Analysis of Extreme Phenotype Bulk Copy Number Variation (XP-CNV) Identified the Association of rp1 with Resistance to Goss\u27s Wilt of Maize

Goss\u27s wilt (GW) of maize is caused by the Gram-positive bacterium Clavibacter michiganensis subsp. nebraskensis (Cmn) and has spread in recent years throughout the Great Plains, posing a threat to production. The genetic basis of plant resistance is unknown. Here, a simple method for quantifying disease symptoms was developed and used to select cohorts of highly resistant and highly susceptible lines known as extreme phenotypes (XP). Copy number variation (CNV) analyses using whole genome sequences of bulked XP revealed 141 genes containing CNV between the two XP groups. The CNV genes include the previously identified common rust resistant locus rp1. Multiple Rp1 accessions with distinct rp1 haplotypes in an otherwise susceptible accession exhibited hypersensitive responses upon inoculation. GW provides an excellent system for the genetic dissection of diseases caused by closely related subspecies of C. michiganesis. Further work will facilitate breeding strategies to control GW and provide needed insight into the resistance mechanism of important related diseases such as bacterial canker of tomato and bacterial ring rot of potato

A time for more booms and fewer busts? Unraveling cereal-rust interactions

Recent advances in our understanding of the nature of resistance genes and rust fungus genomics are providing some insight into the basis of resistance and susceptibility to rust diseases in our cereal crops. Characterized rust resistance genes, for the most part, resemble other resistance genes that interact with effectors intracellularly, but some have unique features. Characterization of rust effectors is just beginning but genomic information and technical advances in rust functional genomics will accelerate their characterization. The ephemeral nature of resistance in past varieties has made the design of cultivars with durable resistance a major focus for geneticists and cereal breeders. This includes strategies for deploying race-specific resistance genes that prolong their effects and methods of predicting which will be difficult for the pathogen to defeat. Identification of resistance genes with race-nonspecific effects is another strategy where recent breakthroughs have been made. Routinely combining the numerous genes required for complex resistance, whether specific or nonspecific, in elite cultivars remains a primary constraint to realizing durable resistance in most programs

Dual host-pathogen small RNA sequencing during wheat stem rust infection

Abstract Objectives RNA sequencing of two organisms in a symbiotic interaction can yield insights that are not found in samples from each organism alone. We present a sequencing dataset focusing on the small RNA fraction from wheat plants (Triticum aestivum) infected with the biotrophic pathogen wheat stem rust fungus (Puccinia graminis f.sp. tritici). Simultaneous small RNA sequencing of this agronomically important crop and its adversary can lead to a better understanding of the role of noncoding RNAs in both plant and fungal biology. Data description Small RNA libraries were constructed from infected and mock-infected plant tissue and sequenced on the Ion Torrent platform. Quality control was performed to ensure sample and data integrity. Using this dataset, researchers can employ previously established methods to map subsets of reads exclusively to each organism’s genome. Subsequent analyses can be undertaken to discover microRNAs, predict small RNA targets, and generate hypotheses for further laboratory experiments

Recombination events generating a novel Rp1 race specificity

Genes at the maize Rp1 rust resistance complex often mispair in meiosis, which allows genes to recombine unequally, creating recombinant haplotypes. Four recombinant haplotypes were identified from progeny of an Rp1-D/Rp1-I heterozygote that conferred a nonparental resistance specificity designated Rp1-I*. Sequence comparisons of paralogs in the recombinant and parental haplotypes demonstrated that all four recombinants were derived from intergenic (between gene) recombination events. The sequence of paralogs in the HRp1-I parental haplotype indicated this haplotype includes 41 or more rp1 genes, at least 31 of which are transcribed. The results indicate that most of the novel resistance specificities that have arisen spontaneously at Rp1 are the result of reassort ment of existing Rp1 genes

Allelic and haplotypic diversity at the rp1 rust resistance locus of maize

The maize Rp1 rust resistance locus is a complex consisting of a family of closely related resistance genes. The number of Rp1 paralogs in different maize lines (haplotypes) varied from a single gene in some stocks of the inbred A188 to >50 genes in haplotypes carrying the Rp1-A and Rp1-H specificities. The sequences of paralogs in unrelated haplotypes differ, indicating that the genetic diversity of Rp1-related genes is extremely broad in maize. Two unrelated haplotypes with five or nine paralogs had identical resistance phenotypes (Rp1-D) encoded in genes that differed by three nucleotides resulting in a single amino acid substitution. Genes in some haplotypes are more similar to each other than to any of the genes in other haplotypes indicating that they are evolving in a concerted fashion

