J. Agric. Res. Vol. 30 No.1, 2017

ALLELOPATHIC EFFECTS OF CURCAS L. AQUEOUS EXTRACT ON EARLY SEEDLING GROWTH OF PARTHENIUM HYSTEROPHORUS L.

Asif ullah Khan*, Faizan Ullah*, Sultan Mehmood*, Muhammad Irshad* and Farhat Ullah Khan**

ABSTRACT:- Jatropha curcasL. ( J. curcas ) is gaining interest as in Pakistan. Therefore, it is necessary to investigate the interaction ofJ. curcas not only with crop but also with weed species before its introduction into agroforestry systems. During present investigation, allelopathic potential ofJ. curcas leaf aqueous extract was investigated on and seedling development of Parthenium hysterophorus L. Extracts were applied at 25%, 20%, 15%, 10% and 5% as seed soaking for 8 hours prior to sowing. Phenolics compounds were found in aqueous extracts and were higher in 25% extract (52.72 mg gallic acid eq. / gm of extract). Higher concentration of the extract (25%) significantly decreased seed germination (%), seed germination index, shoot fresh weight, shoot dry weight, root length, root fresh weight, root dry weight, seedling relative water content and seed vigor index. It was inferred from findings of the present investigation that aqueous extract of J. curcas exhibits inhibitory effects on growth and development of P. hysterophorus and therefore could be a potential source of botanical herbicide in the control of this noxious weed species.

Key Words: Jatropha curcas; Parthenium hysterophorus; Phenolic compounds; Allelopathy

INTRODUCTION 1987; Sangeetha and Baskar, 2015). Allelochemicals affect all functions of The process by which a plant plant life including photosynthesis, species either stimulate or inhibit the respiration, transpiration, resistance growth and development of other nei- and growth (, 1984; Saxena et al., ghboring plant species by pro-ducing 2004). Allelopathy can play important certain chemicals that are released role in sustainable agriculture such into the environment is referred to as as pest control and weed control allelopathy (Rice, 1984; Khattak et (Kohli et al., 1998). Recently scien- al., 2015). The donor plant species tists are sear-ching for biological may affect germination, growth and solutions to minimize the adverse development of the recipient plant effects of synthetic herbicides and species. Allelopathic compounds / insecticides in agriculture production allelochemicals are released into the (Duke and Lydon, 1987). The harmful environment by various means such impacts of allelopathy can be as leaching, volatilization, root exploited for the production of botani- exudation and decomposition of the cal herbicides (Sodaeizadeh et al., plant residue in the (Einhelling, 2009; Sher et al., 2014). * Department of Botany University of Science and Technology Bannu, KP, Pakistan ** Department of Agriculture Science Allama Iqbal Open University ,Islamabad, Pakistan Corresponding author: [email protected]

45 ASIF ULLAH KHAN ET AL.

Weeds are the most aggressive, from field grown plants in the Uni- troublesome and undesirable eleme- versity of Science and Technology nt of the world's vegetation and cause Bannu KP Pakistan; shade dried for 2 reduction in crop yield, wastages of to 3 weeks and were ground finely in resources and human energy. an electric grinder. Extracts were Parthenium hysterophorus (L) belong- prepared by simple maceration tech- ing to family Asteraceae is an invasive nique. The leaf powder (100 g) was weed species of wide spread in Asia, extracted in 200 mL autoclaved Australia and Africa (Evens, 1997; distilled water and left for 48 hr. The Kohli et al., 2006). It is among solution obtained was filtered to noxious weeds threatening crop obtain the aqueous extract. The stock production in different regions of the solution obtained was further diluted world including Pakistan. Control of to make 25%, 20%, 15%, 10% and 5% Parthenium is therefore necessary in solutions. The various solutions for- the country. It is present along road med were stored at 4o C for future use sides, footpaths, cultivated fields, in the experiment. water courses, and other disturbed habitats (Wiesner et al., 2007). Characterization of Aqueous Jatropha curcas L. belongs to Extracts family . It is reported Total phenolics compounds thatJatropha contain 30 to 40 analysis % of oil (Kandpalal and Madan, 1995). Jatropha curcas has gained Determination of total phenolics popularity as biodiesel plant in both was based on the Folin-Ciocalteau developed and developing countries method (Wolfe et al., 2003). Extract of the world. Jatropha seeds oil is non (200 μL) were mixed with freshly edible and is used as biodiesel energy prepared diluted Folin-Ciocalteau (Achten et al., 2008). In Pakistan reagent and 2ml of 7.5% sodium cultivation ofJ. curcas is expanding carbonate. The final mixture was diluted to 7ml with deionized water. rapidly because of government inter- Mixture was kept in dark at ambient est in the resour- condition for 2 h to complete the reac- ces. Therefore it is necessary to tion. The absorbance was measured evaluate interaction ofJ. curcas with at 765nm using a spectrophotometer weed species before its introduction (Hitachis, U-510 Japan). The meas- into the agroforestry system. The pre- urements were compared to a stan- sent study investigated allelopathic dard curve prepared by using various effects ofJ. curcas leaf aqueous concentrations of gallic acid solution. extracts on seed germination and The concentration of total phenolic early seedling growth of Parthenium was expressed as mg gallic acid hyesterophorus. equivalents / g of sample.

