All BGRI Abstracts

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Monitoring the South African leaf rust population through a combined phenotyping and SSR genotyping approach

Selinga Department of Plant Sciences, University of the Free State, South Africa
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Leaf rust is a common wheat disease in South Africa. Annual surveys conducted by the Agricultural Research Council - Small Grain Institute (ARC-SGI) during the last 35 years used infection type (IT) data on a defined differential set to identify individual field isolates. Results from these surveys confirmed that the South African Pt population is affected by both local evolution and foreign introductions. A good correlation was found between avirulence/virulence phenotypes and simple sequence repeat (SSR) genotypes in the South African Pt population. We therefore evaluated whether identification of field isolates by SSR analysis would complement the traditional IT analysis using 47 field isolates collected during the 2013 growing season. Of the 39 phenotyped isolates, 35 were correctly genotyped while three were incorrectly genotyped only because the corresponding race was not included as a control. Five isolates that could not be phenotyped due to non-viable spores were successfully genotyped. The dominant race 3SA145 (North American race annotation CCPS) was represented by nine different genotypes sharing 82% genetic similarity. The SSR data further showed that the field isolates formed part of two distinct lineages with little admixture between them. This study confirmed the supporting value of SSR genotyping to traditional race analysis in monitoring the South African Pt population. 



Wheat cv. Kingbird is introduced to address a new stem rust threat in Ethiopia

Tadesse Kulumsa Agricutural Research Center, Ethiopian Institute of Agricultural Research, Ethiopia

Pgt race TKTTF, virulent for the SrTmp gene present in Ethiopian cv. Digalu and first detected in 2012, caused significant yield losses in Digalu during the 2013 and 2014 seasons. No suitable replacement varieties with significant seed volume were available, and alternate solutions were sought. EIAR, with the support of the DRRW project through CIMMYT-Ethiopia, introduced 5 tonnes of adult plant, rust resistant wheat cv. Kingbird from Kenya. Kingbird was evaluated for agronomic performance at seven locations vs. three checks in 2014, and was also evaluated for stem rust reaction in single-race nurseries (TKTTF, TTKSK, TRTTF and JRCQC). With support from USAID/CIMMYT, seed was concurrently multiplied on 37 ha producing 80 tonnes of seed that was distributed to farmers in 2015. Mean grain yield over locations was 2.76 t ha-1. Mean performance of Kingbird was 3.00 t ha-1 compared to 2.79 t ha-1 for Ogolcho, 2.83 t ha-1 for Biqa and 2.42 t ha-1 for Kakaba. Thus Kingbird gave yield advantages of 5 to 22% over the check varieties. Stem rust severities on Kingbird in the single race nurseries ranged from Tr to 15% and reactions ranged from TMR to SMS. The check varieties rated up to 45% severity with S type reactions. Thus Kingbird was superior in terms of yield potential and stem rust resistance as measured in these trials vs. the check varieties. Stem rust resistance of Kingbird is based on Sr2 and Sr57 and is hypothesized to have at least three additional APR loci. Seedling reactions of Kingbird to races TKTTF and Ug99 are characterized as susceptible. Sr57 is pleiotropic and confers partial resistance to all three rusts, powdery mildew, spot blotch, and BYDV. Based on early maturity, yield performance, and stem rust resistance, Kingbird is recommended for low- to mid-altitude wheat-growing areas of Ethiopia.


Characterization of seedling and adult plant resistance to leaf rust in African wheat germplasm

