Analysis of Glucosinolate Content and Metabolism Related Genes in Different Parts of Chinese Flowering Cabbage

Glucosinolates (GSLs) are important secondary metabolites that play important defensive roles in cruciferous plants. Chinese flowering cabbage, one of the most common vegetable crops, is rich in GSLs and thus has the potential to reduce the risk of cancer in humans. Many genes that are involved in GSL biosynthesis and metabolism have been identified in the model plant Arabidopsis thaliana; however, few studies investigated the genes related to GSL biosynthesis and metabolism in Chinese flowering cabbage. In the present study, the GSL composition and content in three different organs of Chinese flowering cabbage (leaf, stalk, and flower bud) were determined. Our results showed that the total GSL content in flower buds was significantly higher than in stalks and leaves, and aliphatic GSLs were the most abundant GSL type. To understand the molecular mechanisms underlying the variations of GSL content, we analyzed the expression of genes encoding enzymes involved in GSL biosynthesis and transport in different tissues of Chinese flowering cabbage using RNA sequencing; the expression levels of most genes were found to be consistent with the pattern of total GSL content. Correlation and consistency analysis of differentially expressed genes from different organs with the GSL content revealed that seven genes (Bra029966, Bra012640, Bra016787, Bra011761, Bra006830, Bra011759, and Bra029248) were positively correlated with GSL content. These findings provide a molecular basis for further elucidating GSL biosynthesis and transport in Chinese flowering cabbage.

Productivity of camelina and spring rapeseed as affected by varietal characteristics in the Right-Bank Forest-Steppe of Ukraine

Purpose. Determine the oil content and glucosinolate content and productivity of spring camelina and spring rapeseed as affected by varietal characteristics. Methods. The research was conducted in the educational-scientific-industrial complex of Uman National University of Horticulture. Spring rapeseed varieties ‘Belinda’, ‘Aidar’, ‘Heros’, ‘Jerry’ and spring camelina varieties ‘Zevs’, ‘Hirskyi’, ‘Slavutych’, ‘Mirazh’ were studied. Seeds were sown in a row to a depth of 1.5 cm with a seeder CH-16. The sowing rate was 300 seeds/m2, or 5.4 kg/ha. The crop was harvested separately by Sampo-500 combine. Results. In spring rapeseed, the highest yield was marked by variety ‘Aidar’ (2.52 t/ha) and the lowest by ‘Heros’ (2.43 t/ha). In spring camelina, the highest yield was in variety ‘Zevs’ (2.31 t/ha) and lower in ‘Hirska’ (2.15), ‘Mirazh’ (2.27), and ‘Slavutych’ (2.22 t/ha). The highest oil content among rapeseed varieties demonstrated ‘Aidar’ (43.7%). Slightly lower it was in ‘Belinda’ (43.2%), ‘Jerry’ (42.8) and ‘Heros’ (42.6%). Depending on the varietal characteristics, the highest percentage of oil content was found in spring camelina varieties ‘Zevs’ (45.1%). It was lower by 0.5 and 0.9% in ‘Mirazh’ and ‘Slavutych’. The lowest percentage of oil content (43.7%) was determined in camelina variety ‘Hirska. In spring rapeseed varieties, the highest content of glucosinolates was in ‘Aidar’ – 20.5 μmol/g, 0.3 and 0.5 μmol/g less in ‘Belinda’ and ‘Jerry’, and the lowest in ‘Heros’ (19.8 μmol/g). In spring camelina, the highest value was in the variety ‘Zevs’ (22.0 μmol/g), and the lowest in ‘Hirska’ (21.0 μmol/g). ‘Mirazh’ and ‘Slavutych’ contained 21.7 and 21.4 μmol/g of glucosinolatesm respectively. Conclusions. The productivity of oilseed crops varies under the same growing conditions: spring rapeseed, regardless of varietal characteristics, demonstrated higher yield compared to camelina. On average over the years of research and varieties, this indicator was 2.47 t in spring rapeseed, and 2.24 t/ha in spring camelina, which is 0.23 t/ha less. In rapeseed, the oil content of the seeds was 43.07%, the content of glucosinolates varied from 19.8 to 20.5 μmol/g. In spring camelina, the oil content of seeds was 1.33–44.40% higher, the content of glucosinolates ranged from 21.0 to 22.0 μmol/g. The level of profitability of the studied oilseed crops was high and varied as affected by varietal characteristics: in rapeseed from 122 to 141%, and in spring camelina from 182 to 196%.

