Identification and analysis of mechanisms involved in a broad-spectrum disease resistant mutant of the winter wheat cv. Guardian

M66 an X-ray induced mutant of winter wheat (Triticum aestivum) cv. Guardian exhibits broad-spectrum resistance to powdery mildew (Blumeria graminis f. sp. tritici), yellow rust (Puccinia striiformis f. sp. tritici), and leaf rust (Puccinia recondita f. sp. tritici), along with partial resistance to stagnonospora nodorum blotch (caused by the necrotroph Stagonosporum nodorum) and septoria tritici blotch (caused by the hemibiotroph Mycosphaerella graminicola) compared to the parent plant ‘Guardian’. Analysis revealed that M66 exhibited no symptoms of infection following artificial inoculation with Bgt in the glasshouse after adult growth stage (GS 45). Resistance in M66 was associated with widespread leaf flecking which developed during tillering. Flecking also occurred in M66 leaves without Bgt challenge; as a result grain yields were reduced by approximately 17% compared to ‘Guardian’ in the absence of disease. At the seedling stage, M66 exhibited partial resistance. M66, along with Tht mutants (Tht 12, Tht13), also exhibit increased tolerance to environmental stresses (abiotic), such as drought and heat stress at seedling and adult growth stages, However, adult M66 exhibited increased susceptibility to the aphid Schizaphis graminum compared to ‘Guardian’. Resistance to Bgt in M66 was characterized with increased and earlier H2O2 accumulation at the site of infection which resulted in increased papilla formation in epidermal cells, compared to ‘Guardian’. Papilla formation was associated with reduced pathogen ingress and haustorium formation, indicating that the primary cause of resistance in M66 was prevention of pathogen penetration. Heat treatment at 46º C prior to challenge with Bgt also induced partial disease resistance to Blumeria graminis f. sp. tritici in ‘Guardian’ and M66 seedlings. This was characterized by a delay in primary infection, due to increased production of ROS species, such as hydrogen peroxide, ROS-scavenging enzymes and Hsp70, resulting in cross-linking of cell wall components prior to inoculation. This actively prevented the fungus from penetrating the epidermal cell wall. Proteomics analysis using 2-D gel electrophoresis identified primary and secondary disease resistance effects in M66 including detection of ROS scavenging enzymes (4, 24 hai), such as ascorbate peroxidase and a superoxidase dismutase isoform (CuZnSOD) in M66 which were absent from ‘Guardian’. Chitinase (PR protein) was also upregulated (24 hai) in M66 compared to ‘Guardian’.Monosomic and ditelosomic analysis of M66 revealed that the mutation in M66 is located on the long arm of chromosome 2B (2BL). Chromosome 2BL is known to have key genes involved in resistance to pathogens such as those causing stripe rust and powdery mildew. The TaMloB1 gene, an orthologue of the barley Mlo gene, is also located on chromosome 2BL. Sanger sequencing of part of the coding sequence revealed no deletions in the TaMloB1 gene between ‘Guardian’ and M66.

Leaf galls in our native trees and shrubs

Plant galls constitute a branch of study and research which has been to me a subject of much interest for some time. At the start of this work, it was intended to include Plant galls in general, but after some months this was found to be too comprehensive a field and would in fact take a great many years to study fully. Even leaf galls alone, both of herbs and trees provide so large a field of investigation that ultimately I decided to confine my attention to those or our native trees and shrubs. Upon looking up the literature on this subject, it will be found that in nearly all cases, either the gall is described fully and mere mention made or the agent concerned in its production, or vice versa. This state of things is most unsatisfactory, as in studying galls, both the gall-maker and the gall formation must be examined in detail before it is safe to apply nomenclature. This work, therefore, sets out to give accurate and scientific descriptions of both galls and gall-makers. The difficulties encountered are manifold; firstly, our trees are all deciduous, hence, the collecting period is necessarily restricted to that time of the year between the appearance of the buds and the fall of the leaf. Secondly, the rearing of imagines is always difficult, especially in the case or the autumn gall; more will be said on this matter later. Lastly, due to war-time conditions much trouble was experienced in obtaining suitable literature and many invaluable books on this subject were unprocurable. The Plates at the back have all been copied from original material except in the case or the Phytoptid mites which have been sketched with the help of illustrations, the reason for this being the difficulty of making suitable mounts of these minute creatures, Where possible all stages or at least larva and imago have been sketched, together with the host plant and the type of gall-formation produced. Slides have also been made of most larvae and the imagines attached to cards and pinned on to pith or cork in the usual manner.