New wheat-rye 5DS-4RS·4RL and 4RS-5DS·5DL translocation lines with powdery mildew resistance.

Powdery mildew is one of the serious diseases of wheat (Triticum aestivum L., 2 n = 6 × = 42, genomes AABBDD). Rye (Secale cereale L., 2 n = 2 × = 14, genome RR) offers a rich reservoir of powdery mildew resistant genes for wheat breeding program. However, extensive use of these resistant genes may render them susceptible to new pathogen races because of co-evolution of host and pathogen. Therefore, the continuous exploration of new powdery mildew resistant genes is important to wheat breeding program. In the present study, we identified several wheat-rye addition lines from the progeny of T. aestivum L. Mianyang11 × S. cereale L. Kustro, i.e., monosomic addition lines of the rye chromosomes 4R and 6R; a disomic addition line of 6R; and monotelosomic or ditelosomic addition lines of the long arms of rye chromosomes 4R (4 RL) and 6R (6 RL). All these lines displayed immunity to powdery mildew. Thus, we concluded that both the 4 RL and 6 RL arms of Kustro contain powdery mildew resistant genes. It is the first time to discover that 4 RL arm carries powdery mildew resistant gene. Additionally, wheat lines containing new wheat-rye translocation chromosomes were also obtained: these lines retained a short arm of wheat chromosome 5D (5 DS) on which rye chromosome 4R was fused through the short arm 4 RS (designated 5 DS-4 RS · 4 RL; 4 RL stands for the long arm of rye chromosome 4R); or they had an extra short arm of rye chromosome 4R (4 RS) that was attached to the short arm of wheat chromosome 5D (5 DS) (designated 4 RS-5 DS · 5 DL; 5 DL stands for the long arm of wheat chromosome 5D). These two translocation chromosomes could be transmitted to next generation stably, and the wheat lines containing 5 DS-4 RS · 4 RL chromosome also displayed immunity to powdery mildew. The materials obtained in this study can be used for wheat powdery mildew resistant breeding program.

Fu S ，Ren Z ，Chen X ，Yan B ，Tan F ，Fu T ，Tang Z ... -  《-》

Development of Novel Wheat-Rye Chromosome 4R Translocations and Assignment of Their Powdery Mildew Resistance.

Rye ( L.) is an important gene donor for wheat improvement because of its many valuable traits, especially disease resistance. Development of novel wheat-rye translocations with disease resistance can contribute to transferring resistance into common wheat. In a previous study, a wheat-rye T4BL·4RL and T7AS·4RS translocation line (WR41-1) was developed by distant hybridization, and it was speculated that its resistance to powdery mildew, caused by f. sp. (), was derived from rye based on pedigree analysis. To make accurate use of chromosome 4R in wheat improvement, a set of new 4R translocations involving different arm translocations (e.g., 4RS monosomic, 4RL monosomic, 4RL disomic, 4RS monosomic plus 4RL monosomic, 4RS monosomic plus 4RL disomic, and 4RS disomic plus 4RL disomic translocations) was developed from crosses with common wheat. Those translocations were characterized by genomic in situ hybridization and expressed sequence tag simple sequence repeat marker analysis. To confirm the source of powdery mildew resistance, the translocation plants were tested against isolate E09. The results indicated that all translocations with 4RL were resistant at all tested growth stages, whereas those with only 4RS translocation or no alien translocation were susceptible. This further indicated that the powdery mildew resistance of WR41-1 was derived from the alien chromosome arm 4RL. To effectively use 4RL resistance in wheat improvement, two competitive allele-specific PCR markers specific for chromosome arm 4RL were developed to detect the alien chromosome in the wheat genome. These new translocation lines with diagnostic markers can efficiently serve as important bridges for wheat improvement.

Ma P ，Han G ，Zheng Q ，Liu S ，Han F ，Wang J ，Luo Q ，An D ... -  《PLANT DISEASE》

Using the 6RL(Ku) Minichromosome of Rye (Secale cereale L.) to Create Wheat-Rye 6D/6RL(Ku) Small Segment Translocation Lines with Powdery Mildew Resistance.

Long arms of rye ( L.) chromosome 6 (6RL) carry powdery mildew resistance genes. However, these sources of resistance have not yet been successfully used in commercial wheat cultivars. The development of small segment translocation chromosomes carrying resistance may result in lines carrying the 6R chromosome becoming more commercially acceptable. However, no wheat-rye 6RL small segment translocation line with powdery mildew resistance has been reported. In this study, a wheat-rye 6RL minichromosome addition line with powdery mildew resistance was identified, and this minichromosome was derived from the segment between L2.5 and L2.8 of the 6RL chromosome arm. Following irradiation, the 6RL minichromosome divided into two smaller segments, named 6RL and 6RL, and these fragments participated in the formation of wheat-rye small segment translocation chromosomes 6DS/6RL and 6DL/6RL, respectively. The powdery mildew resistance gene was found to be located on the 6RL segment. Sixteen 6RL-specific markers were developed, and their products were cloned and sequenced. Nucleotide BLAST searches indicated that 14 of the 16 sequences had 91⁻100% similarity with nine scaffolds derived from 6R chromosome of L. Lo7. The small segment translocation chromosome 6DL/6RL makes the practical utilization in agriculture of powdery mildew resistance gene on 6RL more likely. The nine scaffolds are useful for further studying the structure and function of this small segment.

