Roi Ben-David

Wild emmer genome architecture and diversity elucidate wheat evolution and domestication

Genomics and domestication of wheat Modern wheat, which underlies the diet of many across the globe, has a long history of selection and crosses among different species. Avni et al. used the Hi-C method of genome confirmation capture to assemble and annotate the wild allotetraploid wheat (Triticum turgidum). They then identified the putative causal mutations in genes controlling shattering (a key domestication trait among cereal crops). They also performed an exome capture–based analysis of domestication among wild and domesticated genotypes of emmer wheat. The findings present a compelling overview of the emmer wheat genome and its usefulness in an agricultural context for understanding traits in modern bread wheat. Science, this issue p. 93 A polyploid wheat genome assembly elucidates wheat domestication history. Wheat (Triticum spp.) is one of the founder crops that likely drove the Neolithic transition to sedentary agrarian societies in the Fertile Crescent more than 10,000 years ago. Identifying genetic modifications underlying wheat’s domestication requires knowledge about the genome of its allo-tetraploid progenitor, wild emmer (T. turgidum ssp. dicoccoides). We report a 10.1-gigabase assembly of the 14 chromosomes of wild tetraploid wheat, as well as analyses of gene content, genome architecture, and genetic diversity. With this fully assembled polyploid wheat genome, we identified the causal mutations in Brittle Rachis 1 (TtBtr1) genes controlling shattering, a key domestication trait. A study of genomic diversity among wild and domesticated accessions revealed genomic regions bearing the signature of selection under domestication. This reference assembly will serve as a resource for accelerating the genome-assisted improvement of modern wheat varieties.

Identification and mapping of PmG16, a powdery mildew resistance gene derived from wild emmer wheat

The gene-pool of wild emmer wheat, Triticum turgidum ssp. dicoccoides, harbors a rich allelic repertoire for disease resistance. In the current study, we made use of tetraploid wheat mapping populations derived from a cross between durum wheat (cv. Langdon) and wild emmer (accession G18-16) to identify and map a new powdery mildew resistance gene derived from wild emmer wheat. Initially, the two parental lines were screened with a collection of 42 isolates of Blumeria graminis f. sp. tritici (Bgt) from Israel and 5 isolates from Switzerland. While G18-16 was resistant to 34 isolates, Langdon was resistant only to 5 isolates and susceptible to 42 isolates. Isolate Bgt#15 was selected to differentiate between the disease reactions of the two genotypes. Segregation ratio of F2-3 and recombinant inbreed line (F7) populations to inoculation with isolate Bgt#15 indicated the role of a single dominant gene in conferring resistance to Bgt#15. This gene, temporarily designated PmG16, was located on the distal region of chromosome arm 7AL. Genetic map of PmG16 region was assembled with 32 simple sequence repeat (SSR), sequence tag site (STS), Diversity array technology (DArT) and cleaved amplified polymorphic sequence (CAPS) markers and assigned to the 7AL physical bin map (7AL-16). Using four DNA markers we established colinearity between the genomic region spanning the PmG16 locus within the distal region of chromosome arm 7AL and the genomic regions on rice chromosome 6 and Brachypodium Bd1. A comparative analysis was carried out between PmG16 and other known Pm genes located on chromosome arm 7AL. The identified PmG16 may facilitate the use of wild alleles for improvement of powdery mildew resistance in elite wheat cultivars via marker-assisted selection.

Identification and characterization of a novel powdery mildew resistance gene PmG3M derived from wild emmer wheat, Triticum dicoccoides

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt) is one of the most important wheat diseases worldwide. Wild emmer wheat, Triticum turgidum ssp. dicoccoides, the tetraploid ancestor (AABB) of domesticated bread and durum wheat, harbors many important alleles for resistance to various diseases, including powdery mildew. In the current study, two tetraploid wheat mapping populations, derived from a cross between durum wheat (cv. Langdon) and wild emmer wheat (accession G-305-3M), were used to identify and map a novel powdery mildew resistance gene. Wild emmer accession G-305-3M was resistant to all 47 Bgt isolates tested, from Israel and Switzerland. Segregation ratios of F2 progenies and F6 recombinant inbred line (RIL) mapping populations, in their reactions to inoculation with Bgt, revealed a Mendelian pattern (3:1 and 1:1, respectively), indicating the role of a single dominant gene derived from T. dicoccoides accession G-305-3M. This gene, temporarily designated PmG3M, was mapped on chromosome 6BL and physically assigned to chromosome deletion bin 6BL-0.70-1.00. The F2 mapping population was used to construct a genetic map of the PmG3M gene region consisted of six simple sequence repeats (SSR), 11 resistance gene analog (RGA), and two target region amplification polymorphism (TRAP) markers. A second map, constructed based on the F6 RIL population, using a set of skeleton SSR markers, confirmed the order of loci and distances obtained for the F2 population. The discovery and mapping of this novel powdery mildew resistance gene emphasize the importance of the wild emmer wheat gene pool as a source for crop improvement.

