Background: Polyphenols are secondary metabolites produced by plants to defend themselves from environmental stressors. We explored the effect of Wolffia globosa ‘Mankai’, a novel cultivated strain of a polyphenol-rich aquatic plant, on the metabolomic-gut clinical axis in vitro, in-vivo and in a clinical trial. Methods: We used mass-spectrometry-based metabolomics methods from three laboratories to detect Mankai phenolic metabolites and examined predicted functional pathways in a Mankai artificial-gut bioreactor. Plasma and urine polyphenols were assessed among the 294 DIRECT-PLUS 18-month trial participants, comparing the effect of a polyphenol-rich green-Mediterranean diet (+1240 mg/polyphenols/day, provided by Mankai, green tea and walnuts) to a walnuts-enriched (+440 mg/polyphenols/day) Mediterranean diet and a healthy controlled diet. Results: Approximately 200 different phenolic compounds were specifically detected in the Mankai plant. The Mankai-supplemented bioreactor artificial gut displayed a significantly higher relative-abundance of 16S-rRNA bacterial gene sequences encoding for enzymes involved in phenolic compound degradation. In humans, several Mankai-related plasma and urine polyphenols were differentially elevated in the green Mediterranean group compared with the other groups (p < 0.05) after six and 18 months of intervention (e.g., urine hydroxy-phenyl-acetic-acid and urolithin-A; plasma Naringenin and 2,5-diOH-benzoic-acid). Specific polyphenols, such as urolithin-A and 4-ethylphenol, were directly involved with clinical weight-related changes. Conclusions: The Mankai new plant is rich in various unique potent polyphenols, potentially affecting the metabolomic-gut-clinical axis.

An outbreak of a new disease infecting tomato plants ( Solanum lycopersicum) which carry the tobamovirus resistance gene Tm-2 occurred in southern Israel during the autumn of 2014. Symptomatic plants showed a mosaic pattern on leaves accompanied by narrowing of leaves and yellow spotted fruit. It was found that the causal agent is a new tobamovirus that was identified earlier in Jordan and named Tomato brown rugose fruit virus (ToBRFV). The virus is fast spreading in many tomato growing-areas in Israel, and since then it was found to be spreading rapidly world-wide. According to tomato growers, ToBRFV induces severe yield loses due to reduction in plant vigor and fruit quality. Here, we assayed the effect ToBRFV has on tomato yield and yield components of four different tomato genotypes, which included two near-isogenic lines, one with and one without the known tobamovirus resistance gene Tm-22, a tolerant genotype and a resistant genotype. Two yield experiments that were carried out at two different geographical locations were conducted. In each experiment, the yield performances of ToBRFV-inoculated plants were compared to those of control non-inoculated plants of the same genotype. It was found that the virus is able to induce a yield reduction of 15-55% regardless if the plants carry the resistance gene Tm-2(2). The resistant and tolerant genotypes also suffered a yield reduction following inoculation with ToBRFV, but the reduction was not statistically significant for the tolerant genotype in both trials, while for the resistant genotype in one trial it was statistically significant and not so in the other trial.

Tomato yellow leaf curl virus (TYLCV), a whitefly-transmitted single-stranded DNA (ssDNA) virus, causes the most important viral disease of tomato worldwide. TYLCV-mediated disease is mainly controlled via extensive insecticide sprays aiming at the whitefly vector. RNA-based vaccination was proven to be a non-transgenic approach leading to efficient plant virus control. In this work, double-stranded RNA (dsRNA) molecules deriving from sequences of the C4 and V2 genes of TYLCV-Mild were produced in vitro and topically applied onto tomato plants along with the virus (via agroinfiltration). DsC4 and dsV2 application reduced disease incidence to 23 and 46 %, respectively, while TYLCV positive control reached 64 %. Bioinformatics analysis of the virus-specific small interfering RNAs (vsiRNAs) from TYLCV-infected tomato revealed 'hot' and 'cold' spots in the TYLCV-Mild genome. Interestingly, the viral C-strand had twofold siRNA reads when compared to that of the V-strand. Overall, vsiRNAs of negative and positive polarity were almost equal (53.5 vs. 46.6 %); vsiRNAs of negative polarity prevailed at the V-strand. Stem-loop RT-PCR validated the presence of six vsiRNAs (hot or cold spots) in TYLCV-Mild-infected and dsRNA-treated tomato. The exogenously applied dsRNA was found to rapidly move systemically in tomato and was detected for 54 days post treatment (dpt). The applied dsRNA molecules were successfully processed by the Dicer-like proteins (DCLs) in tomato since small interfering RNAs (siRNAs) deriving from the dsRNA were detected for at least 54 dpt. This consists the first report of dsRNA-based vaccination applied against a monopartite geminivirus.

