A Phytocomplex Obtained from Salvia officinalis by Cell Culture Technology Effectively Controls the Grapevine Downy Mildew Pathogen Plasmopara viticola.

The negative impact of using conventional fungicides in plant disease protection has increased the interest in safer alternatives such as plant secondary metabolites, generally having a better toxicological profile. However, cultivation conditions and plant material strongly affect the quality and quantity of secondary metabolites obtained from field grown plants, limiting the standardization needed for industrial production. Plant cell culture technology can provide highly homogeneous biomasses with specific chemical characteristics. A phytocomplex with high rosmarinic acid content (10.12% w/w) was obtained from a selected cell line of Salvia officinalis and was tested against the grapevine downy mildew pathogen, Plasmopara viticola. Grapevine leaf discs were sprayed with the phytocomplex at 5 g/L and then inoculated with P. viticola sporangia. Sporulation level on each disc was assessed after 7 days with an image processing software. The phytocomplex reduced by 95% the sporulation level compared to the control and was also more effective than rosmarinic acid alone, used at the same concentration found in the phytocomplex. Persistence of the phytocomplex was also assessed: when applied 5 days before inoculation, it reduced by 90% the sporulation level compared to the control. These results highlight the possibility to take advantage of cell culture techniques to produce safer pesticides with high quality standards.

Transgene pyramiding in wheat: Combination of deoxynivalenol detoxification with inhibition of cell wall degrading enzymes to contrast Fusarium Head Blight and Crown Rot.

Fusarium Head Blight (FHB) and Crown Rot (FCR) are major diseases of wheat crops, causing extensive damages and mycotoxin contamination. In this work, we investigated the possibility to improve resistance to either or both diseases by combining different resistance mechanisms. To this aim, we stacked in the same wheat genotype transgenes controlling the DON-to-D3G conversion by specific UDP-glucosyltransferases (UGT) and the inhibition of cell wall degrading enzymes (CWDEs) by glycosidase inhibitors. We obtained: i) a durum wheat UGT+PMEI double-transgenic line constitutively expressing the HvUGT13248 and AcPMEI genes, coding for a barley UGT and a kiwi pectin methylesterase inhibitor, respectively; ii) a bread wheat UGT+PGIP line, expressing in floral tissues the HvUGT13248 gene and constitutively the PvPGIP2 gene, coding for a bean polygalacturonase inhibiting protein. We observed that both UGT+PMEI and UGT+PGIP plants exhibited increased resistance against Fusarium graminearum in FHB, further reducing by 10-20 % FHB symptoms as compared to the lines carrying the individual transgenes, and of up to 50 % as compared to wild-type plants. On the other hand, double-transgenic UGT+PMEI seedlings exhibited similar FCR symptoms as the UGT single transgenic line after infection with F. culmorum, indicating no contribution of the PMEI transgene to FCR resistance. This result is also supported by the inability of AcPMEI or PvPGIP2, constitutively expressed in durum wheat transgenic lines, to counteract F. graminearum in FCR. We also verified that F. graminearum produces PG and PME activity on infected wheat crown. We conclude that CWDEs inhibition combined with UGT-based DON detoxification contribute in an additive manner to limiting F. graminearum in FHB. Conversely, UGT-based DON detoxification is the only mechanism contributing to resistance observed against FCR. Indeed, the reinforcement of pectin does not enhance resistance against FCR.

The Rare Sugar Tagatose Differentially Inhibits the Growth of Phytophthora infestans and Phytophthora cinnamomi by Interfering With Mitochondrial Processes.

Rare sugars are monosaccharides with limited availability in nature and their biological functions are largely unknown. Among them, tagatose was developed as a low-calorie sweetener and showed beneficial effects on human health. Tagatose is metabolized by only certain microbial taxa and inhibits the growth of important crop pathogens (e.g., Phytophthora infestans), but its mode of action and the microbial responses are unknown. The aim of this study was to understand the tagatose mode of action against Phytophthora spp., with the final aim of developing new plant protection products. Tagatose inhibited P. infestans growth in vitro and caused severe ultrastructural alterations, with the formation of circular and concentric mitochondrial cristae. Decreased ATP content and reduced oxygen consumption rate (OCR) were found in tagatose-incubated P. infestans as compared to the control, with the consequent accumulation of reactive oxygen species (ROS) and induction of genes related to apoptosis and oxidative stress response. On the other hand, tagatose did not, or only slightly, affect the growth, cellular ultrastructure and mitochondrial processes in Phytophthora cinnamomi, indicating a species-specific response to this rare sugar. The mode of action of tagatose against P. infestans was mainly based on the inhibition of mitochondrial processes and this rare sugar seems to be a promising active substance for the further development of eco-friendly fungicides, thanks to its anti-nutritional properties on some phytopathogens and low risk for human health.