德氏兜兰和亨利兜兰成花诱导机制初探

	德氏兜兰和亨利兜兰成花诱导机制初探
	熊进
导师	张石宝
关键词	兜兰 Paphiopedilum 成花启动 Floral inducion 成花诱导 Flower evocation 温度 Temperature
摘要	兜兰属（Paphiopedilum）为兰科植物中极具特色的一个类群，其花型别致、色系丰富、花期较长，是最具市场开发前景的兰花。然而，兜兰属植物开花调控机制不清，开花时间难以精确控制是限制其规模化栽培的重要因素。因此，开展兜兰属植物成花诱导的关键调控途径和主要调控因素的研究有十分重要的意义。兜兰属植物的自然花期多在春季（即春花型）或秋季（秋花型）。本论文以春季开花的德氏兜兰（P. delenatii）和秋季开花的亨利兜兰（P. henryanum）分别作为春花型和秋花型的代表，通过对其顶芽进行形态解剖，观察其成花启动情况以判断是否发生成花诱导，再将成花启动前后的植株叶片分别进行转录组测序，分析筛找参与两种兜兰成花诱导的关键基因和可能代谢途径。此外，课题组前期研究表明，温度可能是影响兜兰属植物在不同季节开花的主要环境因素。因此，本论文还通过温度控制实验，研究德氏兜兰在不同温度培养条件下的成花启动情况。同时，对不同温度处理条件下的德氏兜兰植株叶片进行转录组测序，分析参与温度调控德氏兜兰成花诱导的基因和代谢途径。主要研究结果如下：（1）通过对德氏兜兰和亨利兜兰的顶芽解剖，发现同一培养条件下的德氏兜兰和亨利兜兰成花启动时间几乎一致，都发生于4月中旬。（2）通过对德氏兜兰和亨利兜兰成花启动前后叶片进行转录组测序分析发现，德氏兜兰和亨利兜兰在成花启动前后特有的DEGs分别有1365个和7049个，两种兜兰共有的DEGs共73个。对它们分别进行KEGG富集分析发现，共有的73个DEGs显著富集于“淀粉和糖代谢”、“氮代谢”和“植物昼夜节律”等途径；德氏兜兰特有的DEGs显著富集于“内质网中的蛋白质加工”、“植物激素信号转导”和“植物MAPK信号通路”等途径；亨利兜兰特有的DEGs显著富集到“光合作用天线蛋白”、“碳代谢”和“光合生物的固碳作用”等途径，暗示两种兜兰成花诱导机制有所差异。结合前人相关研究，在成花启动前后检测到的所有DEGs中，共筛选到了70个与成花诱导相关的基因和转录因子编码基因。通过对所有成花诱导相关基因表达模式分析发现，CO-like 13、GI和SOC1等基因在两种兜兰成花启动期表达都上调，而CO-like 1、LF-like和TCP3等基因都下调，暗示它们可能在兜兰成花中具有重要作用。（3）通过解剖观察不同温度培养条件下的德氏兜兰成花启动情况发现，相对平均温度为15/10℃（白天/夜晚）（T1）的温室培养条件，适当升温的21/18℃（T2）能促进德氏兜兰发生成花诱导和成花启动，而较高的温度28/25℃（T3）则抑制德氏兜兰成花诱导和成花启动。（4）通过对不同温度培养条件下的德氏兜兰成花启动前后两个时期的叶片进行转录组测序分析，将已发生成花启动的植株（T1-S0和T2-S0）与未发生成花启动的植株（T3-S0）转录组数据对比发现，T1-S0 vs T3-S0的DEGs有404个，T2-S0 vs T3-S0的DEGs有558个。通过Venn分析发现，T1-S0 vs T3-S0与T2-S0 vs T3-S0的共有的DEGs共有118个。对这118个基因进行KEGG富集分析发现，它们显著富集于“植物激素信号转导”、“植物昼夜节律”、“植物MAPK信号通路”和“光合作用”等与成花诱导相关的途径。在所有响应温度的DEGs中，共筛选到了57个与成花诱导相关的基因和转录因子编码基因，CO-like 13、SOC1和bZIP39等基因在成花启动期表达上调，且它们在T2-S0中的表达量高于T1-S0中，而CO-like 1、CO-like 9和CO-like 10等基因在成花启动前后表达都下调，且它们在T2-S0中的表达量低于T1-S0中，暗示温度可能是通过影响这些基因与途径参与德氏兜兰的成花诱导。综上所述，本论文发现春花型德氏兜兰和秋花型亨利兜兰在同一培养条件下的成花诱导和成花启动时间几乎一致，但它们的成花诱导机制可能有所差异。通过对不同温度培养条件下德氏兜兰的成花启动情况观察发现，适当升温可使德氏兜兰成花诱导和成花启动提前。通过本研究，旨在认识兜兰属植物的成花诱导机制以及温度是通过何种途径和基因参与德氏兜兰的成花诱导，为精确调控兜兰属植物花期提供理论依据。; The genus Paphiopedilum is world-famous orchid with unique flower patterns, colorful flower types and long flowering period, and with high commercial value. However, the lacking of flowering regulation mechanism, and the inaccurate control of flowering time are important factors limiting the large-scale cultivation of Paphiopedilum. Therefore, it is of great significance to investigate the key regulatory pathways and main regulatory factors of the floral induction of Paphiopedilum species. The natural flowering period of Paphiopedilum species is mostly in spring (referred to as spring flowering type) or autumn (autumn flowering type). In this study, the spring flowering P. delenatii and the autumn flowering P. henryanum were used as the representatives of spring and autumn flowering types, respectively. The floral evocation condition was used to determine whether floral induction occurred. Then, the leaves of the plants before and after the floral evocation were used in RNA-sequencing to screen key genes and possible metabolic pathways involved in the flower induction of the two kinds of Paphipedium species, laying the foundation for the floral induction mechanism. In addition, the previous research showed that temperature may be a key environmental factor that affects the flowering of Paphiopedilum species in different seasons. Therefore, we also used temperature-controlling experiments to study the initiation of floral evocation under different temperature cultivation conditions. Meanwhile, RNA-sequencing was performed on the leaves of P. delenatii under different temperature treatments to screen key genes and pathways involved in the temperature-induced floral induction of P. delenatii. The main results are as follows: (1)Anatomical observation of the apical buds of P. delenatii and P. henryanum showed that the floral evocation time of P. delenatii and P. henryanum were all in mid-April under the same culture condition. (2)RNA-sequencing analysis of the leaves before and after the floral evocation of P. delenatii and P. henryanum showed that there were 1365 and 7049 unique differentially expressed genes (DEGs) in P. delenatii and P. henryanum, respectively. Taken together, there are 73 shared DEGs in the two species. KEGG enrichment analysis showed that these 73 shared DEGs were significantly enriched in "Starch and sucrose metabolism", "Nitrogen metabolism" and "Circadian rhythm-plant". P. delenatii specific DEGs were significantly enriched in "Protein processing in endoplasmic reticulum", "Plant hormone signal transduction" and "MAPK signaling pathway-plant". P. henryanum specific DEGs were significantly enriched in "Photosynthesis-antenna proteins", "Carbon metabolism", and "Carbon fixation in photosynthetic organisms". These results indicated that the mechanisms of floral evocation may be different in these two Paphiopedilum species. According to the previous study, 70 flowering related genes and transcription factor encoding genes were se
语种	中文
	2022-05
学位授予单位	中国科学院大学
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/75146
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	熊进. 德氏兜兰和亨利兜兰成花诱导机制初探[D]. 中国科学院大学,2022.

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