Efficacy of colchicine and trifluralin in creating in vitro autotetraploid Gaura lindheimeri engelm. and gray

Gaura lindheimeri is a diploid herbaceous perennial species native to Texas and Louisiana and winter hardy only to USDA hardiness zone 5. A potential source of winter hardiness is G. coccinea Pursh., a polyploid widely distributed in North America; of particular interest are autotetraploid populations of G. coccinea from Minnesota. To facilitate interspecific hybridization, a tetraploid G. lindheimeri would be advantageous. Two G. lindheimeri genotypes, MN selections 443-1 and 01G-02, were treated with two different antimitotic agents at two concentrations, trifluralin - 15 and 30 μM and colchicine - 0.25 and 1.25 mM, along with appropriate controls, to determine the frequency of chromosome doubling. Two-node stem sections were treated for 12, 24, or 48 h and then rooted and grown to flowering. Pollen diameter was measured as an indicator of chromosome doubling in cell layer LII, and morphologic characteristics (days to flower, flower size, plant height, inflorescence height, and plant width) were recorded for all plants. Chromosome doubling was not observed in any plant treated with trifluralin. Based on pollen diameter, genotype 443-1 only had chromosome doubling in the colchicine 1.25 mM concentration when treated for 12 h. All durations of colchicine at 1.25 mM were successful for genotype 01G-02 as well as a small percent treated with colchicine at 0.25 mM treated for 48 h. Autotetraploid plants (2n = 4x = 28) had larger flowers in both genotypes, and autotetraploid derivatives of genotype 01G-02 flowered earlier and were taller than diploid plants. Conformation changes from three-lobed to four-lobed pollen grains were observed when pollen diameter approached that expected of 2n pollen. Visual screening of pollen for conformation changes can quickly determine if chromosome doubling in cell layer LII has occurred. With the autotetraploid G. lindheimeri derived from colchicine application, crosses can be performed with autotetraploid G. coccinea to introgress cold tolerance. Additional breeding can also be done at the tetraploid level to develop new autotetraploid cultivars of G. lindheimeri.