High-resolution mapping of Rsn1, a locus controlling sensitivity of rice to a necrosis-inducing phytotoxin from Rhizoctonia solani AG1-IA.

Rhizoctonia solani is a necrotrophic fungal pathogen that causes disease on many crop-plant species. Anastomosis group 1-IA is the causal agent of sheath blight of rice (Oryza sativa L.), one of the most important rice diseases worldwide. R. solani AG1-IA produces a necrosis-inducing phytotoxin and rice cultivar's sensitivity to the toxin correlates with disease susceptibility. Unlike genetic analyses of sheath blight resistance where resistance loci have been reported as quantitative trait loci, phytotoxin sensitivity is inherited as a Mendelian trait that permits high-resolution mapping of the sensitivity genes. An F(2) mapping population derived from parent cultivars 'Cypress' (toxin sensitive) and 'Jasmine 85' (toxin insensitive) was used to map Rsn1, the necrosis-inducing locus. Initial mapping based on 176 F(2) progeny and 69 simple sequence repeat (SSR) markers located Rsn1 on the long arm of chromosome 7, with tight linkage to SSR marker RM418. A high-resolution genetic map of the region was subsequently developed using a total of 1,043 F(2) progeny, and Rsn1 was mapped to a 0.7 cM interval flanked by markers NM590 and RM418. Analysis of the corresponding 29 Kb genomic sequences from reference cultivars 'Nipponbare' and '93-11' revealed the presence of four putative genes within the interval. Two are expressed cytokinin-O-glucosyltransferases, which fit an apoptotic pathway model of toxin activity, and are individually being investigated further as potential candidates for Rsn1.

Influences from Long-Term Crop Rotation, Soil Tillage, and Fertility on the Severity of Rice Grain Smuts.

False smut (Ustilaginoidea virens) and kernel smut (Neovossia horrida) are diseases of rice (Oryza sativa) that reduce both grain yield and quality. Susceptible rice cultivars are in widespread use on production acreage in the United States, and the effects from crop management practices on smut control are poorly understood. We studied the long-term effects of crop rotation, soil tillage, and fertility level on rice smut severity. The highest levels of false smut observed in this study were on cultivars grown in rotation with soybean, on traditionally tilled soils, with high fertilizer treatments. The highest levels of kernel smut were observed in a rice-soybean rotation with winter wheat grown between summer crops. These rotations are commonly used in rice-growing regions of the southern United States. Using combinations of crop rotation, soil tillage, and fertility rate, several alternative crop-management practices were identified that provided effective control of smuts in susceptible rice cultivars. The most effective method for controlling both false smut and kernel smut was in 3-year rotations of rice, soybean, and corn. Regardless of rotation order or tillage and fertility treatments within the rotations, rotating out of rice for 2 years was the most effective approach for smut control.

Effect of Furrow Irrigation on the Severity of False Smut in Susceptible Rice Varieties.

False smut (Ustilaginoidea virens) is an important emerging disease of rice (Oryza sativa) in the southern United States, where all major rice cultivars and hybrids are susceptible to the disease. False smut susceptibility was evaluated in traditional paddy-rice fields and under furrow-irrigated conditions to determine the effects of alternative agricultural practices on the severity of this disease. Highly effective false smut suppression was observed in furrow-irrigated rice, where the disease was nearly eliminated in susceptible rice entries. False smut suppression was observed for two hybrids and one conventional rice cultivar, demonstrating that suppression was not limited to specific germplasm sources. Kernel smut severity was also monitored, but no effect on this disease was observed from the irrigation treatments. Therefore, suppression of disease severity in nonflooded rice appears to be a phenomenon unique to the rice-false smut pathosystem, which can be exploited to achieve effective field resistance to this disease.

Effect of Cultural Management Practices on the Severity of False Smut and Kernel Smut of Rice.

False smut (Ustilaginoidea virens) and kernel smut (Neovossia horrida) are diseases of rice (Oryza sativa) that reduce both grain yield and quality. False smut is an emerging disease worldwide that is rapidly gaining in importance, whereas kernel smut has historically been a chronic minor disease with sporadic outbreaks that cause considerable losses. Highly effective disease control was obtained for susceptible cultivars by employing conservation tillage (69% reduction in false smut), continuous rice cropping (88% reduction in false smut), and moderate nitrogen fertility rates (34 and 60% reductions in false smut and kernel smut, respectively). Combining these treatments nearly eliminated smuts from cultivars that were fully susceptible under conventional cultivation practices. Furthermore, using a nursery designed to promote smut diseases, two rice hybrids were identified that possessed kernel smut resistance under the most favorable disease conditions. The genetic basis of the resistance is unknown. However, the utility for disease control is great because hybrids occupy significant portions of production rice acreage.

A natural mutation in rc reverts white-rice-pericarp to red and results in a new, dominant, wild-type allele: Rc-g.

The Rc locus regulates pigmentation of the rice bran layer, and selection for the rc allele (white pericarp) occurred during domestication of the crop. White bran is now ubiquitous among cultivated varieties throughout rice growing regions of the world. We identified a new allele that arose by natural mutation within the rc pseudogene of the cultivar 'Wells'. The mutation restored the reading frame of the gene, and reverted the bran layer pigmentation to red (wild-type). By sequencing the Rc locus in plants derived from red seeds, and linkage analysis in a segregating population, we were able to demonstrate that mutation within rc resulted in the new, dominant, wild-type allele Rc-g.

