ABSTRACT
BACKGROUND: Piercing/sucking insect pests in the order Hemiptera causes substantial crop losses by removing photoassimilates and transmitting viruses to their host plants. Cloning and heterologous expression of plantderived insect resistance genes is a promising approach to control aphids and other sap-sucking insect pests. While expression from the constitutive 35S promoter provides broad protection, the phloem-specific rolC promoter provides better defense against sap sucking insects. The selection of plant-derived insect resistance genes for expression in crop species will minimize bio-safety concerns. RESULTS: Pinellia ternata leaf agglutinin gene (pta), encodes an insecticidal lectin, was isolated and cloned under the 35S and rolC promoters in the pGA482 plant transformation vector for Agrobacterium-mediated tobacco transformation. Integration and expression of the transgene was validated by Southern blotting and qRT-PCR, respectively. Insect bioassays data of transgenic tobacco plants showed that expression of pta under rolC promoter caused 100% aphid mortality and reduced aphid fecundity up to 70% in transgenic tobacco line LRP9. These results highlight the better effectivity of pta under rolC promoter to control phloem feeders, aphids. CONCLUSIONS: These findings suggested the potential of PTA against aphids and other sap sucking insect pests. Evaluation of gene in tobacco under two different promoters; 35S constitutive promoter and rolC phloemspecific promoter could be successfully use for other crop plants particularly in cotton. Development of transgenic cotton plants using plant-derived insecticidal, PTA, would be key step towards commercialization of environmentally safe insect-resistant crops.
Subject(s)
Aphids/pathogenicity , Pest Control, Biological , Pinellia/chemistry , Plant Viruses , Tobacco , Blotting, Southern , Polymerase Chain Reaction , Promoter Regions, Genetic , Plants, Genetically Modified , Plant Leaves/chemistry , Transgenes , Disease Resistance , Crop ProtectionABSTRACT
RESUMEN Algunos Bacillus spp. promotores de crecimiento vegetal son microorganismos reconocidos como agentes de control biológico que forman una estructura de resistencia denominada endospora, que les permite sobrevivir en ambientes hostiles y estar en casi todos los agroecosistemas. Estos microorganismos han sido reportados como alternativa al uso de agroquímicos. Sus mecanismos de acción se pueden dividir en: producción de compuestos antimicrobianos, como son péptidos de síntesis no ribosomal (NRPs) y policétidos (PKs); producción de hormonas, capacidad de colonización, formación de biopelículas y competencia por espacio y nutrientes; síntesis de enzimas líticas como quitinasas, glucanasas, protesasas y acil homoserin lactonasas (AHSL); producción de compuestos orgánicos volátiles (VOCs); e inducción de resistencia sistémica (ISR). Estos mecanismos han sido reportados en la literatura en diversos estudios, principalmente llevados a cabo a nivel in vitro. Sin embargo, son pocos los estudios que contemplan la interacción dentro del sistema tritrófico: planta - microorganismos patógenos - Bacillus sp. (agente biocontrolador), a nivel in vivo. Es importante destacar que la actividad biocontroladora de los Bacillus es diferente cuando se estudia bajo condiciones de laboratorio, las cuales están sesgadas para lograr la máxima expresión de los mecanismos de acción. Por otra parte, a nivel in vivo, la interacción con la planta y el patógeno juegan un papel fundamental en la expresión de dichos mecanismos de acción, siendo esta más cercana a la situación real de campo. Esta revisión se centra en los mecanismos de acción de los Bacillus promotores de crecimiento vegetal, expresados bajo la interacción con la planta y el patógeno.
ABSTRACT Some Bacillus spp. plant growth promoters are microorganisms recognized as biological control agents, which form a resistance structure called endospore, which allows them to survive in hostile environments and be in almost all agroecosystems. These microorganisms have been reported as an alternative to the use of agrochemicals. Its mechanisms of action can be divided into: production of antimicrobial compounds, such as non-ribosomal peptides (NRPs) and polyketides (PKs); hormone production, colonization capacity, biofilm formation and competition for space and nutrients; synthesis of lytic enzymes such as chitinases, glucanases, protesases and acyl homoserin lactonases (AHSL); production of volatile organic compounds (VOCs); and induction of systemic resistance (SRI). These mechanisms have been reported in the literature in several studies, mainly carried out in vitro. However, there are few studies that contemplate the interaction within the tritrophic system: plant - pathogenic microorganisms -Bacillus sp. (biocontrol agent), in vivo level. It is important to note that the Bacillus biocontrol activity is different when studied under laboratory conditions, which are biased to achieve maximum expression of the mechanisms of action. On the other hand, at the in vivo level, the interaction with the plant and the pathogen play a fundamental role in the expression of said mechanisms of action, being this closer to the real field situation. This review focuses on the mechanisms of action of the Bacillus promoters of plant growth, expressed under the interaction with the plant and the pathogen.