Bacillus thuringiensis and Lysinibacillus sphaericus: Characterization and use in the field of biocontrol

289 Pages·2017·6.842 MB·English

by  Lidia Mariana Fiuza, Ricardo Antonio Polanczyk, Neil Crickmore (eds.)

Lidia Mariana Fiuza
Ricardo Antonio Polanczyk
Neil Crickmore E ditors
Bacillus
thuringiensis and
Lysinibacillus
sphaericus
Characterization and use
in the fi eld of biocontrol
Bacillus thuringiensis and Lysinibacillus sphaericus
Lidia Mariana Fiuza • Ricardo Antonio Polanczyk
Neil Crickmore
Editors
Bacillus thuringiensis and
Lysinibacillus sphaericus
Characterization and use in the field of
biocontrol
Editors
Lidia Mariana Fiuza Ricardo Antonio Polanczyk
Control Agro Bio Pesquisa e Defesa Plant Protection Department
Agropecuária Ltda Universidade Estadual Paulista
Porto Alegre, Brazil Jaboticabal, Brazil
Instituto Rio Grandense do Arroz - IRGA
Cachoeirinha, Brazil
Neil Crickmore
School of Life Sciences
University of Sussex
Brighton, UK
ISBN 978-3-319-56677-1 ISBN 978-3-319-56678-8 (eBook)
DOI 10.1007/978-3-319-56678-8
Library of Congress Control Number: 2017939599
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Preface
Biological control using biopesticides based on Bacillus thuringiensis and
Lysinibacillus sphaericus has been employed for almost five decades in countries
such as Brazil for the control of agricultural pests and vectors of tropical disease.
Professionals in the academic community have undertaken in-depth studies on a
variety of potential biological control agents including fungi, bacteria, viruses, hel-
minths and others. As a result of these studies commercial products have been
developed for use in the field. A great advantage of these products is that they show
few of the undesirable, nonspecific, activities associated with more traditional
insecticides that affect many arthropods not implicated in crop destruction or the
spread of etiological agents of human or animal disease. Continuing research and
dissemination of results will further the development of new products.
This publication by Springer highlights the scientific and technical progress
made on two of the most important biocontrol agents – the bacteria Bacillus thuring-
iensis and Lysinibacillus sphaericus. Brazil is such a major user of biological con-
trol agents that much of this book concentrates on the characterization, development
and commercialization of these bacteria. Much of the information presented does,
of course, have relevance on a worldwide stage. The seventeen chapters have been
written by a body of highly experienced scholars, professors, scientists and research-
ers. Five of them have been written by experts from outside of Brazil.
In addition to natural selection increasing the range of bacteria effective as bio-
control agents, the contents of this book deal with how studies on the physiology,
biochemistry, general biology, and cellular and molecular biology aspects of the
considered bacteria can help us develop improved products. The characterization
and current classification of Bacillus thuringiensis toxins are addressed, which pro-
vide a useful framework for the optimization of the entomocidal principles required
for their successful use in the control of pests and vectors of human diseases.
Aside from the toxins, the characterization of Bacillus thuringiensis serovars by
AFLP microsatellites, rep-PCR, and more recently genomic sequencing, is
described. Such technologies greatly enhance our ability to identify new potential
products and to understand, and potentially manipulate, the virulence of particular
strains. The expression of insecticidal genes in baculovirus, or in plants, provides an
v
vi Preface
alternative way to study, and employ, the encoded proteins. In addition, understand-
ing the interaction of the insect with the virulence factors of the bacterium, allied to
the spectrum of activity of Bacillus thuringiensis in relation to the main orders of
insect pests, provide useful information for the control of agricultural and agrofor-
estry crops. Another important chapter show the expression of Cry proteins in insect
cells using recombinant baculoviruses has been shown to be an alternative means of
production of these proteins for functional and/or structural studies. Also the novel
strategies include site directed mutagenesis, domain swapping among different Cry
toxins where novel hybrid proteins were constructed containing domains or loop
regions from different Cry proteins that resulted in improved toxicity against
selected insect pests.
