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Herbicidal Activity of Aspergillus niger Metabolites Against Parthenium Weed

Atividade Herbicida dos Metabólitos de Aspergillus niger contra Losna-branca

ABSTRACT:

Metabolites of Aspergillus niger, prepared in malt extract (ME) broth and potato dextrose (PD) broth, were evaluated for their herbicidal activity against a noxious parthenium weed (Parthenium hysterophorus). In laboratory assays, original (X) and diluted (½ X) fungal metabolites significantly reduced germination and seedling growth of weed. However, metabolites prepared in ME broth proved to have greater herbicidal activity than metabolites prepared in other growth medium. Original metabolites prepared in ME broth have completely hinder the germination of parthenium seeds; while those prepared in PD broth have reduced germination by 89% over control. In pot trials, one-week, two-week and three-week-old parthenium seedlings were sprayed three times with original and concentrated (2X) metabolites of A. niger prepared in ME broth. Plants were harvested after 40 days of sowing. One-week treatment plants were most susceptible to fungal metabolites spray, followed by two-week and three-week treatment plants, respectively. Original and concentrated metabolites have significantly reduced shoot biomass of one-week-old plants by 57% and 68%, and root biomass by 50% and 75%, respectively. The present study has come to the conclusion that A. niger metabolites prepared in ME broth can effectively control germination and growth of parthenium.

Keywords:
natural herbicide; Parthenium hysterophorus; noxious weed

RESUMO:

Os metabólitos de Aspergillus niger, preparados em caldo de extrato de malte (ME) e caldo de dextrose de batata (PD), foram avaliados quanto à sua atividade herbicida contra uma erva partênica nociva (Parthenium hysterophorus). Em resenhas de laboratório, metabólitos fúngicos originais (X) e diluídos (½ X) reduziram substancialmente a germinação e o crescimento de plântulas da planta daninha. No entanto, os metabólitos preparados em caldo de ME demonstraram ser superiores na sua atividade herbicida, comparados aos metabólitos preparados em outro meio de crescimento. Os metabólitos originais preparados em caldo ME estagnaram completamente a germinação de sementes de erva partênica, ao passo que os preparados em caldo PD reduziram a germinação em 89%, em relação ao controle. Nos ensaios em vaso, as mudas de erva partênica de uma semana, duas semanas e três semanas foram pulverizadas três vezes com metabólitos originais e concentrados (2X) de A. niger preparados em caldo ME. As plantas foram colhidas após 40 dias de semeadura. As plantas com uma semana de tratamento foram mais suscetíveis à pulverização de fungos metabólitos, seguidas das plantas de duas semanas e de três semanas. Os metabólitos originais e concentrados reduziram significativamente a biomassa de brotos de plantas de uma semana de idade em 57% e 68%, e a biomassa de raízes, em 50% e 75%, respectivamente. O presente estudo conclui que os metabólitos de A. niger preparados em caldo ME podem efetivamente controlar a germinação e o crescimento de ervas partênicas.

