Next generation bacterial inoculants for sustainable

agriculture: advances, challenges, and perspectives

Sergio de los Santos Villalobos

sergio.delossantos@itson.edu.mx
dlsantosv@gmail.com

TECHNOLOGICAL INSTITUTE OF SONORA

July 08, 2023

CURRENT AND FUTURE FOOD SECURITY

www.itson.mx/LBRM
www.itson.mx/COLMENA
MICROBIAL CULTURE COLLECTION:

COLMENA Fungal
strains

Bacterial
http://apps2.itson.edu.mx/colmena/ strains

1600 microbial strains

Trigo
Wheat
Maize
Maíz
13%
Bean
Frijol
3% Asparagus
4% Espárrago
38%
3% Broccoli
Brócoli
5% Safflower
2% Alfalfa
Cártamo
2%
1%
3% Oranges
Alfalfa
Potato
Naranja
26%
Native
Papa
Vegetation
Acacia
de los Santos-Villalobos et al., 2018; de los Santos-Villalobos et al., 2021
A NEW DISEASE IN THE YAQUI
VALLEY

Villa-Rodríguez et al., 2016

BIOCONTROL OF A NEW DISEASE IN THE
YAQUI VALLEY

Villa-Rodríguez et al., 2019

IDENTIFICATION OF BIOLOGICAL CONTROL
AGENTS

Villa-Rodríguez et al., 2019

 Bacillus sp. TE3T: GENOME

Villa-Rodríguez et al., 2021; Valenzuela-Ruíz et al., 2023

Bacillus sp. TE3T: METABOLOME

Villa-Rodríguez et al., 2021

Bacillus sp. TE3T: TRANSCRIPTOME

Villa-Rodríguez et al., 2021

Bacillus sp. TE3T: TRANSCRIPTOME
1800
Valenzuela-Ruíz et al., 2023
1600
1400
1200 881
1000
583 649
800
600
400 750
627 579
200
0
Fase Exponential
exponencialphase
vs. fase
vs. de Fase estacionaria
Stationary phase vs.
vs. fase de
adaptation Fase Stationary
estacionaria vs. fase
phase vs.
adaptación
adaptation phase adaptación
phase exponencial
exponential phase
Sobre-expresados
Up regulated Down regulated
Reprimidos

Exponential phase vs. Stationary phase vs. Stationary phase vs.

adaptation phase adaptation phase exponential phase
Rhizocticin A 92% 78% 0%
Surfactin 30% 0% 0%
Bacillaene 85% 42% 85%
Fengycin 80% 73% 0%
Bacillibactin 100% 85% 100%
Subtilosin A 87% 0% 75%
Bacilysin 0% 0% 0%
Bacillus sp. TE3T: TRANSCRIPTOME

* Rhizocticin A
* Bacillaene
* Fengycin
* Subtilosin A
Bacillibactin

Exponential Phase
Rhizocticin A
Surfactin
Bacillaene
Fengycin
Subtilosin A
Bacillibactin
Time (h)

Bacillibactin

Valenzuela-Ruíz et al., 2023

POLYPHASIC TAXONOMY

Bacillus cabrialesii sp. nov.

TE3T

Morales Sandoval et al., 2021; de los Santos-Villalobos et al., 2019

POLYPHASIC TAXONOMY

Bacillus cabrialesii subsp. cabrialesii

TE3T
Bacillus cabrialesii subsp. tritici
TSO2T

de los Santos-Villalobos et al., 2019; de los Santos-Villalobos et al., 2023

 MUTATION INDUCTION:
Bacillus cabrialesii subsp. cabrialesii TE3T

Gene clusters associated with biocontrol and taxonomic

affiliation?
WHEAT x PGPB INTERACTIONS:
TRANSCRIPTOME

Chaparro-Encinas et al., 2021; Chaparro-Encinas et al., 2022

WHEAT x PGPB INTERACTIONS:
TRANSCRIPTOME
Temperature Bacterial consortium

Stress mitigtion by bacterial consortium

Chaparro-Encinas et al., in preparation

Romero-Silva et al., in preparation
SOIL x BCA INTERACTIONS: METAGENOME

Consortium 3
Consortium 2
Consortium 1
Control

Relative
Relativeabundance
abundance(%):
(%):Fusarium spp.
Fusariumspp.
 SOIL x BCA INTERACTIONS: METAGENOME

Control Consortium 2

Unique Unique
species species

Nitrososphaera = 20 %

Nitrosocosmicus = 10 %

Romero-Silva et al., in preparation; Chávez-Luzania et al., 2023

 BACTERIAL CONSORTIUM ON WHEAT YIELD

Table 3. Effect of the application of a bacterial consortium on wheat CIRNO C2008 grain yield, under different season,
and nitrogen fertilization level.

