AL 09 - Cotton Transformation (Embrapa Cenargen)

Team Leader: Maria Fátima Grossi-de-Sa

Laboratory Activities with INCT PlantStress Biotech

Searching for vital insect-pest genes/molecules (Helicoverpa armigera and Spodoptera frugiperda) by large-scale sequencing of the transcriptome and validation of its expression in vitro.

Selection of new Cry molecules with high toxic activity against S. frugiperda and H. armigera.

Identification of target molecules in silico against pests through the design of specific chemical drugs to select new compounds with potential for the development of new nematicides and insecticides.

Selection of genes/molecules vital for gall nematodes (Meloidogyne spp.) by analysis of their genome.

Selection of potential genes involved in the resistance of contrasting genotypes (peanuts, soybean, rice, cotton, and coffee).

Integration of monocotyledonous transcriptome data (rice, corn, wheat, Musa, and Sorghum) submitted to water deficit, generated by mass sequencing (Illumina – HiSeq) in previous projects.

Integration of transcriptome data from drought-tolerant genotypes of Musa spp., Arachis spp., and cowpea submitted to water deficit combined with biotic stress (Meloidogyne spp. or Mycosphaerella) in bioassays.

Validation of the expression of key genes in the metabolic pathways of the plant response to combined stresses (biotic-biotic, biotic-biotic, abiotic-biotic) by qRT-PCR studies.

Search and validation of novel regulatory sequences (promoters) responsive to biotic and abiotic stresses, in crop plants using transient and stable transformation (soybeans, cotton, and maize).

Sequencing  and selection of small RNAs, mRNAs, and circular RNAs of plants (Arachis, Musa, soybean, pitangueira, cashew tree) submitted to biotic and/or abiotic stresses using the Illumina platform.

Validation of the function of plant genes potentially involved in nematode resistance mechanisms through molecules overexpression or gene silencing (RNAi) strategies.

Validation of the function of nematode genes potentially involved in the parasitism mechanisms through RNAi strategies in model systems.

Assessment and monitoring technology of the biotech assets prospected for intellectual protection.

Development of universal vectors containing all the genetic elements protected (patented) by the different institutions involved in the INCT project.

National and international protection of the use of genes and regulatory sequences prospected, via patents.

Developing of GM soybean, cotton, and maize plants through overexpression of molecules and/or gene silencing strategies for drought tolerance and nematode resistance.

Developing of GM soybean, cotton, and corn plants through overexpressing Bt toxin and dsRNAs sequences applied to H. armigera and S. frugiperda control.

Functional validation of multiple pyramided genes involved in multiple traits, including nematodes and insect-pests (S. frugiperda or H. armigera) resistance and drought tolerance in GM soybean and cotton plants.

Phenotyping (greenhouse and/or field simulation) GM maize, soybean, and cotton plants for drought tolerance and/or resistance to S. frugiperda, H. armigera and Meloidogyne spp.

Laboratory Description

The Plant-Pest Molecular Interaction Laboratory (LIMPP) is coordinated by Dr. Maria Fatima Grossi-de-Sa, in which research interests involve basic and applied sciences, focusing on the plant-pest molecular interactions (pathogens and insect-pests) to develop novel crop protection strategies, mainly for cotton and soybean. The LIMPP group’s research is recognized for their expertise in plant biotechnology using functional genomics, notably working on biotechnological aspects of the RNA interfering (RNAi) mechanism applied to insect-pests and phytonematodes. Other current research interests also encompass the exploration of new plant genetic transformation and genome editing technologies, novel regulatory sequences for genetic engineering of crop plants against insect-pests and phytonematodes controls and drought tolerance, and production of recombinant proteins.

Research Lines

• Search for novel genes/molecules and peptides to be applied in the control insect-pests of cotton and soybean plants.

• Search for the target genes/molecules of plant parasitic nematodes (Meloidogyne spp., Rotylenchus reniformis, Aphelenchoides spp.) to be applied in gene silencing approaches.