Kankwatsa The University of Sydney, Plant Breeding Institute, Australia

Many of the catalogued leaf rust resistance genes in wheat deployed in agriculture have been overcome by variants of Puccinia triticina (Pt), the causal pathogen of leaf rust. Discovery and characterization of new sources of resistance in various germplasms using multipathotype tests and molecular markers could permit future diversification of the genetic base of leaf rust resistance in wheat. In searching for new sources of leaf rust resistance, 140 wheat lines from 14 African countries were tested with 8 Australian Pt pathotypes. Seedling tests revealed that 41% of the lines were susceptible to all pathotypes, 31% were postulated to carry either one of 10 resistance genes (Lr1, Lr2a, Lr3a, Lr13, Lr18, Lr23, Lr24, Lr26, Lr37 or Lr73) or one of five gene combinations (Lr2a+Lr3a, Lr1+Lr13, Lr1+Lr23, Lr1+Lr13+Lr73 and Lr23+Lr73). Twenty-eight percent of the lines were postulated to carry uncharacterized seedling resistance genes. Based on average coefficients of infection (ACI), 101, 25 and 11 lines showed high (ACI 0-19), moderate (ACI 21-38) and low (ACI 41-56) levels adult plant resistance, respectively, whereas three lines were moderately susceptible to susceptible (ACI 63-76). Genotyping of 74-78 lines that were anticipated to carry APR genes, using the molecular markers: csLV34 (linked to Lr34) and KASP SNP markers SNP1G22 and SNPT10 (linked to Lr46 and Lr67), respectively, revealed the presence of Lr34, Lr46 and Lr67 in 11, 22 and 14 wheat lines, respectively. The identities of the APR in the remaining 22 lines are unknown, and potentially represent new resistance sources. Genetic analyses of these uncharacterized APR sources are underway to select single gene lines and allow fine mapping.


Brachypodium distachyon as a model to study nonhost resistance to wheat stripe rust

Gilbert CSIRO Plant Industry, Australia

The model grass Brachypodium distachyon has been used to study nonhost resistance mechanisms to the wheat stripe rust pathogen, Puccinia striiformis f. sp tritici. Numerous B. distachyon accessions were screened with an array of UK and Australian P. striiformis isolates and distinct infection phenotypes identified, ranging from complete resistance to partial susceptibility. Three mapping families were established - BdTR10H x TEK4, BdTR13K x Bd21 and ABR6 x Bd21 - and immunity was dominantly inherited when they were tested with one Australian and three UK isolates. Depending upon the mapping family, between one and three genes for stripe rust resistance were present and designated Yrr1 to Yrr3. Yrr1, which is present in all three families, was effective against all isolates and was fine mapped to a 100 kilobase region containing six candidate genes. Interestingly, no candidate was homologous to a known resistance gene. Yrr2, which is present in the BdTR13K x Bd21 and ABR6 x Bd21 families, is race-specific and was mapped to a 1 megabase region that contains multiple, classic NBS-LRR resistance gene candidates. Yrr3, which is present in the ABR6 x Bd21 family and effective against all isolates, was mapped to a 400 kilobase region also containing NBS-LRR gene candidates. Agrobacterium-mediated transformation of Yrr1 candidates is underway in Brachypodium for complementation, and in common wheat to test for interspecies transfer of characterized resistance.


Identification of naturalized and cultivated Berberis species in South Africa

Keet University of Stellenbosch, South Africa
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While Africa is home to three Berberis species (B. holstii, B. hispanica and B. vulgaris), genera of the family Berberidaceae do not occur naturally in South Africa. However, due to the trade in ornamental plants, a total of 11 Berberis species, 11 cultivars and 8 hybrids were historically and/or are currently cultivated in the country. The current invasive status of most of these species is unknown, but two naturalized Berberis populations were recently discovered. B. julianae was found in the Golden Gate Highlands National Park in eastern Free State province, and B. aristata was found in the Woodbush Forest Reserve in Limpopo province. Since several Berberis species could act as alternate hosts for Puccinia graminis and P. striiformis, a phylogenetic study was conducted to identify both naturalized species, as well as several cultivated specimens. One of the cultivated specimens was identified as B. vulgaris, a species well known for its susceptibility to P. graminis. Knowledge gained from this study will be used to intensify the search for more naturalized Berberis populations, as well as to assess the potential threat to wheat cultivation in the country.


