ArticlePDF Available

Antagonistic Activity Of Endophytic Bacteria Isolated From Mentha Rotundifolia L.

Authors:
Abla Elhartiti at Université Ibn Tofail
Abla Elhartiti
Elhabchi Souad
Elhabchi Souad
Elhabchi Souad
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Abdelhadi Hichar at Université Ibn Tofail
Abdelhadi Hichar
Omar Bazdi
Omar Bazdi
Omar Bazdi
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Show all 5 authorsHide
Download full-text PDF
Read full-text
Download citation

Abstract and Figures

This study is implemented for the isolation, purification and identification of endophytic bacteria which produces antifungal substances from the roots of Mentha rotundifolia L. The 59 obtained bacterial isolates were tested for their antagonistic activity by the dual confrontation against the phytopathogenic fungi Fusarium oxysporum, Aspergillus Niger and Botrytis cinerea. Eight bacterial strains were selected for their strong antifungal activity. These are strains M21, M23, M3a, M4, M14d and M3c which belong to the family Bacillaceae, M12 and M3b which belongs to the family of Pseudomonadaceae. Among these, three bacterial strains namely M21, M23 and M12 induce 70% of inhibition of mycelial growth of phytopathogenic fungi Fusarium oxysporum and Aspergillus Niger while the five bacterial strains M3a, M3c, M3b, M4 and M14d have proved to be effective in inhibiting more than 60% of mycelial growth of Botrytis cinerea. Keywords: Bacteria endophytic, Bacillaceae, Pseudomonadaceae, antagonistic activity, Fusarium oxysporum, Aspergillus niger, Botrytis cinerea
Figures - uploaded by Abla Elhartiti
Author content
All figure content in this area was uploaded by Abla Elhartiti
Content may be subject to copyright.
Content uploaded by Abla Elhartiti
Author content
All content in this area was uploaded by Abla Elhartiti on May 29, 2016
Content may be subject to copyright.
INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 4, ISSUE 12, DECEMBER 2015 ISSN 2277-8616
36
IJSTR©2015
www.ijstr.org
Antagonistic Activity Of Endophytic Bacteria
Isolated From Mentha Rotundifolia L.
Elhartiti Abla, Elhabchi Souad, Hichar Abdelhadi, Omar Bazdi, Ounine Khadija
Abstract: This study is implemented for the isolation, purification and identification of endophytic bacteria which produces antifungal substances from
the roots of Mentha rotundifolia L. The 59 obtained bacterial isolates were tested for their antagonistic activity by the dual confrontation against the
phytopathogenic fungi Fusarium oxysporum, Aspergillus Niger and Botrytis cinerea. Eight bacterial strains were selected for their strong antifungal
activity. These are strains M21, M23, M3a, M4, M14d and M3c which belong to the family Bacillaceae, M12 and M3b which belongs to the family of
Pseudomonadaceae. Among these, three bacterial strains namely M21, M23 and M12 induce 70% of inhibition of mycelial growth of phytopathogenic
fungi Fusarium oxysporum and Aspergillus Niger while the five bacterial strains M3a, M3c, M3b, M4 and M14d have proved to be effective in inhibiting
more than 60% of mycelial growth of Botrytis cinerea.
Keywords: Bacteria endophytic, Bacillaceae, Pseudomonadaceae, antagonistic activity, Fusarium oxysporum, Aspergillus niger, Botrytis cinerea
————————————————————
1 INTRODUCTION
More than two hundred species of dicots plants and
monocots are likely to be attacked by pathogenic fungi
Fusarium oxysporum, Aspergillus Niger and Botrytis
cinerea. Responsible for the significant economic losses on
cultures before and after the harvest [1, 2, 3]. The biological
control is a promising alternative [4]. Indeed, the endophytic
bacteria may be used for landing in the misuse of chemicals
[5]. The objective of this study is to isolate bacteria
endophytes of Mentha rotundifolia L. and to compare them
with mycelium phytopathogenic (F.oxysporume. A.niger and
B.cinerea).
2 MATERIEL ET METHODES
2.1 Isolation and purification of endophytic bacteria
of Mentha rotundifolia L.
The root samples of the plant Mentha rotundifolia L. were
taken at Ouled Amar parcels located in the region of Gharb
Chrarda Beni Hssen, Morocco.Ten grams of roots were
sterilized in the surface with sodium hypochlorite at 1% for
90 seconds, rinsed several times with sterile distilled water,
and ground in the physiological water for 60s. Depositing
0.5 ml of the dilution 1/10 then plated on Petri dishes
containing the agar medium. The incubation is performed at
28 ° C for 48h. The purity of the strains was checked by
successive subculture on agar medium. The purified
bacteria are then stocked at -20 ° C in flasks containing
nutrient broth of 20% of glycerol.
2.2 The antagonist activity of bacterial isolates in
vitro
Bacterial isolates were tested for antagonism against
Fusarium oxysporum, Aspergillus Niger and Botrytis cinerea
on the PDA by the dual culture technique [6].The bacterial
strains are inoculated in rectilinear streaks at opposite ends
of the medium. A cylinder of 4 mm in diameter mycelium
phytopathogenic is deposited in the center of the Petri dish.
The control contains only a phytopathogenic fungi washer.
The petri dish were incubated at 28 ° C. the inhibition of
mycelial growth was observed after five or seven days. The
percentage of the mycelial growth is estimated by the
formula:
(%) = Inhibition= ( – )/ * 100 [7]
- : maximum radial distance fungus growth.