Palavras-chave:
herbicida natural; Parthenium hysterophorus; erva nociva

INTRODUCTION

Parthenium is a noxious annual weed that causes destruction in managed as well as natural ecosystems in many countries in the world (Masum et al., 2013Masum S.M., Hasanuzzaman M., Ali M.H. Threat to Parthenium hysterophorus in agro-ecosystem and its management: a review. Int J Agric Crop Sci. 2013;6:684-97.; Adkins and Shabbir, 2014Adkins S., Shabbir A. Biology, ecology and management of the invasive parthenium weed (Parthenium hysterophorus L.). Pest Manage Sci. 2014;70:1023-9.). Considered one of the most harmful and invasive terrestrial weed (Hussain et al., 2016Hussain N., Abbasi T., Abbasi S.A. Vermicomposting transforms allelopathic parthenium into a benign organic fertilizer. J Environ Manage. 2016;180:180-9.), it can adapt to any type of environmental conditions, invade all types of land, and cause heavy losses to plants by reducing their yield, biodiversity, and disturbing their natural ecosystem (Timsina et al., 2011Timsina B. Impact of Parthenium hysterophorus L. invasion on plant species composition and soil properties of grassland communities in Nepal. Flora. 2011;206:233-40.; Kapoor, 2012Kapoor R.T. Awareness related survey of an invasive alien weed, Parthenium hysterophorus L. in Gautam Budh Nagar district, India. J Agric Technol. 2012;8:1129-40.). It also adversely affects the growth and yield of crop plants (Safdar et al., 2016Safdar M.E. et al. Critical competition period of parthenium weed (Parthenium hysterophorus L.) in maize. Crop Prot. 2016;80:101-7.). The invasive nature of parthenium is due to the presence of parthenin (Verma et al., 2006Verma K.K. Parthenium dermatitis treated with azathioprine weekly pulse doses. Indian J Dermatol Venereol Leprol. 2006;72:24-27.). This weed also causes devastating effects on humans and livestock (Shabbir et al., 2012Shabbir A., Dhileepan K., Adkins S.W. Spread of Parthenium weed and its biological control agent in the Punjab, Pakistan. Pak J Weed Sci Res. 2012;18:581-8.). Direct contact of parthenium plants with humans for a long time causes skin disease, dermatitis, and in severe cases, death (Morin et al., 1997Morin, F. et al. Glyphosate uptake in Catharanthus roseus cells: Role of a phosphate transporter. Pest Biochem Physiol. 1997;58:13‐22.).

Many management practices have been used to reduce losses due to parthenium, but with some inherent limitations in their application (Adkins and Shabbir, 2014Shabbir A. Chemical control of Parthenium hysterophorus L. Pak J Weed Sci Res. 2014;20:1-10.). Among these methods, the use of herbicides has been proved useful against parthenium (Javaid, 2007Javaid A., Shah M.B.M. Phytotoxic effects of aqueous leaf extracts of two Eucalyptus spp. against Parthenium hysterophorus L. Sci Int. 2007;19:303-6.; Shabbir, 2014). However, synthetic herbicides create environmental problems and some resistance of weeds against herbicides. Thus, in this situation there has been a need of safe and eco-friendly weed-management strategies (Teeraraka et al., 2010Teeraraka M., Laosinwattanaa C., Charoenying P. Evaluation of allelopathic, decomposition and cytogenetic activities of Jasminum ofûcinale L. var. grandiûorum L. on bioassay plants. J Bioresour Technol. 2010;101:5677-84.; Akbar et al., 2014Akbar M. et al. Holadysenterine, a natural herbicidal constituent from Drechslera australiensis for management of Rumex dentatus. J Agric Food Chem. 2014;62:368-72.). In recent years, some studies have been carried out to control parthenium and other weeds in laboratory, green houses, and under filed conditions by fungal metabolites (Javaid, 2010Javaid A. Efficacy of some chemical herbicides against Parthenium hysterophorus L. Pak J Weed Sci Res. 2007;13:93-8.; Akbar and Javaid, 2015Akbar M., Javaid A. Management of Rumex dentatus L. (toothed dock) by fungal metabolites under field conditions. Int J Agric Biol. 2015;17:187-92.). A metabolite isolated from Phyllosticta sp. had significant herbicidal activity (Evidente et al., 2008Evidente A. et al. Phyllostictines A-D, oxazatricycloalkenones produced by Phyllosticta cirsii, a potential mycoherbicide for Cirsium biocontrol. J Tetrah. 2008;64:1612-9.). According to Javaid and Ali (2011), metabolites of Trichoderma species effectively controlled Avena fatua.Akbar and Javaid (2012aAkbar M., Javaid A. Herbicidal activity of fungal culture filtrates against Chenopodium album L. and Avena fatua L. J Anim Plant Sci. 2012a;22:977-82.,bAkbar A., Javaid A. Evaluation of herbicidal potential of fungal metabolites against Phalaris minor. Afr J Microbiol Res. 2012b;6:4053-7.) reported that metabolites extracted from Drechsleraspp. showed effective control against weeds of wheat, namely Rumex dentatus, Avena fatua and Phalaris minor. Although metabolites of different fungal groups have been used as natural herbicides against parthenium (Javaid, 2010Javaid A. Herbicidal potential of allelopathic plants and fungi against Parthenium hysterophorus - a review. Allelopathy J. 2010;25:331-44. ; Javaid et al., 2017Javaid A. et al. Management of parthenium weed using metabolites of Alternaria japonica. Planta Daninha. 2017;34, [in press]), literature discussing the use of metabolites of Aspergillus niger as natural herbicide against parthenium is rare. A. niger is a fungus ubiquitous in soil that causes black mould on many fruits and vegetables, and spoils preserved foods (Stratford et al., 2016Stratford M. et al. Mapping the structural requirements of inducers and substrates for decarboxylation of weak acid preservatives by the food spoilage mouldAspergillus niger. Int J Food Microbiol. 2016;157:375-383.). This present study aims to assess the herbicidal potential of metabolites of A. niger against parthenium.

