Targeting the AtCWIN1 Gene to Explore the Role of Invertases in Sucrose Transport in Roots and during Botrytis cinerea Infection

• Published in: Frontiers in plant science 2016-12, Vol.7, p.1899-1899
• Main Authors: Veillet, Florian, Gaillard, Cécile, Coutos-Thévenot, Pierre, La Camera, Sylvain
• Format: Article
• Language: English
• Subjects: Arabidopsis thaliana; Botrytis cinerea; cell wall invertase; CRISPR/Cas9; Life Sciences; Plant Science; Sucrose; sugar transport; Vegetal Biology
• ISSN: 1664-462X; 1664-462X
• DOI: 10.3389/fpls.2016.01899

Cell wall invertases (CWIN) cleave sucrose into glucose and fructose in the apoplast. CWINs are key regulators of carbon partitioning and source/sink relationships during growth, development and under biotic stresses. In this report, we monitored the expression/activity of cell wall invertases in organs behaving as source, sink, or subjected to a source/sink transition after infection with the necrotrophic fungus . We showed that organs with different source/sink status displayed differential CWIN activities, depending on carbohydrate needs or availabilities in the surrounding environment, through a transcriptional and posttranslational regulation. Loss-of-function mutation of the cell wall invertase 1 gene, , showed that the corresponding protein was the main contributor to the apoplastic sucrose cleaving activity in both leaves and roots. The CWIN-deficient mutant exhibited a reduced capacity to actively take up external sucrose in roots, indicating that this process is mainly dependent on the sucrolytic activity of . Using T-DNA and CRISPR/Cas9 mutants impaired in hexose transport, we demonstrated that external sucrose is actively absorbed in the form of hexoses by a sugar/H symport system involving the coordinated activity of AtCWIN1 with several Sugar Transporter Proteins (STP) of the plasma membrane, i.e., STP1 and STP13. Part of external sucrose was imported without apoplastic cleavage into seedling roots, highlighting an alternative -independent pathway for the assimilation of external sucrose. Accordingly, we showed that several genes encoding sucrose transporters of the plasma membrane were expressed. We also detected transcript accumulation of vacuolar invertase (VIN)-encoding genes and high VIN activities. Upon infection, was responsible for all the -induced apoplastic invertase activity. We detected a transcriptional activation of several and genes accompanied with an enhanced vacuolar invertase activity, suggesting that the -independent pathway is efficient upon infection. In absence of , we postulate that intracellular sucrose hydrolysis is sufficient to provide intracellular hexoses to maintain sugar homeostasis in host cells and to fuel plant defenses. Finally, we demonstrated that possesses its own functional sucrolytic machinery and hexose uptake system, and does not rely on the host apoplastic invertases.