-  : radial distance on a line towards the antagonist.
2 Biochemical identification of isolated bacteria of
Mentha rotundifolia L.
The bacterial isolates were identified based on the
characters of the cultural tests, morphological, and
biochemical: the Gram reaction, respiratory-type with
catalase and oxidase etc. as described in the manual of
Bergey's bacteriology 2001 [8].
3 RESULTS AND DISCUSSION
3.1 The antagonistic activity of bacterial isolates in
vitro
From the roots of Mentha rotundifolia L, we isolated and
purified 59 bacterial strains. Among them 17 induce an
inhibition of growth of F.oxysporum, A. Niger and B.cinerea.
(Table I).
__________________
Elhartiti Abla, Elhabchi Souad, Hichar Abdelhadi,
Omar Bazdi, Ounine Khadija
Laboratory of biology and Health, Applied
Microbiology Team; Faculty of Sciences Ibn Tofail
University B.P: 133 14000, Kénitra- MOROCCO.
INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 4, ISSUE 12, DECEMBER 2015 ISSN 2277-8616
37
IJSTR©2015
www.ijstr.org
Table 1: Percentage of inhibition of Fusarium oxysporum (F.o), Aspergillus Niger (A.n) and Botrytis cinerea (B.c) by the
isolated bacteria of Mentha rotundifolia L.
We find that 17, 16 and 12 isolated strains have induce an
inhibition of F.oxysporum, A. Niger and B.cinerea
respectively with a percentage of inhibition higher than 30%.
The strains M21, M23 and M12 are remarkably effective.
They have trained the inhibition of mycelial growth
F.oxysporum and A. Niger with a higher percentage of 70%
(Figure 1 and Figure 2). while the five other isolates M3a,
M3c, M3b, M4 and M14d shown their effectiveness by
inhibiting superior 60% of mycelial growth of B.cinerea
(Figure 3)
Figure 1: Effects of bacterial isolates M23, M21 and M12 on
mycelium growth of Fusarium oxysporum.
Figure 2: Effects of bacterial isolates M23, M21 and the
M12 on the mycelium growth of Aspergillus Niger
strains
% inhibition of mycelial growth
F.o
A.n
B.c
control
0
0
0
M6a
39,47
37,5
11,11
M16c
40,78
31,25
22,22
M14e
57,89
68,75
57,77
M14c
60,52
75
56,36
M7a
64,47
66,25
45,45
M3a
67,10
65
67,27
M3c
63,15
62,50
63,63
M4
67,10
65
60
M14d
53,94
66,25
63,63
M21
75
76,25
56,36
M23
73,68
75
52,72
M12
70,52
77,5
54,54
M3b
67,10
68,75
60
M5b
67,10
57,5
36,36
M2b
60,52
66,25
16,36
M9b
47,36
50
18,18
M3d
30,26
12,25
7,27
INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 4, ISSUE 12, DECEMBER 2015 ISSN 2277-8616
38
IJSTR©2015
www.ijstr.org
Figure 3: Effects of bacterial isolates M3b, M3a, M3c,
M14D and M4 on the mycelium growth of Botrytis cinerea
3 Biochemical identification of endophytic
bacteria isolated from Mentha rotundifolia L.
The 17 strains induced an inhibition of mycelial growth of
phytopathogenic fungi three F. oxysporum, A. Niger and
B.cinerea they are divided in two groups according to their
Gram stain (Tables 2 and 3). They are strictly aerobic bacilli
of oxidase and catalase positive, and they are all capable of
reducing the nitrate to nitrite or ammonia but unable to
produce H2S.
Tableaux 2: biochemical and physiological characters of Gram positive isolates isolated from de Mentha rotundifolia L.
Strains
Test
M4
M6a
M7a
M14c
M14e
M14d
M3c
M16c
M21
M23
Mobility
+
+
+
+
-
+
+
+
+
+
Mannitol
+
+
+
+
+
+
+
-
-
-
Glucose
-
-
-
-
-
-
-
-
-
-
Lactose
+
-
+
+
+
+
+
+
-
+
Methyl red
-
+
-
-
-
-
-
+
-
-
Voges Proskauer
+
-
+
+
+
+
+
-
+
+
Gelatinase
+
-
+
+
+
+
+
-
+
+
casein hydrolysate
+
-
-
+
-
+
-
+
+
+
starch hydrolysate
+
-
-
+
+
+
+
-
+
+
Among the 11 Gram positive strains, nine are sporulation. They belong to Bacillaceae family. The two other bacterial strains are
not acid-resistant and are non-sporulation. They are unable hydrolyze the starch. Which specifies as Corynebacterium xerosis.
