Isolation and Characterization of Microorganisms for Use with Manure and Chemical Fertilizers

Abstract

Isolation and Characterization of Microorganisms for Use with Manure and Chemical Fertilizers Levent Değirmenci1, Özge Kaygusuz İzgördü2, Cihan Darcan3 ORCID: 0000-0001-6608-0398; 0000-0002-3652-4266; 0000-0003-0205-3774 1 Corresponding author: Department of Chemical Engineering, Bilecik Şeyh Edebali University, 11210, Bilecik/Turkey Tel: [phone], e-mail: [e-mail] 2 Biotechnology Application and Research Center, Bilecik Şeyh Edebali University, 11210, Bilecik, Turkey 3 3Department of Molecular Biology and Genetics, Bilecik Şeyh Edebali University, 11210, Bilecik, Turkey Abstract A microbial consortium with the ability to utilize nitrogen of certain chemical fertilizers were established via a series of tests. Results indicated four strains, 3 of which were identified as Bacillus cereus sp and an Alcaligenes faecalis sp. which was the only non-Bacillus member of the consortium. The members of the consortium showed no antagonistic activity against each other implying their successful utilization as components of a commercial fertilizer. Keywords: IAA, siderophore, Bacillus spp., nitrogen fixation, ammonia/nitrite oxidation, MALDI-TOF MS Organik ve Kimyasal Gübre ,le Kullanılmak Üzere Mikroorganizma İzolasyon ve Karakterizasyonu Özet Farklı kimyasal gübrelerin içeriğinde yer alan azotu bitkinin kullanımına sunabilen mikroorganizma karışımı bir dizi testler sonucunda elde edilmiştir. Sonuçların değerlendirilmesiyle mikroorganizma karışımına üç Bacillus Cereus sp. suşu ile karışımın Bacillus sp. olmayan tek üyesi Alcaligenes faecalis sp. seçilmiştir. Mikroorganizma karışımında yer alan suşların birbirlerine karşı antagonistik etki göstermediği dolayısıyla karışımın mikrobiyal gübre olarak ticarileştirilmesinin mümkün olduğu da elde edilen sonuçlar arasındadır. Anahtar Kelimeler: IAA, siderophore, Bacillus spp. azot fiksasyonu, amonyum/nitrit oksidasyonu, MALDI-TOF MS 1. Introduction Chemical fertilizer is gaining an increasing attention and its harmful effects on environment possesses a serious threat [2]. Heavy metal accumulation is another problem during utilization of chemical fertilizers [2].The variation of soil pH, another problem encountered due to excessive fertilizer utilization, has negative impact on phosphorus uptake [3]. Plant growth promoting bacteria when added to soil, operate via certain mechanisms facilitating acquisition of already existing resources in soil [4]. Siderophore producing bacteria also acts as biocontrol agent capable of decreasing phytopathogen population in the flora [4–7]. Certain procedures were developed and/or modified in the present study to obtain a microbial consortium which would act as an enhancer during plant growth. 2. Materials and Method 2.1. Semi-selective isolation procedure as starting-point Semi-selective isolation procedures were conducted according to [12]. 2.2. Standard tests applied to determine the potential of isolates as PGPB. Nitrogen fixation ability, IAA production, phosphate solubility siderophore production were conducted according to [13-18]. 2.3. Specified tests to determine nitrogen evaluation potential of PGP isolates Nitrogen evaluation potential was determined according to [19]. 2.4. Biochemical tests conducted to determine Bacillus sp. in accordance with MALDI-TOF MS analyses evaluated for validation. Biochemical tests were evaluated based on positive results for both caseinase activity and starch hydrolysis [20-22]. MALDI-TOF MS analyses applied for selected strains and an isolate with positive activity towards siderophore production. 3. Results IAA production, Nitrogen fixation performance, Ammonifier selection, phosphate solubility, siderophore production ability, urea degradation, ammonium oxidation and nitrite oxidation abilities of strains were evaluated to establish a microbial consortium. 4. Conclusion and discussion • Media selection could serve as a selection criterion to assemble a certain group of microorganisms. • Tests conducted for isolates could be diversified to establish a consortium with modified properties. • Test procedures could be modified to facilitate establishment of microbial consortium. • Strains with different properties could serve as members of consortium which was the case in our study. • Identification in species level was possible to a certain extent. MALDI-TOF MS analysis presented a brief idea on strains and similar results would have been obtained in the case of applying its costlier alternatives. Hence a relatively simpler procedure is concluded to be better for initial investigation. • Evaluation of results led to investigation of certain prospects including determination of physicotolerant activity of strains and management of Fusarium wilt disease as the topics of future investigations.