MATERIALS AND METHOD Bioassay Seeds of Parthenium hyestero- Preparation of Aqueous Extract of phorus were obtained from National Jatropha curcas Leaf Agriculture Research Center Islama- Fresh and fully expended bad (NARC) Pakistan. Uniform sized ofJatropha curcas were collected seeds were selected and surface

46 ALLELOPATHIC EFFECTS OF JATROPHA CURCAS L. LEAF AQUEOUS EXTRACT sterilized with 0.2% solution of according to method of Mclean mercuric chloride for 2-3 min follo- (1982). The soil used as culture wed by 3-4 times washing with double medium had a pH value of 7.4. distilled water. Pot experiment (8 x 12 cm2 ) was carried out in the glass Parameters Studied house of the Department of Botany The following growth parameters University of Science and Technology were studied. Bannu during May 2014. Pots were The seed germination (%) was filled with autoclaved clay and calculated at 10th day of the experi- (1:1). The calculated quantity (0.5 ml ment when no further germination per seed) ofJatropha curcas extract took place in the control. was applied as seed soaking prior to sowing for 8 h toP. hysterophorus . Seed germination (%) = (Total number The controlled treatment was made of seeds germinated) by soaking seeds of P. hysterophorus /(Total number of seeds in autoclaved distilled water. The see- grown) x 100 ...... …Eq.1 ds were grown in the pots in a glass house. Pots were arranged in com- Germination index = (No of seeds plete randomized design (CRD) with germinated at first count three replicates for each treatment. + No of seeds germinated The following treatments were at final count)/(Days of made first count + days of final count) …....…………Eq2. Treat- Concentrations ments Seed vigor index is determined by multiplying germination percentage T0 Seeds of P. hysterophorus with seedling length of the same seed soaked in autoclaved lot. distilled water T1 Seeds of P. hysterophorus Seed Vigor Index = (Germination (%)) soaked in 5% aqueous x (seedling length) ...Eq3. extract of Jatropha curcas T2 Seeds of P. hysterophorus Equation of Ellis and Roberts soaked in 10% aqueous (1981) was used for the determination extract of Jatropha curcas of mean germination time (MGT). T3 Seeds of P. hysterophorus soaked in 15% aqueous MGT = Dn/ n ….....………...... Eq.4 extract of Jatropha curcas T4 Seeds of P. hysterophorus Where, soaked in 20% aqueous n = the number of seeds, which extract of Jatropha curcas were germination on day D. T5 Seeds of P. hysterophorus D = the number of days counted soaked in 25% aqueous from the beginning of extract of Jatropha curcas germination. Relative water content of seedling The soil used as culture medium was determined according to method in the pots was analyzed for pH of Gao (2000).

47 ASIF ULLAH KHAN ET AL.

RWC (%) = [(FW-DW) / (TW-DW)] × 1). In allelopathic interactions donor 100 …………………………………Eq. 5 plant species inhibits the growth of target plant species through the FW = indicates fresh weight of production of allelochemicals provid- seedlin ing the invader with competitive TW = represents turgid weight of advantage (Callaway and Aschehoug, seedling 2000; Ridenour and Callaway, 2001; DW = represents dry weight of Quddus et al., 2014). Phenolic com- seeding pounds have significant allelopathic potential for application in agricul- The plants in all treatments were ture as herbicides, and harvested 14 days after sowing. After insecticides (Santana et al., 2009; harvest shoot length and root length Hussain et al., 2014). were measured using a common mea- There was significant decrease in suring tape. Shoot fresh weight and seed germination (%) and germina- shoot dry weight, root fresh weight tion index of Parthenium hysteropho- and root dry weight and shoot to root rus in response to higher concen- ratio was determined. tration (25%) of theJ. curcas extract. Mean germination time was not Statistical Analyses significantly affected at 25% of the The data were analyzed statisti- extract. Shoot length was signi- cally by analysis of variance tech- ficantly increased at 5% and 10% of nique (one way ANOVA). The com- the extracts. (Figure 2 A, B, C, D). The parison of treatment means was inhibitory effects of higher concentra- made by least significant differences tions of aqueous extracts of J. curcas (LSD) test (Steel and Torrie, 1984). on seed germination of Parthenium Student Statistix (version 8.1 USA) hysterophorus might be due to the was used for the determination of presence of higher concentration of coefficient of correlation allelopathic compounds that arrested embryo growth. These results were in RESULTS AND DISCUSSION confirmation with those of Rizvi et al. (2000) who investigated allelopathic Phenolics are a group of compo- unds having allelopathic potential in 60 the ecosystem. They have a hydroxyl 50 group (-OH) which is bonded to an aromatic hydrocarbon group (Li et al., 40 2010). Total phenolic compounds 30 were found higher in 25% extract (52.72 mg gallic acid eq. / gram 20 extract) followed by 20% extract Total Phenolics 10 (48.51 mg gallic acid eq. / gram 0 extract), 15% extract (41.77 mg gallic (mg galic acid eq./g extract) 5% 10% 15% 20% 25% acid eq. / gram extract), 10% extract Concentration of extract (38.62 mg gallic acid eq. / gram Figure 1. Total phenolics content of extract) and 5% extract (34.72 mg aqueous extracts of Jatropha gallic acid eq. / gram extract) (Figure curcas leaf

