Salvo Stella A G D, Hirsch Candice N, Buell C Robin, Kaeppler Shawn M, Kaeppler Heidi F
Department of Agronomy, University of Wisconsin, Madison, Wisconsin, United States of America.
Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, United States of America.
PLoS One. 2014 Oct 30;9(10):e111407. doi: 10.1371/journal.pone.0111407. eCollection 2014.
Embryogenic tissue culture systems are utilized in propagation and genetic engineering of crop plants, but applications are limited by genotype-dependent culture response. To date, few genes necessary for embryogenic callus formation have been identified or characterized. The goal of this research was to enhance our understanding of gene expression during maize embryogenic tissue culture initiation. In this study, we highlight the expression of candidate genes that have been previously regarded in the literature as having important roles in somatic embryogenesis. We utilized RNA based sequencing (RNA-seq) to characterize the transcriptome of immature embryo explants of the highly embryogenic and regenerable maize genotype A188 at 0, 24, 36, 48, and 72 hours after placement of explants on tissue culture initiation medium. Genes annotated as functioning in stress response, such as glutathione-S-transferases and germin-like proteins, and genes involved with hormone transport, such as PINFORMED, increased in expression over 8-fold in the study. Maize genes with high sequence similarity to genes previously described in the initiation of embryogenic cultures, such as transcription factors BABY BOOM, LEAFY COTYLEDON, and AGAMOUS, and important receptor-like kinases such as SOMATIC EMBRYOGENESIS RECEPTOR LIKE KINASES and CLAVATA, were also expressed in this time course study. By combining results from whole genome transcriptome analysis with an in depth review of key genes that play a role in the onset of embryogenesis, we propose a model of coordinated expression of somatic embryogenesis-related genes, providing an improved understanding of genomic factors involved in the early steps of embryogenic culture initiation in maize and other plant species.
胚性组织培养系统用于作物的繁殖和基因工程,但应用受到基因型依赖的培养反应的限制。迄今为止,很少有胚性愈伤组织形成所必需的基因被鉴定或表征。本研究的目的是增进我们对玉米胚性组织培养起始过程中基因表达的理解。在本研究中,我们着重介绍了文献中先前认为在体细胞胚胎发生中起重要作用的候选基因的表达。我们利用基于RNA的测序(RNA-seq)来表征高度胚性且可再生的玉米基因型A188未成熟胚外植体在置于组织培养起始培养基后0、24、36、48和72小时的转录组。注释为在应激反应中起作用的基因,如谷胱甘肽-S-转移酶和类萌发素蛋白,以及与激素转运有关的基因,如PINFORMED,在研究中表达增加了8倍以上。与先前在胚性培养起始中描述的基因具有高度序列相似性的玉米基因,如转录因子BABY BOOM、LEAFY COTYLEDON和AGAMOUS,以及重要的类受体激酶,如体细胞胚胎发生受体样激酶和CLAVATA,也在这个时间进程研究中表达。通过将全基因组转录组分析结果与对在胚胎发生起始中起作用的关键基因的深入综述相结合,我们提出了一个体细胞胚胎发生相关基因协调表达的模型,从而更好地理解了参与玉米和其他植物物种胚性培养起始早期步骤的基因组因素。