The results of preliminary trial experimental hybrids of winter rapeseed bred at VNIIMK by economically valuable traits

The purpose of the research was a preliminary estimation of ten experimental hybrids of winter rapeseed (Brassica napus L.) bred in the V.S. Pustovoit All-Russian Research Institute of Oil Crops by the primary economically valuable traits. The research was conducted in the central zone of the Krasnodar region in 2019–2020. The results of the research are presented for the first time. On average for two years, nine of ten the best experimental hybrids increased by seeds yield by 0.32–0.76 t per ha both the standard cultivar Loris and the foreign hybrid Mercure. Middle seed yield of the studied hybrids varied significantly, variance range was from 4.99 t per ha (VN-3 × ОРK-10) to 5.62 t per ha (VN-4 × ОРK-20). Maximal values of seed and oil yields per a unit area were fixed for a hybrid combination VN-4 × ОРK-20. Except that, the hybrid is characterized with high weight of 1000 seeds (4.5 g). A hybrid combination VN-13 × ОРK-10 demonstrated high average by years seeds yield (5.33 t per ha) and oil yield (2.25 t per ha).Oil content in seeds and weight of 1000 seeds of this combination were at the level of the standard cultivar and the foreign hybrid. This hybrid shows the lowest glucosinolate content in seeds – 16.7 µmol/g, that is less that the foreign hybrid Mercure has by 2.3 µmol/g. The preliminary conclusion based on our researches is: the selected hybrids after confirmation of the competitive trial results can be of a great interest for production.

Adaptive reaction of perspective spring rapeseed cultivars in conditions of the central zone of the Krasnodar region

We studied 18 perspective and already registered spring rapeseed cultivars of VNIIMK breeding. Impact of weather conditions in a period flowering – maturing on economically valuable traits was revealed. The cultivars were compared by calculated statistic parameters of adaptivity, environmental plasticity, stability in the conditions of the central zone of the Krasnodar region. The perspective linear cultivars which demonstrated the most stably yield in the different years were selected. The researches were conducted in the V.S. Pustovoit All-Russian Research Institute of Oil Crops, Krasnodar, Russia, in 2017– 2020. The most favorable weather conditions for cultivation of spring rapeseed appeared to be in 2017. Index of environmental condition (Ij) was 1.01. On average for rapeseed cultivar, yield varied from maximal in 2017 (2.83 t per ha) to minimal in 2019 (1.31 t per ha). Oil content in the different years varied from 40.9 to 46.34%, weight of 1000 seeds – from 2.22 to 3.85 g. Glucosinolate content was minimal (14.16 µmol/g) in 2017 and reached maximum (21.48 µmol/g) in 2020. The linear cultivars ВН-LР13, ВНDl 2, and ВН-Dl 1 are characterized with the highest environmental plasticity, having the indicators bi = 1.30; 1.24, and 1.21, respectively. The cultivars Amulet and Ruyan were the most stable: Si² was equal to 0.65 and 0.69, respectively, as well as the main applicant for being a cultivar: ВН-4801, ВН-191, and ВН2478 having the indicators 0.40; 0.62, and 0.80. The perspective cultivars ВН-4801 and ВН-906 (Kenar) are characterized with a valuable combination of increased and stable for years yield. Due to absolute indicator of adaptability, all the studied cultivars are ranking as following: ВН-4801 (113.6%), ВН-Dl 1 (106.3%), ВН-Dl 2 (105.9%), ВН-LР 13 (105.0%), ВН-XT (103.4%), Руян (102.9%), and ВН-906 (101.9%).

Phytopathological and nutraceutical evaluation of cauliflower plants treated with high dilutions of arsenic trioxide

