Institute of Agricultural Research

New parasitoid species for pest control

As an alternative for the indiscriminate use of pesticides, INIA has identified and characterized some native Trichogramma species utilized as biological control agents for agricultural and forestry pests of economic importance. Such species include T. nerudai, T. cacoeciae, T. evanescens, T. pretiosum, identified through the application of taxonomic procedures and molecular tests. At present, certain species are produced industrially, such as T.nerudai, which has been identified as a new scientific specie and was collected on the eggs of the European pine shoot moth (Rhyacionia bouliana). Marcos Gerding (mgerding@inia.cl).

Entomopathogenic fungi to control South American tomato moth.

The South American tomato moth, Tuta absoluta, is a key pest and limiting factor for tomato cultivation in the country, both in the field and under protected conditions. To control the pest, INIA (INIA-Quilamapu, VIII Region) has found native isolations of specific entomopathogenic fungi such as Beauveria bassiana and Metarhizium anisopilae for controlling T. absoluta eggs and larvae.
An entomopathogen-based biopesticide product is presently under formulation that is highly specific and, at the same time, does not cause harm to the environment nor existing beneficial fauna. The use of this biopesticide is complemented with benign insect activity such as Trichogramma sp., Encarsia formosa, Bombus terrestres and Apis mellifera. The benefits of biological control not only involve reduced costs and production efficiency, but also protect fruit quality, human health and the environment in a system of integrated management. Andrés France (afrance@inia.cl).

Developing a biopesticide for controlling the European pine shoot moth

The European pine shoot moth, Rhyacionia buoliana, is a pest that has spread rapidly in Pinus radiata plantations in the central and southern regions of the country. Insect larvae destroy the tree buds, deforming and killing growth points. When damage occurs in apical buds, it causes tree deformity, thus reducing the quality and quantity of wood harvest.? Biological control was introduced in a specific moth parasitoid, Orgilus obscurator, reducing the pest in plantations. Additionally, entomopathogenic fungi Beauveria bassiana, was introduced and has effectively controlled the pest without affecting the O.obscurator. Laboratory and field tests have shown the possibility of combining both organisms, thus not having to resort to chemical control during the establishment phase or in areas where O.obscurator cannot be established. A basic commercial formulation for a field biopesticide that keeps its viability and effectiveness after one year of storage was created. Marcos Gerding (mgerding@inia.cl).

Biological control of native fruit tree weevil (Aegorhinus superciliosus) with native entomopathogenic nematodes.

Entomopathogenic nematodes, native to the Steinernema genera, were identified for the first time and are capable of parasitizing fruit tree weevil larvae as well as other similar pests, both in the field and roots in containerized soil. These nematodes were concentrated in wax moth larvae and evaluated for pest control in laboratories, greenhouses and fields. A dose of 300.000 nematodes/m2, in commercial raspberry and blueberry orchards may limit the occurrence of adults during the season by 71% for Aegorhinus superciliosus, 68% for Asynonychus cervinus and 69% for Otiorhynchus sulcatus. Andrés France (afrance@inia.cl).

Development of strategies aimed at eliminating potato smut (Thecaphora solani) in quarantine areas and their integrated control in endemic areas

The so-called "potato smut" disease caused by Thecaphora solani is one of the most serious problems affecting potato crops. It is considered a quarantine disease in practically all countries in the world, although there is little biological knowledge about the T. solani, its epidemiology and control. Studies at INIA have led to in vitro fungi propagation for the first time, the first induction of galls in potatoes inoculated with in vitro-secured mycelium; the ribosomal ITS1-5.8-ITS2 region of T. solani was completely amplified, purified, cloned and sequenced; region 5' LSU of T.solani was sequenced and analyzed for taxonomic purposes; highly specified monoclonal antibodies for the fungi were obtained; expression of the disease effectively decreased with soil fumigation; production of galls was reduced by 77 to 91% with the use of seed disinfectants and smut-resistant germplasma of S. tuberosum subsp. tuberosum was obtained. A book titled "Potato smut, etiological developments and disease control" has been published, including all existing information on the disease, mostly provided by INIA research. Orlande Andrade (oandrade@inia.cl).

Transcriptional Profiling for characterization of Vitis vinifera in response to the fungal pathogen Botrytis cinerea by using ESTs and microarrays generated in the Genoma Chile Initiative.

