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JXB Advance Access published online on February 27, 2004
Journal of Experimental Botany, doi:10.1093/jxb/erh098
© 2004 by Oxford University Press
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Received November 21, 2003; accepted January 14, 2004
© 2004 Society for Experimental Biology

RESEARCH PAPER

In vitro infection of host roots by differentiated calli of the parasitic plant Orobanche

W. J. Zhou 1, K. Yoneyama 2, Y. Takeuchi 2, S. Iso 2, S. Rungmekarat 2, S. H. Chae 2, D. Sato 2, and D. M. Joel 3*

1 Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China; Center for Research on Wild Plants, Utsunomiya University, Utsunomiya 321-8505, Japan
2 Center for Research on Wild Plants, Utsunomiya University, Utsunomiya 321-8505, Japan
3 Center for Research on Wild Plants, Utsunomiya University, Utsunomiya 321-8505, Japan; Department of Phytopathology and Weed Research, ARO, Newe-Ya’ar Research Center, PO Box 1021, Ramat-Yishay, 30095, Israel

* To whom correspondence should be addressed. E-mail: dmjoel{at}volcani.agri.gov.il.


  Abstract

Root parasites of the genus Orobanche are serious weeds in agriculture. An aseptic infection system of host roots using calli of three Orobanche species was developed for the study of host-parasite interaction. The response of calli to various hormonal combinations was studied, because a requirement for infection is the differentiation of root-like protrusions, which are capable of producing haustorial connections to the host. Infectious root-like protrusions develop under the influence of 0.5-1.0 mg l-1 IAA, and under the combination of 0.2 mg l-1 NAA with 5.0 mg l-1 kinetin. These protocols produced root protrusions with pad-like structures that resembled attachment organs of Orobanche seedlings, and proved effective in parasitizing host roots. Direct contact with the medium inhibited haustorium development and prevented infection. To overcome this problem, certain root portions were isolated from the medium by inserting thin glass plates underneath. Calli were then placed on the raised root portions and successfully infected the roots and developed young Orobanche tubercles with vascular system that directly connected to the host.

Key words: Aseptic infection, Brassica, callus induction, germination, Orobanche, red clover, tomato.


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