Du H ，Tang Z ，Duan Q ，Tang S ，Fu S ... -  《INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES》

Development and molecular cytogenetic identification of a new wheat-rye 4R chromosome disomic addition line with resistances to powdery mildew, stripe rust and sharp eyespot.

A wheat-rye 4R chromosome disomic addition line with resistances to powdery mildew, stripe rust, sharp eyespot and high kernel number per spike was developed and characterized by molecular cytogenetic method as novel resistant germplasm. Rye (Secale cereale L.), a close relative of common wheat, is an important and valuable gene donor with multiple disease resistance for wheat improvement. However, resistance genes derived from rye have successively lost resistance to pathogens due to the coevolution of pathogen virulence and host resistance. Development and identification of new effective resistance gene sources from rye therefore are of special importance and urgency. In the present study, a wheat-rye line WR35 was produced through distant hybridization, embryo rescue culture, chromosome doubling and backcrossing. WR35 was then proven to be a new wheat-rye 4R disomic addition line using sequential GISH (genomic in situ hybridization), mc-FISH (multicolor fluorescence in situ hybridization) and ND-FISH (non-denaturing FISH) with multiple probes, mc-GISH (multicolor GISH), rye chromosome arm-specific marker analysis and SLAF-seq (specific-locus amplified fragment sequencing) analysis. At the adult stage, WR35 exhibited high levels of resistance to the powdery mildew (Blumeria graminis f. sp. tritici, Bgt) and stripe rust (Puccinia striiformis f. sp. tritici, Pst) pathogens prevalent in China, and a highly virulent isolate of Rhizoctonia cerealis, the cause of wheat sharp eyespot. At the seedling stage, it was highly resistant to 22 of 23 Bgt isolates and four Pst races. Based on its disease responses to different pathogen isolates, WR35 may possess resistance gene(s) for powdery mildew, stripe rust and sharp eyespot, which differed from the known resistance genes from rye. In addition, WR35 was cytologically stable and produced high kernel number per spike. Therefore, WR35 with multi-disease resistances and desirable agronomic traits should serve as a promising bridging parent for wheat chromosome engineering breeding.

An D ，Ma P ，Zheng Q ，Fu S ，Li L ，Han F ，Han G ，Wang J ，Xu Y ，Jin Y ，Luo Q ，Zhang X ... -  《-》

Molecular dissection of Secale africanum chromosome 6R(afr) in wheat enabled localization of genes for resistance to powdery mildew and stripe rust.

Introgression of chromatin from Secale species into common wheat has for decades been a successful strategy for controlling the wheat diseases. The wild Secale species, Secale africanum Stapf., is a valuable source for resistance to foliar disease of wheat. A wheat-S. africanum chromosome 6R substitution line displayed resistance to both powdery mildew and stripe rust at the adult-plant stage. Wheat-S. africanum chromosome 6R deletion and translocation lines were produced and identified by sequential non-denaturing fluorescence in situ hybridization (ND-FISH) using multiple Oligo-based probes. Different ND-FISH patterns were observed between S. cereale 6R and S. africanum 6R. With reference to the physical map of the draft genome sequence of rye inbred line Lo7, a comprehensive PCR marker analysis indicated that insertions and deletions had occurred by random exchange between chromosomes 6R and 6R. A survey of the wheat- S. africanum 6R lines for disease resistance indicated that a powdery mildew resistance gene(s) was present on the long arm of 6R at FL0.85-1.00, and that a stripe rust resistance gene(s) was located in the terminal region of 6RS at FL0.95-1.00. The wheat-S. africanum 6R introgression lines also displayed superior agronomic traits, indicating that the chromosome 6R may have little linkage drag in the wheat background. The combination of molecular and cytogenetic methods allowed to precisely identify the chromosome rearrangements in wheat- S. africanum 6R substitution, deletion and translocation lines, and compare the structural difference between chromosomes 6R and 6R. The wheat- S. africanum 6R lines containing gene(s) for powdery mildew and stripe rust resistance could be used as novel germplasm for wheat breeding by chromosome engineering.

Li G ，Tang L ，Yin Y ，Zhang A ，Yu Z ，Yang E ，Tang Z ，Fu S ，Yang Z ... -  《BMC PLANT BIOLOGY》