Person: Pellegrineschi, A.
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Pellegrineschi
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Pellegrineschi, A.
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- Biotechnology applications for wheat improvement at CIMMYT(The Scientific and Technology Research Council of Turkey, 2005) William, H.M.; Singh, R.P.; Trethowan, R.; Van Ginkel, M.; Pellegrineschi, A.; Huerta-Espino, J.; Hoisington, D.A.
Publication - Optimization of an in vitro bioassay for wheat diseases(CIMMYT, 2000) Salgado, M.; Pellegrineschi, A.; Mezzalama, M.; McLean, S.D.; Hoisington, D.A.Alternaria triticina, Fusarium graminearum, Pyrenophora tritici-repentis, Blpolaris sorokinisna (Helminthosporium sativum); Pythium sp., and Rhizoctonia sp. were tested on the basis of being a representative group of important fungal pathogens in order to establish a reliable bloassay system to test eventual fungal resistance in transgenic plants. The inoculum was prepared on V-8 agar medium (in petri dishes) for P. tritici-repents and Pythlum sp., and on PDA (potato dextrose agar) medium for the other fungi. The cultures were incubated at room temperature for 7- 10 days in a culture chamber with a constant standard illumination. The suspensions of conidia or micella were prepared on sterile distilled water with 1 few drops of tween 20 and scrapings of the fungi culture. The inoculum was homogenized by vorlexing the suspension for a few seconds. The concentration used for the conidia solution was adjusted to 106 conidia/ml. Fresh leaf samples from adult plants (heading stage) were sterilized and then dipped In the inoculum suspension. The inoculated leaves were then transferred to water agar medium (1% agar), in 8-well rectangular multi-dishes, at room temperature. The level of resistance of the material to these pathogens was evaluated 4-7 days after inoculation. We believe that the routine use of this protocol (fast, reliable, and inexpensive) could allow the easy identification of resistant/tolerant plants to these diseases. In addition, because of the simplicity of this test, it can be useful for early screening of a large number of individuals, as required for transgenic plant screenings.
Publication - Testing public Bt maize events for control of stem borers in the first confined field trials in Kenya(Academic Journals, 2011) Mugo, S.N.; Mwimali, M.; Taracha, C.; Songa, J.M.; Gichuki, S.T.; Tende, R.; Karaya, H.; Bergvinson, D.J.; Pellegrineschi, A.; Hoisington, D.A.Transgenic maize (Zea mays L), developed using modified genes from the bacterium Bacillus thuringiensis (Bt), controls stem borers without observable negative effects to humans, livestock or the environment, and is now sown on 134 million hectares globally. Bt maize could contribute to increasing maize production in Kenya. Nine public Bt maize events of cry1Ab and cry1Ba genes were tested in confined field trials site (CFTs) to assess the control of four major Kenyan stem borer species. Leaf damage rating, number of exit holes and tunnel length were scored in the field evaluations. Leaf area consumed and mortality rates among stem borers were scored in the leaf bioassays in a Biosafety Level II laboratory, located at the Kenya Agricultural Research Institute (KARI), National Agricultural Research Laboratories (NARL). Field evaluations showed that Bt maize controlled Chilo partellus with mean damage scores of 1.2 against 2.7 for the non-Bt CML216 control. Laboratory bioassays showed high control for Eldana saccharina and Sesamia calamistis, with mean larval mortality of 64 and 92%, respectively. However, substantial control was not observed for Busseola fusca. These results showed that Bt maize could control three of the four major stem borers in Kenya with mortality records of 52.7% for B. fusca, 62.3% for E. saccharina and 85.8% for S. calamistis. Additional Bt genes need to be sought and tested for effective stem borer control in all maize growing ecologies in Kenya.
Publication - Regeneración de maíces blancos subtropicales vía embriogénesis somática(Colegio de Postgraduados, 2007) Hernandez-Garcia, C.M.; Lopez-Peralta, C.; Buenrostro-Nava, M.T.; Cárdenas Soriano, E.; Pellegrineschi, A.Maize (Zea mays L.) is destined mainly for animal feed (yellow maizes) and in a smaller proportion for human consumption (white). The subtropical white maizes are important for humans. However, there are problems of regeneration, via somatic embryogenesis, that limit its genetic transformation. Therefore the objective of the present study was to evaluate somatic embryogenesis in nine lines of subtropical white maize. In the induction of somatic embryogenesis, 1, 2 and 5 mg L−1 of 2,4-D and Dicamba were evaluated. Lines 78, 395 and 444 produced 70.1 to 87.2% of embryogenic calluses, similar to the controls (67.2 to 74.7%). Lines 442 and 332 presented a mean induction rate of calluses of 48.4 to 60.6%, whereas 330, 202, 204 and 331 showed the lowest percentages (3.9 to 26.2%). Lines 330, 331 and the control 216×72 presented the highest percentages with Dicamba, whereas line 442 had its highest percentage with 2,4-D. The induction of the lines varies among doses. Line 395 regenerated 1.11 plants per callus, similar to the three controls (0.74-0.95 plants), while 442, 78 and 332 regenerated 0.67, 0.37 and 0.33. Rooted seedlings (92-99%) and acclimatization (95-100%) were obtained in lines 395, 442, 78 and 332. The number of fertile regenerated plants was 228, 187, 108 and 79 in lines 395, 442, 78 and 332. Four embryogenic lines of subtropical white maize (395, 442, 78 and 332) were identified with regeneration capacity of fertile plants in 17 weeks and with potential for being subjected to genetic transformation.
Publication - Laboratory protocols: CIMMYT applied genetic engineering laboratory(CIMMYT, 1999) Bohorova, N.; Fennell, S.; McLean, S.D.; Pellegrineschi, A.; Hoisington, D.A.This manual provides the theoretical and technical basis required for the successful establishment of maize and wheat somatic cell tissue cultures that are suitable for in vitro genetic manipulation through the use of biolistic-based and Agrobacterium-mediated gene transfer systems for maize and wheat. Achieving effective transfer integration, and expression of transgenes using appropriate gene constructs promotes understanding of the process of gene transfer and the nature of transgenic plants. In addition the manual introduces readers to the basics of plant genome analysis and its application to the analysis of both maize and wheat transgenic plants. Also covered in this manual are the isolation, digestion, electrophoresis, and transfer of DNA to blots; the non-radioactive hybridization and detection technology in use at CIMMYT, and the application of polymerase chain reaction (PCR) technology to the analysis of transgenic plants. To complete the coverage of the topic, information on phenotypic screening of transgenic plants in the biosafety greenhouses is also presented.
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