Dawn Verlin

New sources of rust resistance from alien species : meliorating linked defects and discovery

This paper presents a review of projects undertaken over the past 2 decades aimed at improving the yield and/or quality attributes of translocation lines carrying rust resistance genes from species related to wheat, so as to make these lines more suitable for use in breeding programs. Homeologous recombination between the alien chromosome segments and normal wheat chromosomes was induced in a ph1bph1b background. Lines with shortened alien chromatin were selected using dissociation patterns of molecular-based markers. A new line of bread wheat was developed containing a shortened chromosome 1RS segment carrying rust resistance gene SrR (Secale cereale L.), in which a defect affecting dough-quality appears to have been deleted. In addition, several advanced lines were produced with modified 6Ae#1L chromosome segments with Sr26 (Thinopyrum ponticum), 2S#1 chromosome segments with Sr32, and a previously unnamed gene, a 2S#2 chromosome segment with Sr39 (Triticum speltoides), 4G#1 chromosome segments with Sr37, and 2G#2 chromosome segments with Sr40 (T. timopheevii).

Development of wheat lines carrying stem rust resistance gene Sr39 with reduced Aegilops speltoides chromatin and simple PCR markers for marker-assisted selection

The use of major resistance genes is a cost-effective strategy for preventing stem rust epidemics in wheat crops. The stem rust resistance gene Sr39 provides resistance to all currently known pathotypes of Puccinia graminis f. sp. tritici (Pgt) including Ug99 (TTKSK) and was introgressed together with leaf rust resistance gene Lr35 conferring adult plant resistance to P. triticina (Pt), into wheat from Aegilops speltoides. It has not been used extensively in wheat breeding because of the presumed but as yet undocumented negative agronomic effects associated with Ae. speltoides chromatin. This investigation reports the production of a set of recombinants with shortened Ae. speltoides segments through induction of homoeologous recombination between the wheat and the Ae. speltoides chromosome. Simple PCR-based DNA markers were developed for resistant and susceptible genotypes (Sr39#22r and Sr39#50s) and validated across a set of recombinant lines and wheat cultivars. These markers will facilitate the pyramiding of ameliorated sources of Sr39 with other stem rust resistance genes that are effective against the Pgt pathotype TTKSK and its variants.

A repetitive DNA sequence common to the different B chromosomes of the genus Brachycome

Abstract. Dot-like micro B chromosomes of Brachycome dichromosomatica were analysed for their sequence composition. Southern hybridization patterns of a total micro B probe to genomic DNA from plants with and without micro Bs demonstrated that the micro Bs shared sequences with the A chromosomes. In addition to telomere, rDNA and common A and B chromosome sequences, a new B-specific, highly methylated tandem repeat (Bdm29) was detected. After in situ hybridization with Bdm29 the entire micro B chromosome was labelled and clustering of the condensed micro Bs could be observed at interphase. A high number of Bdm29-like sequences were also found in the larger B chromosomes of B. dichromosomatica and in other Bs within the genus Brachycome.

Isolation of wheat–rye 1RS recombinants that break the linkage between the stem rust resistance gene SrR and secalin

Chromosome 1R of rye is a useful source of genes for disease resistance and enhanced agronomic performance in wheat. One of the most prevalent genes transferred to wheat from rye is the stem rust resistance gene Sr31. The recent emergence and spread of a stem rust pathotype virulent to this gene has refocused efforts to find and utilize alternative sources of resistance. There has been considerable effort to transfer a stem rust resistance gene, SrR, from Imperial rye, believed to be allelic to Sr31, into commercial wheat cultivars. However, the simultaneous transfer of genes at the Sec-1 locus encoding secalin seed storage proteins and their association with quality defects preclude the deployment of SrR in some commercial wheat breeding programs. Previous attempts to induce homoeologous recombination between wheat and rye chromosomes to break the linkage between SrR and Sec-1 whilst retaining the tightly linked major loci for wheat seed storage proteins, Gli-D1 and Glu-D3, and recover good dough quality characteristics, have been unsuccessful. We produced novel tertiary wheat-rye recombinant lines carrying different lengths of rye chromosome arm 1RS by inducing homoeologous recombination between the wheat 1D chromosome and a previously described secondary wheat-rye recombinant, DRA-1. Tertiary recombinant T6-1 (SrR+ Sec-1-) carries the target gene for stem rust resistance from rye and retains Gli-D1 but lacks the secalin locus. The tertiary recombinant T49-7 (SrR- Sec-1+) contains the secalin locus but lacks the stem rust resistance gene. T6-1 is expected to contribute to wheat breeding programs in Australia, whereas T49-7 provides opportunities to investigate whether the presence of secalins is responsible for the previously documented dough quality defects.