BACKGROUND:Rare plants that contain corrinoid compounds mostly comprise cobalamin analogues, which may compete with cobalamin (vitamin B12 (B12)) metabolism. We examined the presence of B12 in a cultivated strain of an aquatic plant: Wolffia globosa (Mankai), and predicted functional pathways using gut-bioreactor, and the effects of long-term Mankai consumption as a partial meat substitute, on serum B12 concentrations.METHODS:We used microbiological assay, liquid-chromatography/electrospray-ionization-tandem-mass-spectrometry (LC-MS/MS), and anoxic bioreactors for the B12 experiments. We explored the effect of a green Mediterranean/low-meat diet, containing 100 g of frozen Mankai shake/day, on serum B12 levels during the 18-month DIRECT-PLUS (ID:NCT03020186) weight-loss trial, compared with control and Mediterranean diet groups.RESULTS:The B12 content of Mankai was consistent at different seasons (p = 0.76). Several cobalamin congeners (Hydroxocobalamin(OH-B12); 5-deoxyadenosylcobalamin(Ado-B12); methylcobalamin(Me-B12); cyanocobalamin(CN-B12)) were identified in Mankai extracts, whereas no pseudo B12 was detected. A higher abundance of 16S-rRNA gene amplicon sequences associated with a genome containing a KEGG ortholog involved in microbial B12 metabolism were observed, compared with control bioreactors that lacked Mankai. Following the DIRECT-PLUS intervention (n = 294 participants; retention-rate = 89%; baseline B12 = 420.5 ± 187.8 pg/mL), serum B12 increased by 5.2% in control, 9.9% in Mediterranean, and 15.4% in Mankai-containing green Mediterranean/low-meat diets (p = 0.025 between extreme groups).CONCLUSIONS:Mankai plant contains bioactive B12 compounds and could serve as a B12 plant-based food source.

AbstractMankai is a cultivated strain of Wolffia globose, an aquatic plant, one of the smallest plants on earth, belonging to the family Lemnaceae, known commonly as Duckweeds. Mankai is being considered as a high-quality substitute for animal protein, and a potential source of vitamin B12(1). In fact, it contains all 9 essential amino acids and its protein profile is extremely close to that of egg. Recently, it has been granted GRAS status, and chosen as a test food in a long-term intervention DIRECT-PLUS (ClinicalTrials.gov identifier (NCT number): NCT03020186) which aims to compare the effect of green-Mediterranean diet (composed with polyphenol enriched food including Mankai) + physical activity (PA) vs. Mediterranen diet + PA vs in 300 volunteers suffering from cardiometabolic disease. Mankai is also rich in iron and has been shown in humans to maintain Iron-Homeostasis and to reverse Anemia in Rats(2). However, molecular characterization of phenolic composition of Mankai plant has not previously been reported. Our initial measurements of total phenolic content determined by Folin-Ciocalteu assay classifies Mankai amongst the foods highest in polyphenols content (8606.7 mg/kg)(3). This poster presents the characterization of Mankai polyphenols according to Vrhovsek et al. 2012 [4] using UHPLC-ESI-MS/MS system, identifying 26 different polyphenols. One of the main advantages of Mankai is its hydroponic cultivation that optimizes yield throughout the year. Light source, water and mineral management can influence the composition of phenolic content. We quantified Mankai polyphenols in 30 different plant batches and 52 different treatments to assess how quality of light may play a major role in the accumulation of secondary plant compounds. Notably, led light seems to enhance the glycated form of luteolin and quercetin compared to sunlight and the aglycone form of luteolin and quercetin are higher in sunlight treatment. The present analysis confirms the high polyphenol status of Mankai, profiles its major polyphenol components and provides new information on how production process in terms of light quality determines polyphenol content.

