Wheat stripe rust, caused by basidiomycete fungus Puccinia striiformis f. sp. tritici (Pst), is a damaging disease worldwide. The recent discovery demonstrated the fungus depends on living wheat and aecial hosts, mainly barberry (Berberis) species, to complete its life cycle. In China, we determined that, under natural conditions, the sexual cycle of Pst occurs based on collections of Pst isolates from the diseased barberry in the past three years. However, no direct evidence to support whether barberry plays a role in spreading inoculums to wheat field to cause stripe rust was detected. In the present study, we recovered 103 Pst samples from natural-infected B. shensiana in the western Shaanxi in spring 2016, and also collected 107 Pst isolates from neighboring wheat fields. Phenotype and genotype of the two Pst populations were tested using a set of Chinese differential hosts for Pst and SSR markers, respectively. The phenotype tests showed that 57 race types produced from the barberry-derived Pst populations, consisting of 58 known races, such as CYR 34, CYR32, G22-14, and Su11-14-3, and 45 new races. Many of the two Pst populations shared the same race types. The genotype tests indicated the barberry-derived Pst population produced a rich genotype, obviously higher than the wheat-derived Pst populations. The seven same genotypes were found on 40 isolates of the former and 26 of the latter. Our results provide evidence to support that sexual cycle of Pst occurs regularly in nature in China and that barberry provides inoculums to neighboring wheat fields, triggering stripe rust infections in the spring. This could be a reason why the Chinese Pst populations represent extreme genetic diversity.
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Pst is highly variable, and new races that overcome newly released resistant cultivars are regular events. The widely virulent race V26 (virulent to Yr26) has a significant potential to cause epidemics in China. In this study teliospores from a single urediniospore isolate of V26 (No. Pinglan 17-7) produced on the Nanjing wheat line 92R137 (Yr26) were induced to germinate and infect Berberis shensiana as a sexual host. One hundred and eighteen single aeciospore (SA) selfed progeny and the V26 parent were typed for pathogenicity on a set of differentials comprising 22 Yrnear-isogenic wheat lines (NILs). Virulence phenotyping was conducted twice for all isolates, and similar results were obtained each time. The V26 isolate (No. Pinglan 17-7) was avirulent on differentials with Yr5, Yr6, Yr8, Yr15, Yr43, YrSp, YrTr1 and virulent on those with Yr1, Yr2, Yr4, Yr7, Yr9, Yr10, Yr17, Yr25, Yr26, Yr27, Yr28, Yr32, Yr44, YrV23, and YrExp2. The progeny were all virulent to Yr1, Yr2 (Kalyansona), Yr7, Yr9, Yr10, Yr17, Yr25, Yr26, YrV23 (Vilmorin 23) and YrExp2, and all avirulent to Yr5, Yr8, Yr15, and YrTr1, suggesting that V26 is homozygous at the corresponding pathogenicity loci. Various segregation ratios were apparent for other Yrgenes (P values ranging from 0.6to 0.09).These included3 avirulent: 1 virulent with respect to Yr6 and Yr43, 1 avirulent : 3 virulent forYr27 and Yr28, 1 avirulent : 15 virulent forYr4, Yr32, and Yr44,and 13 avirulent : 3 virulent for YrSp. Among the 118 progeny，27 of new pathotypes were identified as compared with the avirulence/virulence loci of the parent isolate. A study of the population based on markers and development of a molecular map is in progress.
Berberis holstii, native to the highlands of East Africa, is susceptible to Puccinia graminis and P. striiformis in artificial inoculations. However, it is not known whether these pathogens complete their sexual cycles in the region. In an attempt to understand the role of B. holstii in pathogen variation and epidemiology of wheat stem rust and stripe rust, we investigated the functionality of B. holstii as an alternate host. Natural aecial infections on B. holstii were observed and sampled in August at Mt. Kenya and Narok (Kenya), and June to December at North Shewa (Ethiopia) from 2008. Aeciospores from the collections were inoculated to a panel of cereal species, including Line E and ‘Morocco’ wheat, 'Hiproly’ barley, 'Prolific' rye, and ‘Marvelous’ oat. For the majority of aecial samples, aeciospore viability was lost during shipment and storage; thus inoculations were not successful. Inoculations using relatively fresh samples collected at North Shewa in 2012 and 2014, resulted in stem rust infections on Line E, Prolific, Hiproly, and Marvelous. DNA assays using real-time PCR confirmed the presence of P. graminis in these samples. While it is likely that the pathogen infecting Line E, Prolific and Hiproly is P. graminis f. sp. secalis (Pgs), the inoculation and DNA assays did not provide sufficient resolution to distinguish Pgs from Pgt. Stem rust infections on Marvelous were assumed to involve Pg f. sp. avenae. Experiments are in progress to characterize isolates derived from these samples, and to determine if other rust fungi are present in these samples. Based on these preliminary data, we conclude that P. graminis completes its sexual cycle in Ethiopia. The contribution of the sexual cycle to the observed variation within the Pgt population in the region remains unclear.
