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. 2022 Jul 1;12(1):11153.
doi: 10.1038/s41598-022-15366-5.

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

Md Abuhena #  1 , Jubair Al-Rashid #  1   2 , Md Faisal Azim #  3 , Md Niuz Morshed Khan  2 , Md Golam Kabir  1   2 , Nirmal Chandra Barman  2 , Noorain Munim Rasul  1   2 , Shahina Akter  4 , Md Amdadul Huq  5
Affiliations

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

Md Abuhena et al. Sci Rep. .

Abstract

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.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Media evaluation based on the cell concentration of CW-S.
Figure 2
Figure 2
Pareto chart of the standardized effects for cell concentration of CW–S.
Figure 3
Figure 3
3D response surface displaying the interaction of all those components considered in CCD optimization. (a) Temperature and incubation time. (b) Temperature and pH. (c) Temperature and agitation. (d) Incubation time and pH. (e) Incubation time and agitation. (f) pH and agitation.
Figure 4
Figure 4
Variation in Bacillus subtilis CW-S cell concentration and pH in the formulation and control over time. (a) Viability of cell concentration (CFU/mL). (b) Variability in pH.
Figure 5
Figure 5
The effects of CW-S on minituber production in aeroponic. (a) Plantlet survival rate after transplantation in aeroponic. (b) Stolon number. c) Minituber number. (d) Minituber weight. (e) Root length. Bars without shared letters indicate significant differences between the control and treated ones.
Figure 6
Figure 6
Root and stolon development inside an aeroponic system. (a) Potato plantlets treated with CW–S. (b) Potato plantlets under control.
Figure 7
Figure 7
Two-way ANOVA illustrates the effect of CW-S with different fertilizer doses. (a) Yield per plant. (b) Yield per m2. (c) Canopy cover. (d) Relative density. (e) Under-grade potato percentage. (f) Grade A potato percentage. (g) Grade B potato percentage. (h) Over–grade (industrial-grade) potato percentage. (ns p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001), data are means ± SD (n = 4). Bars without shared letters indicate significant differences.
Figure 8
Figure 8
A comparison of CW-S and control in terms of yield and grade of potatoes. (a) Dose0 and CW-S. (b) Dose1 and CW-S. (c) Dose2 and CW-S. (d) Dose3 and CW-S. (e) A comparison of various grades.
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