Rooting Characteristics of <Emphasis Type="Italic">Solanum chacoense</Emphasis> and <Emphasis Type="Italic">Solanum tuberosum in Vitro</Emphasis>

The precise level of environmental control in vitro may aid in identifying genetically superior plant germplasm for rooting characteristics (RC) linked to increased foraging for plant nitrogen (N). The objectives of this research were to determine the phenotypic variation in root morphological responses of 49 Solanum chacoense (chc), 30 Solanum tuberosum Group Phureja – Solanum tuberosum Group Stenotomum (phu-stn), and three Solanum tuberosum (tbr) genotypes to 1.0 and 0.5 N rate in vitro for 28 d, and identify genotypes with superior RC. The 0.5 N significantly increased density of root length, surface area, and tips. All RC were significantly greater in chc than in either phu-stn or tbr. Based upon clustering on root length, surface area, and volume, the cluster with the greatest rooting values consisted of eight chc genotypes that may be utilized to initiate a breeding program to improve RC in potato.     

Colorimetric Analysis of <Emphasis Type="Italic">Hibiscus</Emphasis> Beverages and their Potential Antioxidant Properties

In food industry, roselle beverages and their subproducts could be functional ingredients since they are an excellent source of bioactive compounds with improved performance due to their important anthocyanins content. The aim of this study was to analyze anthocyanin content and antioxidant properties of aqueous infusions elaborated with color contrasting Hibiscus materials and design a mathematical model in order to predict color-composition relationship. Color measurements of beverages from roselle (Negra, Sudan and Rosa) were made by transmission spectrophotometry, anthocyanins quantification was determined by HPLC, and antioxidant potential was evaluated by in vitro methods (ABTS and FRAP assays). Beverages prepared with particle size minor of 250 μm presented until 4- and 2- times more anthocyanins content and antioxidant capacity respectively, in comparison to beverages prepared with powders with particle size major of 750 μm. Positive correlations among pigments composition and color parameters were found (p?<?0.05), showing that anthocyanins content, antioxidant capacity, C*_ab and h_ab values increased in relation with the smallest particle size of flours. Also, mathematical models were stablished to predict anthocyanin content (r?≥?0.97) and antioxidant capacity (r?≥?0.89) from color data; we propose equations for quick estimation of the antioxidant capacity in the Hibiscus beverages with high anthocyanin content. The obtained models could be an important tool to be used in food industry for pigment characterization or functional compounds with potential health benefits.     

Antioxidant Potential and Health Relevant Functionality of Traditionally Processed <Emphasis Type="Italic">Cassia hirsuta</Emphasis> L. Seeds: An Indian Underutilized Food Legume

The methanolic extract of Cassia hirsuta L. seed materials, an underutilized food legume collected from India, was analyzed for antioxidant activity and health relevant functionality. The methanolic extract of raw seeds contained a total free phenolic content of 15.82?±?1.69 g catechin equivalent/100 g extract DM. Encouraging levels of ferric reducing/antioxidant power (FRAP, 1,446 mmol Fe[II]/mg extract), inhibition of ß-carotene degradation (48.81%) and scavenging activity against DPPH (64.40%) and superoxide (43.78%) radicals were exhibited by the raw samples. Further, 83.11% of α-amylase and 62.79% of α-glucosidase enzyme inhibition characteristics under in vitro starch digestion bioassay were also recorded. Sprouting + oil-frying caused an apparent increase on the total free phenolic content and a significant improvement in the antioxidant and free radical scavenging capacity of methanolic extract of C. hirsuta seeds, while soaking + cooking as well as open-pan roasting treatments showed diminishing effects. The analysis of the phenolic profile revealed the presence of gallic acid, p-coumaric acid and (+)-catechin in the methanolic extract of these seeds.     

Seasonal deviation effects foliar endophyte assemblage and diversity in <Emphasis Type="Italic">Asparagus racemosus</Emphasis> and <Emphasis Type="Italic">Hemidesmus indicus</Emphasis>

Background

Fungal endophytes are the living symbionts which cause no apparent damage to the host tissue. The distribution pattern of these endophytes within a host plant is mediated by environmental factors. This study was carried out to explore the fungal endophyte community and their distribution pattern in Asparagus racemosus and Hemidesmus indicus growing in the study area.

