PH12017000203A1 - Development and utilization of actinomycetes as biocontrol agents against panama wilt causing-fusarium oxysporum tr4 cavendish banana - Google Patents

Development and utilization of actinomycetes as biocontrol agents against panama wilt causing-fusarium oxysporum tr4 cavendish banana Download PDF

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PH12017000203A1
PH12017000203A1 PH12017000203A PH12017000203A PH12017000203A1 PH 12017000203 A1 PH12017000203 A1 PH 12017000203A1 PH 12017000203 A PH12017000203 A PH 12017000203A PH 12017000203 A PH12017000203 A PH 12017000203A PH 12017000203 A1 PH12017000203 A1 PH 12017000203A1
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Philippines
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streptomyces
actinomycetes
against fusarium
fusarium wilt
biocontrol agent
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PH12017000203A
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A Papa Irene
O Zulaybar Teofila
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Univ Of The Philippines Los Banos
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Abstract

The invention describes a microbe-based biopesticide against Fusarium oxysporum TR4 in Cavendish banana. The biopesticide is composed of different species of actinomycetes, Streptomyces misionensis, Streptomyces olivoverticillatus, Streptomyces phaeoluteichromatogenes and Streptomyces abikoensis. The methodology includes the screening of various bacteria for their antagonistic effect against Fusarium and then identifying the bacteria with the most antagonistic effect against Fusarium. When the said actinomycetes were identified and a carrier was established, field testings were done. Result include the reduction of mortality rate among Cavendish bananas from 56.67 pcnt to 33.3 pcnt via preventive treatments. On the other hand, no significant change in mortality was noted via curative treatment. This is noted in the change from 56.67 pcnt mortality rate of the untreated samples to 55.0 pcnt mortality rate of the treated samples through curative method.

