Development of EST-PCR markers specific to the long arm of chromosome 6V of Dasypyrum villosum

Expressed sequence tags-derived polymerase chain reaction(EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6 VL of Dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags(EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6 AL, 6 BL, and 6 DL. By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6 V, it was found that 32(10.77%) primers could amplify specific bands for chromosome 6 V, and 31 could be allocated to chromosome arm 6 VL. These 6 VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6 VL in common wheat background as well as for the selection of useful genes located on 6 VL in breeding programs.

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.

Agronomic characterization and genetic analysis of the supernume-r ary spikelet in tetraploid wheat(Triticum turgidum L.)

The supernumerary spikelets（SS）characters of tetraploid wheat（Triticum turgidum L.）resulting in more spikelets and kernels per spike,thus enhancing sink capacity may contribute to potential wheat yield improvement.In order to investigate the effect of different SS types on agronomic characters and understand the genetic base of SS phenotype in tetraploid wheat,near isogenic lines（NILs）,bh-50 with normal spikelets（NS）,bh-51 with four-rowed spikelets（FRS）,bh-52 with short-ramified spikelets（SRS）,and bh-53 with long-ramified spikelets（LRS）in a Triticum durum cv.ZY1286 genetic background were developed by continuous backcrossing.Agronomic characters showed that the SS phenotype lines,bh-51,bh-52and bh-53 have significant increase in the number of spikelets and grains per spike compared with the NS phenotype line bh-50（P〈0.05）,and bh-53 line showed much more increase than those of bh-51 and bh-52.However,bh-53 had the lowest grain weight and the longest spike development stage than those of other spike phenotypes.These results indicated that the different SS types have different effects on the agronomic and spike characters.Genetic analysis through bh-50/bh-51and bh-51/bh-53 F2 populations showed that a recessive major gene controlled the spike architecture to transform from NS to FRS,and a dominant major gene determined the change of spike phenotype from FRS to RS.DNA sequences of Tt BH/WFZP ortholog on chromosome 2AS revealed that a single nucleotide polymorphism（SNP）substitution happened in the open reading frame（ORF）region of all the SS tetraploid wheat accessions,which may lead to the generation of lateral meristems between glume and lemma during the immature spike development.While the fates of the lateral meristems,developing into lateral spikelets or branched spikelets,may be determined by another major gene.Our results presented here may advance our understanding and knowledge of the genes and genetic pathways determining the spike architecture development in wheat.