Results

Foliar endophytes were isolated for 2 years from A. racemosus and H. indicus at four different seasons (June–August, September–November, December–February, March–May). A total of 5400 (675/season/year) leaf segments harbored 38 fungal species belonging to 17 genera, 12 miscellaneous mycelia sterile from 968 isolates and 13 had yeast like growth. In A. racemosus, Acremonium strictum and Phomopsis sp.1, were dominant with overall relative colonization densities (RCD) of 7.11% and 5.44% respectively, followed by Colletotrichum sp.3 and Colletotrichum sp.1 of 4.89% and 4.83% respectively. In H. indicus the dominant species was A. strictum having higher overall RCD of 5.06%, followed by Fusarium moniliforme and Colletotrichum sp.2 with RCD of 3.83% and 3%, respectively. Further the overall colonization and isolation rates were higher during the wet periods (September–November) in both A. racemosus (92.22% and 95.11%) and H. indicus (82% and 77.11%).

Conclusion

Study samples treated with 0.2% HgCl₂ and 75% EtOH for 30 s and 1 min, respectively, confirmed most favorable method of isolation of the endophytes. Owing to high mean isolation and colonization rates, September–November season proved to be the optimal season for endophyte isolation in both the study plants. Assessing the bioactive potential of these endophytes, may lead to the isolation of novel natural products and metabolites.

     

<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Bio-efficacy of the Essential Oil of Oregano (<Emphasis Type="Italic">Origanum vulgare</Emphasis> Lamiaceae. Ssp. <Emphasis Type="Italic">Hirtum</Emphasis>)

The aim of this study was to investigate the bioactivity of the essential oil isolated from Origanum vulgare L. (EOv). We analyzed the in vivo anti-inflammatory properties in a mouse-airway inflammation model and the in vitro antimicrobial activity, genotoxicity over the anaphase-telophase with the Allium cepa strain and its cytotoxicity/viability in A549 culture cells. In vivo, EOv modified the levels of tumor necrosis factor -α and viable activated macrophages and was capable to mitigate the effects of degradation of conjugated dienes. In vitro, EOv reduced the viability of cultured A549 cells as well as the mitotic index and a number of chromosomal aberrations; however, it did not change the number of phases. We found that EOv presents antimicrobial activity against different Gram (?) and (+) strains, measured by disc-diffusion test and confirmed with a more accurate method, the AutoCad software. We postulate that EOv presents antibacterial, antioxidant and chemopreventive properties and could be play an important role as bioprotector agent.     

The Effects of <Emphasis Type="Italic">Pimpinella anisum</Emphasis> Essential Oils on Young Larvae <Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> Say (Coleoptera: Chrysomelidae)

The effect of essential oil (EO) from anise (Pimpinellia anisum) on the mortality of young larvae of Colorado potato beetles has been studied. In our bioassays, P. anisum EO significantly increased the mortality of the second instar larvae of L. decemlineata. Significantly different values of LD₅₀ and LD₉₀ were established for acute (LD₅₀ = 1.76, and LD₉₀ = 8.29) as well as chronic toxicity (LD₅₀ = 0.45, and LD₉₀ = 1.01). Decrease of both values over experimental period was evident, which showed that the larval mortality was slow and cumulative. The composition of EO used for biological experiments was also assessed. The main component detected in EO from P. anisum was anethole (79.87%), followed by anisaldehyde (7.74%), estragole (5.88%) and β-linalool (1.07%). Within five days, residual concentration of EO decreased from 3.87 mg/g of dry weight immediately after foliar applications to 0.9 mg per g of dry weight. The effect of this slow evaporation could be explained by dominant presence of anethole or by the type of formulation and the addition of oil and tween. Results of our study demonstrate that EO from P. anisum has insecticidal properties that may lead to the development of new organic products for the control of Colorado potato beetles.     

