Gene expression in wheat cv. Toropi in response to Puccinia triticina
Transcrição
Gene expression in wheat cv. Toropi in response to Puccinia triticina
Gene expression in wheat cv. Toropi in response to Puccinia triticina infection Alice Casassola, BSc, MSc Wheat Production Wheat Production Production (thousand tons): PR – 3.978,6 RS – 3.092,9 SC – 227,1 Brazil – 7.667,2 Production estimates CONAB 09/2014 Source: CONAB, 2014 Wheat Production Source: CONAB, 2014 Wheat Consumption Source: CONAB, 2014 Wheat Leaf Rust • Ideal conditions: • 15-20 ºC; • leaf wetness: 20 ºC – 3 hours 10 ºC – 10-12 hours • South region: • Sowing: May to July; • Harvest: September to December. Image: Paulo Roberto da Silva Climate Normals Humidity Mean temperature Source: INMET, 2014 • • Leaf rust is a main problem to the wheat production, causing yield losses of up to 80%. The southern region of Brazil has suitable environment to the disease development. • P. triticina has a high rate of physiological specialization and rapidly overcome resistance genes. Therefore, sources of durable APR have considerable value for Brazilian wheat breeding. • Chaves & Barcellos (2006) identified thirty-eight virulence combinations of the fungus from 5 states of Brazil, grouped in 12 different races. Until 2007, 60 races of the fungus were identified in Brazil (Barcellos & Chaves, 2003). • Arduim et al. (2012) observed the following frequency of races: (B55) < • RS - MDP-MR (B58) = MCP < MCG-MN (B34) < MDT-MR MFH-HT < MDK-MR (B55 4002S). • PR - MFP-CT (B56) < TFK-CS (B49) < TFT-MR (B57) < MDP-MR < MDT-MR < MDK-MR (B55 4002S). Wheat Culture wheat cv. Toropi • Twentieth century: Italian wheat cultivars were introduced in Brazil – Ardito and Mentana – not adapted to the Brazilian environment; • cv. Frontana: 1940 – Fronteira/Mentana: • short cycle; • leaf rust resistance (Lr13 and Lr34); • yellow rust resistance; • intermediate resistance to FHB; • short stature. Wheat Culture wheat cv. Toropi Sousa, 2001 Wheat Culture wheat cv. Toropi Fronteira Mentana Frontana 1971-37 Quaderna A 1940 Toropi 1965 Petiblanco 8 Wheat Culture wheat cv. Toropi • Toropi posses: • increased phosphorous absorption, translocation and distribution; • tolerance to aluminum toxicity; • resistance to FHB; • adult plant resistance to leaf rust – no Lr34. Image: Alice Casassola Timeline • Amarilis Labes Barcellos – PhD – Genética da resistência de planta adulta à ferrugem da folha na cultivar brasileira de trigo Toropi (Triticum aestivum L. em. Thell) – 1994 – UFRGS; • Sandra Patussi Brammer – PhD – Mapeamento de genes de resistência parcial à ferrugem da folha em cultivares brasileiras de trigo (Triticum aestivum L. em Thell) – 2000 – UFRGS; • Paulo Roberto da Silva – Dissertation – Identificação e conversão de marcadores moleculares associados à resistência à ferrugem da folha em trigo – 2001 – UFRGS; Timeline • Paulo Roberto da Silva – PhD – Identificação de marcadores e caracterização de mecanismos moleculares associados à resistência a ferrugem da folha em trigo – 2006 – UFRGS; • Caroline Wesp Guterres – PhD – Histopatologia das reações de resistência de hospedeiro e de não hospedeiro em interações Puccinia x Gramineae – 2011 – UFRGS; • Silvia Barcellos Rosa – PhD – The genetics of leaf rust resistance in the durably resistant wheat cultivar ‘Toropi’ – 2013 – University of Manitoba; • Paula Wietholter – Post-doc – Identificação e caracterização de genes diferencialmente expressos em trigo em resposta à infecção por Puccinia triticina – 2009-2013 – Embrapa Wheat. Timeline • Gerarda Beatriz Pinto da Silva – PhD – Microfenótipos e proteínas associadas à expressão da resistência durável à ferrugem da folha