The genomic complexity of micro B chromosomes of Brachycome dichromosomatica

Abstract. A major sequence component of the micro B chromosome of Brachycomedichromosomatica (2n=4) is the tandem repeat Bdm29, which was found by in situ hybridisation to be distributed along the entire length of the chromosome. A high copy number of this sequence does not occur as a regular feature of the A chromosomes in this species but it was found in infrequent individuals in two wild populations that were analysed. In these instances Bdm29 is localised within heterochromatic, polymorphic segments on the long arm of chromosome 1. The origin of the micro B chromosomes was investigated by determining whether they are related to this A chromosome polymorphism by simple excision and/or integration. Results obtained by using Bdm29, together with a newly isolated repeat sequence, Bdm54, and a number of other sequences known to occur on the micro B chromosome, as probes in in situ hybridisation and Southern analysis demonstrated that the formation of micro B chromosomes is a complex multistep process. The observation that the genomic organisation of the micro B chromosome is unlike anything found on the A chromosomes precludes their origin by simple excision and also indicates that micro Bs do not integrate directly into the A complement to form polymorphic heterochomatic segments.

A monophyletic origin of the B chromosomes of Brachycome dichromosomatica (Asteraceae)

The A and B chromosomes of different karyotype variants (cytodemes A1, A2, A3 and A4) ofBrachycome dichromosomatica were analysed by computer-aided chromosome image analysis and fluorescencein situ hybridisation (FISH). Ribosomal DNA and the B chromosome-specific sequence Bd49 were detected on all B chromosomes. In addition to minor size variation of the Bs, polymorphism of the rDNA and Bd49 position and copy number revealed two major types of B chromosomes. The B chromosomes of all the cytodemes were indistinguishable from each other in length, but that of A3 showed evidence of rearrangements consistent with its long-term geographic isolation. The results presented suggest a monophyletic origin of the B chromosomes ofB. dichromosomatica.

Development of wheat-Aegilops speltoides recombinants and simple PCR-based markers for Sr32 and a new stem rust resistance gene on the 2S#1 chromosome.

Key messageWheat–Aegilops speltoidesrecombinants carrying stem rust resistance genesSr32andSrAes1teffective against Ug99 and PCR markers for marker-assisted selection.AbstractWild relatives of wheat are important resources for new rust resistance genes but underutilized because the valuable resistances are often linked to negative traits that prevent deployment of these genes in commercial wheats. Here, we report ph1b-induced recombinants with reduced alien chromatin derived from E.R. Sears’ wheat–Aegilops speltoides 2D-2S#1 translocation line C82.2, which carries the widely effective stem rust resistance gene Sr32. Infection type assessments of the recombinants showed that the original translocation in fact carries two stem rust resistance genes, Sr32 on the short arm and a previously undescribed gene SrAes1t on the long arm of chromosome 2S#1. Recombinants with substantially shortened alien chromatin were produced for both genes, which confer resistance to stem rust races in the TTKSK (Ug99) lineage and representative races of all Australian stem rust lineages. Selected recombinants were back crossed into adapted Australian cultivars and PCR markers were developed to facilitate the incorporation of these genes into future wheat varieties. Our recombinants and those from several other labs now show that Sr32, Sr39, and SrAes7t on the short arm and Sr47 and SrAes1t on the long arm of 2S#1 form two linkage groups and at present no rust races are described that can distinguish these resistance specificities.

First report of meiotic chromosome number and karyotype analysis of an accession of Trigonella balansae (Leguminosae)

Abstract This study represents the first report of the number of meiotic chromosomes and morphology of the mitotic chromosomes of Trigonella balansae. The haploid complement of chromosomes of T. balansae consists of eight chromosomes (2n = 16). Of these, only the satellited chromosome and possibly a subtelocentric chromosome are identifiable. The remaining six chromosomes of each haploid chromosome complement cannot be reliably identified using arm ratio or relative length measurements.

Transfer to Wheat of Potentially New Stem Rust Resistance Genes from Aegilops speltoides

Stem rust resistance genes have been found in four different sources of Aegilops speltoides. These include diploid accessions AEG357-4 and AEG874-60 and the amphiploids Chinese Spring/Ae. speltoides TA8026 and TS01. Stem rust resistance was mapped to the 2S chromosomes derived from each of these lines. The previously reported 2B-2S#3 translocation derived from AEG357-4 was found to carry two stem rust resistance genes, here temporarily named SrAes2t and SrAes3t. The resistance genes found on the 2S chromosomes each derived from TA8026, TS01 and AEG874-60 are named SrAes4t, SrAes5t and SrAes6t, respectively. Lines carrying genes SrAes2t and SrAes3t are being distributed to wheat breeding programs around the world.