Abstract
Ilex paraguariensis St. Hil. (yerba mate) is a native tree to the Atlantic forest of South America, where Argentina is the world’s largest producer. In a previous study, two strains of Bacillus spp. were isolated from I. paraguariensis St. Hil. seedlings and selected for their ability to promote plant growth. To gain a better understand this, a traceability assay was performed to verify the bioinoculant effects of Bacillus spp. on I. paraguariensis St. Hil. seedlings. A manual search for unique genome sequences of B. altitudinis T5S-T4 and 19RS3 strains was performed to develop strain-specific primers. The traceability assay was performed in a nursery with I. paraguariensis St. Hil. seedlings that were inoculated with B. altitudinis T5S-T4, 19RS3, or a combination of both. Growth parameters, including height, number of leaves, neck diameter, and chlorophyll index, were measured every 15 days in the nursery. Fresh and dry weights of the aerial parts, leaves, roots, and leaf area were measured in the laboratory. The substrate DNA was extracted using a commercial kit. B. altitudinis T5S-T4 and 19RS3 were identified in the substrate 14 and 28 days after the first inoculation, respectively, to day 112 of the assay. The bioinoculation effect of the two-strain combination was confirmed by significant increases in growth parameters, including height, neck diameter, chlorophyll index, and wet weight of aerial parts. The designed and synthesized strain-specific primers allowed the monitoring of the colonization and persistence of B. altitudinis T5S-T4 and B. altitudinis 19RS3 on I. paraguariensis St. Hil seedling substrate inoculated under nursery conditions. The properties of B. altitudinis T5S-T4 and 19RS3 in consortium make them excellent candidates for the formulation of bioinoculants for I. paraguariensis St. Hil.
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Data Availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
The authors would like to thank the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, Argentina) for Iliana J. Cortese and Andrea L. Onetto doctoral fellowships. Also, Gustavo A. Bich, Pedro D. Zapata, María L. Castrillo and Margarita E. Laczeski are CONICET researchers.
Funding
This work was supported by the Instituto Nacional de la Yerba Mate (INYM, Argentina) project “Biofertilizante: validación a campo y estudios de trazabilidad de la utilización de Bacillus sp. como fertilizante para yerba mate” (Res. nº 274/17, Programa de Asistencia al sector yerbatero – PRASY—INYM, Argentina). Yerba mate seedlings were donated by the Fundación Alberto Roth (Santo Pipó, Misiones, Argentina).
This research was founded by project “Proyecto: PICT Start Up 2017–007: BIOFER-YM: Desarrollo de un paquete tecnológico para la fertilización de yerba mate”. ANPCyT—Ministerio de Educación, Cultura, Ciencia y Tecnología. (Res. 595/2017) and project “PDTS 287: Implementación de microorganismos benéficos con aplicaciones biotecnológicas como biofertilizantes y controladores biológicos para cultivos de yerba mate” (adopting institution: Fundación Alberto Roth). Banco del Ministerio de Ciencia y Tecnología.
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The investigation, experiments, formal analysis, and writing of the original draft were performed by Iliana Julieta Cortese. Experiments were performed by Marisa Esther Boycho. Writing, reviewing, and editing were performed by Andrea Liliana Onetto and Gustavo Angel Bich. Pedro Darío Zapata contributed to assessing and investigating. María Lorena Castrillo and Margarita Ester Laczeski conceptualized, investigated, reviewed, edited, supervised, and administered the project. The manuscript has been carefully read and approved by all authors.
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Cortese, I.J., Onetto, A.L., Bich, G.Á. et al. Traceability Assay and Bioinoculant Effects of Two Plant Growth-Promoting Bacillus altitudinis Strains Isolated from Ilex paraguariensis St. Hil.. J Soil Sci Plant Nutr 23, 6798–6812 (2023). https://doi.org/10.1007/s42729-023-01537-4
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DOI: https://doi.org/10.1007/s42729-023-01537-4