RESUMO
Despite the identification of clubroot resistance genes in various Brassica crops our understanding of the genetic basis of immunity to Plasmodiophora brassicae infection in the model plant Arabidopsis thaliana remains limited. To address this issue, we performed a screen of 142 natural accessions and identified 11 clubroot-resistant Arabidopsis lines. Genome-wide association analysis identified several genetic loci significantly linked with resistance. Three genes from two of these loci were targeted for deletion by CRISPR/Cas9 mutation in resistant accessions Est-1 and Uod-1. Deletion of Resistance to Plasmodiophora brassicae 1 (RPB1) rendered both lines susceptible to the P. brassicae pathotype P1+. Further analysis of rpb1 knock-out Est-1 and Uod-1 lines showed that the RPB1 protein is required for activation of downstream defence responses, such as the expression of phytoalexin biosynthesis gene CYP71A13. RPB1 has recently been shown to encode a cation channel localised in the endoplasmic reticulum. The clubroot susceptible Arabidopsis accession Col-0 lacks a functional RPB1 gene; when Col-0 is transformed with RPB1 expression driven by its native promoter it is capable of activating RPB1 transcription in response to infection, but this is not sufficient to confer resistance. Transient expression of RPB1 in Nicotiana tabacum induced programmed cell death in leaves. We conclude that RPB1 is a critical component of the defence response to P. brassicae infection in Arabidopsis, acting downstream of pathogen recognition but required for the elaboration of effective resistance.
Assuntos
Arabidopsis , Brassica , Plasmodioforídeos , Arabidopsis/metabolismo , Doenças das Plantas , Estudo de Associação Genômica Ampla , Brassica/genéticaRESUMO
The hemp aphid Phorodon cannabis Passerini is a well- known (Asia, Europe) or newly emerging (North America) insect. It is a monophagous insect pest causing considerable damage in field and glasshouse cultivations. The aim of this work was to study the effects of meteorological (temperature) and agronomical (herbicide) factors on the biology of the hemp aphid. In one experiment, hemp plants were kept at constant temperatures ranging from 20 to 30 °C, and aphid survival and fecundity were measured. In a related experiment conducted at 20 °C, plants were treated with field-appropriate rates of a selective graminicide containing quizalofop-P-tefuryl (40 gL-1, 4.38%, HRAC group 1), commonly used to control weeds in hemp, and aphid enzyme activity was measured in addition to population parameters. We found that hemp aphids could live, feed and reproduce within the whole studied range of temperatures, demonstrating its great evolutionary plasticity. However, the optimal temperature for development was 25 °C, at which the insect lived and reproduced for 25 and 15 days, respectively, with an average fecundity of 7.5 nymphs per reproduction day. The herbicide treatment increased the activity of superoxide dismutase (SOD), catalase (CAT), ß-glucosidase, S-glutathione transferase (GST), oxidoreductive peroxidase (POD), and polyphenol oxidase (PPO) in the aphids, but only on certain days after treatment, which indicates a mild stress in aphid tissues, related to a higher reproduction and changed feeding behavior; aphids moved from the actively growing tips compared to untreated plants. The results of these experiments are discussed in terms of the impact on the future management of this pest.
RESUMO
The ability of hemp (Cannabis sativa L.) inflorescence extract to counteract lipid oxidation was studied in stripped linseed oil. The ethanolic extract was characterized in terms of terpenes (6.00 mg/mL), cannabidiol (4.99% w/w), phenolic compounds (1.80 mg gallic acid equivalents (GAE)/mL), antiradical, and metal ion-chelating activities (50% effective concentration (EC50) of 2.47 mg/mL and 0.39 mg/mL, respectively). The stripped linseed oil, used as control (CO), was mixed with hemp extract (HO) or α-tocopherol (EO) at a ratio of 0.6% (w/w) and stored for 7 days in darkness at 40 °C. Hemp extract reduced the oxidation and lipolysis processes. At the end of the storage, HO showed a significantly higher level of α-linolenic acid (ALA; 26.64 g/100 g), lower peroxide value (PV) (21.19 meq O2/kg oil), and lower hexanal content (7.67 mmol/kg oil) than those found in the control. In contrast, EO showed a marked lipolysis (the free fatty acids increased by 42.57%) and a noticeable oxidation, since the ALA content decreased by 2.10% and a PV of 50 meq O2/kg oil was observed. This study demonstrates that hemp inflorescences can be used as a source of natural antioxidants in vegetable oils and lipid products to retard their oxidation, especially those characterized by a high degree of unsaturation.
RESUMO
Hemp (Cannabis sativa L.) has become widely used in several sectors due to the presence of various bioactive compounds such as terpenes and cannabidiol. In general, terpenes and cannabidiol content is determined separately, which is time consuming. Thus, a fast gas chromatography with flame ionization detection method was validated for simultaneous determination of both terpenes and cannabidiol in hemp. The method enabled a rapid detection of 29 different terpenes and cannabidiol within a total analysis time of 16 min, with satisfactory sensitivity (limit of detection = 0.03-0.27 µg/mL, limit of quantitation = 0.10-0.89 µg/mL). The inter- and intraday precision (RSD) was <7.82 and <3.59%, respectively. Recoveries at two spiked concentration levels (low, 3.15 µg/mL; high, 20.0 µg/mL) were determined on both apical leaves (78.55-101.52%) and inflorescences (77.52-107.10%). The reproducibility (RSD) was <5.94 and <5.51% in apical leaves and inflorescences, respectively. The proposed and validated method is highly sensitive, robust, fast, and accurate for determination of the main terpenes and cannabidiol in hemp and could be routinely used for quality control.