Genotype Screening of Recipient Resources with High Regeneration and Transformation Efficiency in Chrysanthemum
Genotype Screening of Recipient Resources with High Regeneration and Transformation Efficiency in Chrysanthemum
复制标题
菊花高再生转化效率受体资源基因型筛选
DOI：
10.32604/phyton.2022.018659
复制
发表时间：
2022-01-01
影响因子：
1.7
通讯作者：
Huang, He
中科院分区：
文献类型：
--
作者：
Li, Yajun;Cui, Yumeng;Huang, He
菊花遗传转化是菊花分子育种的关键步骤之一，而菊花的遗传转化依赖于一个优化的再生和转化体系。但不同菊花品种的再生体系各不相同，大多数品种的再生时间较长。为筛选高效再生品种，以8个不同花色和花型的菊花品种的叶片和茎尖薄细胞层（tTCL）为材料，在MS +6-BA（1.0-5.0 mg L ~（-1））+NAA（0.1-1.0 mg L ~（-1））培养基上进行培养。结果表明：叶片外植体在MS + 6-BA 1.0 mg L-1 + NAA 0.5 mg L-1培养基上再生效果最好; tTCL外植体在MS + 6-BA 5.0 mg L-1 + NAA 0.1 mg L-1培养基上再生效果最好。在此基础上，对13个菊花品种进行了再生筛选，最终从21个品种中筛选出3个再生效率较高的品种。其中C1的再生效率最高，叶片外植体培养42 d再生率达80.0%，tTCL外植体培养31 d再生率达100%。此外，我们还建立了C1的转化体系：预培养1d，用OD_（600）= 0.6的农杆菌悬浮液侵染10 min，在含有350 mg L ~（-1）羧苄青霉素和10 mg L ~（-1）卡那霉素的再生培养基中培养，最终获得4.0%的转化率。本研究筛选出了一个具有高效再生和转化体系的菊花新品种，为丰富菊花转基因植株受体的花色品种和花色或花型相关基因的功能研究奠定了基础。
Genetic transformation is one of the key steps in the molecular breeding of chrysanthemum, which relies on an optimal regeneration and transformation system. However, the regeneration system of different chrysanthemum cultivars varies, and the regeneration time of most cultivars is long. To screen cultivars with highly efficient regeneration, leaves and shoot tip thin cell layers (tTCL) from eight chrysanthemum cultivars with different flower colors and flower types were cultured on Murashige and Skoog media (MS) supplemented with 1.0-5.0 mg L-1 6-benzylaminopurine (6-BA) and 0.1-1.0 mg L-1 alpha-naphthaleneacetic (NAA). The results showed that the most efficient regeneration media were MS + 6-BA 1.0 mg L-1 + NAA 0.5 mg L-1 for leaf explants and MS + 6-BA 5.0 mg L-1 + NAA 0.1 mg L-1 for tTCL explants. Subsequently, another 13 chrysanthemum cultivars were screened by using the media, and finally, three cultivars with high regeneration efficiency were obtained from 21 cultivars. Among these, C1 had the highest regeneration efficiency: the regeneration rate of leaf explants reached 80.0% after 42 days of culture, and the regeneration rate of tTCL explants reached 100% after 31 days of culture. Furthermore, we also established the transformation system for C1 as follows: preculturing for one day, infecting with Agrobacterium suspension (OD600 = 0.6) for 10 min, and cultivating in the regeneration medium with 350 mg L-1 carbenicillin and 10 mg L-1 kanamycin, thus ultimately achieving a transformation rate of 4.0%. In this study, a new chrysanthemum cultivar with an efficient regeneration and transformation system was screened, which is beneficial to enrich the flower color of chrysanthemum transgenic plant recipients and to the functional research of flower color or type-related genes.