Grape is a major crop in Chile, generating products such as table grapes for fresh consumption, wines, and "pisco" (liquor). Different cultivars are susceptible to necrotrophic pathogen Botrytis cinerea, which generates severe losses, either in field (cultivation) conditions as well as in post-harvest storage or transporting.
Using the Arabidopsis and tomato models, some plant mechanisms responding to this pathogen have been elucidated. However, there is no information available about biological processes involved in the grape response. In this project, we have designed and evaluated for hybridization experiments, macroarrays with 5.000 unigenes resulting from sequencing aprox. 18 libraries generated from around 100.000 ESTs, from Thompson Seedless (Sultana) and Carménère cultivars. Along these lines, isolated RNAs from different infected tissues (leaves, fruits and inflorescences) from both cultivars, were screened and compared with healthy counterparts, in order to check differences with gene expression levels generated by fungal infection.
Current results have been processed and are under bioinformatic evaluation for metabolic and defense pathways re-construction, and comparative studies with already known model mechanisms. Humberto Prieto (hprieto@inia.cl).

"Development of bumblebee (Bombus) production system for pollination technology of fruit and vegetables"

This project carried out by INIA and associated companies BIOCONTROL (V Region) and MIPAGRO (VI Region) during years 2002-2005, has been of strategic value for the country for its resulting lab-based massive production procedure for the native bumblebee Bombus dahlbomii. Today, a license application for this procedure has been submitted to the Intellectual Property Department of the Chilean Ministry of Economic Affairs.

On the other hand, local units producing the Bombus terrestris bumblebee were identified. Until 2002, this specie was imported from Holland and Israel and used for greenhouse tomato pollination, priced at US$200 for each colony. Today, after the project's production technology was transferred to companies MIPAGRO and BIOCONTROL, the latter are able to produce an average of 2,500 colonies for only US$100 for farmers. The project also revealed pollination capabilities of Bombus. genera insects not only in tomatoes but also blueberries, avocados, strawberries, vegetables grown under plastic and vegetables for seed production.
A strategic alliance has been forged between the companies participating in the project and INIA, through an agreement whereby INIA will provide genetic material (queens) for a period of three years to these companies, ensuring the quality of the material and keeping this information confidential until the companies are solidly established. Patricia Estay (pestay@inia.cl).

Implementation of a traceability system based on molecular markers to improve bovine competition for meat exports.

Traceability of animal origin products has gained relevance as a protection device in the light of the food crisis that arose in the mid-nineties in European Union countries. In view of the Free Trade Agreement signed between Chile and the EU, these countries are an important market for local products.
With the help of this project, various biological samples have been evaluated, such as hair (hair follicle), blood, tissues and/or meat sold in markets, in order to find the best samples for obtaining, purifying and conserving DNA. It was observed that samples of hair follicles and tissue provided the best results. At the same time, the best method for DNA extraction was alkaline lysis with SDS and proteinase K, compared to others evaluated. On a panel of 10 bovine microsatellite markers 3 multiplex reactions have been established (M1:ETH3, BM1824, TGLA112 and TGLA 127, M2: ETH19, BM2113 and ETH225 and M3: SPS115, INRA23 and TGLA126) generating reproducible and reliable results in the various biological samples which were analyzed
The microsatellite system developed for this was capable of discriminating between individual animals of close bovine kinship.
First evaluations of this system within a traceability scheme in the meat production chain led to a 100% discrimination certainty of 25 hair samples with the corresponding samples of medullar channels, confirming the highly discriminating value of this technique and its application to guarantee traceability certification. Ricardo Felmer (rfelmer@inia.cl).

Valuation of sludge from wastewater treatment plants for use as fertilizer

Sludge resulting from wastewater treatment plants has proven to be feasible for fertilization. Plants are able to adequately capture nutrients from the sludge, although it was observed that several years (at least three) of consecutive applications of sludge are required for full results. Our findings confirm the experience of developed countries, especially Europe, over the past 30 years.
The distinctive aspect of this study is that it is the first research project carried out in Chile with sludge produced by the El Trebal plant in the Metropolitan Region, and its calibrated use according to local environmental and technical conditions.
Although its use has proven to be feasible, environmental quality considerations make it inconvenient that sludge should completely replace mineral fertilizers, as this may lead to progressive accumulation of metallic trace elements in the soil, which must be avoided. It was also observed that when sludge applications are introduced in soils, there is no dispersion of pathogen microorganisms. Francisco Tapia (ftapia@inia.cl).