Description

Development and Utilization of Actinomycetes as Biocontrol Agents against Panama Wilt
Causing-Fusarium oxysporum TR4 In Cavendish Banana
TECHNICAL FIELD OF INVENTION
This invention relates to the formulation of a microbe based biocontrol agent against Fusarium oxysporum TR4 composed of various actinomycetes.
BACKGROUND OF THE INVENTION
Cavendish banana is the fourth most important crop worldwide based on production, next to rice, wheat, and maize (Cao, et al, 2005; Akila, et al., 2011;
Thangavellu and Mustaffa, 2009). It is one of the major fruits in the Philippines and is next to coconut in terms of volume of production and export earnings.
One of the economically important diseases affecting barana is Fusarium wilt or
Panama disease. The disease causes yellowing and hanging of leaves, splitting of the pseudostem, significant reduction of fruit yield quality and quantity that ultimately leads to the death of the plant. The manifestation and intensity of the disease varies as affected by the presence of susceptibility genes in the plant genome (Akila, et al, 2011). In pursuit of finding a way to minimize or eradicate the losses due to Fusarium wilt, an array of methods are proposed. One of the methods is the use of naturally occurring microorganisms to regulate the progression of the disease and/or pest (Rodriguez and
Canullo 1992). These biocontrol agents may inhibit the pathogen via production of antimicrobial compounds (antibiosis), competition for iron through siderophore- production, spatial and nutrient competition, strengthening the plant immunity, toxin degradation and/or parasitism (Kumar, 2002).
Several studies cite the use of microorganisms as biocontrol agents. Sibal (1998) reported that single and combined cultures of Bacillus licheniformis strains 1039 and 1042 effectively treated tomatoes infected with Ralstonia solanacearum. Similarly,
Bacillus licheniformis strain 1022 and Bacilus subtilis GC 28 were found to be effective in controlling R. solanacearum (Gloria, 1998). Likewise, Bacillus subtilis has been used as biological control against Rhizoctonia solani (Fiddaman and Rosall, 1995).
Bano and Musarrat (2003) tested Pseudomonas aeruginosa NJ-15 against fungal pathogens Fusarium oxysporum, Trichoderma herizium, Alternaria alternata, and
Macrophomina phasiolina. Singh, et al. (2012) observed that Arthrobotrys oligospora can help in the biological control of nematode Meloidogyne graminicola and Rhizoctonia solani Kuhn that causes root knot in rice and sheath blight in rice, respectively.
Huang, et al. (2011) reported that Bacillus pumilus SQR-N43 is a potential biocontrol agent against Rhizoctonia solani in cucumber. Song, et al. (2013) also screened Bacillus isolates that has potential biocontrol activity against Fusarium cf. incarnatum, the causative agent of ginseng root rot.
Staphylococcus LZ16 from the seawater of East China Sea was found to have biocontrol potential against Magnaporthe oryzae, the causative agent of rice blast in an experiment conducted by Yu, et al. (2013).
Another group of bacteria, called actinomycetes, are also being studied by different researchers and institutions for their biocontrol activity. Rose, et al. (1980), reported that an Streptomyces griseoloalbus, was able to inhibit root pathogens Phellinus weirii, Fomes annosus, and Phytophthora cinnamomi in in vitro experiments. They explained that the antagonistic activity of the organism could be through the production of antimicrobial compounds, and the ability of the actinomycete to rapidly eradicate nonstructural carbohydrates from wood, which is an essential factor for the rapid growth of the hyphae of the pathogens.
Filonow and Lockwood (1985) found in their greenhouse experiments that actinomycetes Actinoplanes missourensis, A. utahensis and Micromonospora sp. used as seed coating has potential in reducing the incidence of phytophthora root rot of soybean caused by Phytophthora megasperam Drechs f. sp. glvcinea Kuan & Erwin via hyperparasitism. Crawford et al. (1993) reported that an isolated actinomycete antagonist coded as WYEC108 has potential to protect lettuce seeds from Pythium ultimum that causes damping-off to the crop. It was noted that there was an observed plant-growth enhancement by the isolate but there was no explanation given for its mechanism. In a separate study, the WYEC108 actinomycete isolate was classified to be Streptomyces lydicus (Yuan and Crawford, 1994).
Chitinolytic bacteria and actinomycetes, Streptomyces viridodiasticus and
Micromonospora carbonacea, were found to have potential against Sclerotinia minor that causes basal drop disease of lettuce (El Tarabily, et al., 2000). The antagonistic activity of the bacteria were their ability to produce compounds such as chitinase and B-1,3- glucanase that cause adverse effects on S. minor.
Several published researches used different species of Streptomyces on the biocontrol studies of different pathogens. A Streptomyces isolate from Nebraska was found to reduce the severity of the root rot on alfalfa and soybean caused by
Phytophthora species (Xiao, et al., 2002). Another Streptomyces isolate associated to
Streptomyces pluricolorescens, was found to have antagontistic effects against tomato pathogens, namely, P. carotovorum brasiliensis, and Ralstonia solanacearum (de
Oliveira, et al., 2010).
Biocontrol products like Mycostop®, Actinovate®, and Actinolron®, which contain spores of Streptomyces as part of their formulation, are available commercially on other countries (de Oliveira, et al., 2010).
Powdered forms of Streptomyces cyaneofuscatus, Streptomyces kamayceticu,
Streptomyces rochei, and Streptomyces flavortricini used as seed coating showed biocontrol activity against Verticillium dahliae, causative agent of verticillium wilt of cotton (Xue, et al, 2012). These organisms was found to produce chitinase, B-1,3- glucosidase, cellulase, and protease (Xue, et al., 2012). Another study conducted by
Bubici, et al. (2013), reported that Streptomyces isolates were able to control corky root of tomato caused by Pyrenochaeta lycopersici in in vitro testing, and even in greenhouse and field-trial testing. In spite of the several reported BCAs studied in other countries, there are no known commercial products in the country that utilizes actinomycetes that are mainly directed for biological control of F. oxysporum subsp. cubense.
SUMMARY OF THE INVENTION
The invention describes a biopesticide against fusarium wilt. The biopesticide is composed of actinomycetes Several bacterial species were isolated from soil, diseased plants, and plant litter of mangrove areas of Quezon, Bataan, and Zambales. The samples > were screened and those with antagonistic effect against Fusarium oxysporum. The bacteria that were observed to have antagonistic effects on the target bacteria were screened further to select the ones most aggressive against the target bacteria. The bacteria that proved most aggressive against the target bacteria was subjected to toxicological evaluation and when proven safe, subjected to field testing with the carrier.
Results include the reduction of mortality rate of the bananas from 56.67% to 33.3% via preventive treatments wherein the biopesticide is applied prior to infection. On the other hand, no significant change in mortality was noted via curative treatment wherein the biopesticide is applied after the plant is infected.
DETAILED DESCRIPTION OF THE INVENTION
Microbial species samples were collected from soil, plant litter, and diseased plant. The bacteria were screened for their biocontrol potential against Fusarium oxysporum TR4. The bacteria with the most antagonistic effect were chosen. Among these were Bacillus species and actinomycetes. The Bacillus species were omitted as a potential biocontrol agent since its growth is hampered by actinomycetes. As a conclusion, the biocontrol agents chosen were four (4) actinomycete species,
Streptomyces misionensis, Streptomyces olivoverticillatus, Streptomyces phaeoluteichromatogenes and Streptomyces abikoensis
Assessments were conducted to test the toxicity of the actinomycetes. Water and oil extracts were prepared from the actinomycete biocontrol agents. Chick embryo assay was done to test the effect of the microbial biocontrol agents on living organisms. A suitable carrier was also prepared. The carrier consists of 3:1 rati o of garden soil and pulverized charcoal. The efficacy of the resulting product, ACTICon, was tested in the field in the province of Davao Del Norte where Panama wilt is prevalent. The Grand nain
Cavendish banana variety, Musa acuminata, was used as plant samples in all field trials.
Two set-ups were done involving the use of ACTICon-the curative and preventive set-up, depending on the manner of applying the fertilizer in reference to infection. Since curative mode was found out to be not effective in the control of Foc TR4, preventive mode of application was used in the field trials in Asuncion, Davao del Norte. +

