and Mohyeldin 2018)
19.338	2-Methyl-Z,Z-3,13-octadecadienol	C₁₉H₃₆O	280.50	(Adibe et al. 2019)
20.179	9,17-Octadecadienal, (Z)-	‎‎C₁₈H₃₂O	264.40	(Adibe et al. 2019)
20.413	Phytol	C₂₀H₄₀O	296.54	(Pejin et al. 2014)
21.008	9,12,15-Octadecatrienoic acid ethyl ester, (Z,Z,Z)-	C₂₀H₃₄O₂	306.48	(Mohy and Mohyeldin 2018)
21.186	Heptadecanoic acid ethyl ester	C₁₉H₃₈O₂	298.50	(Bashir et al. 2019)
23.869	Eicosane	C₂₀H₄₂	282.50	(El-Naggar et al. 2017)

extracted by methanol had antifungal property slightly greater than had those extracted by ethyl acetate and chloroform. This finding is consistent with the findings by Igbinosa et al. (2009) who revealed that, the stem bark methanolic extract of J. curcas inhibited the growth of Escherichia coli, Bacillus subtilis and Proteus vulgaris. Moreover, according to Kalimuthu et al. (2010) the methanolic extract of J. curcas inhibited Pseudomonas, Klebsiella, E. coli and Staphylococcus aureus. Moreover, the J. curcas leaf extracts at the highest concentration significantly reduced late leaf spot disease incidence and severity as compared to the lowest concentration. This finding corresponds with the finding of an investigation by Amah and Aliero (2009) who revealed that disease incidence and severity were reported as being low in plants treated with plant extract at the highest concentration.

Growth parameters varied significantly with the effect of treatments, solvents and concentration. The plants treated with Chlorothalonil and J. curcas had taller shoots and bigger number of leaves per plant at both flowering and maturity compared with the control. In addition, the plants treated with methanolic and chloroform extracts had taller shoots and bigger number of leaves per plant, at both flowering and maturity compared with ethyl acetate extracts. Moreover, yield attributes components varied significantly with J. curcas leaf extracts concentrations, where J. curcas leaf extracts at the highest concentration influenced groundnut yield similar to the standard fungicides (Chlorothalonil). This observation is consistent with the results by Ghewande (1989) who found leaf extracts of Azadirachta indica and Lawsonia inermis effective in managing both groundnut late leaf spot and rust diseases and increased yield by 15–40% under field conditions. For this case, methanolic, ethyl acetate and chloroform leaf extracts of J. curcas were found effective against late leaf spot disease LLS subsequently improved the growth and yield of groundnuts compared with the control treatments.

GC-MS analysis was performed on chloroform, ethyl acetate and methanolic extracts of J. curcas since they exhibited the antifungal activity under in vivo experiment. The GC-MS identified the presence of different phytoconstituents from chloroform), ethyl acetate and methanolic leaf extracts of J. curcas. The qualitative differences of phytochemical constituents observed in this study may be attributed by different solvents employed for extraction. This observation corresponds with the findings by Kordali et al. (2009), who reported that, the spectra solubility of phytochemicals depends on the type of solvent used for extraction. In addition, phytochemical differences could be the result of the habitat for plant growth. This is consistent with Farooq et al. (2007) finding that, the phytochemical compounds composition depends on the plant habitat. The phytochemical analysis revealed the existence of octadecanoic acid; hexadecanoic acid methyl ester (palmitic acid); 9, 12-octadecadienoic acid (Z,Z) methyl ester and phytol in J. curcas leaf extracts. Amongst them hexadecanoic acid; octadecanoic acid methyl ester, and 9, 12-octadecadienoic acid (Z, Z) methyl ester are fatty acids, with the exception of phytol which is diterpene alcohol (Hema et al. 2011; Banaras et al. 2017). According to studies (Belakhdar et al. 2015; Chukwunonye et al. 2015), fatty acids possess antifungal property Table 6: Phytochemical compounds with antifungal activity obtained from methanolic leaf extract of J. curcas