Tables 3: biochemical and physiological characters of Gram-negative isolates isolated from de Mentha rotundifolia L.
strains
Test
M5b
M9b
M2b
M3b
M12
M3d
Citrate
+
+
+
-
+
-
Mobility
+
+
-
+
+
+
Mannitol
-
+
-
+
+
-
Lactose
-
-
-
+
-
-
Methyl red
+
+
+
-
+
-
Voges Proskauer
-
-
-
+
-
+
Gelatinase
+
+
-
+
+
-
casein hydrolysate
-
+
-
+
-
-
starch hydrolysate
-
-
-
+
+
-
The six bacterial Gram-negative isolates belong to
Pseudomonadaceae family. The bacterial isolates M21,
M23, M3a, M4, M14d and M3c belong to the Bacillaceae
family. Among these strains, M21 and M23 induced an
inhibition of mycelial growth greater than 70% of
F.oxysporum and A. Niger while the other strains M3a, M4,
M3c and M14d inhibited 60% of the mycelium growth
B.cinerea. These results are in agreement with those left by
INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 4, ISSUE 12, DECEMBER 2015 ISSN 2277-8616
39
IJSTR©2015
www.ijstr.org
Nourozian [9], Munimbazi and Bullerman [10] who showed
that the Bacillus sp have a strong potential of inhibition of
mycelium growth of many Aspergillus and Fusarium species
Also Sadfi-Zouaoui [11] found that the Bacillus sp. were
antagonistic to B.cinerea. In previous researches, some
strains of Bacillus sp, such as Bacillus subtilis, played an
important role in the biological control of fungal diseases of
post-harvest and the production of antibiotics [12]. M12 and
M3b are Pseudomonadaceae which induce an inhibition
rate higher than 70 % against the phytopathogenic fungi
F.oxysporum and A.niger for M12. Thus, the strain M3b
induced an inhibition superior than 60% against B.cinerea.
This is in agreement with a description that which was made
by several authors who described the successful control of
the Aspergillus flavus by the antagonistic Pseudomonas
fluorescens [13, 14]. Srivastava and Shalni [15] also
reported the antifungal potential of Pseudomonas
fluorescens against the pathogenic fungus, Fusarium sp.
Paez et al [16] have indicated that Pseudomonas putida
and Pseudomonas aeruginosa had great antagonistic
effects on B.cinerea.
4 CONCLUSION
From these results, we can conclude that the bacterial
strains M21, M23, M3a, M4, M14d and M3c belong to the
Bacillaceae family, M12 and M3b belong to the
Pseudomondasea family have an inhibitory effect on the
phytopathogenic fungi Fusarium oxysporum, Niger
Aspergillus and Botrytis cinerea, and therefore can serve as
a biological control agent.
REFERENCES
[1] Rosslenbroich H. J. et Stuebler D. 2000.Botrytis
cinerea – history of chemical control and novel
fungicides for its management. Crop Prot., 19, 557-
561.
[2] Gudelj I., Fitt B. D. L. et van den Bosch F. 2004.
Evolution of sibling fungal plant pathogens in
relation to host specialization, Phytopathol., 94,
789-795.
[3] Hubert J., Stejskal V., Munzbergova Z., Kubatova
A., Vanova M. et Zdarkova E. 2005. Mites and
fungi in heavily infested stores in the Czech
Republic, J. Econ. Entomol., 97, 2144-2153.
[4] Lin L, Qiao YS, Ju ZY, Ma CW, Liu YH, Zhou YJ,
Dong HS 2009. Isolation and characterization of
endophytic Bacillus subtilis Jaas ed1 antagonist of
eggplant Verticillium wilt. Biosci Biotechnol
Biochem 73:1489-1493.
[5] Moenne.Loccoz., Y., Powell, J., Higgins, P.,
McCarthy, J., O´Gara, F.1998. An investigation of
the impact of biocontrol Pseudomonas Fluorescens
F113 on the growth of sugar beet the performance
of subsequent clover-Rhizobium symbiosis.
Appl.Soil.Ecol. 7, pp.225-237.
[6] Zhao LF, Deng ZS, Yang WQ, Cao Y, Wang ET,
Wei GH (2010). Diverse rhizobia associated with
Sophora alopecuroides grown in different regions
of Loess Plateau in China. Syst Appl Microbiol
33:468-477.
[7] Wang S.L., Hsaiao W.J., Chang W.T. et al., 2002.
Purification and characterization of an antimicrobial
chitinase extracellularly produces by monoscus
purpureus CCR 31499 in a shrimp and crab shell
powder medium. J.Aric.Food.Chem., 50: 2249-
2255
[8] Bergey´s Manual of Systematic Bacteriology, 2001,
second ed.
[9] Nourozian, J., H.R. Etebarian and G.
Khodakaramian, 2006. Biological control of
Fusarium graminearum on wheat by antagonistic
bacteria, Songklanakarin J. 19. Sci. and Technol.,
28: 29-38.
[10] Munimbazi, C. and L.B. Bullerman, 1998. Isolation
and partial characterization of antifungal
metabolites of Bacillus pumilus. J. Appl. Microbiol.,
84: 959-968.
[11] Sadfi-Zouaoui N., Essghaier B., Hannachi I.,
Hajlaoui M.R and Boudabous A. 2007. First report
on the use of moderately halophilic bacteria
against stem canker of greenhouse tomatoes
caused by Botrytis cinerea. Ann. Microbiol. 57(3) :
337-339.
[12] Toure, Y., Ongene, M., Jacques, P.,Guiro, A.,
Thonart, P., 2004. Role of lipopeptides produced by
Bacillus subtilis GA1 in the reduction of grey mould
disease caused by botrytis cinerea on apple.
Journal of Applied Microbiology 96, 1151-1160.
[13] Jeffrey, D., L. Palumbo, James, Baker, E. Noreen
and Mahoney, 2006. Isolation of Bacterial
Antagonists of Aspergillus flavus from Almonds,
Microbial Ecol., 52(1): 45-52.
