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Expressed sequence tags and molecular cloning and characterization of gene encoding pinoresinol/lariciresinol reductase from Podophyllum hexandrum

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Title Expressed sequence tags and molecular cloning and characterization of gene encoding pinoresinol/lariciresinol reductase from Podophyllum hexandrum
 
Creator Wankhede, Dhammaprakash Pandhari
Biswas, Dipul Kumar
Rajkumar, Subramani
Sinha, Alok Krishna
 
Subject Podophyllum hexandrum
Suppressive subtraction hybridization
cDNA library
Expressed sequenced tags
Pinoresinol/lariciresinol reductase
RT-PCR
 
Description Accepted date: 18 April 2013
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.
DPW thanks University Grant Commission,
India for fellowships. This work is supported by funding by Department of Biotechnology, India.
 
Date 2015-11-24T11:34:43Z
2015-11-24T11:34:43Z
2013
 
Type Article
 
Identifier Protoplasma, 250(6): 1239-1249
1615-6102
http://172.16.0.77:8080/jspui/handle/123456789/382
http://link.springer.com/article/10.1007%2Fs00709-013-0505-z
10.1007/s00709-013-0505-z
 
Language en_US
 
Publisher Springer