Utilizing Transcriptome of Purple Abelmoschus esculentus for Sequence Analysis on Anthocyanidin Gene
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摘要: 紫色秋葵富含花青素，因此其全株及果实均为紫红色，为获取正常生长状态下紫色秋葵花青素合成代谢相关基因，采用Illumina HiSeq 2500高通量转录组测序技术对紫色秋葵全转录功能基因组测序后组装，再通过Nr、SwissProt和GO等三大数据库进行花青素相关功能基因筛选。三大数据库涉及35个花青素相关Unigene，具体包括19个花青素相关糖基转移酶、6个花青素酰基转移酶家族基因，5个无色花色素双加氧酶（合酶）以及5个花色素相关还原酶。这些花青素相关基因在Nr及SwissProt数据库中涉及较多相关植物，包括拟南芥、可可、树棉、木薯、苹果及葡萄等。聚类分析研究表明：涉及花青素相关四大家族成员的35个Unigene中，无色花色素双加氧酶亲缘关系最近，花青素酰基转移酶次之，花青素相关还原酶亲缘关系稍远，而花青素相关糖基转移酶家族成员亲缘关系最远。本研究获取的紫色秋葵众多花青素相关转移酶、合酶及还原酶等相关Unigene，为进一步研究紫色秋葵或其他植物的花青素合成代谢过程中基因克隆分析等奠定了坚实基础。Abstract: The rich content of anthocyanidin in purple Abelmoschus esculentus renders the characteristic color of the plant and its fruits. This study used the transcriptome of the plant to study the genetics associated with the anthocyanidin biosynthesis pathway under natural conditions. The transcriptome was sequenced using the Illumina HiSeq 2500 platform to isolate 35 anthocyanidin unigenes from the Nr, SwissProt and GO databases. These unigenes belonged to 4 distinct gene types including 19 of anthocyanidin-related glucosyltransferases, 6 of anthocyanidin-related acyltransferases, 5 of leucoanthocyanidin dioxygenases, and 5 of anthocyanidin-related reductases. The genes were annotated by Nr and SwissProt databases to show their associations with a variety of plants, such as Arabidopsis thaliana, Theobroma cacao, Gossypium arboretum, Manihot esculenta, Malus domestica and Vitis vinifera. A clustering analysis indicated that the unigenes had the closest relationship with the leucoanthocyanidin dioxygenase family, followed by the anthocyanin-related acyltransferases and the reductases, while the anthocyanidin-related glucosyltransferases showed the farthest homology. Identification of the 35 anthocyanidin-related unigenes would be useful for the gene cloning and analysis in studying anthocyanin biosynthesis of purple A. esculentus or other plants rich in anthocyanidin.
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图 1 35个花青素相关Unigene亲缘关系分析
Figure 1. Genetic relationship of 35 anthocyanin-related unigenes
表 1 19个花青素相关糖基转移酶信息分析
Table 1. Information on 19 anthocyanin-related glucosyltransferase
