An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars

作者：Khirod Kumar Sahoo, Amit K. Tripathi, Ashwani Pareek, Sudhir Kumar Sopory, Sneh Lata Singla-Pareek · 发表于：Plant Methods · 年份：2011 · DOI：10.1186/1746-4811-7-49 · 被引用次数：226 · 研究领域：Plant tissue culture and regeneration、Plant Molecular Biology Research、Genetic Mapping and Diversity in Plants and Animals

BACKGROUND: Rice genome sequencing projects have generated remarkable amount of information about genes and genome architecture having tremendous potential to be utilized in both basic and applied research. Success in transgenics is paving the way for preparing a road map of functional genomics which is expected to correlate action of a gene to a trait in cellular and organismal context. However, the lack of a simple and efficient method for transformation and regeneration is a major constraint for such studies in this important cereal crop. RESULTS: In the present study, we have developed an easy, rapid and highly efficient transformation and regeneration protocol using mature seeds as explants and found its successful applicability to a choice of elite indica rice genotypes. We have optimized various steps of transformation and standardized different components of the regeneration medium including growth hormones and the gelling agent. The modified regeneration medium triggers production of large number of shoots from smaller number of calli and promotes their faster growth, hence significantly advantageous over the existing protocols where the regeneration step requires maximum time. Using this protocol, significantly higher transformation efficiency (up to 46%) and regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli) were achieved for the four tested cultivars. We have used this protocol to produce hundreds of independent transg...