• Search for target genes/molecules of contrasting soybean genotypes (resistant/susceptible to Meloidogyne spp.) to be applied to the genetic engineering of crop plants using genome editing technologies.

• Search for gene/molecules and small RNAs in pest and drought resistant/tolerant plant genotypes that could be involved in the response to biotic and abiotic stresses (cross-stress).

• Validation of the new prospected biotech assets using overexpression of molecules or gene silencing approaches in plant models.

• Developing the genetic engineering of crop plants (cotton, soybean, sugarcane) using molecules/dsRNAs overexpression, gene silencing, and genome editing approaches for traits that include pest control, increase in biomass, and drought tolerance.

 Our Team

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Carolina Vianna Morgante

Graduated in Biological Sciences from the USP Biosciences Institute (1999), master's and doctorate in Agronomy (Genetics and Plant Breeding) from the University of São Paulo (2003 and 2008, respectively). She is currently a researcher at Embrapa Semiárido. Has experience in Genetics, focusing on Plant Genetics and Molecular.

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Diana Isolda Clotilde Fernandez

She is currently a permanent Senior researcher at the French Institute for Research for Development - Institut de Recherche pour le Développement (IRD, France) and remains until November 2020 at Embrapa-Cenargen. She has experience in Biochemistry, with emphasis on Molecular Biology, acting on the following subjects: Phytopathology, plant-pathogen interactions, plant immunity, nematodes, rust, rice, Coffea arabica, Hemileia vastatrix, Meloidogyne spp..

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Isabela Tristan Lourenço Tessutti

Graduated in Biological Sciences from the University of Brasília and holds a Master's and PhD in Molecular Biology also from UnB. Currently, is an Analyst at Embrapa Genetic Resources and Biotechnology at the Plant-Pest Interaction Laboratory. She has experience in Molecular Biology, Biochemistry and Plant Genetic Engineering, applied to the themes: Plant-Pest molecular interaction; Plant resistance to biotic stress; Plant tolerance to abiotic stresses. The main tools/methodologies applied are: Functional genomics of phytonematodes, insects and plants; Identification, cloning and characterization of genes of phytonematodes, insects, plants and bacteria; Genetic transformation of plants; Gene silencing (RNAi) in phytonematodes and insects; Heterologous protein expression in bacteria; Functional characterization of plant promoters; Spatial and temporal determination of gene expression using real-time PCR.

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Leonardo Lima Pepino de Macedo

Graduated in Biological Sciences from the Federal University of Rio Grande do Norte (2005). The Master's course in Biochemistry was held at the Federal University of Rio Grande do Norte (2007). With a doctorate concluded by the Genomic Science and Biotechnology program at the Catholic University of Brasília (2012). Has experience in Biochemistry and Molecular Biology. Acting on the following topics: Cloning and expression of proteins in heterologous systems; Bioprospecting of proteins with entomotoxic activity (vicillins, lectins, proteinase inhibitors and Cry toxins) aiming at the control of dipterous, lepidopteran and coleopteran insects; and development of gene silencing strategies via interfering RNA (RNAi) for the control of insect pests.

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Maria Eugenia Lisei de Sa

Graduated in Biological Sciences from Faculdades Metodistas Integradas Isabela Hendrix (1981), Master in Agronomy (Phytotechnics) from Universidade Federal do Ceará (1984), PhD in Genetics and Biochemistry from Universidade Federal de Uberlândia (2004) and post-doctorate in Biotechnology Institute de Recherche pour le Développement-França (2013). Are you a researcher (II) at the Minas Gerais Agricultural Research Corporation (EPAMIG) and currently works as a collaborative researcher at Embrapa Genetic Resources and Biotechnology-Cenargen. He has experience in the field of soybean breeding with an emphasis on the development of soybean cultivars with characteristics suitable for human consumption. Develops projects in partnership with Cenargen in the areas of plant defense proteins (proteinase inhibitors, alpha-amylase inhibitors, lectins, defensins, osmotins), plant-pest molecular interaction, development of genetically modified plants for resistance to biotic stress (insects and nematodes) and tolerance to abiotic stress.

