A-genome Arachis species (AA; 2n = 2x = 20) are commonly used as secondary germplasm sources in cultivated peanut breeding, Arachis hypogaea L. (AABB; 2n = 4x = 40), for the introgression of various biotic and abiotic stress resistance genes. Genome doubling is critical to overcoming the hybridization barrier of infertility that arises from ploidy-level differences between wild germplasm and cultivated peanuts. To develop improved genome doubling methods, four trials of various concentrations of the mitotic inhibitor treatments colchicine, oryzalin, and trifluralin were tested on the seedlings and seeds of three A-genome species, A. cardenasii, A. correntina, and A. diogoi. A total of 494 seeds/seedlings were treated in the present four trials, with trials 1 to 3 including different concentrations of the three chemical treatments on seedlings, and trial 4 focusing on the treatment period of 5 mM colchicine solution treatment of seeds. A small number of tetraploids were produced from the colchicine and oryzalin gel treatments of seedlings, but all these tetraploid seedlings reverted to diploid or mixoploid states within six months of treatment. In contrast, the 6-h colchicine solution treatment of seeds showed the highest tetraploid conversion rate (6–13% of total treated seeds or 25–40% of surviving seedlings), and the tetraploid plants were repeatedly tested as stable tetraploids. In addition, visibly and statistically larger leaves and flowers were produced by the tetraploid versions of these three species compared to their diploid versions. As a result, stable tetraploid plants of each A-genome species were produced, and a 5 mM colchicine seed treatment is recommended for A-genome and related wild Arachis species genome doubling.
Lindera benzoin is a dioecious understory shrub native to eastern North America. Northern spicebush is a beautiful shrub with a natural round shrub shape, golden-yellow fall foliage, attractive bright red drupes, and precocious yellow flowers in early spring; however, its market value as an ornamental value has been overlooked. To improve the ornamental values of this under-cultivated nursery crop, breeding for a better compact form, larger leaves, enlarged flower clusters and fruit, and increased stress tolerances could all be beneficial. Polyploidy manipulation is a valuable method to improve such traits for many ornamental plants. This study established the genome doubling method by oryzalin-infused solid agar treatment on young northern spicebush seedlings. The seedlings of two wild populations in North Carolina were collected and used. A total of 288 seedlings were treated with solid agar containing 150 µM oryzalin for 24, 72, and 120 h. The results were sporadic in their survival ratios and tetraploid conversion ratios between different treatments; however, a total of 16 tetraploid L. benzoin plants were produced in this study. The 24-h treatment showed the optimal result, with 7.1% of total treated seedlings or 15.2% of surviving seedlings converted into tetraploids. Tetraploid plants had visible differences in leaf morphology, a statistically significant enlarged stomata size, and reduced stomatal density compared to diploid plants. This research provides ploidy manipulation information for all future breeding processes of L. benzoin and related species.