[14] Palumbo, J.D., T.L. O’Keeffe and H.K. Abbas,
2007. Isolation of maize soil and rhizosphere
bacteria with antagonistic activity Aspergillus flavus
and Fusarium verticillioides, J. Food Protection, 70:
1615-1621.
[15] Srivastava, R. and Shalni, 2008. Antifungal Activity
8. Pseudomonas fluorescence Against Different
Plant Pathogenic Fungi, Electronic J.
Environmental, Agriculture and Food Chemistry, 7:
2789-2796.
[16] Paez, M., Martienz-Nieto, P., Bernal-Castillo, J.,
2005. Siderophore producing Pseudomonas and
pathogenic Rhizoctonia solani and Botrytis cinerea
antagonists. Univ. Sci. 10, 65-74.
... For highlighting the roles of the relationship between endophytic bacteria and plants on bioactive compound production, many studies reported the production capacity of bioactive compounds by endogenous bacteria in medicinal plants. Moreover, these studies also provided new and useful insights needed to inhibit pathogens with a view to human health and other potential medical applications (Abla et al., 2015). ...
... Pseudomonas, Bacillus, and Burkholderia from chili pepper plants (Capsicum annuum) show antifungal activities against Alternaria panax, F. Oxysporum, Botrytis cinerea, Phytophthora capsici, and Colletotrichum acutatum (Haque et al., 2015). Bacillus and Pseudomonas endophytes, isolated from the roots of Metha rotundifolia, inhibited 70% of the mycelial growth in phytopathogenic fungi such as F. oxysporum and Aspergillus niger (Abla et al., 2015). isolated 78 bacterial endophytes from 8 medicinal plants, 10 of which were able to inhibit the growth of Trichoderma koningii, Aspergillus niger, A. avamori, and Fusarium oxysporum (Jalgaonwala and Mahajan, 2011). ...
View
... The five endophytic fungi evaluated for antagonism by dual plate culture techniques (Abla et al. 2015) against test bacterial isolates like Gram-positive bacteria-Staphylococcus aureus (MTCC 902) and Gram-negative bacteria-Pseudomonas aeruginosa (MTCC-4734), Salmonella enterica (MTCC-738) and Xanthomonas campestris (MTCC-2286) were obtained from the Institute of Microbial Technology (IMTECH), Chandigarh, India. Plant pathogenic fungi isolated from the infected tomato, chickpea, and chilli plants (Alternaria alternata, Fusarium oxysporum and Sclerotium rolfsii). ...
In-vitro and in-silico studies on plant growth promotion of endophytic fungi of Ischaemum ciliare Retz.
Article
Full-text available
  • Feb 2024
  • J PLANT PATHOL
The current work sought to characterize the secondary metabolites produced by endophytic fungi from the perennial grass Ischaemum ciliare Retz. and examine in-vitro and in-silico investigations on plant growth promotion. Cochliobolus fallax, Cochliobolus sp., Pestalotiopsis sp., Fusarium oxysporum, and Penicillium islandium were isolated from the aerial part of I. ciliare, and two endophytic fungi, Curvularia protuberata and Pestalotiopsis guepinii, were identified based on ITS rDNA sequence analysis. Furthermore, C. protuberata, F. oxysporum, and P. guepinii demonstrated high antagonistic activity against pathogenic fungal and bacterial strains. They showed significant plant growth promotion in the ethyl acetate extracts of C. protuberata and P. guepinii and the methanolic extract of I. ciliare on Capsicum annuum and Zea mays seedlings in vitro. OHR-LC MS analysis of P. guepinii ethyl acetate extract and I. ciliare methanolic extract identified ten and seventeen potential secondary metabolites, respectively. Furthermore, the docking analysis established the ligand-receptor interaction. It revealed that eight out of ten P. guepinii compounds and nine out of seventeen I. ciliare compounds have a strong binding affinity towards 3OGM, 4LDY, and 6KU3 target proteins. The endophytic fungal and host metabolite research in-vitro and in-silico aids in predicting compounds with bioactive principles, bio-control agents, and plant growth promoters.
View
... Pseudomonas stutzeri and Trichoderma viride possessed the highest antagonistic activity against Botrytis cinerea and Fusraium oxysporium, respectively. These results are supported by similar results reported by [1,35,74]. ...
Seed-endophytes empower Anthemis cotula to expand in invaded range
Article
Full-text available
  • Apr 2023
The seeds serve as a biological vehicle for alien species to colonize new habitats and seed endophytes may enhance their chances of survival and spread in the introduced range. Thus, the endophytic diversity associated with the seeds of alien species and their plant growth-promoting activities need to be investigated in detail. Here we explored the composition and diversity of culturable endophytic microbes inhabiting achenes of Anthemis cotula L. an invasive alien species in Kashmir Himalaya. A total of 812 culturable endophytic isolates were recovered from seeds of A. cotula, comprising 716 bacterial isolates, and 96 fungal isolates. Taxonomic binning resolved the isolates into 39 bacterial OTUs and 22 fungal OTUs belonging to 4 bacterial phyla and 2 fungal phyla. The relative abundance of these culturable seed endophytic taxa differed significantly across the sites (p
View
... Data represented in Table (S3, see supplementary data) revealed that the average growth inhibition percentages of A. alternata, F. oxysporum, V. dahlia, and P. ultimum in presence of 600 mM NaCl were 44.85%, 42.45%, 34.03%, and 16.37%, respectively, and significantly (p ≤ 0.05) different from those recorded in absence of salinity which was 42.38%, 40.07%, 37.09%, and 15.67%, respectively (p ≤ 0.05). Consistent with our results, Abla et al. (2015) isolated bacterial endophytes belonging to Bacillaceae and Pseudomonadaceae from Mentha Rotundifolia L. with in-vitro antagonistic activity against the phytopathogenic fungi Fusarium oxysporum, Aspergillus niger with inhibition percent 70%, and Botrytis cinerea with inhibition percent 60%. On the other hand, bacterial endophytes are associated with nodules of soybean (Glycine max L.) and belonging to Enterobacter, Acinetobacter, Pseudomonas, Ochrobactrum, and Bacillus have an inhibitory effect on the phytopathogen Phytophthora sojae with inhibition percent up to 70% (Zhao et al. 2018). ...
