Expressed sequence tags and molecular cloning and characterization of gene encoding pinoresinol/lariciresinol reductase from Podophyllum hexandrum
Expressed sequence tags and molecular cloning and characterization of gene encoding pinoresinol/lariciresinol reductase from Podophyllum hexandrum
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DOI：
10.1007/s00709-013-0505-z
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发表时间：
2013-12-01
期刊：
影响因子：
2.9
通讯作者：
Sinha, Alok Krishna
中科院分区：
文献类型：
--
作者：
Wankhede, Dhammaprakash Pandhari;Biswas, Dipul Kumar;Sinha, Alok Krishna
鬼臼毒素是一种芳基四氢萘木脂素，是重要的抗癌药物依托泊苷、替尼泊苷和etopophos的来源。桃儿七（Podophyllum hexandrum）的根/根茎是鬼臼毒素的重要来源之一。为了了解参与鬼臼毒素生物合成的基因，合成两个抑制消减杂交文库，分别来自根/根茎和叶，使用高和低鬼臼毒素产生的植物的P. hexandrum。克隆测序鉴定了总共1，141个表达序列标签（EST），产生354个独特的EST。几个独特的EST序列相似的基因参与代谢，压力/防御反应，和信号通路。一些EST还与其他植物物种中参与鬼白毒素生物合成的基因（例如松脂醇/落叶松树脂醇还原酶）表现出高度序列相似性。本研究克隆了中国连翘（P. hexandrum）中松醇/落叶松树脂醇还原酶（PLR）基因的全长编码序列，该基因编码311个氨基酸，与连翘（Forsythia intermedia）和亚麻（Linum spp.）PLR基因序列相似。空间和胁迫诱导的表达模式的PhPLR和其他已知的基因的鬼臼毒素生物合成，开环异落叶松树脂醇脱氢酶（PhSDH），和dirigent蛋白氧化酶（PhDPO）进行了研究。这三个基因都表现出伤害和茉莉酸甲酯诱导的表达模式。本研究为进一步研究桃儿七中鬼臼毒素生物合成的基因组学奠定了基础。
Podophyllotoxin, an aryltetralin lignan, is the source of important anticancer drugs etoposide, teniposide, and etopophos. Roots/rhizome of Podophyllum hexandrum form one of the most important sources of podophyllotoxin. In order to understand genes involved in podophyllotoxin biosynthesis, two suppression subtractive hybridization libraries were synthesized, one each from root/rhizome and leaves using high and low podophyllotoxin-producing plants of P. hexandrum. Sequencing of clones identified a total of 1,141 Expressed Sequence Tags (ESTs) resulting in 354 unique ESTs. Several unique ESTs showed sequence similarity to the genes involved in metabolism, stress/defense responses, and signalling pathways. A few ESTs also showed high sequence similarity with genes which were shown to be involved in podophyllotoxin biosynthesis in other plant species such as pinoresinol/lariciresinol reductase. A full length coding sequence of pinoresinol/lariciresinol reductase (PLR) has been cloned from P. hexandrum which was found to encode protein with 311 amino acids and show sequence similarity with PLR from Forsythia intermedia and Linum spp. Spatial and stress-inducible expression pattern of PhPLR and other known genes of podophyllotoxin biosynthesis, secoisolariciresinol dehydrogenase (PhSDH), and dirigent protein oxidase (PhDPO) have been studied. All the three genes showed wounding and methyl jasmonate-inducible expression pattern. The present work would form a basis for further studies to understand genomics of podophyllotoxin biosynthesis in P. hexandrum.