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ADVANCES IN PGPR RESEARCH:
IDENTIFICATION, DEVELOPMENT
AND APLICATION IN COLOMBIA
BANANA CROPS
Luz Edith Argel Roldán, John Jairo Mira Castillo
MAGDALENA-GUAJIRA
URABÁ
12643 Ha
90´000 boxes/year
34800 Ha
INTRODUCTION
 Microbiological, Chemical and
Physical Soil Degradation
 Fertilizer expensive
 Water and soil contamination
for chemicals fertilizers
 Technological dependence
 Alliance with local universities (Spin off HABA)
 Clean production of bananas
Screening for potential plant
growth promoting
rhizobacteria isolated from
Bananas
M. Sc. Tesis of Camilo Ramírez
(2005)
AIMS
To
know
the
potential
of rhizobacteria associated to banana and
plantain plants to promote growth and plant
health
• Isolate
promissory
bacterias
from
rizosphere
• Physiological and chemical capacity to
promote growth
• Test of promissory isolates
Sampling And Selective Isolation
Rhizobacteria
66
Bacillus spp
105 isolates
38
Pseudomonas
spp
1
Coco variable
Gram KOH
positive
BIOCHEMICALS TESTS
Pseudomonas spp
Activity ACCase
4 isolates
Siderophores
production 11 isolates
Growth Inhibition of Fusarium
ox ysporum f.sp. Cubense
21 isolates
BIOCHEMICALS TESTS…
Bacillus spp.
Pseudomonas spp. &
Bacillus spp.
β-1,3- Glucanases
8 isolates
ISR
C. lindemuthianum /P. vulgaris
9 isolates
Phosphate solubilizing
11 isolates
Quitinases
22 isolates
Design of a production process and
product development based on
Bacillus spp. rhizobacteria to promote
growth in Banana (M usa AAA)
Luisa Fernanda Posada Uribe, María Ramírez Correa,
Luz Edith Argel Roldán, John Jairo Mira Castillo y Valeska Villegas
Escobar
AIM
Develop a biotechnology product based on
rhizobacteria of the genus Bacillus spp. that
promotes root growth and health of banana
plants (Musa AAA)
To isolate, to select and to identify Strain with
potencial PGPR
To Evaluate the effect of the formulations in
greenhouse and field
Cultivar and herbicide application
as a source of variability
1.4E+07
43%
a
Herbicide
Without Herbicide
1.2E+07
CFU /g Root
a
1.0E+07
32%
8.0E+06
6.0E+06
4.0E+06
2.0E+06
b
b
a
72%
b
0.0E+00
G. enano
A
Valery
Cultivar M usa spp.
Hartón
AEFB Collection associated with
Banana and Plantain
TSB + 20% glycerol -80°C
AEFB Collected
837 isolated
PGPR Selection Strategies
In vitro evaluation of PGPR traits (AIA
and antimicrobial metabolites)
Evaluation in a model plant of rapid
growth and easy handling (corn)
Evaluation of selected strains
directly on banana plants
13
Test of PGPR Traits
IAA Production
Salwcosky Solution
562 Isolates
(67%)
C+ UA567 Ramírez (2008)
IAA PRODUCTION
0
µg/mL
275
33%
2-15
µg/mL
404
49%
16-30
µg/mL
105
12%
31-45
µg/mL
43
5%
46-60
µg/mL
10
1%
837 isolates Musa spp.
