Genes mapping to boron tolerance QTL in barley identified by suppression subtractive hybridization

Abstract

Boron tolerance is a quantitative trait controlled by multiple genes. Suppression subtractive hybridization was carried out on root cDNA from bulked boron tolerant and intolerant doubled haploid barley lines grown under moderate boron stress to identify genes associated with boron tolerance. One hundred and eleven clones representing known proteins were found to be up-regulated in the tolerant bulk upon boron stress. Nine clones were genetically mapped to previously reported boron tolerance QTL. These include a clone identical to the boron transporter gene Bot1 and a clone coding for a bromo-adjacent homology domain-containing protein, mapping to the 6H boron tolerance locus and co-segregating with reduced boron intake in a Clipper x Sahara-3771 mapping population. A third clone mapping to the 2H QTL region encoding an S-adenosylmethionine decarboxylase precursor was found to provide tolerance to high boron by heterologous expression. Yeast cells expressing Sahara SAMDC were able to grow on 15 mm boron solid media and maintained cellular boron concentrations at 13% lower than control cells expressing empty vector. The data suggest that an antioxidative response mechanism involving polyamines and the ascorbate-glutathione pathway in Sahara barley may provide an advantage in tolerating high soil concentrations of boron.