Suppressed recombination rate in 6VS/6AL translocation region carrying the Pm21 locus introgressed from Haynaldia villosa into hexaploid wheat

Pm21 is an effective gene for powdery mildew resistance transferred from Haynaldia villosa into common wheat cultivars. No virulence against this gene has been detected so far. A set of 42 powdery mildew isolates collected in Israel and tested in the current study also revealed no virulence against this gene. Pm21 was previously reported to be located on the short arm of 6VS/6AL translocation chromosome. We constructed a high-density genetic map of chromosome 6A, consisting of 28 PCR markers and the Pm21 gene. A comparison with previously published genetic maps of wheat chromosome 6A revealed that the recombination rate in the 6VS/6AL translocation region was poor. We assume that suppressed recombination caused by the alien H. villosa genetic material is the most reasonable explanation for the tight genetic linkage and the inadequacy between the Pm21 genetic map and the Pm21 physical map of 6A. A large number of sequence-tag sites (STS) and simple sequence repeat markers, which co-segregate with or are closely linked to the Pm21 gene, and the conversion of three resistance gene analog markers into new STS markers, provide a reliable and easy-to-use molecular tool for marker-assisted selection of Pm21 in wheat breeding programs. An additional gene, Pm31, previously reported to be derived from Triticum dicoccoides, was mapped into a similar genomic location to Pm21. Screening of the parental lines and the mapping population with Pm21 diagnostic markers clearly confirmed that the donor line of Pm31 is H. villosa and not T. dicoccoides. Therefore, we conclude that Pm21 and Pm31 refer to the same gene, derived from H. villosa, and that the designation of Pm31 as a new Pm gene was erroneous.

AvrPm2 encodes an RNase-like avirulence effector which is conserved in the two different specialized forms of wheat and rye powdery mildew fungus.

Summary There is a large diversity of genetically defined resistance genes in bread wheat against the powdery mildew pathogen Blumeria graminis (B. g.) f. sp. tritici. Many confer race‐specific resistance to this pathogen, but until now only the mildew avirulence gene AvrPm3 a2/f2 that is recognized by Pm3a/f was known molecularly. We performed map‐based cloning and genome‐wide association studies to isolate a candidate for the mildew avirulence gene AvrPm2. We then used transient expression assays in Nicotiana benthamiana to demonstrate specific and strong recognition of AvrPm2 by Pm2. The virulent AvrPm2 allele arose from a conserved 12 kb deletion, while there is no protein sequence diversity in the gene pool of avirulent B. g. tritici isolates. We found one polymorphic AvrPm2 allele in B. g. triticale and one orthologue in B. g. secalis and both are recognized by Pm2. AvrPm2 belongs to a small gene family encoding structurally conserved RNase‐like effectors, including Avr a13 from B. g. hordei, the cognate Avr of the barley resistance gene Mla13. These results demonstrate the conservation of functional avirulence genes in two cereal powdery mildews specialized on different hosts, thus providing a possible explanation for successful introgression of resistance genes from rye or other grass relatives to wheat.

Eco-geographical distribution of Lactuca saligna natural populations in Israel

Crops can be drastically improved by effective utilization of the immense variation in genetic resources for resistance to abiotic and biotic stresses possessed by natural populations of wild relatives. To protect Israels and the worlds lettuce crop, samples of seeds from 562 plants that morphologically seemed to be the wild species Lactuca saligna L. (least lettuce, willow-leaf lettuce) were collected from 41 localities, representing different climatic and edaphic environments throughout Israel. Searching and collecting trips were conducted in September—October of 2004-2006. L. saligna was recorded throughout Israel except for desert areas (e.g., Negev and Judean deserts) and extreme environmental/soil conditions (Dead Sea area). L. saligna was recorded at various altitudes (10 to 1277 m asl) and different habitats and soil types. In all, the taxonomic status of 214 of the accessions was morphologically validated as L. saligna during multiplication of 220 accessions in the greenhouse. In rosette format...

Effect of GA-sensitivity on wheat early vigor and yield components under deep sowing

Establishment of seedlings is a key factor in achievement of uniform field stands and, consequently, stable yields. Under Mediterranean conditions, soil moisture in the upper layer is limited and seedlings may be exposed to frequent dehydration events. The presence of the Reduced height (Rht)-B1b and Rht-D1b semi-dominant dwarfing alleles results in insensitivity to gibberellin (GAI) and, hence, poor emergence from deep sowing. Introduction of alternative dwarfing genes and, thereby, preservation of the gibberellin response (GAR) and coleoptile length, contributes to better emergence from deep sowing. Initially 47 wheat cultivars carrying different Rht alleles were screened for their ability to emerge from deep sowing, and then 17 of them were selected for detailed physiological characterization in the field. The modern wheat lines containing GAI alleles showed significantly lower percentages of emergence from deep sowing than the GAR lines, i.e., 52 and 74%, respectively. Differences in early developmental stages were associated with grain yield, as indicated by a reduction of 37.3% in the modern GAI cultivars. Our results demonstrate the potential of alternative dwarfing genes for improving seedling establishment and grain yields in Mediterranean-like environments.