We are thankful for the justified questions of Dr. Christine Dawczynski and her colleagues regarding the presence of an active B12 vitamin in this specific plant, as this is, indeed, a unique phenomenon, which deserves further studies. Thank you for the opportunity to clarify and to add new accumulated information. Letter to original article by Kaplan et al. 2018 - Protein bioavailability of Wolffia globosa duckweed, a novel aquatic plant, A randomized controlled trialClinical NutritionVol. 38Issue 5PreviewKaplan et al. [1] conducted a randomized controlled human study to compare bioavailability of nutrients from a strain of W. globosa, called Mankai, to soft cheese and peas. Full-Text PDF

Begomoviruses are a major threat to tropical and subtropical tomato production, worldwide. Host resistance provides an effective strategy for reducing yield losses associated with these viruses. Resistance has been introgressed into cultivated tomato (Solanum lycopersicum L.) primarily from a number of wild tomato relatives, and the major resistance genes available to breeders today include Ty-1 (and its allele, Ty-3), Ty-2, Ty-4, ty-5, and a recently-identified gene tentatively named "Ty-6". Important to the effective and efficient deployment of resistant cultivars is an understanding of each resistance allele's effect when used alone and when combined with other resistance genes. Ty-1 and Ty2 are both dominant and, where effective, provide a high level of resistance; whereas the other genes provide partial resistance and are additive to partially dominant or recessive as with ty-5. Two-and three-gene combinations of Ty-3, Ty-4, and ty-5 in semi-isogenic backgrounds provided high levels of resistance to Tomato yellow leaf curl virus (TYLCV). Tomato mottle virus (ToMoV) trials in Florida using the same materials were severe, but indicated that Ty-3 and Ty-4 had small but significant effects in lowering ToMoV disease severity but, ty-5 did not. In two additional populations segregating for both ty-5 and "Ty-6" that were inoculated with ToMoV, ty-5 had no significant effect on resistance. Combining "Ty-6" and ty-5 homozygously provided a high level of resistance to TYLCV while the combination of homozygous ty-5 with heterozygous "Ty-6" was also good, but not as resistant as the former.

The genetic variation and evolutionary mechanisms shaping Cucumber vein yellowing virus (CVYV) populations were investigated by analysis of nucleotide sequences coding for P1b, P1b/P3 and coat proteins (CP) from isolates collected in different countries. The complete genome sequence of isolate ISM from Israel was also determined and compared to those of isolates Jor from Jordan and ALM32 from Spain. This isolate had overall nucleotide identities of 94·23 and 94·96% with ALM32 and Jor, respectively. Nucleotide variation among isolates was not homogeneously distributed, with the 5′ half of the genome being more variable than the 3′ half. A Bayesian phylogenetic tree of the CP showed that CVYV isolates clustered into two main clades: isolates from the Middle East region (Lebanon, Israel and Jordan) clustered in both clades whereas the isolate from Tunisia clustered in clade I and the European isolates clustered as a homogeneous phylogroup in Clade II. A similar topology was observed for P1b but with incongruences with respect to the CP, suggesting genetic exchange among virus isolates, which were confirmed with recombination algorithms. The low genetic diversity within the European phylogroup with respect to the other isolates, neutralist tests and genetic differentiation analyses suggest that the Middle East region is the cradle of CVYV and that a unique virus introduction event occurred in Europe, where the virus spread rapidly. Taken together, these findings indicate a risk of emergence of virulent CVYV isolates in Europe either through migration from the Middle East or by genetic changes of the European isolates.

In autumn 2007, a new disease with unknown etiology was observed in open-field tomato (Solanum lycopersicum) in the Lachish region of Israel. The symptoms included mild mosaic, leaf malformation, and severe stunting of the plants. The causal agent was readily transmitted mechanically from the sap of infected plants to indicator plants. Viral particles were purified from infected plants and cDNA was synthesized from RNA isolated from the particles. Cloning and sequencing of the cDNA showed 95% identity to RNA 3 of Pelargonium zonate spot virus (PZSV). Using reverse-transcription polymerase chain reaction, PZSV was detected in both seed and pollen grains of infected tomato plants. Attempts to disinfect seed by using hydrochloric acid and trisodium phosphate failed to eliminate this PZSV detection. Seed from infected tomato plants gave rise to infected seedlings with a seed-transmission rate of PZSV of 11 to 29%. Pollen grains collected from flowers of infected plants were used to hand pollinate healthy mother tomato plants. Although none of the pollinated mother plants became infected with PZSV, 29% of the seedlings produced from seed harvested from these plants were found to be infected. This is the first demonstration that PZSV is transmitted vertically via both pollen and seed in tomato plants.

