Ferreres Irene, Ortega Mirari, López-Cristoffanini Camilo, Nogués Salvador, Serrat Xavier
Department of Evolutionary Biology, Ecology and Environmental Sciences, Faculty of Biology, University of Barcelona, 08028 Barcelona, Spain.
Plant Biotechnol (Tokyo). 2019 Dec 25;36(4):269-273. doi: 10.5511/plantbiotechnology.19.1022a.
Anther culture is a fast tool to obtain double haploid plant lines for breeding purposes. In rice, this procedure is commonly performed in two steps: i) induction of calli from anthers and ii) regeneration of plantlets from calli. It has been stated that genotype highly influences the anther culture efficiency, so the media used in each step should be optimized for each variety. In this study, we tested different media modifications of an efficient protocol optimized for a medium sized grain temperate japonica NRVC980385, used as a control, in a long grain temperate japonica rice variety (NRVC20120346), and two long grain tropical japonica varieties (303012 and 303013). We found that the addition of 150 mg l colchicine to the induction medium worked best for all genotypes except for NRVC20120346, whose best induction was obtained with the colchicine-free medium. Referring to regeneration, increased gelling agent in the medium provided the best rates in NRVC980385, improving our former NRVC980385-optimized anther culture protocol. Sorbitol fortified regeneration medium worked the best in the case of the long grain varieties. The presence of colchicine in the induction medium was also related to a higher obtention of double haploid plantlets. This study highlights that genotype is a key factor in the performance of rice anther culture. It has set a first anther culture study on long grain japonica varieties and optimizes the anther culture protocol for temperate japonica medium grain NRVC980385 with the use of colchicine and other additives that increase osmotic stress.
花药培养是一种用于育种目的获取双单倍体株系的快速方法。在水稻中，该过程通常分两步进行：i）从花药诱导愈伤组织；ii）从愈伤组织再生植株。据报道，基因型对花药培养效率有很大影响，因此每个步骤所使用的培养基应针对每个品种进行优化。在本研究中，我们测试了针对中等粒型温带粳稻NRVC980385优化的高效方案的不同培养基改良方法，将其用作对照，应用于长粒型温带粳稻品种（NRVC20120346）以及两个长粒型热带粳稻品种（303012和303013）。我们发现，除NRVC20120346外，在诱导培养基中添加150 mg/l秋水仙碱对所有基因型效果最佳，而NRVC20120346在无秋水仙碱的培养基中诱导效果最佳。关于再生，培养基中增加胶凝剂在NRVC980385中提供了最佳比率，改进了我们之前针对NRVC980385优化的花药培养方案。在长粒型品种的情况下，添加山梨醇的再生培养基效果最佳。诱导培养基中秋水仙碱的存在也与更高的双单倍体植株获得率相关。本研究强调基因型是水稻花药培养表现的关键因素。它首次对长粒型粳稻品种进行了花药培养研究，并通过使用秋水仙碱和其他增加渗透胁迫的添加剂，优化了针对温带粳稻中等粒型NRVC980385的花药培养方案。
