异型花柱植物莕菜两型花间差异基因的克隆及表达分析

异型花柱是受遗传控制的花的多态现象，包括二型花柱和三型花柱。异型花柱是自然界中较为典型的植物性系统之一，也是进化生物学工作者研究植物进化和适应问题的经典范例。自达尔文以来，异型花柱的起源、演化及其遗传控制机制等方面一直是进化生物学研究领域的热点，但目前对异型花柱分子遗传基础却知之甚少。水生植物莕菜为典型的花柱二型性植物并具有诸多研究二型花柱分子遗传机制的有利条件，是研究该问题的理想材料。本项目拟在前期对莕菜不同花型转录组测序的基础之上，选择其长、短花型间差异显著的基因，并进行PCR扩增测序及荧光定量PCR验证其可靠性；同时对这些差异基因进行不同花型间不同组织器官中时空表达模式研究，确定长、短花柱型植株中特异表达的基因，并对这些基因进行分子克隆和结构分析；建立特异表达基因与莕菜不同花型形态建成间的联系，为后续全面深入揭示花柱二型性的分子遗传基础提供基础。

Heterostyly (including distyly and tristyly) is a genetic controlled floral polymorphism. Heterostyly is one of the classic sexual systems and has been a model system in the study of plant evolution and adaptation. Since Darwin, the origin, evolution and genetic mechanisms of heterostyly has been a hot spot of evolutionary biology, but little is known of the molecular genetic basis of the heterostyly. The aquatic plant Nymphoides peltata (Menyanthaceae) is a typical distylous plant and is an ideal material for studying the molecular genetic basis of the distyly. In the our previous research, the transcriptome sequencing experiments have been conducted on the materials from the long- and short-morph flower of N. peltata. Based on the transcriptome sequences, firstly in this project we are to validate gene fragments reliability by using PCR amplification and Real-time quantitative PCR techniques, and then to study the temporal and spatial expression patterns of these identified genes in different floral organs. After indentifying the genes showing differential expression between morphs in developing flowers of N. peltata, we then to clone and sequence these genes and to study the gene structure and protein function. The aims are to link the differential expressed genes to the floral morphogenesis, which could provide a basis for the subsequent in-depth study on the molecular genetic basis of distyly.