To eliminate the complex conditions found in the intact plant, Shen et al. [52] used the detached rice leaf sheath, which is a very active part of the rice seedling, for a simple analysis of proteins in wound-response stress.
Therefore, we used detached leaf bioassays [ 36], a method that measures one of the major components of late leaf spot resistance as defined for cultivated peanut.
Exogenous application of JA to attached and detached leaves promotes leaf senescence in Arabidopsis but does not induce leaf senescence in the JA-insensitive mutant coi1, suggesting that the JA-signaling pathway is required for JA to promote leaf senescence [ 14].
A mycelial agar plug 3 mm in diameter was cut from the growing margins of a 6-day-old F. verticillioides culture and applied to the adaxial surface of a leaf detached from 6-week-old in vitro plants (n = 12 per line; repeated twice).
Detached leaf drying assay was reported to be an easy assay for testing large numbers of plants for their dehydration stress avoidance [ 60].
Two methods of inoculation were applied: i) seedling inoculation and ii) detached leaf inoculation.
Detached leaf assays were first performed with fully expanded leaves collected from the pTRV2- SlHUB1- and pTRV2- SlHUB2-infiltrated plants.
Detached leaf assays were also performed to study symptom development on both intact and wounded foliar tissues inoculated with Fg or Fc.
Detached leaf assays have been used previously to identify components of resistance related to FHB but these experiments, although using unwounded inoculation, were carried out using Microdochium majus, a non-toxin producing FHB species [ 40].
Resistance data were generated using three different inoculation methods, i.e. high-throughput in vitro assays [ 17], detached leaf assays [ 18] and multi-year field trials [ 19] (Table 1).
Detached leaves or leaf segments are generally considered to yield reliable estimates of host resistance that correspond to those of whole plants (Nicot et al. 2002).
