Fig 1.
Symptoms of R. solani AG8 infection of wheat.
A) T. aestivum cv Wyalkatchem seedlings photographed two-week post-infection with AG8 or mock treatment. B) Shoot weight of the wheat cultivars, Chinese Spring and Wyalkatchem following two-week post-infection with AG8 or a mock treatment. Stars indicates significant differences for mock vs R. solani infection (Student’s t-test, P <0.05).
Table 1.
List of number of wheat genes induced >2 in response to R. solani and categorised into functional subsets.
Fig 2.
Analysis of selected wheat genes in response to R. solani AG8 using RT-qPCR.
One day old wheat seedlings were infected with R. solani AG8 for 2, 3 or 4 days, and RNA from root and shoot tissue were tested for relative RNA expression for A) GLP1 (Germin), B) UGE1, C) PR10 and D) TaGlu (glucan endo-1,3-beta-glucosidase GII precursor). RNA expression was normalised to the ADP-ribosylation factor R wheat gene. Stars indicates significant differences for mock vs R. solani infection (log transformed Student’s t-test, P <0.05).
Fig 3.
ROS measurement in R. solani infected wheat.
A) Roots of wheat seedlings that were mock infected and stained with DAB, B) Roots of wheat seedlings that were R. solani AG8 infected and stained with DAB, C) Wheat seedlings infected with mock (left side) or R. solani (right side) and stained with NBT, D) Biochemical analysis of mock and R. solani treated wheat roots using titanium sulphate to measure H2O2. Units are expressed as μmol hydrogen peroxide/fwt and the AG8 treated plants were shown to be significantly different to mock treated (Student’s t-test, P <0.05) E) Biochemical analysis of mock and R. solani treated wheat roots using NBT to measure relative superoxide anions per fresh weight. The relative superoxide anion amount was shown to be significantly different to mock treated (Student’s t-test, P <0.05).
Fig 4.
Analysis of R. solani genes using RT-qPCR in response to wheat infection.
Three day wheat seedlings were infected with R. solani AG8 for 4 or 7 days, and RNA from R. solani adjacent to the wheat was extracted. Relative RNA expression of noxA, noxB, noxC, noxD, TmpL, SOD and OAH was measured. RNA expression was normalised to the R. solani ribosomal protein gene. Stars indicates significant differences of infection vs vegetative R. solani (Student’s t-test, P <0.05).