MATERIAL AND METHODS

Culture of A. niger, was procured from First Fungal Culture Bank of Pakistan, Institute of Agriculture Sciences, University of the Punjab, Lahore, Pakistan. Subculturing of procured fungal strain was done under laboratory conditions on 2% malt extract agar medium (2 g malt extract, 2 g agar in 100 mL water). After thorough shaking, growth medium was placed in an autoclave at 121 oC for 30 minutes and poured in sterilized Petri plates aseptically after cooling to 70 oC. Fungal culture was inoculated and Petri plates were incubated for one week at 25 oC and stored at 4 oC.

For preparation of fungal metabolites, two liquid growth medium viz. potato dextrose broth (PDB) and malt extract broth (MEB) were used. An amount of 200 mL of each of PDB and MEB was autoclaved in 250 mL volumetric flasks. Flasks were inoculated with 5 mm discs of the selected fungal species and incubated at 25 oC for 15 days. Thereafter, metabolites were filtered through sterilized muslin cloth, followed by filtration through sterilized Whattsman filter paper No. 1. Filtered metabolites were centrifuged for 5 minutes at 600 rpm and then filtered through Millipore filter paper (Akbar et al., 2014Akbar M. et al. Holadysenterine, a natural herbicidal constituent from Drechslera australiensis for management of Rumex dentatus. J Agric Food Chem. 2014;62:368-72.).

Parthenium seeds were collected from University of the Punjab Lahore, Pakistan. Seeds were dried in an electric oven at 45 oC. Dried seeds were rubbed for the removal of seed shells. Seeds were surface sterilized with 1% solution of sodium hypochlorite for 10 minutes followed by several washings with distilled water. Pre-sterilized Petri plates (9 cm diameter) were lined with sterilized filter papers. Twenty five surface sterilized seeds were arranged in each Petri plate. Two milliliters of original (X) and diluted (½ X) filtered metabolites of the fungal species prepared in malt extract and potato dextrose broths were poured in each Petri plate. Diluted concentration was made by adding sterilized distilled water in an appropriate amount (Akbar and Javaid, 2013Akbar M., Javaid A. Prospects of using fungal metabolites for the management of Rumex dentatus, a problematic weed of wheat. Int J Agric Biol. 2013;15:1277-82.). In control treatment, distilled water was used instead of fungal metabolites. Treatments were replicated 4 times. The experiment was conducted in a completely randomized design in a growth chamber maintained at 25 oC for 10 h light period daily. Harvest was taken after two weeks and data on germination of seeds, root, and shoot length and plants fresh and dry weights were noted (Javaid and Ali, 2011).

