Cytogenetic and molecular markers for detectingAegilops uniaristatachromosomes in a wheat background

Abstract

Aegilops uniaristata has many agronomically useful traits that can be used for wheat breeding. So far, a Triticum turgidum - Ae. uniaristata amphiploid and one set of Chinese Spring (CS) - Ae. uniaristata addition lines have been produced. To guide Ae. uniaristata chromatin transformation from these lines into cultivated wheat through chromosome engineering, reliable cytogenetic and molecular markers specific for Ae. uniaristata chromosomes need to be developed. Standard C-banding shows that C-bands mainly exist in the centromeric regions of Ae. uniaristata but rarely at the distal ends. Fluorescence in situ hybridization (FISH) using (GAA)8 as a probe showed that the hybridization signal of chromosomes 1N-7N are different, thus (GAA)8 can be used to identify all Ae. uniaristata chromosomes in wheat background simultaneously. Moreover, a total of 42 molecular markers specific for Ae. uniaristata chromosomes were developed by screening expressed sequence tag - sequence tagged site (EST-STS), expressed sequence tag - simple sequence repeat (EST-SSR), and PCR-based landmark unique gene (PLUG) primers. The markers were subsequently localized using the CS - Ae. uniaristata addition lines and different wheat cultivars as controls. The cytogenetic and molecular markers developed herein will be helpful for screening and identifying wheat - Ae. uniaristata progeny.

Characterization of a New Wheat-Aegilops biuncialis1Mb(1B) Substitution Line with Good Quality-associated HMW Glutenin Subunit

“…Reliable alien chromosome-specific molecular markers not only are very useful for detecting alien chromatin in wheat background, but also enable us to screen and identify translocations and recombinants from large progenies (Gong et al 2014). Up-to-now, very few chromosome-specific molecular markers have been developed in Aegilops species (Gong et al 2014).…”

“…Up-to-now, very few chromosome-specific molecular markers have been developed in Aegilops species (Gong et al 2014). Moreover, only 2 simple sequence repeat (SSR) markers GWM44 and GDM61 (Schneider and Molnár-Láng 2012) and some gene-based conserved orthologous set (COS) markers (Molnár et al 2013) were obtained in Ae.…”

Molecular and Cytogenetic Characterization of a Powdery Mildew-Resistant Wheat-<b><i>Aegilops mutica</i></b> Partial Amphiploid and Addition Line

“…Although chromosome C-banding requires precise timing and temperature control and accurate concentrations of acid, alkali, and salt, it is a very useful tool for the identification of genetic stocks of wheat-alien species [Liu et al, 2011a, b;Gong et al, 2014]. Friebe et al [1996a] established the standard C-banding karyotype of Ae.…”

“…Markers specific for alien chromosomes have proven very convenient for tracing alien-derived chromatin in a wheat background [Liu et al, 2011a, b;Gong et al, 2014]. In the tribe Triticeae, many alien chromosome-specific markers have been developed, including those for Secale africanum [Liu et al, 2008], Dasypyrum breviaristatum [Yang et al, 2006], Ae.…”

“…A total of 153 pairs of expressed sequence tag-sequence tagged site (EST-STS) primers were synthesized. The primer design, PCR protocol, PCR product digestion, and gel separation followed Gong et al [2014].…”