Wang Yan, Wang Yiting, Li Kunfeng, Song Xijiao, Chen Jianping
State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Ministry of Agriculture Key Laboratory of Biotechnology in Plant Protection, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences Hangzhou, China.
Agriculture Experiment Station, Zhejiang University Hangzhou, China.
Front Plant Sci. 2016 Dec 22;7:1897. doi: 10.3389/fpls.2016.01897. eCollection 2016.
Plant browning is a recalcitrant problem for culture and often leads to poor growth of explants and even failure of tissue culture. However, the molecular mechanisms underlying browning-induced physiological processes remain unclear. is considered one of the most difficult genera for tissue culture owning to its severe browning. In the present study, intact aseptic plantlets of Hayata previously obtained by ovary culture, were used to explore the characteristics and molecular mechanism of the browning response. Successive morphological and anatomical observations after cutting showed that the browning of was not lethal but adaptive. transcriptome and digital gene expression (DGE) profiling using Illumina high-throughput sequencing were then used to explore molecular regulation after cutting. About 7.5 million tags of transcriptome were obtained and 58,073 unigenes were assembled and annotated. A total of 6,431 differentially expressed genes (DEGs) at three stages after cutting were identified, and the expression patterns of these browning-related genes were clustered and analyzed. A number of putative DEGs involved in signal transduction and secondary metabolism were particularly studied and the potential roles of these cutting-responsive mRNAs in plant defense to diverse abiotic stresses are discussed. The DGE profiling data were also validated by quantitative RT-PCR analysis. The data obtained in this study provide an excellent resource for unraveling the molecular mechanisms of browning processes during tissue culture, and lay a foundation for future studies to inhibit and eliminate browning damage.
植物褐变是培养过程中一个难以解决的问题，常常导致外植体生长不良甚至组织培养失败。然而，褐变诱导的生理过程背后的分子机制仍不清楚。由于其严重的褐变现象，[具体植物名称]被认为是组织培养中最难培养的属之一。在本研究中，使用先前通过子房培养获得的完整无菌[具体植物名称]组培苗，来探究褐变反应的特征和分子机制。切割后的连续形态学和解剖学观察表明，[具体植物名称]的褐变并非致命，而是一种适应性反应。随后利用Illumina高通量测序技术对[具体植物名称]进行转录组和数字基因表达（DGE）分析，以探究切割后的分子调控机制。获得了约750万个[具体植物名称]转录组标签，并组装和注释了58,073个单基因。共鉴定出切割后三个阶段的6,431个差异表达基因（DEG），并对这些与褐变相关基因的表达模式进行了聚类和分析。特别研究了一些参与信号转导和次生代谢的假定DEG，并讨论了这些切割响应mRNA在植物抵御多种非生物胁迫中的潜在作用。DGE分析数据也通过定量RT-PCR分析进行了验证。本研究获得的数据为揭示[具体植物名称]组织培养过程中褐变过程的分子机制提供了极好的资源，并为未来抑制和消除褐变损伤的研究奠定了基础。