Metabolites prepared in malt extract broth exhibited good herbicidal activity in Petri plate experiment so they were selected for further pot experiment. Field soil was filled in 10 cm plastic pots. Twenty seeds of parthenium were sown in each pot. Extra seedlings were removed from each pot after germination and five uniform seedlings were left in each pot for taking accurate data. Pots were divided into three groups to carry out foliar spray on 1 week, 2 week and 3 week old plants. Two different concentrations of metabolites viz. original (X) and concentrated (2X) were used. Concentrated metabolites were prepared by evaporating the water in an electric oven at 45 oC. Both (original and concentrated) concentrations were sprayed on pot grown plants 4 times with a 5 day intervals. Distilled water was sprayed on plants of control treatment. Experiment was conducted under natural conditions.

Parthenium plants were harvested after 45 days of sowing. Plants were carefully uprooted and roots were separated from shoots and washed to remove soil particles. Various vegetative growth parameters like shoot growth and root growth, dry weight of seedlings were determined. Data were analyzed by ANOVA followed by LSD test at 5% level of the significance using computer software Statistics 8.1.

RESULTS AND DISCUSSION

Laboratory bioassays

Both original and diluted metabolites of A. niger significantly reduced germination of parthenium seeds. However, the inhibitory effect of original metabolites was far better than the diluted ones. Moreover, metabolites prepared in malt extract broth were more inhibitory to germination than those prepared in potato dextrose. There was 99% and 21% reduction in germination due to original and diluted metabolites of A. niger prepared in malt extract broth, compared to 89% and 14% reduction due to metabolites prepared in potato dextrose broth, respectively (Figure 1A). Earlier, Javaid et al. (2014Javaid A. et al. Screening of Aspergilli for herbicidal activity of their culture filtrates against Parthenium weed. Pak J Weed Sci Res. 2014;20:137-44.) reported that metabolites of other Aspergillus species namely A. fumigatus, A. terreus, A. flavus, A. parasiticus and A. spelunceus reduced germination of parthenium in up to 28%.

Figure 1
Effect of original (X) and diluted (½X) metabolites of Aspergillus niger prepared in malt extract broth and potato dextrose broth on germination and shoot growth of parthenium seedlings in laboratory bioassays.

The effect of fungal metabolites, prepared in the two growth media, on various parameters of shoot and root growth was similar to that on seed germination. In general, the effect of fungal metabolites prepared in malt extract broth was more severe on shoot and root growth than those prepared in potato dextrose broth. There was 99.4%, 99.3% and 99.4% reduction in length, fresh weight and dry weight of shoot due to original metabolites prepared in malt extract broth. Whereas these parameters were reduced by 97%, 95% and 98%, respectively, due to original metabolites prepared in potato dextrose broth (Figure 1B-D). Likewise, length, fresh weight and dry weight of root were reduced by 99.2%, 99.4% and 99.1% due to original metabolites prepared in malt extract broth and 97.3%, 98.8% and 87.7% due to original metabolites prepared in potato dextrose broth, respectively (Figure 2A-C). Variation in herbicidal activity of fungal metabolites prepared in different growth medium has also been reported in some previous studies. Javaid et al. (2013Javaid A. et al. Effect of culture medium on herbicidal potential of metabolites of Trichoderma species against Parthenium hysterophorus. Int J Agric Biol. 2013;15:119-24.) found that metabolites of various Trichoderma species prepared in M-1-D medium showed greater herbicidal activity against germination and growth of parthenium than those prepared in malt extract broth. Similar variable herbicidal activity was found in metabolites of Drechslera species prepared in different growth media against Rumex dentatus (Akbar and Javaid, 2010Akbar M., Javaid A. Management of some problematic weeds of wheat by metabolites of Drechslera spp. prepared in malt extract. Pak J Weed Sci Res. 2010;16:145-51., 2013). Recently, Javaid et al. (2017) reported that metabolites of Alternaria japonica prepared in malt extract broth and potato dextrose broth showed different herbicidal activities against parthenium. Variation in herbicidal activities of fungal metabolites prepared in different growth media might be associated with production of different herbicidal constituents or different quantities of the same herbicidal compound in different growth medium.