Introduction: This research aimed at verifying the effects of highly diluted (HD) treatments on cauliflower (Brassica oleracea L.) plants both healthy and inoculated by the fungus Alternaria brassicicola, causing the dark leaf spot disease. In vitro spore germination assays (A), growth chamber experiments (B) and field trials (C) were performed. Material and Methods: (A): spore suspensions were prepared in HD treatments and their inhibiting effect on germination was recorded microscopically after incubation at 25°C for 5 h. (B): the same treatments were tested in plants artificially inoculated with the fungus. The infection level on leaves was blindly evaluated by a previously defined infection scale. (C): the field was divided into plots according to a complete randomized block design. In the first trial (i), plants were artificially inoculated and weekly treated; the infection level was evaluated on cauliflower heads. The second trial (ii) was performed on the same field with the aim to induce a natural infection, mediated by infected crop residues. Measurement endpoints concerned the evaluation of some physiological parameters along with the glucosinolate content on cauliflower heads. Results: (A): arsenic trioxide (As 35x and 35x diluted 1:5000) and Cuprum 5x induced highly significant inhibition of germination rate (-60%) vs. control. (B): As 35x and Cu 3 g/l induced a significant decrease of mean infection level (-50%). (C): in (i), a significant reduction of disease symptoms on heads was recorded for As 35x and Cu 3 g/l (-45%). In (ii) natural fungal infection did not occur due to dry weather conditions; physiological and nutraceutical analyses of healthy heads demonstrated that As 35x induced a significant increase of both head size and glucosinolate content. Discussion: Some evidences on the efficacy of arsenic, at different decimal and centesimal HD, in fungal and viral disease control were previously reported [1]. In the present study the efficacy of HD arsenic in dark leaf spot control in field has been shown for the first time: since fungal inoculation was performed on the leaves before flowering, we can hypothesize that this treatment induced an increase of plant resistance to fungal infection. Conclusions: This research showed the possibility of using HD arsenic in agriculture (“agrohomeopathy”), as it increased both plant resistance to fungal infection and the content of glucosinolates, ie secondary metabolites involved in plant resistance mechanisms [2] and considered as “plant food protection agents” [3]. Acknowledgments: Authors declare there is no conflict of interest. This research has been supported by Marche Region. A grateful acknowledgement to Dr. Leonardo Valenti for his support to this research. The authors thank Laboratoires Boiron srl for the grant awarded to one of the author Dr. Grazia Trebbi. Finally, authors are grateful to Cemon srl for financial support of glucosinolate analyses. We had full access to all the data in this study and we take responsibility for the integrity of the data and the accuracy of the data analysis Keywords: cauliflower, arsenic trioxide, Alternaria brassicicola, glucosinolates References [1] Betti L, Trebbi G, Majewsky V, Scherr C, Shah-Rossi D, Jäger T, et al. Use of homeopathic preparations in phytopathological models and in field trials: a critical review. Homeopathy 2009; 98: 244-266. [2] Ménard R, Larue J-P, Silué D, Thouvenot D. Glucosinolates in cauliflower as biochemical markers for resistance against downy mildew. Phytochemistry 1999; 52: 29-35. [3] Talalay P, Fahey JW. Phytochemicals from Cruciferous plants protect against cancer by modulating carcinogen metabolism. J Nutr. 2001; 131:3027S- 3033S.

Glucosinolates and Isothiocyantes in Processed Rapeseed Determined by HPLC-DAD-qTOF

Glucosinolates are well known as natural antimicrobials and anticarcinogenic agents. However, these compounds can lose their properties and transform into antinutrients, depending on processing conditions. In addition, the bitterness of some glucosinolate in rapeseed meal can affect the likability of the final product. Therefore, it is important to identify and determine each glucosinolate and its derived form, not just the total glucosinolate content, in order to evaluate the potential of the final rapeseed protein product. This study provides a comprehensive report of the types and quantities of glucosinolates and their derived forms (isothiocyanates) associated with different rapeseed processing conditions. Glucosinolates and isothiocyanates were determined by HPLC-DAD-qTOF. In our study, the enzymatic degradation of glucosinolates by myrosinase was the main factor affecting either glucosinolate or isothiocyanate content. Other factors such as pH seemed to influence the concentration and the presence of glucosinolates. In addition, process parameters, such as extraction time and separation technology, seemed to affect the amount and type of isothiocyanates in the final protein extracts. Overall, both determined intact glucosinolates and their derived forms of isothiocyanates can give different types of biological effects. More studies should be performed to evaluate the impact of glucosinolates and isothiocyanates on human health.

Efficacy of Different Forms of Green Manure Crops to Reduce Verticillium dahliae in Different Soils

The efficacy of green manure crops to reduce the number of Verticillium dahliae microsclerotia in different soils was investigated. Green manures tested were Indian mustard with a high glucosinolate content and sorghum-sudangrass as biocidal plants, and Indian mustard with a low glucosinolate content and rye as non-biocidal plants. The green manure plants were applied in fresh, dried, and ensilaged form. When applied as fresh plants, the glucosinolate content determining the biocidal activity of Indian mustard was only important in loam soil but not in sandy loam soil. In the latter soil, the non-biocidal rye had significantly higher efficacy than the Indian mustard. Volatiles released by fresh and dried, but not ensilaged, Indian mustard with a high glucosinolate content strongly decreased the number of living V. dahliae microsclerotia. When the same green manure crops were added to sandy loam and clay loam soil, the effect of the high glucosinolate content Indian mustard in fresh and dried form disappeared, whereas the ensilaged green manure crops had the highest efficacy. This effect was based on the increase of the soil microbial activity and the Streptomyces population size, which were negatively correlated with the number of living V. dahliae microsclerotia in the soil.