Biotechnological application of plant growth‑promoting endophytic bacteria isolated from halophytic plants to ameliorate salinity tolerance of Vicia faba L.
Article
Full-text available
  • Oct 2021
Salinity is a major problem afecting crop production all over the world. A wide range of adaptation strategies are required to overcome this problem. Endophytic bacteria can build a symbiotic association with their host to improve host plant salt tolerance. In this study, eighteen bacterial endophyte strains were isolated from two native halophytic plants Arthrocnemum macrostachyum and Spergularia marina, and identifed as Bacillus, Brevibacillus, Agrobacterium, and Paenibacillus. These endophytic strains exhibit plant growth-promoting activities including phosphate solubilizing, ammonia production, biocontrol of phytopathogen, extracellular enzymatic activities, and indole-3-acetic acid production under normal and salinity stress. A pot experiment was conducted under feld conditions to alleviate the harmful efects of soil salinity on bean (Vicia faba L.) by inoculating their seeds with the most potent bacterial isolates Bacillus subtilis (AR5) and Bacillus thuringiensis (BR1). Salinity treatments induced a signifcant decrease in both growth parameters and metabolic activities, while the activity of antioxidant enzymes and proline content was signifcantly increased. However, salinity stress induced higher contents of Na+ and decreased contents of N+, P3+, K+, Ca2+, Mg2+, and K+:Na+, it was found that treatment with B. subtilis (AR5) and B. thuringiensis (BR1) individually or in a combination mitigated the efect of salt stress and improved the plant height, shoot dry weights, proline contents, enzymes activities as well enhanced the accumulation of mineral nutrients in shoot plants. Our results concluded that treatment with co-inoculation of B. subtilis (AR5) and B. thuringiensis (BR1) exerted the greatest efect in alleviating the harmful efect of soil salinity stress and can be used as a suitable bio-approach to reclaim salinity-stressed soils
View
... found in this study by Pleban et. al., (1995) were inoculated in to cotton plant to study in vivo efficiency. B. cereus, B. subtilis and B. pumilus reduced 51%, 46% and 56% disease incidence by Rhizoctonia solaniI and 79%, 72% and 26% disease incidence by Sclerotium rolfsii. Pleban et. al., (1995) did not use Pseudomonas for this purpose. However, Abla et. al., (2015) isolated 59 bacterial strains from roots of Mentha rotundifolia L. mostly belonging to Pseudomonadaceae family. In addition to the antagonism to F. oxysporum, the endophytes could also restrict Aspergillus Niger and Botrytis cinerea with inhibition of more than 60%. ...
ASSOCIATION OF MICROBIAL DIVERSITY WITH POST HARVEST CROPS AND BIOPROSPECTING OF ENDOPHYTIC MICROORGANISMS FOR MANAGEMENT
Chapter
Full-text available
  • Jul 2019
While thinking about the bigger scenario of global food security, loss of food materials during post harvest period is bigger challenge. About one third of food products get wasted before reaching the consumers stomach and this loss is big in tropical and less developed countries. Damage of harvested food crops and stored crop produce by microbial infestation shares a quite big space in this regard. Crops may be infected by pathogenic microflora in pre-harvest stage and then may flourish and damage the food materials in post harvest period especially in the storage. Most of the time these microbial association leads to the deterioration of food crops like fruits, vegetables and cereal grains 264 which causes great economic loss. Microbial pathogens like Botrytis, Penicillium, Sclerotium, Colletotrichum, Dothiorella, Phytophthora, Pythium, Rhizopus and Xanthomonas cause various rots, spots in many fruits and vegetables. Aspergillus, Penicillium, Fusarium species are largely involved in the production of mycotoxins mostly in cereal grains. Long term control of the post harvest pathogens with the application of synthetic chemicals is never the best of idea. Endophytic microorganisms (bacteria, fungi and actinomycetes) which live asymptomatically inside plant tissues possess the potential to inhibit a broad spectrum of these pathogens. Antagonistic activities of endophytes like Muscodor, Trichoderma, Penicillium, Phoma, Phaeosphaeria, Bacillus, Pseudomonas, Streptomyces, etc. have been well explored by many researchers throughout the world. The current review has highlighted the enormous potential of this important microbial group to control the post harvest pathogens including storage pathogens. The research gaps in this area has also been indicated while the need for carrying out advanced research and devising suitable management strategies using endophytes has been brought forward.