Figure 2
Effect of original (X) and diluted (½X) metabolites of Aspergillus niger prepared in malt extract broth and potato dextrose broth on root growth of parthenium seedlings in laboratory.

Pot trials

Analysis of variance showed that the effect of age of host at the time of start of fungal metabolites application (W), concentrations of the metabolites (C) as well as W x C were significant for all the studied parameters of root and shoot growth of parthenium (Table 1). In general, herbicidal activity of the fungal metabolites was reduced with increase in age of host plant. 1 week old parthenium seedlings were highly susceptible to foliar application of fungal metabolites. There was 58% and 76% decrease in shoot length, and 57% and 68% decrease in shoot dry weight due to foliar spray of original and concentrated fungal metabolites on 1 week old plants, respectively. On the other hand, original and concentrated fungal metabolites reduced shoot length by 36% and 58%, and shoot dry weight by 30% and 40% over control, respectively, in 2 week old plants. 3 Week old parthenium plants were least susceptible to fungal metabolites where there was 13% and 20% reduction in shoot length, and 8% and 35% suppression in shoot dry weight was recorded over control due to original and concentrated fungal metabolites, respectively (Figure 3). The effect of fungal metabolites on root growth of various ages of parthenium plants was generally similar to that on various parameters of root growth. Root growth in 1 week old plants was the most susceptible to foliar application of metabolites where 71% and 84% reduction in root length, and 50% and 75% reduction in root dry weight over control was recorded due to original and concentrated fungal metabolites, respectively (Figure 4). The results of present study are in line with the findings of earlier studies which also showed that herbicidal activity of fungal metabolites decreased with the age of the weed plants possibly because of increased resistance in old plants (Javaid et al., 2011Javaid A., Ali S. Alternative management of a problematic weed of wheat Avena fatua L. by metabolites of Trichoderma spp. Chilean J Agric Res. 2011;71:205-11., 2013, 2016).

Table 1
Analysis of variance (ANOVA) for the effect of age of host plant (1, 2, 3 weeks) and concentrations of fungal metabolites (original and concentrated) on various parameters of shoot and root growth of pot grown parthenium seedlings

Figure 3
Effect of foliar spray with original (X) and concentrated (2X) metabolites of Aspergillus niger prepared in malt extract broth on shoot growth of pot grown parthenium plants of different ages.

Figure 4
Effect of foliar spray with original (X) and concentrated (2X) metabolites of Aspergillus niger prepared in malt extract broth on root growth of pot grown parthenium plants of different ages.

This present study concludes that metabolites of A. niger prepared in malt extract broth possessed pronounced herbicidal activity against parthenium weed. The herbicidal activity of these metabolites is gradually reduced with the aging of the host plant.