48 ALLELOPATHIC EFFECTS OF JATROPHA CURCAS L. LEAF AQUEOUS EXTRACT

60 25 a A B a 50 20 a

b b b 40 b ab b tion time 15

tion (%) ab ab 30

10 20

Germina b 5 10 Mean germina

0 0 T0 T1 T2 T3 T4 T5 T0 T1 T2 T3 T4 T5

Treatment Treatment

1.8 14 C a a 1.6 ab D 12 ab 1.4 abc bc 10 c b 1.2 b b 8 1 b b tion index b 6 0.8

0.6 4

Germina Shoot length (cm) 0.4 2 0.2 0 T0 T1 T2 T3 T4 T5 0 T0 T1 T2 T3 T4 T5

Treatment Treatment Treatments: T0- control, T1- 5% aqueous extract, T2-10% aqueous extract , T3-15% aqueous extract , T4- 20% aqueous extract, T5- 25% aqueous extract

Figure 2. Effect of Jatropha curcas aqueous leaf extract on (A) Germination % (B) Mean germination time (C) Germination index (D) Shoot length of Parthenium hysterophorus. Means sharing a common English letter are statistically similar effects of on growth of and Torabi-Sirchi, 2011; Knox et al., wild . The decrease in seed germi- 2014). After seed germination, the nation (%) can be correlated with the effects of aqueous extracts on shoot existence of phenolic compounds in length were stimulatory. This might aqueous extracts which prevented be because those allelochemicals had the action of gibberellins as they are a short lasting effect when applied as main factors for seed germination; seed soaking prior to sowing. The hence it is supposed that phenolic results established that allelopathic compounds limited performances of effects ofJ. curcas leaf; in course of natural growth regulators (Saffari time decreased. These results con-

49 ASIF ULLAH KHAN ET AL.

1.6 a 0.02 A a B 0.018 1.4 ab ab b 0.016 bc c 1.2 0.014 c bc 0.1 0.012 cd cd 0.8 0.01 d 0.008 0.6 0.006 0.4

Shoot dry weight (g)

Shoot fresh weight (g) 0.004 0.2 0.002

0 0 T0 T1 T2 T3 T4 T5 T0 T1 T2 T3 T4 T5

Treatment Treatment

0.050 a 0.007 C a D 0.045 0.006 0.040 ab abc 0.035 0.005 b

0.030 0.004 c bc bc 0.025 c c 0.003 0.020 d 1.115 0.002 c

Root dry weight (g)

Root Fresh weight0.010 (g) 0.001 0.005

0 0 T0 T1 T2 T3 T4 T5 T0 T1 T2 T3 T4 T5

Treatment Treatment

Treatments: T0- control, T1- 5% aqueous extract, T2-10% aqueous extract , T3-15% aqueous extract , T4- 20% aqueous extract, T5- 25% aqueous extract Figure 3. Effect of Jatropha curcas aqueous leaf extract on (A) Shoot fresh weight (B) Shoot dry weight (C) Root fresh weight (D) Root dry weight of Parthenium hysterophorus. Means sharing a common English letter are statistically similar firmed the previous findings of inhibition of seedling growth and the Prasanta and Bhowmika (2003) who effect is dose dependant (Rice, 1984). have reported similar effects of Results of the present investigation different wheat straw extracts on showed that inhibitory effects of J. wheat culture. The increase in shoot curcas extracts on growth attributes length can be attributed to the of Parthenium hysterophorus were presence of nitrogenous and other concentration dependant (Figure 3 A, compounds present in the extracts. B, C, D). Shoot fresh weight, shoot dry Allelochemicals suppress the mitotic weight, root fresh weight and root dry activity of young cells, resulting in weight were decreased by aqueous

