Developing Adventitious Root Meristems Induced by Layering for Plant Chromosome Preparation

Chromosome numbers and morphology are important characteristics of a species and its evolution. Root tips are the most commonly used tissue as a source of actively dividing cells for chromosome visualization in plants. Previously, rapidly growing root tips were collected from germinating kernels or from seedlings growing in pots or fields. However, the use of adventitious roots (ARs) derived from aerial tissue as meristems for chromosome visualization has always been overlooked. Here, we successfully induced ARs in 12 materials that were investigated, with the exception of <i>Sorghum nitidum</i>. Using ARs meristem we obtained high-quality chromosome spreads for <i>Morus alba</i>, <i>Broussonetia papyrifera</i>, <i>Lolium multiflorum</i>, <i>Sorghum sudanense</i>, <i>S</i>. <i>propinquum</i>, <i>S</i>. <i>bicolor</i> × <i>S</i>. <i>sudanense</i>, <i>Zea mays</i>, <i>Z</i>. <i>mexicana</i>, <i>Glycine max</i>, <i>Medicago sativa</i>, and <i>Brassica napus</i>. The results reported here demonstrate that layering is an alternative and effective method for producing meristematic cells for high-quality chromosome preparation in plant species producing ARs. For species that produce ARs by layering, this protocol is particularly valuable for the development of cost-effective and high-throughput non-invasive cytogenetic studies.