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Maria Fátima Grossi de Sa

Maria Fatima Grossi-de-Sa has a bachelor's degree in Biological Sciences - biomedicine modality from the University of Brasilia (1979), master's degree in Biological Sciences (Molecular Biology) from the University of Brasilia (1982), doctorate (Doctorat Et Sciences) in Molecular Biology from the Université Paris VII-France (1987), and postdoctoral fellow at the Plant Genetic System-Ghent-Belgium (1988) and at the University of California of San Diego (1995-1996). She is currently Lead Researcher at the Research Group at EMBRAPA Genetic Resources and Biotechnology (since 1989) and professor at the Catholic University of Brasília (since 2004). She has a CNPq productivity scholarship (level 1A), Member of the CAPES International Advisory Committee (since 2007), full member (Agrarian Sciences) of the Brazilian Academy of Sciences (elected in 2011) and a member of the World Academy of Science -TWAS (elected in 2014).She is president of the Brazilian Society of Biotechnology - SBBIOTEC (2008-until now). Among other awards, he received the Scopus 2010 Award (Elsevier/Capes) and served as Coordinator of the Biotechnology Area at CAPES and Substitute Member of CTC-ES (2007-2014). She has experience in the field of Molecular Biology, with emphasis on Plant Genetic Engineering and Plant Molecular Biology, acting on the themes: plant defense proteins (proteinase inhibitors, alpha inhibitors -amylases, lectins, defensins, osmotins), insecticidal proteins, molecular plant-pest interaction, development of GM plants for resistance to biotic stress and tolerance to abiotic stress and development of biopharmaceuticals.

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Maria Cristina Mattar da Silva

Graduated in Biological Sciences from Universidade Estadual Paulista Júlio de Mesquita Filho (1984) and from Universidade de Brasília (1987), master's degree in Biological Sciences (Molecular Biology) from Universidade de Brasília (1992) and doctorate in Biological Sciences (Molecular Biology) from University of Brasília (2002). Researcher at Embrapa Genetic Resources and Biotechnology since 1989. Specialist in plant molecular biology, he works in the field of plant biotechnology aiming to resist biotic and abiotic stress. Develops research focusing on the following themes: Evolution of molecules in vitro for selection of variants with improved activity, molecular studies of plant-pest interaction aimed at resistance to insects. Member of the Brazilian Society of Biotechnology.

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Wagner Alexandre Lucena

Graduated in Biological Sciences from the Federal University of Pernambuco (1994), master's degree in Genetics from the Federal University of Pernambuco (1998) and doctorate from UFRGS (2012). He is currently a researcher at the Brazilian Agricultural Research Corporation. Has experience in the field of Agronomy, with emphasis on Molecular Vegetal Biology, acting on the following topics: Prospection of genes of agricultural interest. Structural bioinformatics (DM and molecular modeling). Construction of synthetic cry genes. Transgenic cotton for insect pest control and fiber quality. Insect-pest transcriptome. Bacillus thuringiensis, Spodoptera frugiperda, Anthonomus grandis, Heliothis virescens, Pectinophora gossypiella and Alabama argillaceae.

Contact

Maria Fatima Grossi de Sá

EMBRAPA Genetic Resources and Biotechnology
W5 Norte Avenue (end) - P.O. Box 02372 - Postal Code 70770-917 - Brasília, DF - Brazil 
E-mail: fatima.grossi@embrapa.br 
Phone:+55 61 3448-4705 

Embrapa Genetic Resources and Biotechnology

Parque Estação Biológica - PqEB - W5 Norte Avenue (end)

Phone: +55 (61) 3448-4705

inctplantstress@gmail.com