Retention time (min)	Compound name	Molecular formula	Molecular weight (g/mol)	References
7.539	1,2,3-Propanetriol monoacetate	C₅H₁₀O₄	134.13	(Teoh and Mashitah 2012)
9.273	Methyl salicylate	C₈H₈O₃	152.15	(Essien et al. 2015)
10.549	2-Undecanone	C₁₁H₂₂O	170.29	(Bisht and Chanotiya 2011)
10.841	Indole	‎C₈H₇N	117.15	(Sumiya et al. 2017)
10.898	Decanoic acid methyl ester	C₁₁H₂₂O₂	186.29	(Belakhdar et al. 2015)
11.121	2-Methoxy-4-vinylphenol	C₉H₁₀O₂	150.17	(Guo et al. 2008)
11.287	Tert-hexadecanethiol	C₁₆H₃₄S	258.50	(Yang et al. 2016)
11.653	Phenol, 2,6-dimethoxy-	C₈H₁₀O₃	154.16	(Yang et al. 2016)
11.813	Tetradecane	‎C₁₄H₃₀	198.39	(Begum et al. 2016)
11.905	Cyclotetradecane	C₁₄H₂₈	196.37	(Afrouzan et al. 2018)
11.991	Pentanoic acid ethyl ester	C₇H₁₄O₂	130.18	(Sumiya et al. 2017)
12.248	2-Propenoic acid 3-phenyl-, methyl ester	C₁₀H₁₀O₂	162.18	(Umaiyambigai et al. 2017)
12.334	Diphenyl ether	C₁₂H₁₀	170.21	(Zhang et al. 2018)
13.198	Pentadecane	C₁₅H₃₂	212.41	(Zhang et al. 2015)
13.272	Tridecane	C₁₃H₂₈	184.36	(Yuan et al. 2012)
14.503	Hexadecane	C₁₆H₃₄	226.44	(Oliveira et al. 2014)
16.706	Heptadecane	C₁₇H₃₆	240.47	(Musa et al. 2015)
16.797	17-Pentatriacontene	C₃₅H₇₀	490.93	(Zhang et al. 2015)
16.889	1-Nonadecene	C₁₉H₃₈	266.50	(Asong et al. 2019)
17.015	E-15-Heptadecenal	C₁₇H₃₂O	252.43	(Begum et al. 2016)
17.192	8-Hexadecenal 14-methyl-,	C₁₇H₃₂O	252.40	(Aja et al. 2014)
17.787	Cyclopentadecane	C₁₅H₃₀O	210.40	(Nakashima et al. 2014)
18.474	Hexadecanoic acid methyl ester	C₁₇H₃₄O₂	270.45	(Belakhdar et al. 2015)
18.777	1-Octadecene	C₁₈H₃₆	252.48	(Omoruyi et al. 2014)
18.868	2-Methyl-Z, Z-3, 13-octadecadienol	C₁₉H₃₆O	280.49	(Phatangare et al. 2017; Adibe et al. 2019)
18.983	Oleic acid	C₁₈H₃₄O₂	282.46	(Walters et al. 2004)
19.486	9,17-Octadecadienal, (Z)-	‎C₁₈H₃₂O	264.40	(Adibe et al. 2019)
19.836	2-Methyl-Z,Z-3,13-octadecadienol	C₁₉H₃₆O	280.28	(Adibe et al. 2019)
20.288	9, 12-Octadecadienoic acid (Z,Z)-methyl ester	C₁₉H₃₄O₂	294.47	(Chukwunonye et al. 2015)
20.413	Phytol	C₂₀H₄₀O	296.0	(Hema et al. 2011)
20.556	Octadecanoic acid methyl ester	C₁₉H₃₈O₂	298.50	(Banaras et al. 2017)
21.129	Behenic alcohol	C₂₂H₄₆O	326.60	(Chandrasekaran et al. 2011)
21.186	Octadecanoic acid ethyl ester	C₂₀H₄₀O₂	312.53	(Mohy and Mohyeldin 2018)
21.380	3,7,11,15-Tetramethyl-2-hexadecen-1-ol	C₂₀H₄₀O	296.53	(Mohy and Mohyeldin 2018)
22.096	9,17-Octadecadienal, (Z)-	‎C₁₈H₃₂O	264.40	(Chukwunonye et al. 2015)
23.875	Eicosane	‎CH	282.50	(Shirani et al. 2017)
24.241	Docosanoic acid methyl ester	C₂₃H₄₆O₂	354.61	(Aida et al. 2016)

against diverse mycological pathogens. Since the fungal tissue is lipophilic in nature the fatty acids will attract the absorption of the fungus more easily (Inouye et al. 1999). Moreover, even the minor phytochemical components possibly contributed to antifungal effect by working synergistically with major compounds as reported by (Marino et al. 2001). The possession of these important phyto-compounds with antifungal properties in J. curcas leaf extracts signifies they are effective against fungal pathogens including P. personata.

Conclusion

The study concludes that the methanolic, ethyl acetate and chloroform leaf extracts of J. curcas contain important antifungal phytoconstituents such as hexadecane; n-hexadecanoic acid; phenol, 2,4 bis (-dimethylethyl); phytol and hexadecanoic methyl ester, which are responsible for the control of late leaf spot disease. Hence, methanolic, ethyl acetate and chloroform leaf extracts of J. curcas can be used as substitute bio-pesticides for inhibiting late leaf spot disease on groundnut.

Acknowledgements

We acknowledge DAAD (German Academic Exchange) and CREATES, Tanzania for their financial support.

Author Contributions

MF developed and planned the study, MF, EM and MC statistically analysed the data MF, PN and EM, interpreted the results and MF made write up.

Conflict of Interest

Authors declared no conflicts of interest.

Data Availability

The research data can be obtained through concerting the corresponding author.

Ethics Approval

The ethical approval was obtained from the Tropical Pesticide Research Institute under Herbarium section, Arusha.

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