Journal of Agronomy & Agricultural Science Category: Agriculture Type: Research Article

Screening of Herbicidal Potential of Crude Extract of Isolated Fungal Strains against some Noxious Weeds of India: A Preliminary Evaluation

Ajay Kumar Singh1* and Akhilesh Kumar Pandey1
1 Department Of Biological Science, Mycological Research Laboratory, Madhya Pradesh, India

*Corresponding Author(s):
Ajay Kumar Singh
Department Of Biological Science, Mycological Research Laboratory, Madhya Pradesh, India
Tel:+91 9704700060,
Email:drajay2009@gmail.com

Received Date: Oct 23, 2019
Accepted Date: Nov 07, 2019
Published Date: Nov 14, 2019

Abstract

Herbicidal potential of crude extract (Cell Free Culture Filtrate) of nine fungal strains against six problematic weeds of Madhya Pradesh viz. Parthenium hysterophorus, Lantana camara, Xanthium strumarium, Cassia tora, Hyptis suaveolens, Sida acuta and Antigonon leptopus was determined by employing shoot cut and seedling bioassays. Crude extract obtained from 21 days old fermented broth of Fusarium oxysporum FGCCW#43, F. moniliforme FGCCW#16 and F.roseum #55 showed excellent herbicidal activities against these all weeds, whereas CFCF from Phoma herbarum FGCCW#54 showed strong mortality against Parthenium hysterophorus. Findings of these studies revealed a new perspective for weeds control and their use in crops will prospect, potential fungal species and their crude extract that could be used as a novel, lucrative source of natural herbicides in the future.

Keywords

Cell free; Culture filtrate; Fungal strains; Natural herbicide; Screening

INTRODUCTION

Weeds are one the most serious cause of economic losses in agricultural production. The exploitation of synthetic herbicides for weed control has been increasing; however, their heavy application in crop fields has resulted in environmental and medical problems. Exploitation of secondary metabolites or biorationals of both pathogenic and non-pathogenic fungi in weed management have attracted the attention of scientists in recent years. Several comprehensive reviews have been published on biological, technological and economical feasibility of microbial products as herbicides [1,2]. Potential herbicidal properties of host specific toxin, Maculosin produced by Alternaria alternata, against spotted knapweed [3] and Cornexistin from Paceilomyces variotii [4] against several monocotyledonous and dicotyledonous weeds are well known and used as alternative means in developing eco-friendly herbicides.

Natural herbicides, probably obtained from microorganisms are eco-friendly, biodegradable and less toxic are now considered as one of the best alternative to existing weed management strategies [5,6] Fungi are well recognized for their ability to produce diverse biologically active metabolites including herbicides [7]. Therefore, screening for fungal products with herbicidal activity has been one of the most interesting features in weed management research. The main objective of this study was to determine herbicidal potential of cell free culture filtrates obtained from 21 days old fermented broth of nine different isolates of fungi against seven prominent weeds viz., Parthenium hysterophorusLantana camara, Xanthium strumariumCassia tora, Hyptis suaveolensSida acuta and Antigonon leptopus recovered earlier from various places of Madhya Pradesh, India. Activities of the CFCF were measured under control laboratory conditions.

METHODOLOGY

Fungi investigated

The test strains were obtained from Fungal Germplasm Collection Centre, Department of Biological Sciences, R. D. University, Jabalpur. These were isolated earlier from infected/infested parts of weeds collected from various places of M.P. The cultures were maintained by subculturing on PDA (Potato- 200gms; Agar- 18gms; Dextrose-20gms; D.W.-1000ml) slants at 28±1°C in tightly capped culture tubes.

Preparation of Cell Free Culture Filtrate (CFCF)

250ml Erlenmeyer’s flasks containing 200ml, modified richards broth were seeded with 5mm disc of inoculum separated from 7 days old cultures grown on PDA medium. Inoculated flasks were incubated at 28±1°C in a BOD incubator (Remi, India) and the Cell Free Culture Filtrate (CFCF) was extracted after 21 days [8].

