Glucosinolates in Sisymbrium officinale (L.) Scop.: Comparative Analysis in Cultivated and Wild Plants and in Vitro Assays with T2Rs Bitter Taste Receptors.

Sisymbrium officinale (L.) Scop., commonly known as "hedge mustard" or "the singer's plant" is a wild plant common in Eurasian regions. Its cultivation is mainly dedicated to herboristic applications and it has only recently been introduced into Italy. The active botanicals in S. officinale are glucosinolates, generally estimated by using UV or high-performance liquid chromatography (HPLC). Using both techniques, we measured the total glucosinolates from S. officinale in different parts of the plant as roots, leaves, seeds, and flowers. A comparison was made for cultivated and wild samples, and for samples obtained with different pre-treatment and fresh, frozen, and dried storage conditions. Cultivated and wild plants have a comparable amount of total glucosinolates, while drying procedures can reduce the final glucosinolates content. The content in glucoputranjivin, which is the chemical marker for glucosinolates in S. officinale, has been determined using HPLC and a pure reference standard. Glucoputranjivin and two isothiocyanates from S. officinale have been submitted to in vitro assays with the platform of bitter taste receptors of the T2Rs family. The results show that glucoputranjivin is a selective agonist of receptor T2R16.

Isothiocyanates and Glucosinolates from Sisymbrium officinale (L.) Scop. ("the Singers' Plant"): Isolation and in Vitro Assays on the Somatosensory and Pain Receptor TRPA1 Channel.

Sisymbrium officinale (L.) Scop. is a wild common plant of the Brassicaceae family. It is known as "the singers' plant" for its traditional use in treating aphonia and vocal disability. Despite its wide use in herbal preparations, the molecular mechanism of action of S. officinale extracts is not known. The plant is rich in glucosinolates and isothiocyanates, which are supposed to be its active compounds. Some members of this family, in particular allylisothiocyanate, are strong agonists of the transient receptor potential ankyrin 1 (TRPA1) channel, which is involved in the somatosensory perception of pungency as well as in the nociception pathway of inflammatory pain. This study aims to isolate the glucosinolates and isothiocianates from fresh S. officinale to identify the major components and test their activity in in vitro assays with a cloned TRPA1 channel. Samples of cultivated S. officinale have been extracted and the active compounds isolated by column chromatography, HPLC and PTLC. The main components glucoputranjivin, isopropylisothiocyanate and 2-buthylisothiocianate have been tested on TRPA1. The glucosinolates glucoputranjivin and sinigrin turned out to be inactive, while isopropylisothiocyanate and 2-buthylisothiocyanate are potent agonists of TRPA1, with an EC50 in the range of the high potency natural agonists identified so far for this somatosensory channel.

Involvement of miRNAs in Metabolic Herbicide Resistance to Bispyribac-Sodium in Echinochloa crus-galli (L.) P. Beauv.

Several mechanisms involved in weed herbicide resistance are unknown, particularly those acting at the epigenetic level, such as the capacity of small-non-coding RNAs (sncRNAs) to target messenger RNAs of genes involved in herbicide detoxification. The transcription of these sncRNAs is stimulated by epigenetic factors, thereby affecting gene expression. This study was carried out in order to evaluate, for the first time in Echinochloa crus-galli (L.) P. Beauv. (barnyardgrass), the capacity of miRNAs to regulate the expression of genes associated with bispyribac-sodium detoxification. The expression profiles of eight miRNAs with a high degree of complementarity (≥80%) with mRNAs of genes involved in herbicide detoxification (CYP450, GST and eIF4B) were determined by qRT-PCR before and after herbicide spraying. Five of the miRNAs studied (gra-miR7487c, gma-miR396f, gra-miR8759, osa-miR395f, ath-miR847) showed an increased expression after herbicide application in both susceptible and resistant biotypes. All the miRNAs, except gra-miR8759, were more highly expressed in the herbicide-resistant biotypes. In specimens with increased expression of miRNAs, we observed reduced expression of the target genes. The remaining three miRNAs (ata-miR166c-5p, ath-miR396b-5p and osa-miR5538) showed no over-expression after herbicide treatment, and no difference in expression was recorded between susceptible and resistant biotypes. Our results represent a first overview of the capacity of miRNAs to regulate the expression of genes involved in bispyribac-sodium detoxification in the genus Echinochloa. Further research is required to identify novel miRNAs and target genes to develop more focused and sustainable strategies of weed control.