The applicability of three series of EST-based-PCR primers of barley to ten wild species covering wide range of variation in Triticeae was examined in this study. The first series were randomly chosen from the pool of barley EST primers, while the other two series were pre-screened and showed polymorphic fragments between barley and wheat in the previous study. The percentage of primers that produced single clear band in the studied species ranged from 40 to 76%. The frequency of amplification and the polymorphism agreed well with the phylogenetic distance from barley. Number of the amplified polymorphic markers using the pre-screened primer sets was significantly higher than that of using the randomly chosen ones. A large number (79-572) of DNA markers for the different alien species used in this study were obtained from the 1,165 EST-based-PCR primers of barley examined. These markers will be valuable tool in identifying the alien chromosomes added to wheat genome.
P-17 Y. Saito1, M. Kishii2, T. Abe3,
H. Tanaka1, H. Tsujimoto1 (1Faculty of
Agriculture, Tottori University, 2CIMMYT, 3Riken)
Chromosome engineering in wheat by heavy ion beam irradiation
Triticeae species are genetic resources for the improvement of wheat and barley. We have introduced the chromosomes of wild species to common wheat. The next steps necessary to be solved are to find the location of the useful gene and introduce the chromosome segment to a wheat chromosome. Here we demonstrate heavy ion beam as a useful method to break chromosome. We irradiated nitrogen (N) and neon (Ne) ion beams to the dry seeds of common wheat cv. Chinese Spring (CS) and of the disomic addition line with the chromosome J of Leymus racemosus (Kishii et al. 2004). The chromosomes were observed by acetocarmine staining and gez֌nomic in situ hybridization to the chromosomes in the root tip cells. We found that LD50 after one month was 150 and 90 Gy in N and Ne, respectively. Chromosome observation by GISH revealed occurrence of about 20 chromosome breakage by irradiation of 30 Gy of Ne ion beam, which is eight times higher than the value observed by acetocarmine staining.
P-18 M. Ito, M. Kishii, A. A. Hagras, H. Tanaka,H. Tsujimoto
(Faculty of Agriculture, Tottori University)
Production of Psathyrostachys huashanica chromosome addition lines
and finding its use for breeding.
Psathyrostachys huashanica Keng (2n=2x=14,NhNh) is a perennial wild species distantly related to wheat. It is endemic in Huashan branches of Qinling mountains in China. It has yellowish green thin leaves and 40-60 cm in height, early-maturing, drought and poor soil tolerance and resistance to many diseases such as stem rust, leaf rust, black rust, and powdery mildew. It is also known to be resistance to damping-off that is one of the obstacles to continuous cropping of wheat. We crossed wheat (Triticum aestivum cv. Chinese Spring) with the pollen of P. huashanica and produced the chromosome addition lines. We selected monosomic addition line in all chromosomes and are selecting dlisomic chromosome addition lines from the selfed offspring. We selected P. huashanica-specific PCR markers from the barley EST markers developed by Sate et al. and identified the added chromosomes.