In selected chapters, many interesting aspects are highlighted, such as reports of
biotechnological studies linked to genetically modified plants containing new
Bacillus thuringiensis proteins, and the beneficial interactions between
Bt-bioinsecticides and parasitoids and predators in agricultural ecosystems. Also
interesting is the work presented on a range of microorganisms with biopesticidal
action, other than the two main protagonists of this book, which the authors propose
as new alternatives as active agents in the field aimed at targets such as Acromyrmex
spp., Nasutitermes ehrhardt and Rhizoctonia solani. This also includes the potential
of using synergistic interactions of Bacillus thuringiensis and Bacillus subtilis with
Purpureocillium lilacinus, or even the Nuclear Polyhedrosis virus, among others.
Concerns over mosquito resistance to the entomopathogenic bacteria Bacillus
thuringiensis and Lysinibacillus sphaericus are revisited with information on the
current state of the art. Finally, aspects related to the role of the Brazilian govern-
mental company “Empresa Brasileira de Pesquisa Agropecuária”, which is focused
on agriculture and which develops products for application at the field level, as well
as studies to combat new pests, are addressed.
At the end of this preface, I am sure that the plethora of authors are convinced
that they have done their best to make this book a reality, and that they have contrib-
uted greatly to the work, which will hopefully fulfill the curiosity of an increasing
number of readers and students interested in this area.
Leon Rabifnovitch, PhD
Head of Bacterial Physiology Laboratory
Fundação Oswaldo Cruz-Instituto Oswaldo Cruz;
Manguinhos-Rio de Janeiro, Brazil
Contents
1 Bacillus thuringiensis Characterization: Morphology, Physiology,
Biochemistry, Pathotype, Cellular, and Molecular Aspects . . . . . . . . . . 1
Leon Rabinovitch, Adriana Marcos Vivoni, Vilmar Machado,
Neiva Knaak, Diouneia Lisiane Berlitz, Ricardo Antonio Polanczyk,
and Lidia Mariana Fiuza
2 The Biology, Ecology and Taxonomy of Bacillus thuringiensis
and Related Bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Ben Raymond
3 Bacillus thuringiensis Toxin Classification . . . . . . . . . . . . . . . . . . . . . . . 41
Neil Crickmore
4 Insecticidal Proteins from Bacillus thuringiensis and Their
Mechanism of Action . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Alejandra Bravo, Sabino Pacheco, Isabel Gómez, Blanca Garcia-Gómez,
Janette Onofre, and Mario Soberón
5 Effect of Bacillus thuringiensis on Parasitoids and Predators . . . . . . . 67
Sergio Antonio De Bortoli, Alessandra Marieli Vacari,
Ricardo Antonio Polanczyk, Ana Carolina Pires Veiga,
and Roberto Marchi Goulart
6 Characterization of Bacillus thuringiensis Using Plasmid
Patterns, AFLP and Rep-PCR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Fernando Hercos Valicente and Rosane Bezerra da Silva
7 New Sequencing Technologies and Genomic Analysis
Applied to Bacillus thuringiensis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Roberto Franco Teixeira Correia, Anne Caroline Mascarenhas dos Santos,
Raimundo Wagner de Souza Aguiar, Bergmann Morais Ribeiro,
and Fernando Lucas Melo
vii
viii Contents