Trichoderma hamatum Increases Productivity, Glucosinolate Content and Antioxidant Potential of Different Leafy Brassica Vegetables

Brassica crops include important vegetables known as “superfoods” due to the content of phytochemicals of great interest to human health, such as glucosinolates (GSLs) and antioxidant compounds. On the other hand, Trichoderma is a genus of filamentous fungi that includes several species described as biostimulants and/or biological control agents in agriculture. In a previous work, an endophytic strain of Trichoderma hamatum was isolated from kale roots (Brassica oleracea var. acephala), describing its ability to induce systemic resistance in its host plant. In the present work, some of the main leafy Brassica crops (kale, cabbage, leaf rape and turnip greens) have been root-inoculated with T. hamatum, having the aim to verify the possible capacity of the fungus as a biostimulant in productivity as well as the foliar content of GSLs and its antioxidant potential, in order to improve these “superfoods”. The results reported, for the first time, an increase in the productivity of kale (55%), cabbage (36%) and turnip greens (46%) by T. hamatum root inoculation. Furthermore, fungal inoculation reported a significant increase in the content of total GSLs in cabbage and turnip greens, mainly of the GSLs sinigrin and gluconapin, respectively, along with an increase in their antioxidant capacity. Therefore, T. hamatum could be a good agricultural biostimulant in leafy Brassica crops, increasing the content of GSLs and antioxidant potential of great food and health interest.

Role of Glucosinolates in the Nutraceutical Potential of Selected Cultivars of Brassica rapa

Brassica rapa L. subsp. rapa (turnip greens), a traditionally consumed vegetable, is well-known due to its high content of glucosinolates, which are secondary metabolites with a positive biological activity for human health. Our hypothesis has been based on the relation between B. rapa glucosinolate content and its healthy properties, and our aim is to establish guidelines for safe B. rapa vegetable consumption. Three B. rapa cultivars (143N5, 143N7 and 163N7) have been characterized by HPLC analysis of purified extracts from leaf samples in order to determine their glucosinolate content and to relate this content to beneficial effects on DNA protection, lifespan extension and chemoprevention. In order to ascertain the heath properties in vitro and in vivo, toxicity activities were assayed in the Drosophila melanogaster and leukaemia cell models; genomic safety was also assessed in both models using genotoxicity, fragmentation and comet assay. The Drosophila model has also been used to study the antioxidative activity and the longevity induction. Our results showed a relationship between B. rapa glucosinolate content and its safety and benefices in its consumption. Gluconapin, the main B. rapa glucosinolate, was directly related with these wholesome effects. The relevant conclusion in the present research is focused on B. rapa cultivar 163N7 due to its high gluconapin content and low progoitrin content, which exert anti-cancer and DNA protection properties and could be recommended as being safe and healthy for human consumption.

Analysis of Glucosinolate Content, Composition and Expression Level of Biosynthesis Pathway Genes in Different Chinese Kale Varieties

The content and component of glucosinolates in edible stems and leaves of eight Chinese kale varieties from Japan and eight varieties from China were determined by HPLC-MS. Simultaneously, the expression levels of glucosinolate biosynthesis pathway genes from four varieties with high and low total glucosinolate contents were analyzed by the qRT-PCR method. Four types of aliphatic glucosinolates (A-GLSs: GRA, SIN, GNA and GER) and indole glucosinolates (I-GLSs: 4-HGBS, GBS, 4-MGBS and NGBS) were detected in the stems and leaves of 16 varieties, and no aromatic glucosinolates (R-GLSs) were detected. A-GLSs account for more than 80.69% of the total content of total glucosinolates (T-GLSs), in which GNA and GRA are the main components of stems and leaves. Among Japanese varieties, QB1 has higher content of A- and T-GLSs, while that of XLB was lower; however, the corresponding varieties were ZH and DSHH in Chinese varieties. Among the above four varieties, the expression levels of SOT16, CYP83B1, SOT17, CYP83A1 and MAM1 genes were significantly higher in the varieties with higher GLSs; the expression levels of SOT16 and CYP83B1 were consistent with the content of I-GLSs; and SOT17, CYP83A1 and MAM1 expression levels were consistent with A-GLSs content. At the same time, the expression levels of SOT16 and CYP83B1 in the leaves were higher than those in the stems. CYP83A1 and MAM1 genes were less expressed in the leaves than in the stems of lower content varieties. It is speculated that these genes may be the key genes regulating GLS biosynthesis in Chinese kale.