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.
In the northeastern United States, outside the boundaries of the 20th century federal barberry eradication zone, both common barberry (Berberis vulgaris) and Japanese barberry (B. thunbergii) are found in great abundance, to the extent that both are considered invasive species. Much less common and relatively less studied is their interspecific hybrid, B. ×ottawensis, which has been produced in the ornamental horticultural industry but which also occurs naturally. Since B. vulgaris is a competent host of Puccinia graminis and B. thunbergii is not, B. ×ottawensis presents a unique system for characterizing the genetic mechanism(s) underlying what appears to be non-host resistance to P. graminis in B. thunbergii. In this study, a natural population of about 1,000 individuals (mixed B. vulgaris, B. thunbergii, and B. ×ottawensis) in Sheffield, MA, was investigated. While wide morphological variation was observed among and within the populations of all three species at the site, the most pronounced variation was observed among B. ×ottawensis individuals. A subset of the population was selected for genotyping by sequencing (GBS) and evaluated for reaction to P. graminis via controlled inoculations. The response was found to segregate clearly among B. ×ottawensis individuals; and GBS was shown to be a viable means of generating molecular markers in these species, despite the lack of a reference genome. These results suggest that P. graminis resistance in B. thunbergii can be genetically mapped, and mapping populations are currently under development to accomplish this goal. The genomic resources developed in this work may facilitate both barberry surveillance efforts and ornamental barberry testing programs. Furthermore, knowledge of the genetics of response to P. graminis in the alternate host has the potential to inform efforts in breeding for stem rust resistance in wheat.
The Himalayan and near Himalayan region of Pakistan, China and Nepal was recently identified as the center of diversity of Pst. The Pakistani Himalayan populations were shown to be recombinant and possibly maintained through sexual reproduction on the alternate host, Berberis spp. To examine the role of Berberis spp. in supporting Puccinia spp. in the Himalayan region of Pakistan, 274 pycnial/aecial-infected Berberis leaves and 16 grass samples with uredinial infections were collected in the region from 2012 to 2014. Amplification of infected grass and Berberis spp. samples with EF, ITS region, and β-tubulin primers and subsequent species identification based on comparisons of the sequences to sequences in GenBank identified at least five Puccinia spp. viz., P. brachypodii, putative P. coronata-loli and P. coronati-agrostis, P. striiformis f. sp. dactylis (P. striiformoides), and P. striiformis on Berberis and grasses. This infers a role of Berberis as alternate hosts to Puccinia spp. in the Himalayan region of Pakistan, and in contributing to the overall diversity of these species in the region. Microsatellite characterization of Pst samples collected on wheat in 2013 and 2014 indicated an overall high diversity and recombinant population structure in the region. However, the low frequency of wheat-infecting P. striiformis isolates obtained from Berberis spp. necessitates ongoing investigation.
Identifying and tracking new races of wheat rust pathogens in a timely manner is important for early warning of disease potential in wheat-growing regions. Ecuador, located in northwestern South America, serves as a strategic monitoring location for rust fungi between the wheat production areas of North and South America. New races are likely to occur more often when the fungus is in the proximity of the alternate Berberis species host. More than 30 Berberis species have been reported in Ecuador, most of them endemic. However, most herbarium collections correspond to types, have only been found once, and/or date back 20 years or more. Therefore, the current status of diversity in Berberis spp. in Ecuador is largely unknown. Our goal is to collect Berberis species in Ecuador, document their distribution, prepare herbarium specimens, and identify the species morphologically and genetically. We will use this information to establish the relationships of neotropical Berberis species with Berberis in other parts of the world, and determine the pathogenicities of various rust fungi associated with them. Preliminary results show that the Ecuadorian Berberis spp. are phylogenetically distinct from those of Argentina and Brazil. To date, the rust fungi on Ecuadorean Berberis do not infect wheat. We have identified three potentially new rust fungal species based on DNA analysis.