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Summary of Puccinia Striiformis Race Analysis 2016 AARHUS AU UNIVERSITY Report for Puccinia striiformis race analyses and molecular genotyping 2016, Global Rust Reference Center (GRRC), Aarhus University, Flakkebjerg, DK- 4200 Slagelse, Denmark. Mogens Støvring Hovmøller, Julian Rodriguez-Algaba, Tine Thach, Annemarie Fejer Justesen & Jens Grønbech Hansen, 2 February, 2017. Email: [email protected] Key highlights • Two highly virulent races associated with rust epidemics in Europe and East Africa/Central Asia, respectively, were detected for the first time across large areas: o A new race detected in Italy and Morocco was genetically closely related (but not identical) to isolates of the same race in northern Europe. o A new race in Ethiopia and Uzbekistan, previously detected in Afghanistan, 2012 and 2013, was assigned a unique genetic lineage. • Warrior(-) was the most prevalent race in Europe, now also present in several countries in eastern Europe and western Asia, along with the Warrior race. • A new race variant of Warrior(-) shared virulence phenotype with isolates of the original Warrior race and could only be separated by genotyping. • SSR genotyping was fully implemented as part of the race surveys. Genotyping showed high correlation with race phenotyping, in addition to providing results for samples which could not be recovered. • Summary of race phenotyping results from GRRC (2008-2016) are available online (http://www.wheatrust.org/) The race typing of non-European isolates are based on agreements between Aarhus University, CIM- MYT, ICARDA, Cornell University and FAO, to facilitate race analyses of P. striiformis isolates infect- ing wheat and other cereals, mainly from Africa and Asia. The analyses of European isolates were supported by RAMP, Ministry of Environment and Food of Denmark (2013-2016), Jordbruksverket, Sweden (2008-2017) and RUSTFIGHT, Danish Strategic Research Council (2012-2016). A summary of the results can be shared within relevant countries and organizations providing that the authors of this report are acknowledged along with funding sources, i.e. “Global Rust Reference Center: Research funded by: Aarhus University, Denmark; Ministry of Environment and Food of Denmark; CIMMYT; ICARDA; FAO”. Results from previous years are available as pdf files from the GRRC home page, and results are available on maps and charts. The site is being continuously updated by improved analytical tools, and country representatives can upon request get access to and download detailed results nationally. Report of yellow rust races 2016: Global Rust Reference Center, Aarhus University, Denmark www.wheatrust.org 2 We have assigned common names based on the genetic lineage of significant races demonstrating epidemic potential. Important lineages have been named Pst followed by a digit. Race variants were designated by the additional virulence observed or (-) in case a new variant had fewer virulences than the first defined race within the considered lineage. Race names already adopted by the farming com- munity in Europe were maintained, e.g., Warrior and Kranich, which are named according the wheat variety where they caused the first confirmed epidemic outbreaks, and Triticale2015 causing severe epidemics in 2015 in multiple triticale varieties. A more comprehensive justification and rationale for the naming of significantP. striiformis races and genetic groups is in progress. The new mapping tools allow the user to highlight particular countries, years and/or races, which can be shown by geograph- ical coordinates, if this information has been provided. Submission and preparation of samples Prior to submission of rust infected leaf samples, a request must be sent by e-mail to GRRC to obtain an import permit issued. This permit must be enclosed with any sample submission. Information about details of collector (person), host variety, sampling date, location, and disease severity of the field plot from where samples were taken, should be provided. The details of sampling preparation are given at http://wheatrust.org/submission-of-isolates/. On this page you will also find a YouTube video, where Dr Ali, University of Peshawar, Pakistan, demonstrates the ideal sampling procedures at a Borlaug Global Rust Initiative training workshop, Nepal, 2015. Focus sampling areas in 2017 outside Europe will be selected in collaboration with staff at ICARDA, CIMMYT, NARCs in Africa and Asia, and FAO, with a focus on high risk epidemic areas. Since 2011, GRRC also accepted samples of stem rust (P. graminis tritici) as agreed upon within BGRI projects. A total of 174 samples of yellow rust Table 1. Number of samples of rust infected wheat handled in 2016, Africa and Asia infected leaves from 9 countries in Af- Inoculated Not Race typing Additional rica and Asia were submitted and en- Country Failed (in progres) inoculated completed recovered Total Afghanistan 1 1 8 4 14 tered for recovery using susceptible Azerbaijan 6 2 8 seedlings of Cartago, Morocco