do trigo – 2013 (in progress) – UFRGS; • Alice Casassola – PhD – Gene expression in wheat in response to Puccinia triticina – 2012 (in progress) – UPF; • Andréia Caverzan – Post-doc – Estudo do transcriptoma de trigo durante o processo de infecção com Puccinia triticina, agente causal da ferrugem da folha – 2013 (in progress) – Embrapa Wheat. Timeline Barcellos, A.L.; Roelfs, A.P.; Moraes-Fernandes, M.I.B. 2000. Inheritance of adult plant leaf rust resistance in the Brazilian wheat cultivar Toropi. Plant Dis, 84(1): 90-93. Timeline • Barcellos et al. (2000) • • • • • adult plant leaf-rust resistance reaction of F2, F1 and parental (Toropi x IAC13) was determined; F2 indicated two recessive genes in cv. Toropi for APR to race LCG-RS; Its resistance is independent of Lr12, Lr13 and Lr22b; F2 Toropi/ThatcherLr34 indicated two recessive genes plus one dominant gene; Only one of the recessive resistance genes is responsible for the reduction in the pustule size. Timeline • Zoldan et al. (2002) • Toropi has no seedling leaf-rust resistance genes. Timeline • Da-Silva PhD (2006) • Subtractive suppression hybridization (SSH) 1hpi – 3 lines resistant and 3 lines susceptible – produced 800 clones and 200 were sequenced; • 69% of the 200 has known function: protein synthesis and processing, energy production; amino acid metabolism, signal transduction and transport, transcriptional regulation, cytoskeleton and resistance proteins; • 25 sequences were analyzed via qPCR and in silico monossomic lines: none were located in 1A and 4D chromosomes. Timeline • Rosa et al. (2011) – WIT (Women in Triticum) Award Year 2011: • • • • • • University of Manitoba, Winnipeg - Agriculture and Agri-Food Canada; Toropi had good leaf rust resistance in the range of 0 to 10MS during three years in Canada and 0 in New Zealand; Segregation data indicated the presence of one seedling resistance gene, a dominant racespecific adult plant resistance and two complementary race non-specific APR genes in Toropi; The seedling and race-specific APR genes do not confer good resistance in the field in Canada, but seem to be effective in New Zealand; Resistance conferred by the complementary race non-specific APR was effective in both locations; Toropi showed resistance to stem rust and stripe rust. Timeline • Da-Silva et al. (2012) • • • • • Toropi has two recessive genes with additive effects; trp1 – 1AS chromossome; trp2 – 4DS chromossome; XPPcgMac3 (3 %) and XPPcgMac6 (5 %). SP Brammer PhD thesis Agriculture and Agri-Food Canada/Cereal Research Centre – Winnipeg; XPPcgMac3 was converted to SCAR (“sequence characterized amplified regions”) – assisted selection. Timeline • Wesp-Guterres et al. (2013) • Toropi shows a pre-haustorial resistance phenotype; • It restricts the formation of the primary infection structures; stomatal appressoria and sub-stomatal vesicles. Timeline Toropi always showing lower established infection events than BRS 194. WespGuterres et al. (2013) Timeline • Silva PhD thesis (2013) • Among the 95 segregating lines from Toropi/IAC13, they observed that some progenies are more resistant than Toropi. Images: Gerarda Beatriz Pinto da Silva Timeline • Caverzan Post-Doc (2013) • Will evaluate the gene expression of the more resistant and more susceptible segregating line from Toropi/IAC13 via RNA-Seq in response to P. triticina. Gene Expression in wheat in response of Puccinia triticina infection • University of Passo Fundo; Faculty of Agronomy and Veterinary Medicine; 2012-2015; • Sandwich PhD: NIAB; • Scholarship: CAPES. • General Objective: • To identify, characterize and validate differential expressed sequences involved in the adult plant resistance