Differentiation Among Blumeria graminis f. sp. tritici Isolates Originating from Wild Versus Domesticated Triticum Species in Israel.

Israel and its vicinity constitute a center of diversity of domesticated wheat species (Triticum aestivum and T. durum) and their sympatrically growing wild relatives, including wild emmer wheat (T. dicoccoides). We investigated differentiation within the forma specialis of their obligate powdery mildew pathogen, Blumeria graminis f. sp. tritici. A total of 61 B. graminis f. sp. tritici isolates were collected from the three host species in four geographic regions of Israel. Genetic relatedness of the isolates was characterized using both virulence patterns on 38 wheat lines (including 21 resistance gene differentials) and presumptively neutral molecular markers (simple-sequence repeats and single-nucleotide polymorphisms). All isolates were virulent on at least some genotypes of all three wheat species tested. All assays divided the B. graminis f. sp. tritici collection into two distinct groups, those from domesticated hosts and those from wild emmer wheat. One-way migration was detected from the domestic wheat B. graminis f. sp. tritici population to the wild emmer B. graminis f. sp. tritici population at a rate of five to six migrants per generation. This gene flow may help explain the overlap between the distinct domestic and wild B. graminis f. sp. tritici groups. Overall, B. graminis f. sp. tritici is significantly differentiated into wild-emmer and domesticated-wheat populations, although the results do not support the existence of a separate f. sp. dicocci.

Distinct domains of the AVRPM3A2/F2 avirulence protein from wheat powdery mildew are involved in immune receptor recognition and putative effector function

Summary Recognition of the AVRPM3A2/F2 avirulence protein from powdery mildew by the wheat PM3A/F immune receptor induces a hypersensitive response after co‐expression in Nicotiana benthamiana. The molecular determinants of this interaction and how they shape natural AvrPm3 a2/f2 allelic diversity are unknown. We sequenced the AvrPm3 a2/f2 gene in a worldwide collection of 272 mildew isolates. Using the natural polymorphisms of AvrPm3 a2/f2 as well as sequence information from related gene family members, we tested 85 single‐residue‐altered AVRPM3A2/F2 variants with PM3A, PM3F and PM3FL 456P/Y458H (modified for improved signaling) in Nicotiana benthamiana for effects on recognition. An intact AvrPm3 a2/f2 gene was found in all analyzed isolates and the protein variant recognized by PM3A/F occurred globally at high frequencies. Single‐residue alterations in AVRPM3A2/F2 mostly disrupted, but occasionally enhanced, the recognition response by PM3A, PM3F and PM3FL 456P/Y458H. Residues enhancing hypersensitive responses constituted a protein domain separate from both naturally occurring polymorphisms and positively selected residues of the gene family. These results demonstrate the utility of using gene family sequence diversity to screen residues for their role in recognition. This approach identified a putative interaction surface in AVRPM3A2/F2 not polymorphic in natural alleles. We conclude that molecular mechanisms besides recognition drive AvrPm3 a2/f2 diversification.

Agronomic and genetic characterization of wild emmer wheat ( Triticum turgidum subsp . dicoccoides ) introgression lines in a bread wheat genetic background

During a long evolutionary history across a range of environmental conditions in the Near East Fertile Crescent, wild emmer wheat (Triticum turgidum subsp. dicoccoides) has accumulated a wealth of genetic diversity and adaptations to multiple biotic and abiotic stress conditions. The joint effects of the domestication and subsequent selection in man-made agro-ecosystems have considerably reduced the genetic variation in cultivated crop species as compared to their wild progenitors. The wild emmer gene pool harbors a rich allelic repertoire for improving numerous agronomically important traits. A set of advanced wild emmer wheat introgression lines (ILs) was previously established on a bread wheat genetic background. It combines genetic diversity from the wild progenitor as well as the genetic background of the cultigen. Based on field evaluations, the set of ILs (divided hierarchically into different families based on the genetic background) has exhibited high phenotypic variance in agronomically important traits. Analyses of variance (ANOVA) have shown that the effect of the hierarchical genetic factors was significant in the vast majority of the traits. Similarly, characterizations by molecular markers of known traits have shown that the genetic profile differs between ILs from different genetic backgrounds. Multivariate principal component analysis has revealed interesting associations between vegetative and reproductive traits and illustrated the impact of some of these traits on ILs distribution in the PC1 and PC2. The exploitation of the wild emmer wheat ILs as a tool for reintroduction of alleles that were lost during domestication and possible implications for wheat breeding are discussed.