View
... The isolates showed corresponding antagonistic activity against the fungal pathogens; A. alternata, F. oxysporum, P. ultimum and V. dahlia in a growth media amended with 600 mM NaCl. Abla et al., 2015, isolated bacterial endophytes belonging to Bacillaceae and Pseudomonadaceae from Mentha Rotundifolia L. with in vitro antagonistic activity against the phytopathogenic fungi Fusarium oxysporum, Aspergillus niger and Botrytis cinereal. Isolated endophytes inhibited the growth of Fusarium oxysporum and Aspergillus niger by 70 %, while inhibited the growth of Botrytis cinereal by more than 60 %. ...
PLANT GROWTH PROMOTING ACTIVITIES OF ENDOPHYTIC BACTERIA ASSOCIATED WITH THE HALOPHYTE ARTHROCNEMUM MACROSTACHYUM (MORIC) K. KOCH. PLANT. BY
Article
Full-text available
  • Jan 2018
Plant growth and production is negatively affected by soil salinity. However, there is an increasing interest to use plant growth and promoting bacterial (PGPB) endophytes to enhance plant growth under salinity stress. During our study, endophytic bacteria isolated from halophyte plants were used as inoculant to stimulate plant growth and increase the tolerance of non-halophytic crops to salinity. In present study, two endophytic bacteria belonging to Bacillus sp. were isolated from the halophyte Arthrocnemum macrostachyum growing in salty soils. These endophytes were examined for having plant growth promoting (PGP) properties such as enzymatic activities, ammonia and indole acetic acid production. In addition, effect the inoculation of these endophytes on germinating seeds of maize (Zea mays L.) under concentration of 600 mM NaCl was studied. The results showed that, bacterial inoculation of plants mitigated the effects of high salinity on plant growth and physiological performance. Endophytic bacteria isolated from A. macrostachyum seem to play an important role in plant salt tolerance and it could be an adequate tool to enhance plant growth under salinity stressed conditions.
View
... Pathogenic fungi used are Aspergillus sp., Fusarium sp., and Penicillium sp. Agar plug method was done based on Abla et al. (2015). Pathogenic fungi were grown in Potato Dextrose Agar (PDA) (Oxoid) supplemented with chloramphenicol (Kimia Farma) at 30 o C for 7 days. ...
View
Identification of endophytic bacteria in medicinal plants and their antifungal activities against food spoilage fungi
Article
  • Aug 2019
Endophytes are fungi or bacteria living in the intracellular parts of the plants. In this study, 12 medicinal plants were examined for the presence of endophytic bacteria. Antifungal activities of the isolates were determined against Aspergillus flavus PTCC 5006, Penicillium citrinum PTCC 5304, Aspergillus fumigatus PTCC 5006, Fusarium oxysporum MTCC 284, and Rhizopus stolonifer. In order to determine the metabolites characteristics of these isolates, catalase and proteolytic enzyme treatments were assessed. Moreover, approximate molecular weights of the antifungal substance were measured by fractionation method and the volatile compounds were determined by using GC-mass spectroscopy. Finally, 16s rRNA gene sequencing confirmed the strain of the bacteria. Twenty-one endophytic bacteria, out of a total of 82 isolates, showed antifungal activities against all five spoilage fungi. The results of the PCR assay revealed two species: Bacillus pumilus and B. safensis. Proteolytic enzyme activities and the fractionation of the supernatants proved that more than one compound was responsible for the antifungal activities. This compound could be proteins, peptides, and other low-molecular compounds, such as Butanal, 3-methyl-, Propene, 2-butene, 2-heptanone, 6-methyl-5-methylene-, and 6-oxabicyclo[3.1.0] hexane, which all were identified in the headspace of the GC-mass spectroscopy.
View
ISOLATION AND CHARACTERIZATION OF ENDOPHYTIC BACTERIA FROM THE LEAF EXPLANTS OF Avicennia marina (Forsk.)
Conference Paper
Full-text available
  • Sep 2016
Mangroves have important role in maintaining coastal area ecosystem as well as in protecting it from abrasion. The overexploitation of mangroves is of today's concern because it threatened the sustainability of the ecosystems. Avicennia marina (Forsk.) is one of mangroves that grow in Ekowisata Mangrove Wonorejo Surabaya. The previous aim of the experiment was to clone this species by plant tissue culture method in order to provide a sufficient number of seedlings for reforestation. However, there were a lot of obstacles in obtaining the sterile explants because the contaminants that live inside the leaf samples (endophytic). The isolation of endophytic bacteria was done after the surface sterilized leaf discs were cultured on MS and PCA media. The characterization was conducted biochemically based on Bergey's manual of Determinative Bacteriology. The results confidently showed that there were two genera of endophytic bacteria inside the leaves of Avicennia marina (Forsk.), i.e. Bacillus and Corynebacterium. However, there was also another genus but still not sure yet whether it was Shigella or Yersinia. The results were beneficial to further investigate the strategy to be able to obtain sterile leaf explants of Avicennia marina (Forsk.).