8 Expression of Bacillus thuringiensis Toxins in Insect Cells . . . . . . . . . . 99
Bergmann Morais Ribeiro, Érica Soares Martins,
Raimundo Wagner de Souza Aguiar,
and Roberto Franco Teixeira Corrêa
9 Bacillus thuringiensis: Different Targets and Interactions . . . . . . . . . 111
Lidia Mariana Fiuza, Diouneia Lisiane Berlitz,
Jaime Vargas de Oliveira, and Neiva Knaak
10 Specificity and Cross-order Activity of Bacillus thuringiensis
Pesticidal Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
Kees van Frankenhuyzen
11 The American Bacillus thuringiensis Based
Biopesticides Market . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173
Ricardo Antonio Polanczyk, Kees van Frankenhuyzen,
and Giuliano Pauli
12 Mass Production, Application and Market Development
of Bacillus thuringiensis Biopesticides in China . . . . . . . . . . . . . . . . . . 185
Lin Li, Zhenmin Chen, and Ziniu Yu
13 The Role of Embrapa in the Development of Tools
to Control Biological Pests: A Case of Success . . . . . . . . . . . . . . . . . . . 213
Rose Gomes Monnerat, Glaucia de Figueiredo Nachtigal,
Ivan Cruz, Wagner Bettiol, and Clara Beatriz Hoffman Campo
14 Bacillus Entomopathogenic Based Biopesticides
in Vector Control Programs in Brazil . . . . . . . . . . . . . . . . . . . . . . . . . . 223
Clara Fátima Gomes Cavados, Wanderli Pedro Tadei,
Rosemary Aparecida Roque, Lêda Narcisa Regis,
Claudia Maria Fontes de Oliveira, Helio Benites Gil,
and Carlos José Pereira da Cunha de Araujo-Coutinho
15 Resistance of Mosquitoes to Entomopathogenic
Bacterial-Based Larvicides: Current Status and Strategies
for Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 239
Maria Helena Neves Lobo Silva-Filha
16 The Importance of Bacillus thuringiensis in the Context
of Genetically Modified Plants in Brazil . . . . . . . . . . . . . . . . . . . . . . . . 259
Deise Maria Fontana Capalbo and Marise Tanaka Suzuki
17 Resistance of Spodoptera frugiperda to Bacillus
thuringiensis Proteins in the Western Hemisphere . . . . . . . . . . . . . . . 273
Samuel Martinelli, Renato Assis de Carvalho,
Patrick Marques Dourado, and Graham Phillip Head
Chapter 1
Bacillus thuringiensis Characterization:
Morphology, Physiology, Biochemistry,
Pathotype, Cellular, and Molecular Aspects
Leon Rabinovitch, Adriana Marcos Vivoni, Vilmar Machado, Neiva Knaak,
Diouneia Lisiane Berlitz, Ricardo Antonio Polanczyk,
and Lidia Mariana Fiuza
Abstract In this publication, “Bacillus thuringiensis and Lysinibacillus
sphaericus – characterization and use in the field of biocontrol,” this chapter can be
seen as a brief general and historical introduction to the central theme of the book,
where data on the cellular physiology, biochemical, genetic, molecular, and toxico-
logical aspects of the bacterium, B. thuringiensis (Bt), are reported. This predomi-
nant entomopathogenic prokaryote was discovered and denominated Bt around a
century ago, between 1902 and 1911. From the microbiological point of view, this
bacterium is ubiquitous, Gram-positive, produces ellipsoidal but predominantely
cilindrical endospores (central to paracentral) and contains a parasporal inclusion
body called crystal or δ-endotoxin. The crystal is constituted of Cry proteins with
molecular weight between 30 kDa and 140 kDa, which are coded by cry genes.
