Professional Documents
Culture Documents
OF PAGES 7
Please cite this article in press as: Pe rez-Garca A, et al. Plant protection and growth stimulation by microorganisms: biotechnological applications of Bacilli in agriculture, Curr Opin Biotechnol
(2011), doi:10.1016/j.copbio.2010.12.003
Available online at www.sciencedirect.com
Plant protection and growth stimulation by microorganisms:
biotechnological applications of Bacilli in agriculture
Alejandro Pe rez-Garca, Diego Romero and Antonio de Vicente
The increasing demand for a steady, healthy food supply
requires an efcient control of the major pests and plant
diseases. Current management practices are based largely on
the application of synthetic pesticides. The excessive use of
agrochemicals has caused serious environmental and health
problems. Therefore, there is a growing demand for new and
safer methods to replace or at least supplement the existing
control strategies. Biological control, that is, the use of natural
antagonists to combat pests or plant diseases has emerged as
a promising alternative to chemical pesticides. The Bacilli offer
a number of advantages for their application in agricultural
biotechnology. Several Bacillus-based products have been
marketed as microbial pesticides, fungicides or fertilisers.
Bacillus-based biopesticides are widely used in conventional
agriculture, by contrast, implementation of Bacillus-based
biofungicides and biofertilizers is still a pending issue.
Address
Departamento de Microbiologa, Facultad de Ciencias, Universidad de
Ma laga, Instituto de Hortofruticultura Subtropical y Mediterra nea,
Boulevard Louis Pateur-Campus Universitario de Teatinos s/n, 29071
Ma laga, Spain
Corresponding author: Pe rez-Garca, Alejandro (aperez@uma.es)
Current Opinion in Biotechnology 2011, 22:17
This review comes from a themed issue on
Food biotechnology
Edited by Oscar Kuipers and Tjakko Abee
0958-1669/$ see front matter
# 2010 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.copbio.2010.12.003
Introduction
Insects and fungi affecting crops and post-harvested
fruits and vegetables are major threats to food production.
They have led to important economic losses worldwide,
particularly over the past few decades as agricultural
production has intensied. To face these problems, pro-
ducers have become increasingly dependent on agro-
chemicals. However, intensive use of these compounds
in conventional crop management has led to the emer-
gence of frequent problems of pesticide resistance in
insect pests and microbial pathogens and has also caused
serious problems affecting not only human health but also
the quality of the environment. Therefore, there is an
increasing demand by growers and consumers for new
environmentally friendly methods to replace, or at least
supplement, the existing chemical-based strategies
thereby achieving safer and more effective pest and
disease control.
Biological control, that is, the use of natural antagonistic
organisms to combat pests or suppress plant diseases,
offers an interesting alternative to the use of chemicals
[1
,2
,8].
The toxicity of the Cry proteins have traditionally been
explained by the formation of transmembrane pores or ion
channels that lead to osmotic cell lysis [7
]. In a recent
study, a more precise trimeric building block model for
Cry toxins ion channel formation has been proposed
www.sciencedirect.com Current Opinion in Biotechnology 2011, 22:17
based on sequence conservation and mutagenesis data
[9]. In addition to this, Cry toxin monomers also seem to
promote cell death in insect cells through a mechanism
involving an adenylyl cyclase/PKA signalling pathway
[10]. However, despite this entomopathogenic potential,
controversy has arisen regarding the pathogenic lifestyle
of B. thuringiensis. Recent reports claimthat B. thuringiensis
requires the co-operation of commensal bacteria within
the insect gut to be fully pathogenic [11,12]. In clear
opposition, genomic and proteomic studies have been
argued as the most solid data to convincingly demonstrate
that B. thuringiensis is a primary pathogen rather than a
soil-dwelling saprophyte [13,14,15
]. The ability of
various Bacillus strains to control fungal soilborne, foliar
and postharvest diseases has been attributed mostly to
iturins and fengycins [28
,30
].
Surfactin and fengycin lipopeptides have recently been
identied as bacterial determinants responsible for eli-
citation of ISR in the host plant [34
]. It is yet unclear
whether the induction of the ISR response by lipopep-
tides requires specic receptors in the plant membrane.
It is postulated that some lipopeptides may induce a
disturbance or transient channelling in the plasma mem-
brane, which in turn activates a cascade of molecular
events leading to enhanced defence [35
]. Nevertheless,
these ndings open up a new area of research to further
exploit these potential benecial effects of bacilli and
especially to gain more insight on the key structural
features and constituents of lipopeptides involved in
the induction of plant defence responses [36
].
The efcient protection of plants by biocontrol agents
requires their proper establishment in the host plant.
Chemotaxis, motility and growth are essential players
in this process. Lipopeptides can also inuence the eco-
logical tness of the producing strain contributing to plant
colonisation and persistence in the plant environment.
Thus, surfactin seems to be essential for swarming moti-
lity in B. subtilis [37]. In planta, the secretion of surfactin
and the formation of a stable, extensive biolm that
occurs upon root colonisation by B. subtilis has been
shown to be crucial for disease suppression [38]. Similar
results have been obtained with surfactin-decient
mutants of B. subtilis, which show disorganised biolm
formation and reduced biocontrol ability of fungal and
bacterial diseases after application on leaves (H Zeriouh
et al., unpublished). Moreover, a recent study attributes to
surfactin the role of triggering signal molecule for extra-
cellular matrix formation in functional and robust biolms
of B. subtilis [39
].