N level, Bacterial Grain yield, Grain yield increment

Season
kg ha-1 Consortium (BC) ton ha-1 by BC, %
Inoculated 5.78 d
0 28.4
Un-inoculated 4.50 e
1 Inoculated 8.20 a
2016-2017 130 10.7
Un-inoculated 7.41 bc
Inoculated 7.89 ab
250 8.2
Un-inoculated 7.29 bc
Inoculated 6.73 c
0 30.7
Un-inoculated 5.15 de
2
2017-2018 130
Inoculated 8.3 a
18.9
Un-inoculated 6.98 c
Inoculated 8.02 ab
250 13.4
Un-inoculated 7.07 c

Letters indicate differences between parameter means using the Tukey test, p ≤ 0.05.

Nitrogen use efficiency (15N) = 30%

 STRATEGIES TO INCREASE
CONCENTRATION OF CELLS-SPORES

Biorreactor assays
Parámetro/ Tratamiento B(T1- 𝟑𝑪𝒂)₂-𝟐𝑷𝑫 B(T1- 𝟑𝑪𝒂)₂-𝟐𝑷𝑫 , 𝑪 BT1-𝑷𝑫 , 𝟑𝑪𝒂 − 𝐅𝐞𝐞𝐝
µ𝑪𝒆𝒍/𝒎𝑳 (𝒉−𝟏 ) 0.39 ± 0.00 a 0.36 ± 0.03 a 0.39 ± 0.02 a
Td𝑪𝒆𝒍/𝒎𝑳 (h) 1.79 ± 0.00 a 1.92 ± 0.17 a 1.78 ± 0.11 a
X 𝒎𝒂𝒙 (CFU/mL) 8.33 ± 0.35 𝑥109 b 8.70 ± 0.50 𝑥109 b 1.17 ± 0.06 𝑥1010 a
µ𝒔𝒑 (𝒉−𝟏 ) 0.29 ± 0.00 c 0.49 ± 0.02 a 0.38 ± 0.01 b
Td𝒔𝒑 (h) 2.36 ± 0.01 a 1.43 ± 0.06 c 1.84 ± 0.02 b
X𝒎𝒂𝒙 𝒔𝒑 (Spores/mL) 4.04 ± 0.02 𝑥109 b 7.20 ± 0.00 𝑥109 a 2.93 ± 1.26 𝑥109 b
Sporulation % 48.55 ± 2.28 b 80.45 ± 0.00 a 24.96 ± 9.66 c
Data presented based on their mean (n=2) ± range. Data analyzed with the analysis of variance ANOVA, using fisher's
LSD test (P <0.05). Different literals indicate significant differences.

Villarreal-Delgado et al., in preparation

 STRATEGIES TO INCREASE
CONCENTRATION OF CELLS-SPORES

Roja-Padilla et al., 2023

BIOFORMULATION OF BACTERIAL CONSORTIA

Roja-Padilla et al., 2023

BIOFORMULATION OF BACTERIAL CONSORTIA:
OMIC SCIENCES

2.500 1.40E+05

1.20E+05
2.000
1.17E+05
1.00E+05

CFU/mL or spores/mL
Absorbance (630 nm)

1.500
8.00E+04

6.00E+04
1.000

4.00E+04

0.500
2.00E+04

0.000 0.00E+00
2.20E+03
0 5 10 15 20 25
Hours
Díaz-Rodríguez et al., In preparation
BIOFORMULATION OF BACTERIAL CONSORTIA:
OMIC SCIENCES

Production Formulation

Cells or spores

Metabolites Scale-up

Additives

Medium optimization Feasibility

and controlled culture Cost-benefit
conditions Viability
Díaz-Rodríguez et al., In preparation
 KEY POINTS FOR
NEW AGRO-BIOTECHNOLOGIES

Economically viable

Socially justifiable

Optimal use of resources

New
Technologies

Environmental
improvement

Feasible functionality

Díaz-Rodríguez et al., 2021

MICROBIAL INOCULANTS

Chávez - Díaz et al., 2020

COLLABORATIONS
MEMBERS OF LBRM - COLMENA
MEMBERS OF LBRM - COLMENA
MEMBERS OF LBRM - COLMENA
RESEARCH PROJECTS AND PUBLICATIONS

ResearchGate Facebook
Next generation bacterial inoculants for sustainable
agriculture: advances, challenges, and perspectives

Sergio de los Santos Villalobos

sergio.delossantos@itson.edu.mx
dlsantosv@gmail.com

TECHNOLOGICAL INSTITUTE OF SONORA

July 08, 2023