Cyto-molecular characterization of rDNA and chromatin composition in the NOR-associated satellite in Chestnut (Castanea spp.).

The American chestnut (Castanea dentata, 2n = 2x = 24), once known as the "King of the Appalachian Forest", was decimated by chestnut blight during the first half of the twentieth century by an invasive fungus (Cryphonectria parasitica). The Chinese chestnut (C. mollissima, 2n = 2x = 24), in contrast to American chestnut, is resistant to this blight. Efforts are being made to transfer this resistance to American chestnut through backcross breeding and genetic engineering. Both chestnut genomes have been genetically mapped and recently sequenced to facilitate gene discovery efforts aimed at assisting molecular breeding and genetic engineering. To complement and extend this genomic work, we analyzed the distribution and organization of their ribosomal DNAs (35S and 5S rDNA), and the chromatin composition of the nucleolus organizing region (NOR)-associated satellites. Using fluorescent in situ hybridization (FISH), we have identified two 35S (one major and one minor) and one 5S rDNA sites. The major 35S rDNA sites are terminal and sub-terminal in American and Chinese chestnuts, respectively, originating at the end of the short arm of the chromosome, extending through the secondary constriction and into the satellites. An additional 5S locus was identified in certain Chinese chestnut accessions, and it was linked distally to the major 35S site. The NOR-associated satellite in Chinese chestnut was found to comprise a proximal region packed with 35S rDNA and a distinct distal heterochromatic region. In contrast, the American chestnut satellite was relatively small and devoid of the distal heterochromatic region.

Sorghum bicolor x S. halepense interspecific hybridization is influenced by the frequency of 2n gametes in S. bicolor.

Tetraploid johnsongrass [Sorghum halepense (L.) Pers.] is a sexually-compatible weedy relative of diploid sorghum [Sorghum bicolor (L.) Moench]. To determine the extent of interspecific hybridization between male sterile grain sorghum and johnsongrass and the ploidy of their progeny, cytoplasmic (CMS), genetic (GMS) and chemically induced male sterile lines of Tx623 and Tx631 were pollinated with johnsongrass pollen. At maturity 1% and 0.07% of the developing seeds of Tx623 and Tx631 respectively were recovered. Ninety-one percent of recovered hybrids were tetraploid and two percent were triploid, the tetraploids resulting from 2n gametes present in the sorghum female parent. Their formation appears to be genotype dependent as more tetraploids were recovered from Tx623 than Tx631. Because a tetraploid sorghum x johnsongrass hybrid has a balanced genome, they are male and female fertile providing opportunities for gene flow between the two species. Given the differences in 2n gamete formation among Tx623 and Tx631, seed parent selection may be one way of reducing the likelihood of gene flow. These studies were conducted in controlled and optimum conditions; the actual outcrossing rate in natural conditions is expected to be much lower. More studies are needed to assess the rates of hybridization, fitness, and fertility of the progeny under field conditions.