On the other hand, this bacterial species synthesizes several enzymes and
toxins that give them a wide adaptation to natural habitats. Bt strains have been
L. Rabinovitch • A.M. Vivoni
Fundação Oswaldo Cruz – Instituto Oswaldo Cruz,
Manguinhos, Rio de Janeiro, RJ, Brazil, CEP.: 21045-900
V. Machado
Pós Doc – Programa de Pós Graduação em Ciências e Biotecnologia – UNOESC,
Videira, SC, Brazil
N. Knaak • L.M. Fiuza (*)
Control_Agro_Bio Pesquisa e Defesa Agropecuária Ltda,
Porto Alegre, RS, Brazil, CEP.: 91230-140
Instituto Rio Grandense do Arroz – IRGA, Cachoeirinha, RS, Brazil, CEP.: 94930-030
e-mail: [email protected]
D.L. Berlitz
Control_Agro_Bio Pesquisa e Defesa Agropecuária Ltda,
Porto Alegre, RS, Brazil, CEP.: 91230-140
R.A. Polanczyk
Departamento de Proteção de Plantas, Universidade Estadual Paulista,
Jaboticabal, CEP 14884-900 SP, Brazil
© Springer International Publishing AG 2017 1
L.M. Fiuza et al. (eds.), Bacillus thuringiensis and Lysinibacillus sphaericus,
DOI 10.1007/978-3-319-56678-8_1
2 L. Rabinovitch et al.
studied and, over time, characterized and described as toxic and specific for
Lepidoptera, Diptera, Coleoptera, Nematoda, Protozoa, Trematoda, Acari,
Hymenoptera, Hemiptera, Orthoptera, Isoptera, Mallophaga, and among other tar-
get pests. Globally, 82 Bt serovars sometimes called subspecies were described until
1999, which currently correspond to more than 700 cry genes distributed in about
70 classes. The nomenclature review of cry genes, which encode Bt Cry proteins,
has been published by Crickmore et al. and has been constantly updated on the
website: http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/.
Keywords Bacteria • Bacillus spp. • B. thuringiensis • Physiology • Biochemistry
• Genetics • DNA • Toxicology
1.1 The Bacterium Bacillus thuringiensis Berliner, 1911
Among prokaryotes, bacteria of the genus Bacillus (family Bacillaceae) have been
used in the microbial control of pests. In this genus, the species Bacillus thuringien-
sis (Bt) is distinguished by its biopesticide potential.
Bt was discovered by Ishiwatta in 1902 in reared Bombyx mori L. (Lepidoptera,
Bombycidae) in Japan. Later it was reisolated by Berliner, in 1911, from Ephestia
kuehniella Zeller (Lepidoptera, Pyralidae) larvae in Thuringia, which gave rise to its
current name (De Barjac and Bonnefoi 1968). According to Van Frankenhuyzen
(1993), the first biological control trials of Bt were conducted against Ostrinia nubi-
lalis Hübner (Lepidoptera, Pyralidae) between 1920 and 1930 in Europe. Between
1930 and 1940, several trials were carried out with other lepidopteran species in
Europe and in the USA. Regarding the biological control of insects today, Bt is the
mostly used microorganism worldwide (Lacey et al. 2015).
Bt is a ubiquitous bacterium with a large enzyme complement, which allows it to
be found in a variety of sites, such as: soil, insects and their habitats, stored prod-
ucts, plants, forest, and aquatic environments. It can remain latent in the environ-
ment even in adverse conditions for its development (Azevedo et al. 2000; Fiuza
2001). On the other hand, 18 Bt strains were isolated from Simulium sp. larvae and
adult and of B. sphaericus (Cavados et al. 2001). Larvae were collected in different
rivers of states  of Rio de Janeiro and São Paulo.  Between strains obtained, only two
were shown to belong to serotype H-14, B. thuringiensis serovar israelensis. Nine
strains were “autoagglutinating” ones and seven B. thuringiensis serovar
Oswaldocruzi (H-39) were also indentified and it was the first report of the isolation
of entomopatogenic Bacillus from Simulium sp. larvae and adult collected in Brazil
(Cavados et al. 2001). Its entomopathogenic activity is highly dependent on the
parasporal inclusion body that forms during sporulation, which consists of Cry pro-
teins that are encoded by cry genes (Höfte and Whiteley 1989; Schnepf et al. 1998).
According to the literature, parasporal inclusion bodies (crystals) are composed
of proteins of varying quantity and quality according to the bacterial strain (Hofte
and Whiteley 1989). Strains containing these crystals were measured as being toxic