View
Siderophore producing Pseudomonas as pathogenic Rhizoctonia solani and Botrytis cinerea antagonists
Article
Full-text available
  • Jan 2005
RESUMEN Pseudomonas aeruginosa, Pseudomonas putida biovar B, Pseudomonas marginalis y Burkholderia cepacia, aisladas de rizosfera y filosfera de plantas de rosa y alstroemeria, identificadas por ensayos bioquímicos y cultivadas en medio King B, mostraron propiedades antagónicas contra los patógenos (se usó medio PDA agar par el cultivo) Rhizoctonia solani y Botrytis cinerea. Estas propiedades coincidieron con la presencia de un sideróforo, sustancia polar con bandas de absorción en 260 nm y 402 nm. Se observó incremento del crecimiento longitudinal de las plantas, medido sobre el tallo central, por influen- cia de P. putida biovar B, P. aeruginosa y P. marginalis. El crecimiento de rizomas (a: 0.05) fue notorio bajo la influencia de P. marginalis. Palabras clave: Alstroemeria, Antagonismo, Filosfera, Fito-patógeno, Pseudomonas sp, Sideróforo. ABSTRACT Pseudomonas aeruginosa, Pseudomonas putida biovar B, Pseudomonas marginalis and Burkholderia cepacia, isolated from the rhizosphere and the phyllosphere of rose and alstroemeria plants, identified by biochemical assays, and cultured in King B medium, showed antagonistic properties, in vitro, against the pathogens Rhizoctonia solani and Botrytis cinerea (PDA agar medium was used for culturing). These properties coincided with the presence of siderophore, a polar substance with bands of absorption at 260 nm and 402 nm. Longitudinal plant growth enhancement, measured on the central stem of alstroemeria, was observed under the influence of P. putida biovar B, P. aeruginosa and P. marginalis. Rhizome growth (a: 0.05) was notable under the influence of P. marginalis.
View
Biological control of Fusarium graminearum on wheat by antagonistic bacteria
Article
Full-text available
  • Mar 2006
Bacillus subtilis strains 53 and 71, Pseudomonas fluorescens biov1 strain 32 and Streptomyces sp. Strain 3 were evaluated as potential biological agents for control of fusarium head blight (FHB) caused by Fusarium graminearum. Mycelial growth of the pathogen was reduced by cell free and volatile metabolites of bacterial antagonists by 37%-97%. Streptomyces sp. Strain 3 reduced disease severity of FHB 21 d after inoculation. The yield of wheat from plants treated with Streptomyces sp. strain 3 and F. graminearum was significantly greater than in controls inoculated with the pathogen alone. Treatments with Streptomyces sp. alone increased the yield of wheat compared to the uninoculated controls.
View
First report on the use of moderately halophilic bacteria against stem canker of greenhouse tomatoes caused by Botrytis cinerea
Article
  • Sep 2007
Experiments were conducted to study the effect of moderately halophilic bacteria isolated from different Tunisian Sebkhas (hypersaline soils), on stem canker caused byBotrytis cinerea on tomato plants grown under greenhouse conditions. Treatments performed with moderately halophilic isolates ofBacillus subtilis J9 andHalomonas sp. K2-5 significantly reduced stem lesion expansion byB. cinerea on tomato plants under greenhouse conditions. The use of such bacteria may constitute an important alternative to synthetic fungicides, which failed to suppress the development of the fungal pathogen.
View
Antifungal activity of Pseudomonas fluorescens against different plant pathogenic fungi
Article
  • May 2008
Antifungal activity of different strains of Pseudomonas fluorescens were tested against some plant pathogens such as Alternaria cajani, Curvularia lunata, Fusarium sp., Bipolaris sp. and Helminthosporium sp. in in vitro. Different concentrations (1000, 2000, 3000, 4000 and 5000 μg/mL) of Pseudomonas fluorescens were used and maximum spore germination of fungus was inhibited at 4000 and 5000 μg/mL. The result indicated that all the strains of Pseudomonas fluorescens presented a most significant value against Alternaria cajani and Curvularia lunata. Out of the five strains studied, the best result was shown by A-5, which showed almost complete inhibition against pathogenic fungi such as Curvularia lunata and Fusarium sp. at 4000 and 5000 μg/mL while strain L-5 was resistant against Fusarium sp. and Helminthosporium sp. at 5000 μg/mL. Among the fungus tested, bacterial strains C-03 and Pf4-1 were found to be more sensitive to Fusarium sp. and Helminthosporium sp.
View
An investigation of the impact of biocontrol Pseudomonas fluorescens F113 on the growth of sugarbeet and the performance of subsequent clover-Rhizobium symbiosis
Article
  • Mar 1998
  • APPL SOIL ECOL
The biocontrol agent Pseudomonas fluorescens F113 produces the antimicrobial metabolite 2,4-diacetylphloroglucinol and controls the extent of Pythium-mediated damping-off of sugarbeet. A field site without damping-off pressure in 1994 was chosen to investigate the effect of a spontaneous rifampicin-resistant mutant of F113 (i.e., F113Rif) on growth of sugarbeet and subsequent clover, with the objective of developing a database for future risk assessments of genetically-improved derivatives of F113. The inoculant established itself at about 6 log CFU per root system of seed-inoculated sugarbeet for 3 months after inoculation in 1994. In the absence of damping-off disease, the F113Rif treatment did not influence sugarbeet germination and had no effect on root yield, sugar yield, and root quality (contents in α-amino nitrogen, potassium and sodium, alkalinity of root tissue, purity of juice from extracted roots, sugar extractability). In the second year, the site was sown with uninoculated red clover, and F113Rif was found at low population levels in the rhizosphere (2.0 log CFU per root system at the first harvest). The F113Rif treatment had no effect on nodulation of red clover by the resident rhizobia and did not influence foliage dry matter and total nitrogen at the two harvests carried out in 1995. Overall, inoculation of sugarbeet seeds with the phloroglucinol-producing pseudomonad F113Rif had no apparent ecological impact at the level of the sugarbeet crop and the subsequent clover-Rhizobium symbiosis.