REFERENCES

  • Adkins S., Shabbir A. Biology, ecology and management of the invasive parthenium weed (Parthenium hysterophorus L.). Pest Manage Sci. 2014;70:1023-9.
  • Akbar M., Javaid A. Management of some problematic weeds of wheat by metabolites of Drechslera spp. prepared in malt extract. Pak J Weed Sci Res. 2010;16:145-51.
  • Akbar M., Javaid A. Herbicidal activity of fungal culture filtrates against Chenopodium album L. and Avena fatua L. J Anim Plant Sci. 2012a;22:977-82.
  • Akbar A., Javaid A. Evaluation of herbicidal potential of fungal metabolites against Phalaris minor Afr J Microbiol Res. 2012b;6:4053-7.
  • Akbar M., Javaid A. Prospects of using fungal metabolites for the management of Rumex dentatus, a problematic weed of wheat. Int J Agric Biol. 2013;15:1277-82.
  • Akbar M. et al. Holadysenterine, a natural herbicidal constituent from Drechslera australiensis for management of Rumex dentatus J Agric Food Chem. 2014;62:368-72.
  • Akbar M., Javaid A. Management of Rumex dentatus L. (toothed dock) by fungal metabolites under field conditions. Int J Agric Biol. 2015;17:187-92.
  • Evidente A. et al. Phyllostictines A-D, oxazatricycloalkenones produced by Phyllosticta cirsii, a potential mycoherbicide for Cirsium biocontrol. J Tetrah. 2008;64:1612-9.
  • Hussain N., Abbasi T., Abbasi S.A. Vermicomposting transforms allelopathic parthenium into a benign organic fertilizer. J Environ Manage. 2016;180:180-9.
  • Javaid A. Efficacy of some chemical herbicides against Parthenium hysterophorus L. Pak J Weed Sci Res. 2007;13:93-8.
  • Javaid A. Herbicidal potential of allelopathic plants and fungi against Parthenium hysterophorus - a review. Allelopathy J. 2010;25:331-44.
  • Javaid A., Shah M.B.M. Phytotoxic effects of aqueous leaf extracts of two Eucalyptus spp. against Parthenium hysterophorus L. Sci Int. 2007;19:303-6.
  • Javaid A., Ali S. Alternative management of a problematic weed of wheat Avena fatua L. by metabolites of Trichoderma spp. Chilean J Agric Res. 2011;71:205-11.
  • Javaid A. et al. Effect of culture medium on herbicidal potential of metabolites of Trichoderma species against Parthenium hysterophorus Int J Agric Biol. 2013;15:119-24.
  • Javaid A. et al. Screening of Aspergilli for herbicidal activity of their culture filtrates against Parthenium weed. Pak J Weed Sci Res. 2014;20:137-44.
  • Javaid A. et al. Management of parthenium weed using metabolites of Alternaria japonica Planta Daninha. 2017;34, [in press]
  • Kapoor R.T. Awareness related survey of an invasive alien weed, Parthenium hysterophorus L. in Gautam Budh Nagar district, India. J Agric Technol. 2012;8:1129-40.
  • Masum S.M., Hasanuzzaman M., Ali M.H. Threat to Parthenium hysterophorus in agro-ecosystem and its management: a review. Int J Agric Crop Sci. 2013;6:684-97.
  • Morin, F. et al. Glyphosate uptake in Catharanthus roseus cells: Role of a phosphate transporter. Pest Biochem Physiol. 1997;58:13‐22.
  • Safdar M.E. et al. Critical competition period of parthenium weed (Parthenium hysterophorus L.) in maize. Crop Prot. 2016;80:101-7.
  • Shabbir A., Dhileepan K., Adkins S.W. Spread of Parthenium weed and its biological control agent in the Punjab, Pakistan. Pak J Weed Sci Res. 2012;18:581-8.
  • Shabbir A. Chemical control of Parthenium hysterophorus L. Pak J Weed Sci Res. 2014;20:1-10.
  • Stratford M. et al. Mapping the structural requirements of inducers and substrates for decarboxylation of weak acid preservatives by the food spoilage mouldAspergillus niger Int J Food Microbiol. 2016;157:375-383.
  • Teeraraka M., Laosinwattanaa C., Charoenying P. Evaluation of allelopathic, decomposition and cytogenetic activities of Jasminum ofûcinale L. var. grandiûorum L. on bioassay plants. J Bioresour Technol. 2010;101:5677-84.
  • Timsina B. Impact of Parthenium hysterophorus L. invasion on plant species composition and soil properties of grassland communities in Nepal. Flora. 2011;206:233-40.
  • Verma K.K. Parthenium dermatitis treated with azathioprine weekly pulse doses. Indian J Dermatol Venereol Leprol. 2006;72:24-27.

Publication Dates

  • Publication in this collection
    2018

History

  • Received
    02 Aug 2016
  • Accepted
    19 Aug 2016
Sociedade Brasileira da Ciência das Plantas Daninhas Departamento de Fitotecnia - DFT, Universidade Federal de Viçosa - UFV, 36570-000 - Viçosa-MG - Brasil, Tel./Fax::(+55 31) 3899-2611 - Viçosa - MG - Brazil
E-mail: rpdaninha@gmail.com