against leaf rust of the cultivar Toropi. • 2012-2015. • • Gene Expression in wheat in response of Puccinia triticina infection • Team: • Alice Casassola, BSc, MSc; • Sandra Patussi Brammer, PhD, Supervisor, Embrapa Wheat; • Márcia Soares Chaves, PhD, Co-supervisor, Embrapa Wheat; • Lesley Boyd, PhD, Co-Suvervisor, NIAB; • Collaborators: José Antônio Martinelli (UFRGS), Magali Ferrari Grando (UPF), Francesca Stefanato (NIAB), Andréia Caverzan, Antonio Nhani Junior, Alceu José Vicari and Marcos Roberto Copetti (Embrapa Wheat), Paula Regina Kuser Falcão and Adhemar Zerlotini (Embrapa CNPTIA). Gene Expression in wheat in response of Puccinia triticina infection • • Toropi inoculated (MDT-MR) x mock: 0, 6, 12 and 24 hpi (three biological replicates) – Embrapa Wheat (2012/2013); Sandwich PhD – RNA-Seq 58 million bp per library – TGAC/NIAB (July 2013 – January 2014); • Comparison to the available Chinese Spring (CS42) transcriptome (Bowtie and Blast): 58-59% were common – Embrapa Wheat/CNPTIA; • Reference transcriptome was built via de novo assembly – Embrapa Wheat/CNPTIA; • Differential expression were performed (RSEM and EdgeR) – Embrapa Wheat/CNPTIA. Gene Expression in wheat in response of Puccinia triticina infection • 10,181 DE contigs: • 9,156 were common to CS42; • 1,025 were unique to Toropi and expressed only in the inoculated libraries. Conclusions We believe that these results will be of interest to the broader plant-pathogen research community, while providing knowledge to guide further functional studies on the adult plant resistance to leaf rust on wheat. Thanks We thank Embrapa, CNPq, FAPERGS, NIAB and BBRSC for their help and funding. Especially to CAPES for the scholarship granted. References • • • • • Arduim, G.S.; Reis, E.M.; Barcellos, A.L.; Turra, C. 2012. In vivo sensitivity reduction of Puccinia triticina races, causal agent of wheat leaf rust, to DMI and QoI fungicides. Summa Phytopathol, 38(4): 306-311. Barcellos, A.L.; Roelfs, A.P.; Moraes-Fernandes, M.I.B. 2000. Inheritance of adult plant leaf rust resistance in the Brazilian wheat cultivar Toropi. Plant Dis, 84(1): 90-93. Barcellos, A.L.; Chaves, M.S. Epidemias -de ferrugem da folha em cultivares brasileiras de trigo - alterações na população do patógeno de 1993 a 2002. In: SEMINARlO INTERNACIONAL RESITENCIA A ROY AS EM TRlGO, 3., 2003, La Estanzuela.Resumenes ... La Estanzuela: INIA,2003. p. 13. Chaves, M.S.; Barcellos, A.L. Especialização Fisiológica de Puccinia triticina no Brasil em 2002. Fitopatol Bras, 31(1): 57-62. Da-Silva, P.R.; Brammer, S.P.; Guerra, D.; Milach, S.C.K.; Barcellos, A.L.; Baggio, M.I. 2012. Monosomic and molecular mapping of adult plant leaf rust resistance genes in the Brazilian wheat cultivar Toropi. Gen Mol Res 11(3): 2823-2834. References • • • • Rosa, S.B.; McCallum, B.; Brule-Babel, A. Quantitative resistance conferring durable leaf rust resistance in wheat cultivar Toropi. In: BRGI Technical Workshop, 2011, Minnesota. Poster Abstracts: BRGI, 2011. p.184. Sousa, C.N.A. de. Contribuição das cultivares de trigo de Strampelli para o melhoramento de trigo no Brasil/Cantídio Nicolau Alves de Sousa - Passo Fundo: Embrapa Trigo, 2001. 5 p. html, 2 tab. (Embrapa Trigo. Documentos Online; 7). Disponível: http://www.cnpt.embrapa.br/biblio/p_do07.htm Wesp-Guterres, C.; Martinelli, J.A.; Graichen, F.A.S.; Chaves, M.S. 2013. Histopathology of durable adult plant resistance to leaf rust in the Brazilian wheat variety Toropi. Eur J Plant Pathol, 137(1): 181-196. Zoldan, S.M.; Barcellos, A.L. Postulation of genes (Lr) for resistance to leaf rust in Brazilian wheat cultivars. Fitopatol Bras, 27(5): 508-516. Thanks for your attention! [email protected]