In addition, phytases eliminate chelate-forming phytate,
which is known to bind nutritionally important minerals
(such as Zn
2+
, Fe
2+
, Ca
2+
). Something similar happens to
iron. In the soil, the most prevalent form of iron is Fe
3+
,
which is relatively insoluble compared with the more
reduced form Fe
2+
and less readily taken up by plants
andmicroorganisms. Bacillus species, suchas B. megaterium,
can reduce metals, potentially increasing the bioavailabil-
ity of iron [43].
Direct plant growth promotion by Bacillus involves the
modulation of plant development through the production
of phytohormones [44]. Thus, several Bacillus species are
capable of producing auxin that might stimulate root
proliferation and nutrient uptake [45]. For example, in
B. amyloliquefaciens the biosynthesis of indole-3-acetic
acid (IAA) is responsible for plant growth promotion,
which in turn is strictly dependent on the presence of
tryptophan, one of the main compounds present in plant
root exudates [46]. Similarly, the inoculation of plants
with cytokinin-producing B. subtilis or B. megaterium
strains has a benecial effect on plant growth [47,48
].
In B. pumilus, however, plant growth promotion has been
associated with production of either gibberellin or ABA
and jasmonic acid [49,50]. Finally, volatile organic com-
pounds from B. subtilis have been shown to trigger growth
promotion in Arabidopsis by regulating auxin homeostasis,
thus providing a new paradigm as to how these bacteria
promote plant growth [51
].
There are a few commercial Bacillus-based bionematicidal
formulations but their implementation is still very limited.
Furthermore, novel functions of bacilli such as quorum
quenching have been demonstrated to restrain bacterial
infections. Many bacterial pathogens have evolved cell
cell communication (quorum-sensing) mechanisms to
regulate expression of virulence factors. Key components
in these regulation systems are N-acyl homoserine lactones
(AHLs), which act as signal molecules. Some Bacillus
species such as B. thuringiensis are able to break down
AHLs (quorumquenching) by production of N-acyl homo-
serine lactone lactonases, which open the lactone ring of
AHLs, signicantly silencing bacterial virulence [53
].
Therefore, exploitation of these activities could expand
the phytoprotection possibilities of bacilli also as microbial
bactericides.
Bacillus-based products have great potential for use in
integrated pest management (IPM) systems; unfortu-
nately, relatively little work has been undertaken on their
integration with other IPM tools such as cultural prac-
tices, host resistance, chemical control, and other bio-
logical control agents. Formulation and application
methods are key issues inuencing the efcacy of com-
mercial products [54
], high-throughput studies
can be undertaken to gain knowledge about the biocon-
trol competence of these agents. As newprogress on these
topics is made, new and better Bacillus-based formu-
lations will be developed. Therefore, we anticipate a
more relevant role for these microorganisms in promoting
plant health in the 21st century agriculture.
Acknowledgements
The authors gratefully acknowledge past and ongoing support for their work
on the use of bacilli as biocontrol agents from Plan Nacional de I + D + I of
the Ministerio de Ciencia e Innovacio n, Spain (AGL2001-1837; AGL2004-
0656; AGL2007-65340; AGL2010-21848), co-nanced with FEDER funds
(European Union).
References and recommended reading
Papers of particular interest, published within the annual period of
review, have been highlighted as:
of special interest
of outstanding interest
1.
Roh JY, Choi JY, Li MS, Jin BR, Je YH: Bacillus thuringiensis as a
specic, safe, and effective tool for insect pest control. J
Microbiol Biotechnol 2007, 17:547-559.
This is a comprehensive reviewon B. thuringiensis as a tool for insect pest
control. In this paper the authors describe the most important milestones
in the history of B. thuringiensis, from the rst isolation and description to
the most recent introduction of Cry genes in crop plants.
8. Kumar S, Chandra A, Pandey KC: Bacillus thuringiensis (Bt)
transgenic crop: an environmentally friendly insect-pest
management strategy. J Environ Biol 2008, 29:641-653.
9. Torres J, Lin X, Boonserm P: A trimeric building block model for
Cry toxins in vitro ion channel formation. Biochim Biophys Acta
2008, 1778:392-397.
10. Zhang X, Candas M, Griko NB, Taussig R, Bulla LA Jr: A
mechanism of cell death involving an adenylyl cyclase/PKA
signaling pathway is induced by the Cry1Ab toxin of Bacillus
thuringiensis. Proc Natl Acad Sci USA 2006, 103:9897-9902.
11. Broderick NA, Raffa KF, Handelsman J: Midgut bacteria required
for Bacillus thuringiensis insecticidal activity. Proc Natl Acad
Sci USA 2006, 103:15196-15199.
12. Broderick NA, Robinson CJ, McMahon MD, Holt J, Handelsman J,
Raffa KF: Contributions of gut bacteria to Bacillus
thuringiensis-induced mortality vary across a range of
Lepidoptera. BMC Biol 2009, 7:11.
13. Gohar M, Gilois N, Graveline R, Garreau C, Sanchis V, Lereclus D:
A comparative study of Bacillus cereus. Bacillus thuringiensis
and Bacillus anthracis extracellular proteomes. Proteomics
2005, 5:3696-3711.
14. Han CS, Xie G, Challacombe JF, Altherr MR, Bhotika SS, Bruce D,
Campbell CS, Campbell ML, Chen J, Chertkov O et al.:
Pathogenomic sequence analysis of Bacillus cereus and
Bacillus thuringiensis isolates closely related to Bacillus
anthracis. J Bacteriol 2006, 188:3382-3390.
15.