View
Botrytis cinerea - History of chemical control and novel fungicides for its management
Article
  • Sep 2000
  • CROP PROT
Infections caused by Botrytis cinerea are of considerable economic importance in grapes, vegetables and berries world-wide, especially when grown under protection. Newly introduced botryticides are the anilinopyrimidines pyrimethanil, cyprodinil and mepanipyrim, the phenylpyrrol fludioxonil and the hydroxyanilide fenhexamid. Biochemical studies indicate that the anilinopyrimidines inhibit the biosynthesis of methionin by blocking cystathionine-β-lyase. The target site of fludioxonil seems to lie in the osmoregulatory signal transmission pathway. Although the exact biochemical target of fenhexamid is not yet characterized, there are clear indications that it is different from all other botryticides known. Therefore, fenhexamid is an effective tool for anti-resistance management strategies.
View
Diverse rhizobia associated with Sophora alopecuroides grown in different regions of Loess Plateau in China
Article
  • Oct 2010
A total of seventy-five symbiotic bacterial strains isolated from root nodules of wild Sophora alopecuroides grown in different regions of China's Loess Plateau were characterized. Based on the combined RFLP patterns, thirty-five genotypes were defined among the rhizobia and they were classified into nine genomic species, including Mesorhizobium alhagi and M. gobiense as the main groups, as well as Agrobacterium tumefaciens, M. amorphae, Phyllobacterium trifolii, Rhizobium giardinii, R. indigoferae, Sinorhizobium fredii and S. meliloti as the minor groups according to the 16S rRNA and recA gene analyses. Five and three lineages of nodA and nifH were found, respectively, in these strains, implying that the symbiotic genes of the S. alopecuroides rhizobia had different origins or had divergently evolved. Results of correspondence analysis showed that there was a correlation between rhizobial genotypes and the geographic origins. Possible lateral transfer of the recA and 16S rRNA genes between the P. trifolii and A. tumefaciens strains, and that of symbiotic genes (nodA, nifH) between different genera, was shown by discrepancies of the phylogenetic relationships of the four gene loci. These results revealed diverse rhizobia associated with wild S. alopecuroides grown in different regions of China's Loess Plateau, and demonstrated for the first time the existence of symbiotic A. tumefaciens strains in root nodules of S. alopecuroides.
View
Isolation and Characterization of Endophytic Bacillius subtilis Jaas ed1 Antagonist of Eggplant Verticillium Wilt
Article
  • Aug 2009
  • BIOSCI BIOTECH BIOCH
An endophytic bacterial strain, Jaas ed1, was isolated from the interior of eggplant (Solanum melongena L.) stems in Jiangsu Province, China. According to the morphological and physiological characteristics and phylogenetic analysis of the 16s rDNA sequence, it was identified as Bacillius subtilis. Strain Jaas ed1 and its cell-free filtrate had strong antifungal activity against Verticillium dahliae. The strain was an internal colonizer within the eggplant without any harmful side effects to the plant. In greenhouse experiments, the strain cell suspension effectively controlled Verticillium wilt of eggplant, and its control efficiency was far more significant than that of the cell-free filtrate after inoculation of V. dahliae.
View
Evolution of Sibling Fungal Plant Pathogens in Relation to Host Specialization
Article
  • Aug 2004
  • PHYTOPATHOLOGY
ABSTRACT Sibling plant pathogens can be grouped according to their host rangesthe following groups: group 1, sibling pathogens with nonoverlapping host ranges; group 2, sibling pathogens with both overlapping and nonoverlapping host ranges; and group 3, sibling pathogens with overlapping host ranges. Using the adaptive dynamics methodology, we investigated the evolution of sibling pathogens in relation to host specialization for groups 1 to 3. In particular, we focused on the role of multiple host niches and a trade-off in infectivity of pathogens to these hosts on the evolutionary outcome. We have shown that this ecological mechanism can explain only the evolution of sibling pathogens in group 1 and that other ecological and epidemiological mechanisms must be responsible for the evolution of sibling pathogens in the other two groups.
View
Isolation and partial characterization of antifungal metabolites of Bacillus pumilus
Article
  • Jul 1998
Antifungal metabolites produced by Bacillus pumilus in Potato Dextrose Broth (PDB) were isolated from culture supernatant fluid by precipitation with ammonium sulphate. The antifungal metabolites inhibited mycelial growth of many species of Aspergillus, Penicillium and Fusarium. They also inhibited production of aflatoxins, cyclopiazonic acid, ochratoxin A and patulin. The metabolites were heat-stable and remained active after sterilization at 121 degrees C for 15 min. Their activity was stable over a wide range of pH (2-10). The metabolites were resistant to hydrolysis by various proteases, peptidases and other enzymes. They were also resistant to denaturation by many protein-denaturing detergents except Nonidet P-40. The metabolites were soluble in water and relatively polar organic solvents. Chromatographic bioassay revealed that a crude precipitate of the metabolites contained only one compound with antifungal activity. The active compound did not form a fluorescent derivative with fluorescamine suggesting that the compound is either a cyclic polypeptide or a non-peptidic compound.
View