编码 FPKM 核苷酸 非冗余蛋白数据库(Nr) 蛋白质序列数据库(SwissProt) 基因本体论数据库(GO) 基因 匹配物种 基因 匹配物种 C80207 0.96 423 UDP糖基转移酶 可可 花青素3，2-O-双糖基转移酶 矮牵牛 花青素3-O-糖基转移酶 C4554 0.99 610 未知功能蛋白 中粒咖啡 UDP-糖基转移酶 拟南芥 花青素3-O-糖基转移酶 C74868 1.13 429 未知功能蛋白 树棉 花青素3-O-糖基转移酶 草莓 转移酶活性 C72801 1.27 391 UDP糖基转移酶 可可 UDP糖基转移酶 拟南芥 花青素3-O-糖基转移酶 C68173 1.34 325 UDP糖基转移酶 可可 花青素3-O-糖基转移酶 葡萄 转移酶活性 c16602 1.35 857 花青素3-O-糖基转移酶 可可 花青素3-O-糖基转移酶 木薯 未注释到信息 c17288 1.58 578 UDP糖基转移酶 可可 脱落酸β-葡糖基转移 赤豆 花青素3-O-糖基转移酶 c62098 1.94 587 UGT蛋白 可可 UDP糖苷环烯醚萜糖基转移酶 长春花 花青素3-O-糖基转移酶 c1778 2.61 372 黄酮类7-O糖基转移酶 可可 花青素3, 5-O-糖基转移酶 美女樱 UDP糖基转移酶 C43129 3.42 612 唐-葡萄糖转移酶 可可 花青素3-O-糖基转移酶 木薯 花青素3-O-糖基转移酶 C40678 3.61 717 UDP糖基转移酶 可可 花青素3, 5-O-糖基转移酶 月季 对苯二酚葡糖基转移酶 C44130 4.75 743 超氧化物歧化酶铜伴侣蛋白 甜叶菊 花青素3-O-糖基转移酶 木薯 未注释到信息 C58548 8.23 519 唐-葡萄糖转移酶 可可 花青素3-O-糖基转移酶 木薯 花青素3-O-糖基转移酶 C49793 13.3 1700 UDP糖基转移酶 可可 花青素3-O-糖基-2-O-木糖基转移酶 拟南芥 转移酶活性 C39786 13.32 568 UDP糖基转移酶 可可 UDP糖基转移酶 拟南芥 花青素3-O-糖基转移酶 C50927 17.72 1939 UDP糖基转移酶 可可 UDP糖基转移酶 拟南芥 花青素3-O-糖基转移酶 c18432 21.18 833 UDP糖基转移酶 可可 UDP-糖基转移酶 拟南芥 花青素3-O-糖基转移酶 c16536 60.47 1856 UDP糖基转移酶 可可 马钱苷元糖基转移酶 栀子花 花青素3-O-糖基转移酶 C48231 342.33 1744 UDP糖基转移酶 可可 东莨菪内酯葡糖基转移酶 土豆 花青素3-O-糖基转移酶 
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表 2 6个花青素相关酰基转移酶信息分析
Table 2. Information on 6 anthocyanin-related acyl-transferase
编码 FPKM 核苷酸 非冗余蛋白数据库(Nr) 蛋白质序列数据库(SwissProt) 基因本体论数据库(GO) 基因 匹配物种 基因 匹配物种 C73355 1.03 400 酰基转移酶 可可 花青素3-O-糖基-6-O-香豆酰转移酶 拟南芥 未注释到信息 C28010 3.56 1664 酰基转移酶 可可 花青素3-O-糖基-6-O-香豆酰转移酶 拟南芥 转移酶 c17535 10.89 370 酰基转移酶 可可 花青素3-O-糖基-6-O-香豆酰转移酶 拟南芥 转移酶 C37660 17.41 401 酰基转移酶 可可 花青素3-O-糖基-6-O-香豆酰转移酶 拟南芥 转移酶 C43671 37.23 1566 酰基转移酶 可可 花青素3-O-糖基-6-O-香豆酰转移酶 拟南芥 转移酶 C49043 89.64 949 酰基转移酶 可可 花青素3-O-糖基-6-O-香豆酰转移酶 拟南芥 转移酶
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表 3 5个花青素相关双加氧酶信息
Table 3. Information on 5 anthocyanin-related dioxygenase
编码 FPKM 核苷酸 非冗余蛋白数据库(Nr) 蛋白质序列数据库(SwissProt) 基因本体论数据库(GO) 基因 匹配物种 基因 匹配物种 c3740 1.93 353 无色花色素双加氧酶 树棉 阿魏酰辅酶A羟化酶 拟南芥 未注释到信息 C20762 1.93 896 戊二酸铁双加氧酶 可可 无色花色素双加氧酶 拟南芥 氧化还原酶 c25316 3.97 940 无色花色素双加氧酶 树棉 无色花色素双加氧酶 苹果 铁离子结合 C36803 4.06 755 无色花色素双加氧酶 树棉 无色花色素双加氧酶 苹果 黄酮醇合酶 C42576 46.02 1588 戊二酸铁双加氧酶 可可 无色花色素双加氧酶 苹果 未注释到信息
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表 4 5个花青素相关还原酶信息分析
Table 4. Information on 5 anthocyanin-related reductase
编码 FPKM 核苷酸 非冗余蛋白数据库(Nr) 蛋白质序列数据库(SwissProt) 基因本体论数据库(GO) 基因 匹配物种 基因 匹配物种 c23823 0.89 524 二氢黄酮醇还原酶 树棉 花色素还原酶 拟南芥 氧化还原酶 C26348 3.6 1587 结合NADαβ折叠酶 可可 花色素还原酶 拟南芥 花色素还原酶 C38710 3.85 1091 花色素还原酶 葡萄 二氢黄酮醇还原酶 雏菊 未注释到信息 C59615 4.49 393 二氢黄酮醇还原酶 树棉 花色素还原酶 拟南芥 二氢黄酮醇还原酶 C28410 18.9 1831 无色花色素还原酶 可可 无色花色素还原酶 三叶草 无色花色素还原酶
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