Optimization of industrial (3000 L) production of Bacillus subtilis CW-S and its novel application for minituber and industrial-grade potato cultivation.

A commercial plant probiotic product was developed employing Bacillus subtilis CW-S in submerged fermentation. The effects of molasses and urea on cell growth were investigated with the goal of low-cost manufacturing. Plackett-Burman and Central-Composite Design (CCD) were utilized to optimize production parameters to maximize productivity. The stability of the formulated product and its efficacy in cultivating minituber in aeroponics and industrial-grade potatoes in the field were assessed. The results showed that the medium BS10 (molasses and urea) produced satisfactory cell density (7.19 × 108 CFU/mL) as compared to the control (1.51 × 107 CFU/mL) and BS1-BS9 (expensive) media (1.84 × 107-1.37 × 109 CFU/mL). According to validated CCD results, optimized parameters fitted well in pilot (300 L; 2.05 × 109 CFU/mL) and industrial (3000 L; 2.01 × 109 CFU/mL) bioreactors, resulting in a two-fold increase in cell concentration over laboratory (9.84 × 108 CFU/mL) bioreactors. In aeroponics, CW-S produced excellent results, with a significant increase in the quantity and weight of minitubers and the survival rate of transplanted plantlets. In a field test, the yield of industrial-grade (> 55 mm) potatoes was increased with a reduction in fertilizer dose. Overall, the findings suggest that CW-S can be produced commercially utilizing the newly developed media and optimized conditions, making plant probiotics more cost-effective and accessible to farmers for crop cultivation, particularly in aeroponic minituber and industrial-grade potato production.

A bio-sustainable approach for reducing Eucalyptus tree-caused agricultural ecosystem hazards employing Trichoderma bio-sustained spores and mycorrhizal networks.

The presence of the exotic Eucalyptus tree in crop-growing soil and the accumulation of its undecomposed leaves is a significant ecological hazard. The waxy coating on the leaves and the phenolic compounds takes a long time to break down under normal conditions. It is necessary to explore various fungi that can degrade these leaves for an eco-friendly solution to this problem. In this study, spores of nine native Trichoderma strains were produced on wheat agar using a lactic acid-induced sporulation strategy (LAISS). Trichoderma biosustained spores and Serendipita indica (SI) spores were applied to a rice field with accumulated Eucalyptus leaves under continuous ponding (CP) and alternate flooding and wetting conditions (AFW). Among the strains, TI04 (Trichoderma viride) and TI15 (Trichoderma citrinoviride) showed faster (5 days) and massive sporulation (1.06-1.38 × 1011 CFU/g) in LAISS. In vitro, TI04 and TI15 biosustained on Eucalyptus leaves and improved rice seedling growth and SI infection under greenhouse conditions. In the rice-field experiment, Trichoderma-treatment had a threefold yield (percentage) increase from control, with TI04 (CP) increasing the yield by 30.79, TI04 (AFW) by 29.45, TI15 (CP) by 32.72, and TI15 (AFW) rising by 31.91. Remarkably, unfilled grain yield significantly decreased in all the Trichoderma treatments. Under AFW conditions, TI04 and TI15 showed a higher pH increase. Furthermore, TI04 and TI15 under AFW had higher water productivity (t ha-1 cm-1) of 0.0763 and 0.0791, respectively, and the highest rates (percentage) of SI colonization of 86.36 and 83.16, respectively. According to the findings, LAISS-produced Trichoderma spores can be applied to break down persistent wastes and restore agricultural ecosystems through increased mycorrhizae networking.

Sand Particle Size and Phosphorus Amount Affect Rhizophagus irregularis Spore Production Using In Vitro Propagated Spore as a Starter Inoculum in Rhizosphere of Maize (Zea mays) Plantlets.

Microbial inoculants, particularly arbuscular mycorrhizal (AM) fungi, have great potential for sustainable crop management. In this study, monoxenic culture of indigenous R. irregularis was developed and used as a tool to determine the minimum phosphorus (P) level for maximum spore production under the in vitro conditions. This type of starter AM fungal inoculum was then applied to an in vivo substrate-based mass-cultivation system. Spore production, colonization rate, and plant growth were examined in maize (Zea mays L.) plant inoculated with the monoxenic culture of R. irregularis in sand graded by particle size with varying P levels in nutrient treatments. In the in vitro culture, the growth medium supplemented with 20 µM P generated the maximum number of spores (400 spores/mL media) of R. irregularis. In the in vivo system, the highest sporulation (≈500 spores g-1 sand) occurred when we added a half-strength Hoagland solution (20 µM P) in the sand with particle size between 500 µm and 710 µm and omitted P after seven weeks. However, the highest colonization occurred when we added a half-strength Hoagland solution in the sand with particle sizes between 710 µm and 1000 µm and omitted P after seven weeks. This study suggests that substrate particle size and P reduction and regulation might have a strong influence on the maximization of sporulation and colonization of R. irregularis in sand substrate-based culture.