Development of a Host-Induced RNAi System in the Wheat Stripe Rust Fungus Puccinia striiformis f. sp. tritici

Rust fungi cause devastating diseases of wheat and other cereal species globally. Genetic resistance is the preferred method to control rusts but the effectiveness of race-specific resistance is typically transient due to the genetic plasticity of rust populations. The advent of RNA interference (RNAi) technology has shown promise for the engineering of resistance to some biotrophic pathogens in plants by altering the expression of essential pathogens\u27 genes. Gene fragments from the rust fungi Puccinia striiformis f. sp. tritici or P. graminis f. sp. tritici were delivered to plant cells through the Barley stripe mosaic virus system, and some reduced the expression of the corresponding genes in the rust fungus. The ability to detect suppression was associated with the expression patterns of the fungal genes because reduction was only detected in transcripts with relatively high levels of expression in fungal haustoria. The results indicate that an in planta RNAi approach can be used in functional genomics research for rust fungi and that it could potentially be used to engineer durable resistance. © 2011 The American Phytopathological Society

Additional file 1: of Small RNAs from the wheat stripe rust fungus (Puccinia striiformis f.sp. tritici)

Bioinformatics Pipeline. Graphical summary of the bioinformatic pipeline used to obtain putative Puccinia striiformis small RNAs (Pst-sRNAs). The total sRNA library consists of mostly wheat reads (green) with a small fraction of stripe rust reads. After mapping to the stripe rust genome, discarding reads present in uninfected controls, and discarding sequences matching wheat miRNA and protein-coding sequences, the library is increasingly enriched for stripe rust reads (orange). Sequences discarded at each step are shown as arrows to the right. (PDF 36 kb

Characterization of a tryptophan 2-monooxygenase gene from Puccinia graminis f. sp. tritici involved in auxin biosynthesis and rust pathogenicity

The plant hormone indole-3-acetic acid (IAA) is best known as a regulator of plant growth and development but its production can also affect plant-microbe interactions. Microorganisms, including numerous plant-associated bacteria and several fungi, are also capable of producing IAA. The stem rust fungus Puccinia graminis f. sp. tritici induced wheat plants to accumulate auxin in infected leaf tissue. A gene (Pgt-IaaM) encoding a putative tryptophan 2-monooxygenase, which makes the auxin precursor indole-3-acetamide (IAM), was identified in the P. graminis f. sp. tritici genome and found to be expressed in haustoria cells in infected plant tissue. Transient silencing of the gene in infected wheat plants indicated that it was required for full pathogenicity. Expression of Pgt-IaaM in Arabidopsis caused a typical auxin expression phenotype and promoted susceptibility to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000

A maize resistance gene functions against bacterial streak disease in rice

Although cereal crops all belong to the grass family (Poacea), most of their diseases are specific to a particular species. Thus, a given cereal species is typically resistant to diseases of other grasses, and this nonhost resistance is generally stable. To determine the feasibility of transferring nonhost resistance genes (R genes) between distantly related grasses to control specific diseases, we identified a maize R gene that recognizes a rice pathogen, Xanthomonas oryzae pv. oryzicola, which causes bacterial streak disease. Bacterial streak is an important disease of rice in Asia, and no simply inherited sources of resistance have been identified in rice. Although X. o. pv. oryzicola does not cause disease on maize, we identified a maize gene, Rxo1, that conditions a resistance reaction to a diverse collection of pathogen strains. Surprisingly, Rxo1 also controls resistance to the unrelated pathogen Burkholderia andropogonis, which causes bacterial stripe of sorghum and maize. The same gene thus controls resistance reactions to both pathogens and nonpathogens of maize. Rxo1 has a nucleotide-binding site-leucine-rich repeat structure, similar to many previously identified R genes. Most importantly, Rxo1 functions after transfer as a transgene to rice, demonstrating the feasibility of nonhost R gene transfer between cereals and providing a valuable tool for controlling bacterial streak disease