Crosstalk between CBL-CIPK and SA signaling pathways in wheat-Puccinia striiformis f. sp. tritici pathosystem

Guo State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, PR China

Intracellular calcium changes during plant–pathogen interaction are essential early events leading to both local and systemic acquired resistances. Salicylic acid, a critical messenger, is also required for both responses. However, the relationship between the CBL-CIPK and SA signaling pathways during wheat–Pst interaction is unclear. In this study, we isolated seven wheat CBL and 11 wheat CIPK genes and designated them as TaCBL1, 2, 3, 4, 6, 7, 9 and TaCIPK2, 5, 7, 9, 10, 14, 15, 17, 23, 31, 32. Some wheat CBLs and CIPKs were functionally characterized. Concurrently, wheat TaNPR1 as a master regulator of SA-mediated host response during Pst infection was functionally characterized. Silencing of TaCBL4, TaCIPK10 and TaNPR1 permitted increased rust development in a wheat variety that was resistant to Pst pathotype CYR23. Decreased levels of salicylic acid (SA) were observed in TaCBL4- and TaCIPK10-silenced wheat plants. Yeast two-hybrid and biomolecular fluorescence complementation (BiFC) revealed that TaCIPK10 interacted with both TaCBL4 and TaNPR1. These results suggest that a TaCBL4-TaCIPK10-TaNPR1 complex is involved in innate immunity of wheat to Pst.


Responses of some Turkish winter durum wheat genotypes in preliminary yield trials to stem, leaf and stripe rusts

Mert Central Research Institute for Field Crops, Turkey

Durum wheat is second important crop after bread wheat and it was grown as spring and winter type in Turkey. Rusts are the most important diseases limiting durum wheat production in Turkey. The aim of the study was determining of the resistance of the 232 Turkish winter durum wheat genotypes in preliminary yield trials developed by the Central Research Institute for Field Crops (CRIFC) to rusts.  For this purpose, adult plant and seedling test were conducted for yellow rust while only seedling test were conducted for leaf and stem rust. Evaluations were carried out at the research facilities of CRIFC at İkizce and Yenimahalle in Ankara in the 2014 season. For adult plant reactions; the genotypes were inoculated with local Pst populations (virulent on Yr2,6,7,8,9,25,27,Sd,Su,Avs). Stripe rust development on each entry were scored using the modified Cobb scale when the susceptible check cv. Little Club had reached 80S infection severity in June, 2014. Coefficients of infections were calculated and values below 20 were considered to be resistant. For seedling test; the seedling was inoculated with local Pgt (avirulent on Sr24, Sr26, Sr27, and Sr31), Pt (avirulent on Lr9, Lr19, Lr24, and Lr28) and Pst populations. Stripe, leaf and stem rust development on each entry were scored after 14 days with 0-4 and 0-9 scale for leaf-stem rust and yellow rust, respectively. In seedling stage, 141(65%), 41(18%), and 114 (49%) genotypes were resistant to local Pgt, Pt, and Pst populations, respectively. In adult plant test, 21 (9%) genotypes were resistant to Pst.  The resistance genotypes to stem, leaf, and stripe rust were determined with this research.


Achieving durable rust resistance in wheat through deployment of major and minor genes

Thapa Agriculture Botany Division, Nepal Agricultural Research Council (NARC), Nepal

Stripe rust and leaf rust have been major constraints to wheat production in Nepal since the 1960s. Several rust epidemics causing hardship for Nepalese wheat growers were due to race changes. Breeding for rust resistance was initiated with establishment of the National Wheat Research Program in 1972, but concerted searches for durable resistance came later with the introduction of wheat genetic resources from CIMMYT, Mexico. The early wheat varieties Nepal 297, Siddhartha, Vinayak, BL1473, BL 1022 and Annapurna series with leaf rust and stripe rust resistance genes Lr13, Lr23, Lr26 and Yr9, and Yr27 in the 1970s and 1980s succumbed to new races within a few years of release. However, Bhrikuti (CMT/COC75/3/PLO/FURY/ANA) with both major and minor gene combinations (Lr10, Lr14a, Lr26/Yr9/Sr31+ and Lr34/Yr18) and released in 1994 was unaffected by Yr9 virulence in 1997 and Yr27 virulence in 2004. This variety with >20 years of leaf rust and stripe rust protection continues to be the most popular wheat variety in Nepal. Three other varieties, Gautam (Siddhartha/Ning8319//Nepal 297) released in 2004, WK 1204 (SW89-3064/Star) released in 2007, and Pasang Lhamu (PGO/SERI) released in 1997 with Lr16, Lr26/Yr9/Sr31, Lr34/Yr18, Lr46/Yr29, Yr7, and Sr2 also remain resistant. The Ug99 resistant varieties Vijay (NL748/NL837), Danphe(KIRITATI//2*PBW65/2*SERI.1B) and Tilottama (Francolin#1 = Waxwing*2/Vivitsi) also possesses APR to the three rusts. Nepalese wheat researchers work closely with the CIMMYT Global Wheat Program and DRRW/BGRI to utilize knowledge and APR germplasm. Strong networks for participatory varietal selection involving women farmers in both the hills and terai help in faster adoption and in establishing varietal diversity. In summary, Nepalese wheat breeders have successfully used APR in protecting wheat crops.


