Workshop on Biotic and Abiotic Stress Tolerance in Plants
:
the Challenge for the 21st Century
Cana Brava Resort Ilhéus-Bahia, Brazil 6th-8th November 2013
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S01P09RNAseq analysis of different Hevea genus inoculated by Microcyclus ulei
L.R. Salgadoa, D.M. Koopb, M. Magalhãesc, A.T.R. de Vasconcelosc, W.A Silva Jra, D. Garciad.
a- Departamento de Genética/FMRP/USP. Laboratório de Genética Molecular e Bioinformática. Rua
Tenente Catão Roxo, 2501, CEP 14.051- 140, Ribeirão Preto, São Paulo, Brazil. b- Universidade Estadual
de Santa Cruz, Laboratório de Genômica e Biologia Molecular, Rod. Ilhéus-Itabuna BR 415, Km 16, Ilhéus, Bahia CEP 45.662-000, Brazil. c- Laboratório Nacional de Computação Científica, av. Getulio Vargas, n°
333 Quitandinha, CEP 25.651-075, Petrópolis, Rio de Janeiro, Brazil. d- CIRAD, BIOS, UMR-AGAP, TA
A96/03, Av. Agropolis, 34.398 Montpellier, France. Email :lrippel@usp.br
The rubber tree, Hevea is native from the Brazilian Amazon region and it is responsible for almost all natural rubber produced in the world, a strategic raw material for a variety of products. One of major challenges for rubber tree cultivation is the adaptation to biotic stress caused by several leaf pathogens. Microcyclus ulei, causal agent of the South American leaf blight (SALB) is responsible of massive losses of leaves by infection, leading to a low latex production and sometimes leading to plant death. The main proposed strategies to avoid the M. ulei damaging on plantations are to cultivate productive and SALB resistant genotypes. In Hevea, quantitative and qualitative resistance to M. ulei can be found in cultivars and but also between genus. By transcriptomic comparison of RNAseq of three genus of Hevea (H. brasiliensis cv. MDF180 and cv. PB314, H. pauciflora cv. PA31 and H. benthamiana cv. F4542), our purpose was to identify genes differentially expressed and characteristic of each genus.
Hevea leaf lesions and non-inoculated controls were collected 24, 48, 96 and168 hpi. For each cultivar
inoculated or non-inoculated, total RNA of time samples were pooled and enriched in mRNA. The eight cDNA libraries were sequenced with 454 XL+ sequencing kit and produced a total 4,225,291 reads de
novo assembled on 28,349 contigs. A tBLASTx against a NCBI plant RefSeq was performed and 15,442
sequences were successfully annotated. A differential expression analysis was conducted to each library with the NOIseq 2.0 software (Bioconductor) and for a cutoff of q score of 0.9, we identified in F4542 cultivar (quantitative resistance) 89 up-regulated and 32 down-regulated genes, in MDF180 cultivar (quantitative resistance), 117 up- and 71 down-regulated genes, in PA31cultivar (quantitative resistance), 72 up- and 12 down-regulated genes and in PB314 cultivar (susceptible) 455 up-regulated and 169 down-regulated genes. This preliminary analysis provides a first look at the host and parasite transcriptome of Hevea spp. Further analyses are underway to distinguish the host genes from those of the parasite and identify with more accuracy the genes differentally expressed in each Hevea genus during the infection.
Work supported by CAPES/CNPq S01P10
Protein profile in 2D-SDS-PAGE of sweet orange infected with the Citrus tristeza virus (CTV)
M.S. Dória1; C.J. Barbosa2, A.S. Gesteira,2 A.O. Sousa1 and C.P. Pirovani1
1Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz – UESC, Ilhéus, Bahia, Brasil.
2Embrapa Mandioca e Fruticultura – Cruz das Almas Bahia, Brasil.
Email : miledoria01@gmail.com
The citriculture is important worldwide mainly because the citrus fruits are among those which are the most consumed in natura but also used in several industrial processes. Despite its first position as main exporter of concentrated and frozen orange juice in the world, the Brazil suffers of production losses due to several microbial diseases. Among them, the Citrus Tristeza caused by Citrus tristeza virus (CTV), a phloem-limited virus, affects the fruit yield in the São Paulo and Bahia States, the two main Brazilian production areas. The interaction plant-CTV is complex and depends of the virus variants – which induce a large scale of symptom severity – as well as of the scion/rootstock combination used in the producing area. To assess the plant molecular changes occurring through the Citrus Tristeza disease, a proteomic analysis using 2-D electrophoresis was performed in two varieties of sweet orange, "Westin" and "Pera C21", infected (or not - control) by the CTV. Bark stem containing, among other, the phloem tissue was harvested on each variety (infected or not) and submitted to protein extraction using the ADP method followed by a phenol/SDS-dense extraction. About 500 mg of proteins were separated on 13 cm strips (pH 3-10NL) and then on SDS-PAGE before staining with Coomassie colloidal blue. 2-D gel images were analyzed with the Image Master 2D Platinum 7.0 software. In the "Westin" variety, 618 and 604 spots were detected in infected and non infected samples, respectively;