50 ALLELOPATHIC EFFECTS OF JATROPHA CURCAS L. LEAF AQUEOUS EXTRACT

6 A 80 a a a B 5 70 a a

4 tin 60 a

b 50 3 b b b 40 b b 2 30

Root length (cm) 20 1 Shoot/Root ra 10 0 T0 T1 T2 T3 T4 T5 0 T0 T1 T2 T3 T4 T5 Treatment Treatment

90 a ab 60 C D a a 80 ab 50 70 bc b 60 c 40 ab b c 50 30 40

20 30

Seed vigor index d 20 10 10 0 T0 T1 T2 T3 T4 T5 0 T0 T1 T2 T3 T4 T5

Relative water content of seeding (%) Treatment Treatment

Treatments: T0- control, T1- 5% aqueous extract, T2-10% aqueous extract , T3-15% aqueous extract , T4- 20% aqueous extract, T5- 25% aqueous extract

Figure 4 Effect of Jatropha curcas aqueous leaf extract on (A) Root length (B) Shoot / root ratio (C) Seed vigor index (D) Seedling relative water content of Parthenium hysterophorus. Means sharing a common English letter are statistically similar extracts at all the concentrations. at all the concentrations. Seedling However, the most effective concen- relative water content was signi- trations were 25% and 20% (T4 and ficantly decreased (70%) by appli- T5). cation of higher concentration (25%) Root length was significantly in- of the extract as compared to control hibited by extracts at all the concen- (Figure 4 A, B, C, D). trations with a concomitant increase The reduction in shoot fresh in shoot / root ratio as compared to weight and dry weight, root fresh control (p>0.05). Seed vigor index was weight and dry weight of Parthenium not significantly affected by extracts hysterophorus could be attributed to

51 ASIF ULLAH KHAN ET AL. the presence of phenolics compounds herbicides. in aqueous extracts ofJ. curcas . Root is very sensitive indicator of phyto- LITERATURE CITED toxic activity. During present investi- gation, root growth of Parthenium Achten, W. M. J., L. Verchot , Y. J. hysterophorus was inhibited by Franken , E. Mathijs , V. P. Singh, aqueous extracts ofJ. curcas . R. Aerts and B. Muys . 2008. Leachates, exudates and residues of a Jatropha bio-diesel production large number of plant species have and use. . 32: been reported for allelopathy effects 1063–1084. (Alam and Islam, 2002). Phenolic Alam, S. M. and E. U. Islam. 2002. allelochemicals adversely affect root Effect of aqueous extract of Leaf, growth by inhibiting the process of stem and root of nettle leaf goose- mitosis in the root tips, alter cell ultra foot and NaCl on germination and structure and then interfere with the seedling growth of rice. Pak. J normal growth processes of the entire .Sci. Technol. 1: 47-52. plant (Cruz et al., 1998; Mondal et al., Barkosky, R. R. and F. A. Einhellig. 2015). The Jatropha leaf was found 2003. Allelopathic interference of as a good source of natural phenolics plant-water relationships by (Khattak et al., 2015). Batish et al. para-hydroxy benzoic acid. Bot. (2008) found that caffeic acid affected Bull. Acad. Sin. 44: 53-58. the growth and morphology of mong Batish, D.R.M., H.P.Singh, S.Kaur, bean. Decrease in relative water R..K. Kohli and S.S. Yadav. 2008. content of Parthenium hysterophorus Caffeic acid affects earlier growth, seedlings could be attributed to the and morphogenetic response of presence of higher phenolics content hypocotyl cuttings of mung bean in the aqueous extracts of J. curcas (Phaseolus aureus ).J.Plant. because Phenolic acids results in the Physiol. 165: 297-305. induction of water stress conditions Callaway, R. M. and E. T. Aschehoug. in plants (Barkosky and Einhellig, 2000. Invasive plants versus 2003). their new and old neighbors: A mechanism for exotic plant CONCLUSION invasion. Sci. 290: 521-523. The findings of the present inves- Cruz,O.R.,A. Anaya and B. E. tigation established that higher con- Hernandez-Bautista. 1998. centrations of theJ. curcas leaf Effects of allelochemical stress aqueous extracts suppressed the produced by sicyosdeppei on seed germination and growth of seedling root ultrastructure of Parthenium hysterophorus. There- Phaseolous valgaris and fore, it is inferred thatJ. curcas leaf Cucurbita ficifolia. J. Chem. can yield adequate amount of Ecol. 24: 2039-2057. allelochemicals to effectively control Duke, S. O. and J. Lydon .1987. Parthenium hysterophorus. Future Herbicide from natural compo- studies on the isolation and puri- unds. Weed Technol. 1: 122-128. fication of the effective compounds Evans, H. C. 1997. Parthenium fromJ. curcas leaf could lead to the hysterophorus: A review of its production of some novel botanical weed status and the possibilities

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