Early leaf responses of cell physiological and sensor-based signatures reflect susceptibility of wheat seedlings to infection by leaf rust.

Leaf rust caused by Puccinia triticina Erikss. can have devastating effects on wheat (Triticum aestivum L.), causing severe economic losses. This comprehensive study serves to facilitate our understanding of the impact of carbohydrate and antioxidant metabolism in association with sensor-based phenotyping and leaf rust stress responses in wheat seedlings. After 24 h of inoculation (hai) very susceptible variety to leaf rust (Ficko) increased cell-wall invertase (cwInv; EC 3.2.1.26), compared to other varieties that significantly increased cwInv later. This could mean that the Ficko variety cannot defend itself from leaf rust infections once symptoms have started to develop. Also, Ficko had significantly decreased amounts of cytoplasmic invertase (cytInv; EC 3.2.1.26) at 8 hai. The downregulation of cytInv in susceptible plants may facilitate the maintenance of elevated apoplastic sucrose availability favoring the pathogen. The significant role of vacuolar invertase (vacInv; EC 3.2.1.26) in moderately resistant varieties was recorded. Also, a significant decrease of glucose-6-phosphate dehydrogenase (G6PDH; EC 1.1.1.49) and UDP-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) in moderately resistant varieties might restrict normal development of leaf rust due to reduced sugar. During plant-pathogen interaction, when the invader spreads systemically throughout the plant, the main role of ascorbate peroxidase (APX; EC 1.11.1.11) activity in one moderately resistant variety (Olimpija) and catalase (CAT; EC 1.11.1.6) activity in another moderately resistant variety (Alka) is to protect the plant against oxidative damage in the early stages of infection. Non-invasive phenotyping with a sensor-based technique could be used as a rapid method for pre-symptomatic determination of wheat leaf rust resistance or susceptibility.

Role of the Colletotrichum acutatum sesquiterpene synthase CaTPS in the biosynthesis of sesquiterpenoids.

Colletotrichum acutatum is a major fungal pathogen of fruit crops, which causes severe yield losses in strawberry production. A potential key factor in plant-pathogen interactions is fungal sesquiterpenoids which have mycotoxic and phytotoxic activities. The first committed step in sesquiterpenoid biosynthesis is performed by sesquiterpene synthases (TPS). Only a few TPSs have been functionally characterized from filamentous fungi and none from the genus Colletotrichum. Despite being an important fungal pathogen to agriculture, it is poorly understood at the molecular and chemical levels. The terpenoid biochemistry in Coll. acutatum strain SA 0-1 was studied and one Coll. acutatum TPS (CaTPS) was successfully cloned and characterized in yeast. CaTPS catalyses the biosynthesis of multiple sesquiterpenoids. The two major products are ß-caryophyllene and an unidentified sesquiterpenoid along with α-humulene as one of the minor sesquiterpenoid products. These products were also secreted by the fungus in strawberry fruit medium along with several other sesquiterpenoids indicating other TPSs are active during in vitro growth. ß-Caryophyllene and α-humulene are known cytotoxic products important for ecological interactions and are produced by SA 0-1. Interestingly, a gene expression analysis using quantitative real-time PCR revealed a significant increase in expression of CaTPS during strawberry fruit infection, thus indicating that it could be involved in fruit infection. This is, we believe, the first characterization of TPS in Colletotrichum spp. and terpenoid profiles of Coll. acutatum, which could facilitate studies on the role of terpenoids in the ecology of Coll. acutatum.