Genetic characterization of slow rusting resistance to leaf rust in durum wheat cv. Bairds

Lan International Maize and Wheat Improvement Center (CIMMYT), México

The CIMMYT durum Bairds is susceptible to leaf rust (LR) at the seedling stage but shows an adequate level of slow rusting adult plant resistance (APR) in Mexican field environments. A recombinant inbred line (RIL) population developed from a cross of Bairds and the susceptible parent Atred#2 was phenotyped for LR response at Ciudad Obregon, Mexico, during 2013, 2014 and 2015 under artificial epidemics created with Pt race BBG/BP. Genetic analysis indicated that 3-4 additive genes conferred LR resistance. The RILs and parents were also genotyped with the 50K diversity arrays technology (DArT) sequence system and 93 SSR markers. A genetic map comprising 1,150 markers was used to map the resistance loci. Inclusive composite interval mapping analysis detected four quantitative trait loci (QTL) on chromosomes 1BL, 2BC (centromere region), 5BL and 6BL. These QTL, designated as QLr.cim-1BL, QLr.cim-2BC, QLr.cim-5BL and QLr.cim-6BL, explained 20.1-60.7%, 6.4-13.1%, 4.3-11.2%, and 7.1-28.0%, respectively, of the variation in leaf rust severity. QLr.cim-1BL was close to the previously reported APR gene Lr46, whereas QLr.cim-6BL, detected in all three seasons, is a new resistance locus in durum wheat. The four QTL combined showed a significant additive effect on resistance with a disease severity of 18-20%, whereas RILs carrying the individual QTL showed mean leaf rust severities ranging from 56 to 98%. Three QTL, except for QLr.cim-2BC, were derived from Bairds. The final LR severity of Bairds ranged from 15-25% across three years. This cultivar can be used as a source for complex APR in durum wheat breeding.


Diversity of resistance genes in candidate cultivars for planting in the overwintering area of central Shaanxi province, China

Han State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, P. R. China

China is the largest stripe rust epidemic area in the world. Central Shaanxi, as an important stripe rust overwintering zone for the disease serves as a “bridge” for the pathogen, where early sown wheat infected during the previous autumn provides inoculum for spring epidemics in more eastern regions. Studies of resistance and Yr-gene distribution among local candidate cultivars provide valuable insights into the influence of host genotype on selection of the rust pathogen population. A total of 183 local advanced lines from 2009 to 2011 were tested for seedling resistance with 12 Pst races in the greenhouse, and with mixed races at Tianshui in Gansu province. Gene postulations were based on the seedling response data and molecular markers. Four (2.2%) entries were resistant at all growth stages; 15 (8.2%) were resistant as adult plants; 164 (89.6 %) were susceptible to one or more races at the seedling or adult stages; and 40 were resistant to the currently prevalent races CYR32 and/or CYR33, but susceptible to at least one of the potentially important races Su11-4, Su11-5 and Su11-7, V26/CM42 and V26/Gui22. All entries showed seedling stage susceptibility at Tianshui. Postulated genes included Yr7, Yr9, Yr10, Yr17, Yr18, and Yr24/Yr26. Yr5, Yr15 and Yr61, currently effective against all Chinese races, were not present. Although advanced wheat lines bred in Shaanxi may be diverse our results show that most of them are highly susceptible to one or more prevalent or low frequency races in Shaanxi or adjacent Gansu. This situation indicates that Shaanxi farmers should be using partial adult plant resistances to reduce inoculum levels and hence reduce the amount of primary inoculum spread to more easterly wheat growing areas.