JXB Advance Access originally published online on March 7, 2005
Journal of Experimental Botany 2005 56(414):1229-1237; doi:10.1093/jxb/eri117
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Published by Oxford University Press [2005] on behalf of the Society for Experimental Biology.
RESEARCH PAPER |
Application of Bruchin B to pea pods results in the up-regulation of CYP93C18, a putative isoflavone synthase gene, and an increase in the level of pisatin, an isoflavone phytoalexin*
1Horticultural Crops Research Unit, USDA-ARS, Corvallis, OR 97330, USA
2Department of Horticulture, Oregon State University, Corvallis, OR 97331, USA
3USDA-ARS, Chemicals Affecting Insect Behavior Laboratory, Building 007, BARC-W, Beltsville, MA 20705, USA
Present address and to whom correspondence should be sent: Department of Crop and Soil Science, Oregon State University, Corvallis, OR 97331, USA. Fax: +1 541 737 0909. E-mail: lol.cooper{at}oregonstate.edu
Bruchins, mono and bis (3-hydroxypropanoate) esters of long chain 
,
-diols, are a recently discovered class of insect elicitors that stimulate cell division and neoplasm formation when applied to pods of peas and certain other legumes. Differential display analysis resulted in the identification of an mRNA whose level was increased by the application of Bruchin B to pea pods. The corresponding amplification product was cloned and sequenced and a full length cDNA sequence was obtained. This cDNA and the gene from which it was derived were assigned the name CYP93C18 based upon sequence similarities to the cytochrome P450 mono-oxygenase CYP93C subfamily, which contains isoflavone synthase genes from legumes. RNA gel blots and quantitative RT-PCR demonstrated that expression of CYP93C18 increased within 8 h of bruchin treatment to a maximum of 100–200-fold of the level in untreated pods, and then declined. The up-regulation of CYP93C18 was followed by an increase in the level of the isoflavone phytoalexin, pisatin. Pisatin was detectable in the bruchin-treated pods after 16 h and reached a maximum between 32 h and 64 h. This, the first report of induction of phytoalexin biosynthesis by an insect elicitor, suggests that Bruchin B not only stimulates neoplasm formation, but also activates other plant defence responses.
Key words: Bruchin, CYP93C18, cytochrome P450, insect elicitor, Neoplastic pod, phytoalexin, pisatin, Pisum sativum
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