Cesari Adriana B, Paulucci Natalia S, Yslas Edith I, Dardanelli Marta Susana
INBIAS, Instituto de Biotecnología Ambiental y Salud, CONICET, Rio Cuarto, Argentina.
Departamento de Biología Molecular, Universidad Nacional de Río Cuarto, Ruta Nacional 36, Km, 601, Rio Cuarto, Argentina.
Appl Microbiol Biotechnol. 2020 Dec;104(23):10145-10164. doi: 10.1007/s00253-020-10910-7. Epub 2020 Oct 6.
Immobilizarion of PGPR for agricultural applications aims to provide temporary physical protection from stressful environmental conditions and the gradual release of cells for successful root colonization, release the cells gradually. In this work, we immobilized Bradyrhizobium sp. SEMIA6144 or Azospirillum brasilense Az39 cells in 2% alginate beads prepared by ionic gelation process, and then stored up to 12 months at 4 °C. Alginate matrix showed interaction with the immobilized bacteria (FTIR), allowed a constant release of cells, and improved their viability and capability to interact with Arachis hypogaea. Cell number into beads reached 10 CFU.bead; however, viability decreased from 4 months of storage for Az39, while it was maintained up to 12 months for SEMIA6144, showing a low metabolic activity measured by the MTT assay. Adhesion of SEMIA6144 and Az39 from new beads to peanut root was 11.5% and 16%, respectively, higher than non-immobilized bacteria. Peanut inoculation with 12 months storage SEMIA6144 beads significantly increased root length and biomass at 30 days of growth, and under restrictive water condition (RWC), nodulation and total plant N content increased compared with liquid inoculation. Our results demonstrate that immobilization of SEMIA6144 and Az39 in alginate matrix is a potential alternative to enhance peanut growth even under RWC. KEY POINTS: • Alginate encapsulation enhances viability of SEMIA6144 or Az39 under storage at 4 °C for 1 year. • Alginate beads 2% ensure the gradual release of the microorganisms. • Cells from beads stored for long periods present chemotaxis and adhesion to peanut root. • Peanut inoculation with 1-year-old SEMIA6144 beads improves nodulation and growth in RWC.
将用于农业的植物根际促生菌(PGPR)固定化旨在提供临时物理保护,使其免受恶劣环境条件影响,并使细胞逐步释放以成功定殖于根部。在本研究中,我们通过离子凝胶化过程将慢生根瘤菌属SEMIA6144或巴西固氮螺菌Az39细胞固定在2%的海藻酸钠珠中,然后在4℃下储存长达12个月。海藻酸钠基质与固定化细菌存在相互作用(傅里叶变换红外光谱法),能使细胞持续释放,并提高其活力以及与花生相互作用的能力。珠中的细胞数量达到10 CFU/珠;然而,Az39的活力从储存4个月起下降,而SEMIA6144的活力则维持长达12个月,通过MTT法测定显示其代谢活性较低。新珠中的SEMIA6144和Az39对花生根的粘附率分别比未固定化细菌高11.5%和16%。用储存12个月的SEMIA6144珠接种花生,在生长30天时显著增加了根长和生物量,并且在水分限制条件(RWC)下,与液体接种相比,结瘤和全株氮含量增加。我们的结果表明,将SEMIA6144和Az39固定在海藻酸钠基质中是一种潜在的替代方法,即使在水分限制条件下也能促进花生生长。要点:• 海藻酸钠包封可提高SEMIA6144或Az39在4℃储存1年时的活力。• 2%的海藻酸钠珠可确保微生物的逐步释放。• 长期储存的珠中的细胞对花生根具有趋化性和粘附性。• 用储存1年的SEMIA6144珠接种花生可改善水分限制条件下的结瘤和生长。