To determine the effects of plant age on the expression of genetic resistance to tomato yellow leaf curl virus (TYLCV), six TYLCV-resistant and two susceptible tomato varieties were inoculated at 14, 28 or 45 days after sowing (DAS). Inoculation at 14 and 28 DAS was performed in the greenhouse, and the plants were transplanted to the field at 30 DAS. Inoculation at 45 DAS was performed in the field, by covering the target plants with polypropylene ("Agril") sheets and releasing viruliferous whiteflies under them. Resistance was assayed mainly by comparing yield components of inoculated plants to those of control, non-inoculated plants of the same variety. Symptom severity and plant height were also followed. Plant age at inoculation had no effect on disease-severity scores of the susceptible varieties, and little or no effect on those of the resistant varieties. In contrast, plant age at inoculation had a significant effect on the yield of all varieties tested. All varieties suffered a significant yield reduction due to inoculation with TYLCV; the lowest yield was produced by plants inoculated at 14 DAS. A smaller TYLCV-induced yield reduction (yield increase of 50 to 100%, depending on the variety's resistance level), was achieved following inoculation at 28 DAS. A further reduction in yield loss (yield increase of 30 to 40%) was achieved following inoculation at 45 DAS. Our results clearly demonstrate the occurrence of age-related (or mature-plant) resistance in tomato plants to TYLCV.

Tomato (Solanum lycopersicum) production in tropical and subtropical regions of the world is limited by the endemic presence of Tomato yellow leaf curl virus (TYLCV). Breeding programmes aimed at producing TYLCV-resistant tomato cultivars have utilized resistance sources derived from wild tomato species. So far, all reported breeding programmes have introgressed TYLCV resistance from a single wild tomato source. Here, we tested the hypothesis that pyramiding resistances from different wild tomato species might improve the degree of resistance of the domesticated tomato to TYLCV. We have crossed TYLCV-resistant lines that originated from different wild tomato progenitors, Solanum chilense, Solanum peruvianum, Solanum pimpinellifolium, and Solanum habrochaites. The various parental resistant lines and the F-1 hybrids were inoculated in the greenhouse using viruliferous whiteflies. Control, non-inoculated plants of the same lines and hybrids were exposed to non-viruliferous whiteflies. Following inoculation, the plants were scored for disease symptom severity, and transplanted to the field. Resistance was assayed by comparing yield of inoculated plants to those of the control non-inoculated plants of the same variety. Results showed that the F-1 hybrids between the resistant lines and the susceptible line suffered major yield reduction because of infection, but all hybrids were more resistant than the susceptible parent. All F-1 hybrids resulting from a cross between two resistant parents, showed a relatively high level of resistance, which in most cases was similar to that displayed by the more resistant parent. In some cases, the hybrids displayed better levels of resistance than both parents, but the differences were not statistically significant. The F-1 hybrid between a line with resistance from S. habrochaites and a line with resistance from S. peruvianum (HAB and 72-PER), exhibited the lowest yield loss and the mildest level of symptoms. Although the resistance level of this F-1 hybrid was not statistically different from the level of resistance displayed by the 72-PER parent itself, it was statistically better than the level of resistance displayed by the F-1 hybrids between 72-PER and any other resistant or susceptible line.