Claims (5)

q + I) CLAIMS
1. An actinomycete-based biocontrol agent against Fusarium wilt, composed of: Mowe mixture of actinomycetes; and 1 <¢ # ie 190 a carrier;
>
2. Anactinomycete-based biocontrol agent against Fusarium wilt according to claim 1 poverty wherein the Fusarium strain is Fusarium oxysporum TR4,; loge bree 2 <_
3. Anactinomycete-based biocontrol agent against Fusarium wilt according to claim 1 lev Y . . y . ; y : a) wherein the actinomycetes are Streptomyces misionensis, Streptomyces olivoverticillatus, Streptomyces phaeoluteichromatogenes and Streptomyces abikoensis; Vo 2 i
10.
4. Anactinomycete-based biocontrol agent against Fusarium wilt according to claim 1 ever" wherein the carrier is a 3:1 ratio of soil and charcoal, respectively; and =: tr TET ve
5. An actinomycete-based biocontrol agent against Fusarium wilt according to claim 1 loveltyy wherein the product can be used against Fusarium wilt in Cavendish banana, Fie 8 Looe ve 02 Ted } Heep fore on 4 Can & rae ES =! Hac si - * “eo L Fuge, My TY Va, - - Agere ol civ dao SL Cd oo Sa Soden! of Foor vo Sm foe Lh - ! wegone lic 0 Lr od Are . A “qo , Clrcon of Smdong ol welaceos,, Ge le gu Taepenn i Lo Fubhgene VAs U4 Co Gell ¥ oe ve cfiactyg + : Fore Cn 74 < : ’ wee d cc h See vd ’ Fo
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PH12017000203A 2017-07-18 2017-07-18 Development and utilization of actinomycetes as biocontrol agents against panama wilt causing-fusarium oxysporum tr4 cavendish banana PH12017000203A1 (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113801808A (en) * 2021-07-08 2021-12-17 北京林业大学 Streptomyces albocongensis and application thereof
CN115948301A (en) * 2023-01-06 2023-04-11 中国热带农业科学院热带生物技术研究所 Oligosaccharide compound microbial inoculum for preventing and controlling banana wilt and application thereof
CN116179640A (en) * 2023-01-04 2023-05-30 中国热带农业科学院热带生物技术研究所 Method for efficiently screening antagonistic bacteria of banana vascular wilt
CN116716190A (en) * 2023-06-02 2023-09-08 云南省农业科学院农业环境资源研究所 Chaetomium spirochaeta 201 and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104403976A (en) * 2014-12-09 2015-03-11 南京农业大学 Antagonistic actinomycete for preventing and controlling continuously cropped strawberry root rot and preparation method of biocontrol agent of antagonistic actinomycete

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104403976A (en) * 2014-12-09 2015-03-11 南京农业大学 Antagonistic actinomycete for preventing and controlling continuously cropped strawberry root rot and preparation method of biocontrol agent of antagonistic actinomycete

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Biological control of Fusarium oxysporum f.sp. cubense using non-pathogenic F. oxysporum endophytes (Belgrove, A.) *
Disease preventing and growth promoting effects of Streptomyces aureoverticillatus strain HN6 on banana (Lanying, W. et al.) *
Isolation and characterization of endophytic streptomycete antagonists of fusarium wilt pathogen from surface-sterilized banana roots (Cao, L. et al.) *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113801808A (en) * 2021-07-08 2021-12-17 北京林业大学 Streptomyces albocongensis and application thereof
CN113801808B (en) * 2021-07-08 2022-12-09 北京林业大学 Streptomyces albocongensis and application thereof
CN116179640A (en) * 2023-01-04 2023-05-30 中国热带农业科学院热带生物技术研究所 Method for efficiently screening antagonistic bacteria of banana vascular wilt
CN116179640B (en) * 2023-01-04 2023-10-24 中国热带农业科学院热带生物技术研究所 Method for efficiently screening antagonistic bacteria of banana vascular wilt
CN115948301A (en) * 2023-01-06 2023-04-11 中国热带农业科学院热带生物技术研究所 Oligosaccharide compound microbial inoculum for preventing and controlling banana wilt and application thereof
CN115948301B (en) * 2023-01-06 2023-10-24 中国热带农业科学院热带生物技术研究所 Oligosaccharin composite microbial inoculant for preventing and controlling banana vascular wilt and application thereof
CN116716190A (en) * 2023-06-02 2023-09-08 云南省农业科学院农业环境资源研究所 Chaetomium spirochaeta 201 and application thereof
CN116716190B (en) * 2023-06-02 2024-04-05 云南省农业科学院农业环境资源研究所 Chaetomium spirochaeta 201 and application thereof

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