An efficient approach to finding Siraitia grosvenorii triterpene biosynthetic genes by RNA-seq and digital gene expression analysis.
An efficient approach to finding Siraitia grosvenorii triterpene biosynthetic genes by RNA-seq and digital gene expression analysis.
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DOI：
10.1186/1471-2164-12-343
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发表时间：
2011-07-05
期刊：
影响因子：
4.4
通讯作者：
Qiu D
中科院分区：
文献类型：
--
作者：
Tang Q;Ma X;Mo C;Wilson IW;Song C;Zhao H;Yang Y;Fu W;Qiu D
罗汉果（Siraitia grosvenorii）是原产于我国南方的一种多年生草本植物，在桂林市分布最广。它的果实含有甜，肉质，可食用的果肉，广泛用于传统中药。果实提取物中的主要生物活性成分是被称为罗汉果苷的葫芦烷型三萜皂苷。其中罗汉果苷V的甜度是蔗糖的近300倍。然而，对S. grosvenorii，尤其是路径的后期步骤。本研究采用等量的S.使用Illumina/Solexa平台对开花后50天和70天的罗汉果果实进行测序。从cDNA文库中产生了超过48，755，516个高质量读段，其被组装成43，891个单基因。从头组装和缺口填充产生了43，891个单基因，平均序列长度为668个碱基对。共有26，308条（59.9%）独特序列被注释，其中11，476条独特序列被京都基因和基因组百科全书指定为特定代谢途径。从我们的文库中鉴定了参与罗汉果苷骨架合成的所有已知酶的cDNA序列。此外，总共鉴定了85个细胞色素P450（CYP 450）和90个UDP-葡糖基转移酶（UDPG）单基因，其中一些似乎编码负责罗汉果苷骨架转化为各种罗汉果苷的酶。采用Solexa测序技术对罗汉果果实发育的3个重要阶段进行了数字基因表达谱（DGE）分析，根据其表达模式，筛选出7个CYP 450和5个UDP基因，作为罗汉果皂苷生物合成的候选基因。基于下一代测序技术的RNA-seq和DGE分析的组合被证明是用于鉴定编码负责非模式植物中新型次级代谢物生物合成的酶的候选基因的有力方法。7个CYP 450和5个UDPGs被选为参与罗汉果苷生物合成的潜在候选者。本研究获得的转录组数据为理解S.罗汉果。
Siraitia grosvenorii (Luohanguo) is an herbaceous perennial plant native to southern China and most prevalent in Guilin city. Its fruit contains a sweet, fleshy, edible pulp that is widely used in traditional Chinese medicine. The major bioactive constituents in the fruit extract are the cucurbitane-type triterpene saponins known as mogrosides. Among them, mogroside V is nearly 300 times sweeter than sucrose. However, little is known about mogrosides biosynthesis in S. grosvenorii, especially the late steps of the pathway. In this study, a cDNA library generated from of equal amount of RNA taken from S. grosvenorii fruit at 50 days after flowering (DAF) and 70 DAF were sequenced using Illumina/Solexa platform. More than 48,755,516 high-quality reads from a cDNA library were generated that was assembled into 43,891 unigenes. De novo assembly and gap-filling generated 43,891 unigenes with an average sequence length of 668 base pairs. A total of 26,308 (59.9%) unique sequences were annotated and 11,476 of the unique sequences were assigned to specific metabolic pathways by the Kyoto Encyclopedia of Genes and Genomes. cDNA sequences for all of the known enzymes involved in mogrosides backbone synthesis were identified from our library. Additionally, a total of eighty-five cytochrome P450 (CYP450) and ninety UDP-glucosyltransferase (UDPG) unigenes were identified, some of which appear to encode enzymes responsible for the conversion of the mogroside backbone into the various mogrosides. Digital gene expression profile (DGE) analysis using Solexa sequencing was performed on three important stages of fruit development, and based on their expression pattern, seven CYP450s and five UDPGs were selected as the candidates most likely to be involved in mogrosides biosynthesis. A combination of RNA-seq and DGE analysis based on the next generation sequencing technology was shown to be a powerful method for identifying candidate genes encoding enzymes responsible for the biosynthesis of novel secondary metabolites in a non-model plant. Seven CYP450s and five UDPGs were selected as potential candidates involved in mogrosides biosynthesis. The transcriptome data from this study provides an important resource for understanding the formation of major bioactive constituents in the fruit extract from S. grosvenorii.
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影响因子：
7
作者：
Li, Ruiqiang;Zhu, Hongmei;Wang, Jun
通讯作者：
Wang, Jun
影响因子：
--
作者：
Lin GP;Jiang T;Hu XB;Qiao XH;Tuo QH
通讯作者：
Tuo QH
锯齿石杉和红刺蛇 454-EST 的比较揭示了参与石松生物碱生物合成和发育调控的推定基因
影响因子：
5.3
作者：
Luo H;Li Y;Sun C;Wu Q;Song J;Sun Y;Steinmetz A;Chen S
通讯作者：
Chen S
影响因子：
7.2
作者：
Achnine, L;Huhman, DV;Dixon, RA
通讯作者：
Dixon, RA
DOI：
10.1007/bf01932442
发表时间：
1975-01-01
期刊：
EXPERIENTIA
影响因子：
--
作者：
LEE, CH
通讯作者：
LEE, CH