Tomato production in the Mediterranean region and elsewhere is under constant threat of the whitefly-transmitted begomoviruses that cause Tomato yellow leaf curl disease (TYLCD). Sequencing has indicated that the generic 'TYLCV' includes a large number of viruses and strains. We studied the distribution of the tomato yellow leaf curl disease-associated viruses in Egypt, Israel, Jordan and Lebanon. Two simple and reliable multiplex PCR protocols (mPCR) were developed that allowed the detection of Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl Sardinia virus (TYLCSV) species, in addition to Tomato yellow leaf curl virus-Mild (TYLCV-Mld) strain. PCR products were sequenced to confirm their identity. The full-length genomes were also sequenced of TYLCV from Egypt, Jordan and Lebanon, TYLCV-Mld strain from Jordan and Lebanon, and Tomato yellow leaf curl Sardinia virus-Sicily from Israel. This is the first time that TYLCSV-Sic has been detected in Israel and Jordan, and the first report of TYLCV-Mld strain from Egypt and Lebanon.

在辣椒(Capsicum annuum)种内进行抗CMV QTL分析。从感病品种Maor与抗病品种Perennial杂交获得180个F3。在美国和以色列的三个试验中用两个病毒株系接种。大部分RFLP和AFLP标记被用作构建遗传图谱,区间分析被用作QTL检测。有4个QTL与抗CMV显著相关。检测到了两个标记对CMV抗性的双基因互作,没有检测到单基因效用。3个试验中检测到控制表现型变异(cmv11.1)的QTL,其变异最大百分比为16%～33%,该QTL与双基因互作有关。这个QTL与L位点连锁,并证实该QTL与抗TMV有关。在Perennial上早期非常有趣的观察到抗CMV感TMV的现象。来自于不相关的种群的一个高世代的回交育种株系3990,被选择用来分析对CMV的抗性,标记覆盖整个基因组,检测到来自Perennial的基因渗入。这些基因渗入的区域中包括4个与抗CMV相关的QTL。在两个基因区域的标记被鉴定与抗CMV的QTL相关,同时也与控制果实重量的QTL相关,另外的育种观察也证实对CMV的抗性来源于Perennial和小果型品种。

For Electron Beam Direct Write (EBDW) a systematic investigation of defect density using a Negevtech 3100 darkfield inspection system was performed. A special defect learning pattern for memory applications with coverage of 50% was designed and printed partially on 300mm wafers using chemically amplified positive and negative E-Beam resists. By optical defect measurements post litho it was possible to inspect 50nm dense lines to characterize the exposure system as well as the used resist process. Using this method a large exposed area in millimeter range can be inspected and an overview on exposure quality can be gained in a reasonable amount of time. Particle measurements were performed additionally to distinguish between particles and exposure issues. By using darkfield measurements, process related issues like development problems and resist residuals can be found, as well as writing issues like shot butting and write field stitching can be quickly determined and controlled with this method. In this paper, the measurement methodology is described as well as the effect of writer imperfections on the darkfield images. A pareto analysis is performed and shows the frequency of occurrence of different defects. Measures to reduce defects - especially on the tool side - are given. The method is feasible to use in a regular check to control tool and process performance.

Members of the insulin family of hormones are generally not regarded as gender-speciWc, although there is sporadic evidence for the possible involvement of insulin pathways in sexual diVerentiation. In crustaceans, sexual diVerentiation is controlled by the androgenic gland (AG), an organ unique to males. To date, attempts to identify active AG factors in decapods through either classical puriWcation methods or sequence similarity with isopod AG hormones have proven unsuccessful. In the present study, the Wrst subtractive cDNA library from a decapod AG was constructed from the red-claw crayWsh Cherax quadricarinatus. During library screening, an AG-speciWc gene, expressed exclusively in males even at early stages of maturation and termed Cq-IAG (C. quadricarinatus insulin-like AG factor), was discovered. In situ hybridization of Cq-IAG conWrmed the exclusive localization of its expression to the AG. Following cloning and complete sequencing of the gene, its cDNA was found to contain 1445 nucleotides encoding a deduced translation product of 176 amino acids. The proposed protein sequence encompasses Cys residue and putative cleaved peptide patterns whose linear and 3D organization are similar to those of members of the insulin/insulin-like growth factor/relaxin family and their receptor recognition surface. The identiWcation of Cq-IAG is the Wrst report of a pro-insulin-like gene expressed in a decapod crustacean in a gender-speciWc manner. Its expression in a male-speciWc endocrine gland controlling sex diVerentiation supports the notion that insulin may have evolved in the context of regulating sexual diVerentiation.