基于基因多态性、蛋白质组学和化学活性成分研究益智遗传多样性

With wild Alpinia oxyphylla Mique resources have been severely damaged and efficacy of cultivar medicinal also began to degenerate, investigate and collect Alpinia oxyphylla Mique of different geographical populations and genetic diversity of germplasm resources is particularly important. This study was based on polymorphism, proteomics and chemical active substances to study the genetic diversity of Alpinia oxyphylla Mique. First, ITS and AFLP molecular markers were used to analyze genomic DNA polymorphism of Alpinia oxyphylla Mique in different geographical populations and phylogenetic tree was constructed and screened to obtain a representative Alpinia oxyphylla Mique core germplasm bank. Secondly, the two-dimensional electrophoresis combined with MALDI-TOF MS technique was used to study Alpinia oxyphylla Mique core germplasm and get proteomics data, to analyze the existence of genetic differences at the protein level. Finally, the data of chemical active substances were used to explore to affect the gene and protein of Alpinia oxyphylla Mique core germplasm quality. Systems biology clustering method was used to process the data of polymorphism, proteomics and chemical active substances, to study Alpinia oxyphylla Mique of genetic diversity, the genetic diversity of Alpinia oxyphylla Mique was to understand at gene and protein levels, to explore the genetic background of different geographical populations of Alpinia oxyphylla Mique quality differences, clarify genetic diversity, to explore the genetic background of different geographical populations of educational quality differences, genuineness of the formation mechanism of Alpinia oxyphylla Mique resources is clarified, to provide a scientific basis for the protection, identification and evaluation of Alpinia oxyphylla Mique core germplasm resources.

随着野生益智资源不断受到严重破坏以及栽培品种药材功效的逐渐退化，调查和收集不同地理居群的益智种质资源，研究其遗传多样性尤为重要。本研究基于基因多态性、蛋白质组学和化学活性成分相关联的三位一体技术来研究益智遗传多样性。首先利用ITS和AFLP分子标记技术分析不同地理居群益智基因多态性，构建系统发育树并筛选获得具有代表性的益智核心种质资源库。其次采用双向电泳结合MALDI-TOF MS技术研究益智核心种质资源获得蛋白质组学数据，分析其在蛋白质水平上存在的遗传差异。最后结合化学活性成分数据来探究影响益智核心种质资源品质的基因和蛋白。运用系统生物学聚类方法将基因多态性、蛋白质组学和化学活性成分数据有机结合研究益智的种质资源遗传多样性，将从基因和蛋白质水平弄清其遗传多样性，以探索不同地理居群益智品质差异的遗传背景，阐明益智资源的道地性形成机制，为其核心种质资源保护、鉴定和评价提供科科学依据。

目前海南野生益智资源不断受到严重破坏以及栽培品种药材功效的逐渐退化，调查和收集不同地理居群的益智种质资源研究其遗传多样性尤为重要。本项目共收集了海南15个不同地理居群的益智样品。首先，采用了ITS、SRAP、AFLP三种分子标记技术分析了其遗传多样性，研究表明不同地理居群的益智种质具有遗传差异，遗传多样性主要集中在益智种群内，种群间的基因流动不高；其次采用转录组学和蛋白质组学阐明了益智不同组织部位和不同发育时期的果实中关于黄酮类化学活性成分功能基因FLS、ANS、DFR和ANR的调控机制；另外，对益智叶绿体基因组进行测序，阐明了益智在姜科系统发育中的地位，并发现了NADH-脱氢酶基因（ndhB和ndhC），光系统II相关基因（psbZ）和ATP合成酶基因（atpE和atpF）在喜阳和喜阴姜科植物中分子水平的进化差异。结合以上数据阐述南药益智的遗传多样性与药材道地性，为南药益智种质资源保护、鉴定和评价及GAP种植提供科学依据。该项目培养硕士研究生2名，发表期刊论文6篇，其中SCI论文1篇，待发表SCI论文3篇。

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