and Eritrea 7 1 8 Anja (Table 1). A total of 84 isolates Ethiopia 10 1 6 7 19 43 Iraq 23 1 24 from these regions were recovered Morocco 9 6 15 and multiplied. The recovery rates var- Pakistan 14 1 15 ied greatly from case to case empha- Tanzania 1 3 1 5 Uzbekistan 4 11 13 14 42 sizing the importance of appropriate Total 2016 68 1 21 43 41 174 sample handling and preservation and submission without delay. Similarly, 275 samples were submitted from 12 European countries (Table 2). A total of 193 isolates were recovered from these samples. Multiple factors may influence viability Table 2. Number of samples of rust infected wheat handled in 2016, Europe and recovery rates, e.g., 1) emerging Not Race typing Additional crop senescence at time of sampling, Country Failed inoculated completed recovered Total 2) delayed time between sample col- Croatia 4 1 6 11 Denmark 23 67 24 114 lection and arrival at GRRC and 3) Finland 7 1 2 10 non-favorable condition after sam- Hungary 3 3 6 pling, i.e., during preparation and post- Italy 12 5 17 Latvia 4 1 2 7 age. Certain couriers may use radia- Lithuania 1 1 tion during the handling of parcels, Norway 5 13 13 31 which may result in poor recovery or Spain 11 5 16 Sweden 16 11 21 48 failure. Additional cycles of multipli- UK 1 1 2 cation were often needed to obtain Ukraine 8 4 12 sufficient amount of spores for storage Total 2016 70 12 117 76 275 and race analyses, which were con- Report of yellow rust races 2016: Global Rust Reference Center, Aarhus University, Denmark www.wheatrust.org 3 ducted according to Thach et al. (2015) and Sørensen et al. (2016). The procedure of genotyping of isolates based on DNA extraction from infected leaves (single lesions) was generally successful, and proved highly useful for generating results in case of poor recovery and for confirming genetic purity and assignment of races to specific genetic lineages. 2016 results A subset of recovered isolates were multiplied during 2-3 cycles to produce spores for race typing on wheat differential lines carrying resistance genes to P. striiformis. A combination of lines from ‘World’ and ‘European’ differential sets and NILs in an Avocet background gave a high resolution in terms of virulence determination despite that additional previously unreported resistance genes were detected in a number of differential lines including some of the Avocet NILs. For commonly used resistance genes like Yr1, Yr2, Yr6, Yr7, Yr8, Yr9, Yr17, Yr25, Yr27, Yr32 and Yr(Sp), respectively, two differ- ential lines were included to confirm new races. DNA was extracted from recovered isolates or from selected incoming samples of infected leaves in case recovery was unsuccessful. The race typing re- sults in Table 3 were confirmed by SSR genotyping, allowing unique differentiation of genetic lineage of the different races (data not shown). Two important new races having unique molecular genotypes were detected across wide areas in Europe/North Africa and East Africa/Central Asia, respectively (Tables 3 and 4). One of the races, temporarily designation “Pst(new)” was detected in Morocco, Italy and northern Europe, where sam- pling was done systematically in 2016. The race was most prevalent in Morocco, a non-epidemic situ- ation during the drought in winter and spring in that area, and on Sicily in a severe epidemic situation, where yellow rust until recent years was considered insignificant. Isolates of this new race in Sicily and Morocco had a 100% genotype match. The same race was detected at low frequency in northern Europe and all these isolates were diverging by a single marker from the genotype detected in Sicily/ Morocco. Another race was detected for the first time in East Africa in the autumn of 2016, after be- ing first detected in Afghanistan in 2012 and 2013 (temporarily designated “AF2012”). The race was prevalent in epidemics in Ethiopia, where a series of varieties became severely affected by yellow rust. All Isolates of the race shared molecular genotype, including isolates collected in Afghanistan and Uzbekistan in 2016, where yellow rust epidemics were widespread. The PstS5 lineage with a complex virulence profile was prevalent in Central Asia, including Uzbeki- stan, which is in accordance with results from previous years, and the aggressive PstS2 with virulence to Yr27 was again detected across wide areas (Walter et al., 2016). The PstS6 race, which has previ- ously been prevalent in East Africa, was in 2016 also detected in Afghanistan. In Europe, Warrior(-) was by far the most prevalent race, now being detected in several countries in East Europe and West Asia along with the original Warrior race (Tables 3 and 4) (Hovmøller et al., 2016) A new variant of Warrior(-) with additional virulence to variety ‘Ambition’ was detected in several countries, i.e., regular race typing cannot distinguish this new variant from isolates of the original War- rior race, emphasizing the advantage of integrating SSR genotyping and race typing activities.
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