黔东南地区高花青素蓝莓品种鉴选及花青素重要基因克隆与功能分析

The natural conditions in Qiandongnan are suitable for the growth of blueberry, so the region has introduced and developed blueberry industry. Blueberry is one of the world's most recognized high-quality fruit in the processing chain. The most important development is anthocyanin. Therefore, the selection of blueberry anthocyanin varieties and the mechanism of anthocyanin synthesis have become a research hotspot. . This project intends to carry out the following research work with blueberries with good adaptability in Qiandongnan region: 1) Screening of blueberry varieties with excellent growth performance in southeastern Yunnan, and selecting high anthocyanin varieties through indicators such as yield, quality and growth characteristics.2) Qualitative and quantitative study on anthocyanins of different blueberry varieties by high performance liquid chromatography, and establish chromatographic fingerprints of anthocyanins.3) Compare the antioxidant activities of anthocyanins in different blueberry varieties, and determine the relationship between anthocyanin components and antioxidant capacity of different blueberry qualities.4) Cloning and functional analysis of promoters of blueberry anthocyanin synthesis related genes, which laid a foundation for the analysis of blueberry anthocyanin synthesis mechanism and variety improvement.. The development of this project will lay the foundation for the selection of high anthocyanin blueberry varieties and the study of anthocyanin synthesis mechanism in Qiandongnan. It will serve as a guiding significance for the development of blueberry varieties and germplasm improvement in Qiandongnan ..

黔东南地区自然条件适合蓝莓的生长，因此该地区已大力引进与发展蓝莓产业。蓝莓是世界公认的加工链最长的精品水果之一，其中最具开发价值的是花青素，因此蓝莓高花青素品种鉴选及花青素合成机理研究已成为研究热点。. 本项目拟以黔东南地区适应性良好的蓝莓为材料，开展以下研究工作：1）筛选黔东南地区生长表现优异的蓝莓品种，通过产量、品质和生长特性等指标鉴选高花青素品种。2）采用高效液相色谱法对不同蓝莓品种的花青素进行定性定量研究，建立花青素色谱指纹图谱。3）比较不同蓝莓品种花青素的抗氧化活性，探明不同蓝莓品质的花青素成分与抗氧化能力之间的关系。4）蓝莓花青素合成相关基因启动子克隆与功能分析，为探明蓝莓花青素合成机制及品种改良奠定基础。本项目的开展将对黔东南地区高花青素蓝莓品种的鉴选及花青素合成机制的研究奠定基础，将为黔东南地区发展蓝莓品种和种质改良起到指导意义。

巴尔德温的生长适应性较好、产量较高，灿烂的花青素抗氧化活性最强，夏普兰成熟期花青素含量最高，以上三个品种在我省具有极大开发利用价值。代谢组分析显示灿烂、粉蓝和莱克西花青素成分由13种花青素成分组成，果实不同的发育阶段13种花青素表达量变化较大，开发花青素的深加工药用价值可根据果实发育期进行适时采收。灿烂、粉蓝和莱克西成熟期均松香花青素 O-己糖苷含量最低，而灿烂和莱克西的牵牛花色素 3-O-葡萄糖含量最高，粉蓝的矢车菊素 O-丁香酸含量最高。蓝莓花青素的自由基清除能力与果实的成熟程度呈正相关关系，果实越成熟，自由基清除能力就越强。在成熟果实中，灿烂的DPPH自由基清除能力、羟自由基清除能力、ABTS·+自由基清除能力、总抗氧化活性及还原能力都高于其他4个品种。. 从蓝莓中鉴定了15种候选花青素生物合成相关MYB（ABRM）蛋白，包括12种R2R3 MYB和3种1R MYB。亚细胞定位预测结果表明，除VcRVE8（定位于叶绿体和细胞核）外，所有蓝莓ABRM均为细胞核定位。蓝莓ABRM基因的外显子数量变化很大，在1到8之间。VcMYB、VcMYB6、VcMYB23、VcMYBL2和VcPH4在蓝莓果实中大量表达，并且在所有蓝莓ABRM中，VcMYB在转色期、紫果期和成熟期果实中表达最高。VcMYB-1的瞬时过表达促进了绿色水果中花青素的积累。. 蓝莓基因组数据中识别出六种与花青素生物合成相关的bHLH。这些bHLH都属于bHLH-MYC_N结构域，具有DNA结合位点，它们均为G盒结合蛋白。bHLH蛋白质亚细胞定位预测均位于细胞核。VcAN1 gDNA含有8个内含子，而其5个均含有7个内含子。VcAN1在老叶、茎和蓝果中高度表达，并且随着蓝莓果实的成熟，其表达增加。VcAN1不仅在果实中和叶中都参与花青素的生物合成调控。VcbHLH1-1在幼叶和茎中表达最高，在绿色果实中表达最低。VcbHLH1-2在茎中高表达，但在果实中低表达，特别是在红色果实中。发表论文2篇，获得发明专利授权1项，培养学生7名。