The Differential Susceptibility of Vetch (Vicia spp.) to Orobanche aegyptiaca: Anatomical Studies

Vetches (Vicia spp .) are important forage legumes in the MiddleEast and Mediterranean regions. Most vetches are highly susceptibleto the root holoparasites Orobanche aegyptiaca and O. crenata,suffering severe quality and yield losses. However, purple vetch(V. atropurpurea) has shown good resistance to Orobanche. Microscopicstudies were performed to reveal anatomical differences of host-parasiteinteractions between susceptible and resistant vetch. SusceptibleV. sativa‘Yovel’ and resistant V. atropupurea‘Popany’were grown in association with O. aegyptiaca seeds, on glassmicrofibre sheets in a polyethylene-bag system. Whole root observationsusing stereoscopic microscopy detected necrotic lesions surroundingthe attachments of Orobanche radicles on resistant vetch roots.Hand-cut sections examined under the light microscope revealedthat in both susceptible and resistant vetch genotypes the Orobancheseeds germinated, attached and penetrated the host root epidermisand cortex. A reddish-brown secretion was observed in the apoplastat the interface between the parasite haustorium and the hosttissues, including the cell walls of the resistant vetch xylemvessels. Fixed and embedded sections observed under the lightmicroscope showed that in the susceptible genotype the parasitehaustorium penetrated through the endodermis into the host vascularcylinder, successfully forming a continuum with host vascularvessels. However, in the resistant genotype, the parasite haustoriumwas blocked at the root endodermis layer. The blockage was coupledwith secretion of unidentified material, thus preventing theparasite from establishing. These findings indicate that mechanicaland possibly chemical barriers are responsible for the hostdefence mechanism(s) conferring resistance of V. atropurpureato O. aegyptiaca. Copyright 2000 Annals of Botany Company Broomrape, Vicia atropurpurea, Vicia sativa, parasitic plants, plant resistance, tissue sections.     

High levels of expression of full length human pro-alpha 2(V) collagen cDNA in pro-alpha 2(V)-deficient hamster cells

A full length cDNA encoding human pro-alpha 2(V) collagen was constructed. Partial sequencing of the cDNA and primer extension analysis of mRNA from fibroblasts found that pro-alpha 2(V) mRNA differs from the mRNAs of other fibrillar collagens in the increased length of its 5'-untranslated region. The pro-alpha 2(V) cDNA was placed downstream of the human cytomegalovirus immediate early promoter/regulatory sequences for expression studies in cultured Chinese hamster lung cells. These cells have been shown previously to synthesize large quantities of pro-alpha 1(V) homotrimers as their only collagenous product. Transfection resulted in a number of clonal cell lines that express human alpha 2(V) RNA at levels comparable to, and in some cases greater than, levels found in normal human skin fibroblasts. Pro-alpha 2(V) chains produced in the majority of clonal lines were of sufficient quantity to complex all available endogenous pro-alpha 1(V) chains. Chimeric heterotrimers, composed of hamster alpha 1(V) and human alpha 2(V) chains in a 2:1 ratio, were stable to pepsin digestion and were found predominantly associated with the cell layer. Surprisingly, pro-alpha 2(V) chains, in excess to pro-alpha 1(V) chains, were found in the extracellular matrix and, in much greater abundance, in media. These chains were pepsin sensitive, indicating that pro-alpha 2(V) chains can be secreted as nonstable homotrimers or as free chains.