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
•
•
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•
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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
•
•
•
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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!
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