Keywords

• IAA
• siderophore
• Bacillus spp.
• nitrogen fixation
• ammonia/nitrite oxidation
• MALDI-TOF MS

Organik ve kimyasal gübre ile kullanılmak üzere mikroorganizma izolasyon ve karakterizasyonu

Abstract

Isolation and characterization of microorganisms for use with manure and chemical fertilizers Levent Değirmenci1, Özge Kaygusuz İzgördü2, Cihan Darcan3 ORCID: 0000-0001-6608-0398; 0000-0002-3652-4266; 0000-0003-0205-3774 1 Corresponding author: Department of Chemical Engineering, Bilecik Şeyh Edebali University, 11210, Bilecik/Turkey Tel: [phone], e-mail: [e-mail] 2 Biotechnology Application and Research Center, Bilecik Şeyh Edebali University, 11210, Bilecik, Turkey 3 3Department of Molecular Biology and Genetics, Bilecik Şeyh Edebali University, 11210, Bilecik, Turkey Abstract A microbial consortium with the ability to utilize nitrogen of certain chemical fertilizers were established via a series of tests. Results indicated four strains, 3 of which were identified as Bacillus cereus sp and an Alcaligenes faecalis sp. which was the only non-Bacillus member of the consortium. The members of the consortium showed no antagonistic activity against each other implying their successful utilization as components of a commercial fertilizer. Keywords: IAA, siderophore, Bacillus spp., nitrogen fixation, ammonia/nitrite oxidation Organik ve kimyasal gübre olarak kullanım potansiyeline sahip mikroorganizmarın İzolasyon ve karakterizasyonu Özet Farklı kimyasal gübrelerin içeriğinde yer alan azotu bitkinin kullanımına sunabilen mikroorganizma karışımı bir dizi testler sonucunda elde edilmiştir. Sonuçların değerlendirilmesiyle mikroorganizma karışımına üç Bacillus Cereus sp. suşu ile karışımın Bacillus sp. olmayan tek üyesi Alcaligenes faecalis sp. seçilmiştir. Mikroorganizma karışımında yer alan suşların birbirlerine karşı antagonistik etki göstermediği dolayısıyla karışımın mikrobiyal gübre olarak ticarileştirilmesinin mümkün olduğu da elde edilen sonuçlar arasındadır. Anahtar Kelimeler: IAA, siderophore, Bacillus spp. azot fiksasyonu, amonyum/nitrit oksidasyonu 1. Introduction Chemical fertilizer is gaining an increasing attention and its harmful effects on environment possesses a serious threat [2]. Heavy metal accumulation is another problem during utilization of chemical fertilizers [2].The variation of soil pH, another problem encountered due to excessive fertilizer utilization, has negative impact on phosphorus uptake [3]. Plant growth promoting bacteria when added to soil, operate via certain mechanisms facilitating acquisition of already existing resources in soil [4]. Siderophore producing bacteria also acts as biocontrol agent capable of decreasing phytopathogen population in the flora [4–7]. Certain procedures were developed and/or modified in the present study to obtain a microbial consortium which would act as an enhancer during plant growth. 2. Materials and Method 2.1. Semi-selective isolation procedure as starting-point Semi-selective isolation procedures were conducted according to [12]. 2.2. Standard tests applied to determine the potential of isolates as PGPB. Nitrogen fixation ability, IAA production, phosphate solubility siderophore production were conducted according to [13-18]. 2.3. Specified tests to determine nitrogen evaluation potential of PGP isolates Nitrogen evaluation potential was determined according to [19]. 2.4. Biochemical tests conducted to determine Bacillus sp. in accordance with MALDI-TOF MS analyses evaluated for validation. Biochemical tests were evaluated based on positive results for both caseinase activity and starch hydrolysis [20-22]. MALDI-TOF MS analyses applied for selected strains and an isolate with positive activity towards siderophore production. 3. Results IAA production, Nitrogen fixation performance, Ammonifier selection, phosphate solubility, siderophore production ability, urea degradation, ammonium oxidation and nitrite oxidation abilities of strains were evaluated to establish a microbial consortium. 4. Conclusion and discussion • Media selection could serve as a selection criterion to assemble a certain group of microorganisms. • Tests conducted for isolates could be diversified to establish a consortium with modified properties. • Test procedures could be modified to facilitate establishment of microbial consortium. • Strains with different properties could serve as members of consortium which was the case in our study. • Identification in species level was possible to a certain extent. MALDI-TOF MS analysis presented a brief idea on strains and similar results would have been obtained in the case of applying its costlier alternatives. Hence a relatively simpler procedure is concluded to be better for initial investigation. • Evaluation of results led to investigation of certain prospects including determination of physicotolerant activity of strains and management of Fusarium wilt disease as the topics of future investigations.

Keywords

• IAA
• siderophore
• Bacillus spp. azot fiksasyonu
• amonyum/nitrit oksidasyonu

References

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