Sensitivity to a Phytotoxin from Rhizoctonia solani Correlates with Sheath Blight Susceptibility in Rice.

ABSTRACT Sheath blight is one of the most important and intractable diseases of rice (Oryza sativa) where limited control has been achieved using traditional approaches. Quantitative inheritance, extraneous traits, and environmental factors confound genetic analysis of host resistance. A method was developed to isolate and utilize a phytotoxin from Rhizoctonia solani to investigate the genetics of sheath blight susceptibility. Infiltration of the toxin preparation into plant leaves induced necrosis in rice, maize, and tomato. Using 17 rice cultivars known to vary in sheath blight resistance, genotypes were identified that were sensitive (tox-S) and insensitive (tox-I) to the toxin, and a correlation (r = 0.66) between toxin sensitivity and disease susceptibility was observed. Given the broad host range of R. solani, genotypes of host species may be both tox-S and tox-I. A total of 154 F(2) progeny from a cross between Cypress (tox-S) and Jasmine 85 (tox-I) segregated in a 9:7 ratio for tox-S/tox-I, indicating an epistatic interaction between two genes controls sensitivity to the toxin in rice. This work provides the means to genetically map toxin sensitivity genes and eliminate susceptible genotypes when developing sheath blight-resistant rice cultivars.

Structural variation and evolution of a defense-gene cluster in natural populations of Aegilops tauschii.

Genetic mapping and sequencing of plant genomes have been useful for investigating eukaryotic chromosome structural organization. In many cases, analyses have been limited in the number of representatives sampled from specific groups. The degree of intraspecific genome diversity remains in question. The possibility exists that a single model genome may have limited utility for identifying genes in related members of the species or genus. Crop improvement programs have particular interests in disease resistance genes that are harbored by wild relatives of modern cultivated crops. These genes are evolutionarily dynamic and under selective pressure by a broad range of pathogenic organisms. Using resistance gene analogs as models for gene evolution, intraspecific genome comparisons were made among populations of wild diploid wheat (Aegilops tauschii). We observed that deletion haplotypes are occurring frequently and independently in the genome. Haplotypes are geographically correlated and maintenance of gene complements in localized populations indicates selective advantage. Furthermore, deletion haplotypes are not detrimental to plant health, since genes without adaptive value in alternate environments are eliminated from the genome. Deletion haplotypes appear to be a common form of allelic variation in plants, and we address the consequences on genome restructuring and gene evolution.

Map-based cloning of leaf rust resistance gene Lr21 from the large and polyploid genome of bread wheat.

We report the map-based cloning of the leaf rust resistance gene Lr21, previously mapped to a gene-rich region at the distal end of chromosome arm 1DS of bread wheat (Triticum aestivum L.). Molecular cloning of Lr21 was facilitated by diploid/polyploid shuttle mapping strategy. Cloning of Lr21 was confirmed by genetic transformation and by a stably inherited resistance phenotype in transgenic plants. Lr21 spans 4318 bp and encodes a 1080-amino-acid protein containing a conserved nucleotide-binding site (NBS) domain, 13 imperfect leucine-rich repeats (LRRs), and a unique 151-amino-acid sequence missing from known NBS-LRR proteins at the N terminus. Fine-structure genetic analysis at the Lr21 locus detected a noncrossover (recombination without exchange of flanking markers) within a 1415-bp region resulting from either a gene conversion tract of at least 191 bp or a double crossover. The successful map-based cloning approach as demonstrated here now opens the door for cloning of many crop-specific agronomic traits located in the gene-rich regions of bread wheat.

A bacterial artificial chromosome contig spanning the major domestication locus Q in wheat and identification of a candidate gene.

The Q locus played a major role in the domestication of wheat because it confers the free-threshing character and influences many other agronomically important traits. We constructed a physical contig spanning the Q locus using a Triticum monococcum BAC library. Three chromosome walking steps were performed by complete sequencing of BACs and identification of low-copy markers through similarity searches of database sequences. The BAC contig spans a physical distance of approximately 300 kb corresponding to a genetic distance of 0.9 cM. The physical map of T. monococcum had perfect colinearity with the genetic map of wheat chromosome arm 5AL. Recombination data in conjunction with analysis of fast neutron deletions confirmed that the contig spanned the Q locus. The Q gene was narrowed to a 100-kb segment, which contains an APETALA2 (AP2)-like gene that cosegregates with Q. AP2 is known to play a major role in controlling floral homeotic gene expression and thus is an excellent candidate for Q.

Analysis of 106 kb of contiguous DNA sequence from the D genome of wheat reveals high gene density and a complex arrangement of genes related to disease resistance.

Vast differences exist in genome sizes of higher plants; however, gene count remains relatively constant among species. Differences observed in DNA content can be attributed to retroelement amplification leading to genome expansion. Cytological and genetic studies have demonstrated that genes are clustered in islands rather than distributed at random in the genome. Analysis of gene islands within highly repetitive genomes of plants like wheat remains largely unstudied. The objective of our work was to sequence and characterize a contiguous DNA sequence from chromosome IDS of Aegilops tauschii. An RFLP probe that maps to the Lr21 region of IDS was used to isolate a single BAC. The BAC was sequenced and is 106 kb in length. The contiguous DNA sequence contains a 46-kb retroelement-free gene island containing seven coding sequences. Within the gene island is a complex arrangement of resistance and defense response genes. Overall gene density in this BAC is 1 gene per 8.9 kb. This report demonstrates that wheat and its relatives do contain regions with gene densities similar to that of Arabidopsis.