Ralstonia solanacearum EAP009
Fusarium oxysporum EAP004
Qualitative
153 (18.3%) inhibition halos
27 (3.2%) inhibición halos
Quantitative
28 (3.3%) ]Inhibition diametre de
= ó > a los C+
Inhibition Halos 5-12 mm
C+: B. subtilis UA321 Ramírez (2008)
Rhapsody B. subtilis QST713
Isolates
Code
B. amyloliquefaciens
EA-CB0158
B. subtilis
EA-CB1121
EA-CB0575
B. pumilus
EA-CB0177
B. altitudinis
EA-CB0686
B. cereus
EA-CB0131
B. megaterium
EA-CB0784
Photos
Photos
Villegas (2010)
A
0,90
30
Peso
aéreo (g)
Shoot
dryseco
weitght
Shoot dry weight
a
0,80
25
ab
0,70
abc
abc
abc
abc
0,60
20
c
c
0,50
15
0,40
0,30
10
EACB0158 (25%)
EA-CB0177 (14%)
0,20
5
0,10
0,00
EA-CB0158 EA-CB0177 EA-CB0131 EA-CB1121 EA-CB0686 EA-CB0784
PSA (g)
% Incremento
C-
EA-CB0575
0,78
0,67
0,65
0,62
0,61
0,61
0,56
0,48
25
14
0
0
0
0
0
0
0
Isolates
Aislado
bacteriano
B
a
0,40
Peso seco raíz (g)
Root dry weight
0,45
45
40
a
ab
0,35
ab
35
ab
ab
0,30
30
b
0,25
0,20
Control
0,15
b
25
20
EA-CB158
15
0,10
10
0,05
5
0,00
PSR
% Incremento
EAEAEAEACB0177 CB0784 CB0158 CB1121
0,41
39
0,37
32
0,34
26
0,34
26
EAEACB0686 CB0131
0,33
24
Aislado
bacteriano
Isolates
0,30
24
C-
EACB0575
0,25
0
0,24
0
0
EA-CB0177 (39%)
EA-CB0784 (32%)
9,0
8,4
20
ab
ab
ab
Longitud
aérea (cm)
Shoot longitude
(cm)
8,0
abc
18
abc
bc
c
7,0
16
14
6,0
12
5,0
10
4,0
8
3,0
6
2,0
4
1,0
2
0,0
LA (cm)
% Incremento
0
EACB0158
EACB1121
EACB0177
EACB0686
EACB0131
EACB0575
EACB0784
C-
8,4
18
8,2
16
8,1
15
7,9
13
7,5
8
7,4
7
7,3
5
6,9
0
Bacterian
isolate
Aislado
bacteriano
Uninoculated C-
Inoculated 158
LA
EA-CB0158 (18%)
EA-CB01121 (16%)
EA-CB0177 (15%)
EA-CB0686 (13%)
Measurements
Microorganisms
Pseudostem
B. cereus EA-CB0131
Root
B. subtilis EA-0575
Fresh weight
B. pum ilus EA-0784
Dry weight
B. subtil is EA-1121
TREATMENT
Formulated
Strain
Pesta
EACB 0131
EACB 0575
EACB 0 784
EACB 1121
0
Talc
EACB 0131
EACB 0575
EACB 0 784
EACB 1121
0
Absolute Control
Whitout inoculation
0
20
plants/treatments
Reinoculation
Monthly
Measurements
Foliar Emit
Longitude
Stem diameter
Flowering
Weigth Raceme
Experimental field
AUGURA, CarepaAntioquia. Lote 4.
TREATMENT
PRODUCTION TIME PROGENY
(months)
T131
P131
T575
P575
T784
P784
T1121
P1121
T0
P0
Absolut Control
7.06 abc
7.26 abcd
7.41 abcde
6.8 a
6.92 abc
8 de
6.94 abc
7.54 abcde
7.67 cde
7.67 cde
8.07 e
1.58 MONTHS OF
DIFFERENCE
1.266 MONTHS
OF DIFFERENCE
TREATMENT
T131
P131
T575
P575
T784
P784
T1121
P1121
T0
P0
Absolute Control
leaves
Bacota- bellota
Stem
Progeny
Corm
PRODUCTION TIME
FLOWERING (months)
7.05 abc
7.05 abc
7.25 abc
6.68 a
7.25 abcd
7.63 de
6.85 ab
7.29 bcd
7.45 cd
7.4 cd
8.26 e
Strains EACB-0158, EACB-0177, EACB-0575,
EACB-0784 and EACB-1121 were the best in vitro
in corn plants
Strains P575, P784, T575, T784 Showed the
best results in banana plants in greenhouse
Bioformulation P575 showed the best
response in growth promotion and reduction
time of the crop in the field. (6 weeks)
CONCEPTUAL DESIGN OF PRODUCTION
PROCESS OF Bacillus subtilis EA-CB0575
FOR PROMOTING GROWTH IN Musa AAA
Luisa Fernanda Posada Uribe
M.Sc. Candidate
(2012)
AIMS
To design conceptually the production process of the
strain Bacillus subtilis EACB-0575 to promote growth
in Musa AAA
 To develop a Bacillus subtilis culture medium and to
optimize the physico chemical factors involved in plant
growth at bioreactor level.
 To Scale of biomass production
 To Evaluate the effect in promoting growht in greenhouse
 To Desgin conceptually the production process
OUR NEXT STEP
BIOPROSPECTING
OF TROPICAL
BIODIVERSITY
TEST OF
PROMISSORY
ISOLATES
FORMULATION
OF
BIOPESTICIDES
CO- DEVELOPMENT
Biological Solutions for Agriculture
CLEAN
AGRICULTURE
Thank You
[email protected]

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