Extraction of CFCF

The metabolized broth was passed through a pre-weighed what man filter paper no.1 under aseptic conditions and was centrifuged at 400xg for 15-20 min. The pellet was thrown and the supernatant was again filtered in vacuo by microfiltration using sterile micro filters, 0.45µm pore size, Minisart (Sartorius, Gottingen, Germany) making it cell free [9]. Thus cell free culture filtrate was obtained.

General Bioassays: Bioassay procedure

The potential metabolites of recovered isolates were assessed through following bioassays:

Shoot cut bioassay: 30-35 days old seedlings were grown in pots containing soil: Sand: Peat (1:1:1) inside a plant growth chamber (Yorco, India). These were taken and an inclined cut was made at the tip in sterilized water. They were then dipped in different dilutions i.e., 25%, 50%, 75% and 100% of the CFCF in test vials. These were then incubated under daylight or artificial illumination (3.5 x 104 erg/cm2/s). The vials were sealed with cotton buds and the effects of different days old fermented broths and different dilutions of the toxic metabolites were observed on the shoots of the test weed after 48 h at room temperature [10].

Seedling bioassay: Seedlings of the target weeds were raised in pots containing soil: Sand: Peat (1:1:1). 15-20 days old seedlings were treated with different days old CFCF (7, 14, 21, 28 days) and also with different dilutions (25%, 50%, 75% & 100%) of the phytotoxic metabolites and were then incubated. Phytotoxicity of CFCF of various strains was determined by employing shoot cut and seedling bioassay tests in accordance to Thapar et al., [11]. Phytotoxicity was determined following the visual method suggested by Abbas et al. [12], and Buckle and Sanders on a rating scale of 0-5.

RESULTS AND DISCUSSION

Data recorded in tables 1&2 showed that toxicity of CFCF varied significantly amongst various fungal strains. CFCF obtained from Fusarium oxysporum FGCCW#43 showed strongest herbicidal potential (100%) against Parthenium hysterophorus, Lantana camara, Xanthium strumarium, Cassia tora, Hyptis suaveolens, Sida acuta and Antigonon leptopus while CFCF from F.moniliforme FGCCW#16 and F.roseum FGCCW#55 ranked second and third respectively. CFCF of these strains showed broad-spectrum herbicidal property. Production of herbicidal metabolites viz., fumonisins, Fusaric acid, moniliformin, trichothecenes by Fusarium spp. have also been recorded by many workers [12,13]. Similarly, CFCF obtained from fermented broth of Phoma sp FGCCW #54 also exhibited broad spectrum herbicidal activity and was responsible for 100% damage to P.hysterophorus. CFCF of Sclerotium rolfsii FGCCW#08 caused significant damage (>50%) to P. hysterophorus but failed to exhibit herbicidal property against the other target weeds. CFCF of Alternaria alternata FGCCW#32; Colletotrichum dematium FGCCW# 09; Curvularia lunata FGCCW#25 and Myrothecium roridum FGCCW# 03 did not cause any appreciable damage to the target weeds. Variation in phytotoxicity due to toxin has also been recorded by other workers [10,12,14-17]. Quereshi et al. [18], recorded significant herbicidal activity in CFCF obtained from 21 days old fermented broth of Phoma sp FGCCW#54 against Parthenium. Variations in phytotoxicity of CFCF of S.rolfsii isolates against P.hysterophorus have also been reported by Pandey et al., [19]. A.alternata FGCCW#32 and C.lunata FGCCW#25 failed to cause considerable damage against target weeds, however in contrast, to this, Saxena and Pandey [7] and Thapar et al. [11], have reported appreciable phytotoxicity of secondary metabolites from Alternaria isolate LC#110 and LC#104 against Lantana camara and C.lunata Ph#38 against Parthenium respectively.
 

