Sporogenous midgut bacteria of the leafroller, Pandemis pyrusana (Lepidoptera: Tortricidae)

The composition of the microbial flora within the midgut of Pandemis pyrusana Kearfott was analyzed by using the polymerase chain reaction (PCR) and culturing techniques. Universal 16S ribosomal RNA gene-specific eubacterial primers were used in PCR, and the resulting products cloned and sequenced....

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Bibliographic Details
Published in:Environmental entomology 1997-12, Vol.26 (6), p.1475-1481
Main Authors: McKillip, J.L. (Mississippi State University, Mississippi State.), Small, C.L, Brown, J.L, Brunner, J.F, Spence, K.D
Format: Article
Language:English
Subjects:
16S RDNA
ADN
BACILLUS LICHENIFORMIS
BACTERIA
Biological and medical sciences
Biological control
CLOSTRIDIUM
CLOSTRIDIUM RAMOSUM
CLOSTRIDIUM SPIROFORME
Control
DNA
FLORA MICROBIANA
FLORE MICROBIENNE
Fundamental and applied biological sciences. Psychology
GENETIC TRANSFORMATION
IDENTIFICACION
IDENTIFICATION
INTESTIN
INTESTINES
INTESTINOS
ISOLATION
MICROBIAL FLORA
NUCLEOTIDE SEQUENCE
PANDEMIS
Phytopathology. Animal pests. Plant and forest protection
Protozoa. Invertebrates
RIBOSOMAS
RIBOSOME
RIBOSOMES
SECUENCIA NUCLEOTIDICA
SEQUENCE HOMOLOGY
SEQUENCE NUCLEOTIDIQUE
SPOREFORMING BACTERIA
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
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Summary:The composition of the microbial flora within the midgut of Pandemis pyrusana Kearfott was analyzed by using the polymerase chain reaction (PCR) and culturing techniques. Universal 16S ribosomal RNA gene-specific eubacterial primers were used in PCR, and the resulting products cloned and sequenced. High homology (86-95%) was obtained between the cloned products and Clostridium ramosum and Clostridium spiroforme when compared with computer database sequence information. Initial culturing steps aimed at endospore-forming bacteria yielded reproducible growth from a bacterium that matches most closely with Bacillus licheniformis when analyzed by partial 16S ribosomal DNA sequencing and biochemical/cultural profiling. This isolate was transformed with an Escherichia coli-Bacillus subtilis shuttle vector and reintroduced into the insect, where it remained stable under nonselective conditions for at least 9 d after initial ingestion. This confirmed the ability of the genetically altered bacterium to survive within the leafroller midgut in the midst of competing microorganisms and perhaps to serve as a suitable vehicle for biological control
ISSN:0046-225X
1938-2936
DOI:10.1093/ee/26.6.1475