Tested strains

 

Phytotoxicity (%)*

Parthenium hysterophorus

Lantana camara

Hyptis suaveolens

Xanthium strumarium

Cassia tora

Sida actua

Antigonon leptopus

Alternaria alternata
FGCCW#32

19±3

4±4

3±2

3±4

6±3

15±3

6±4

Colletotrichum dematium
FGCCW#09

50±4

17±1

0

0

0

0

0

Curvularia lunata
FGCC#25

37±4

30±2

40±1

32±1

21±2

22±1

30±1

Fusarium oxysporum
FGCC#43

100

100

100

100

100

100

100

Fusarium moniliforme
FGCCW#16

98±1

98±3

98±2

98±1

98±2

98±2

98±3

Fusarium roseum
FGCCW#43

96±4

96±2

98±2

98±1

98±3

98±3

96±4

Myrothecium roridum
FGCCW#55

0

10±1

9±3

0

2±2

0

0

Phoma herbarum
FGCCW#54

100

65±2

61±2

69±3

68±1

58±3

69±2

Sclerotium rolfsii
FGCCW#08

58±2

33±2

30±1

0

19±3

8±1

0

Table 1: The phytotoxicity activity of Cell free culture filtrate of selected isolated fungi in shoot cut bioassay.
Fermentation medium: Modified Richard’s Broth. Incubation Temperature: 28±1°C
* Data are presented as Means ±SD from three replicates.
 

Tested strains

 

Phytotoxicity (%)*

Parthenium hysterophorus

Lantana camara

Hyptis suaveolens

Xanthium strumarium

Cassia tora

Sida actua

Antigonon leptopus

Alternaria alternata
FGCCW#32

17±3

5±4

4±2

6±4

9±3

15±3

6±4

Colletotrichum dematium
FGCCW#09

51±2

18±1

0

0

0

0

0

Curvularia lunata
FGCC#25

35±1

30±1

38±2

30±1

25±1

21±1

30±1

Fusarium oxysporum
FGCC#43

100

100

100

100

100

100

100

Fusarium moniliforme
FGCCW#16

98±2

98±1

98±4

98±3

98±2

98±1

98±3

Fusarium roseum
FGCCW#43

96±1

96±3

96±3

96±4

96±3

96±2

96±4

Myrothecium roridum
FGCCW#55

0

9±1

10±

0

12±2

0

0

Phoma herbarum
FGCCW#54

100

65±2

61±3

69±2

68±1

58±4

69±2

Sclerotium rolfsii
FGCCW#08

60±4

35±3

30±1

0

19±2

4±1

0

Table 2: The phytotoxicity activity of Cell free culture filtrate of selected isolated fungi in seedling bioassay.
Fermentation medium: Modified Richard’s Broth. Incubation Temperature: 28±1°C
* Data are presented as Means ±SD from three replicates

The above findings clearly indicate that among nine strains screened, CFCF of Fusarium spp. and Phoma sp. FGCCW#54 have significant broad-spectrum herbicidal potential against the test weeds viz Parthenium hysterophorus, Lantana camara, Xanthium strumarium, Cassia tora, Hyptis suaveolens, Sida acuta and Antigonon leptopus. Thus, it can be concluded that these strains could be used for production of herbicidal compounds. However, further investigations are needed to determine mode, mechanism and site of action and develop suitable formulations and production technologies before its large-scale application.

ACKNOWLEDGEMENT

We are grateful to the Head, Dept of Biological Sciences, R.D. University, Jabalpur for laboratory facilities. Financial assistance received from Council of Scientific and Industrial Research (CSIR) New Delhi is also thankfully acknowledged.

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Citation: Singh AK, Pandey AK (2019) Screening of Herbicidal Potential of Crude Extract of Isolated Fungal Strains against some Noxious Weeds of India: A Preliminary Evaluation. J Agron Agri Sci 2: 015.

Copyright: © 2019  Ajay Kumar Singh, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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