Anti-proliferative Effect of <Emphasis Type="Italic">Melissa officinalis</Emphasis> on Human Colon Cancer Cell Line

Melissa officinalis L. (Lamiaceae) is consumed as a traditional herbal tea in the Mediterranean region. The cytotoxic effect of the 50% ethanolic and aqueous extract, determined by the MTT and NR assays, was evaluated in vitro on Human Colon Cancer Cell Line (HCT-116), using Triton 10% as positive control. The 50% ethanolic extract showed significant differences after 72 h of treatment, reducing cell proliferation to values close to 40%, even the lowest dose tested (5 μg/ml). In the MTT assay, the same extract caused the lowest cell viability with 13% at a concentration of 1,000 μg/ml after 72 h of treatment, being a value lower than Triton 10%. The antioxidant activity was also confirmed evaluating the capacity of the extracts to scavenge ABTS and DPPH radicals, and IC₅₀ values were highly correlated with the total phenolic and flavonoid content. Bioassay guided fractionation led to the isolation of an anti-proliferative compound, rosmarinic acid. Its structural elucidation was performed by HPLC/DAD/ESI/MS analysis. High dose of rosmarinic acid (1,000 μg/ml) was clearly cytotoxic against HCT-116 cells, with a significant decrease in cell number since the earliest time point (24 h).     

Effective Management of the <Emphasis Type="Italic">Phthorimaea operculella</Emphasis> (Zeller) Using PVA Nanofibers Loaded with <Emphasis Type="Italic">Cinnamomum zeylanicum</Emphasis> Essential Oil

Phthorimaea operculella (Zeller) is one of the most common insect pests of cultivated potato in tropical and subtropical regions. In this research, a potential strategy to improve the insecticidal activity of plant essential oils for the effective management of P. operculella was studied. The insecticidal and residual effects of nanofiber oil (NFO) and pure essential oil (PEO) of Cinnamomum zeylanicum were assessed on PTM under laboratory conditions. The nanofibers were made by the electrospinning method using polyvinyl alcohol (PVA) polymer. The morphological characteristics of the nanofibers were evaluated by scanning electron microscopy and Fourier transform infrared spectroscopy. The chemical constituents of cinnamon essential oil (EO) were detected by GC/MS. Fumigant toxicity of NFO and PEO were evaluated on different growth stages (egg, male and female adults) of P. operculella. SEM and FTIR analyses confirmed the presence of EO on the nanofiber structure. The yield of the EO from C. zelanicum on the nanofibers was 1.86%. GC/MS analysis showed that cinnamaldehyde was the primary constituent (69.88%) of cinnamon EO. LC₅₀ values of C. zelanicum EO and NFO were 4.92 and 1.76 μl/l air for eggs, 0.444 and 0.212 μl/l air for female adults, and 0.424 and 0.192 μl/l air for male adults, respectively. Fumigant bioassays revealed that NFO was more toxic than C. zeylanicum oil against at all stages of P. operculella. The residual effect of PEO and NFO was evaluated against the egg stage of the P. operculella. NFO lost insecticidal effectiveness 47 days after application, while the efficacy of PEO decreased 15 days after application. Our results suggest that NFO of C. zeylanicum can be used as an effective new tool for the management of P. operculella.     

Maturity-Adjusted Resistance of Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Cultivars to Verticillium Wilt Caused by <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Verticillium wilt is a fungal disease of potato caused by two species of Verticillium, V. dahliae and V. albo atrum. The pathogen infects the vascular tissue of potato plants through roots, interfering with the transport of water and nutrition, and reducing both the yield and quality of tubers. We have evaluated the reaction of 283 potato clones (274 cultivars and nine breeding selections) to inoculation with V. dahliae under greenhouse conditions. A significant linear correlation (r = 0.4, p < 0.0001) was detected between plant maturity and partial resistance to the pathogen, with late maturing clones being generally more resistant. Maturity-adjusted resistance, that takes into consideration both plant maturity and resistance, was calculated from residuals of the linear regression between the two traits. Even after adjusting for maturity, the difference in the resistance of clones was still highly significant, indicating that a substantial part of resistance cannot be explained by the effect of maturity. The highest maturity-adjusted resistance was found in the cv. Navajo, while the most susceptible clone was the cv. Pungo. We hope that the present abundance of data about the resistance and maturity of 283 clones will help potato breeders to develop cultivars with improved resistance to V. dahliae.     

Water and air temperature impacts on rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>) phenology

Air temperature (T_a) is commonly used for modeling rice phenology. However, since the growing point of rice is under water during the vegetative and the early part of the reproductive period, water temperature (T_w) is likely to have a greater influence on crop developmental rates than T_a during this period. To test this hypothesis, we monitored T_w, T_a, and crop phenology in three commercial irrigated rice fields in California, USA. Sampling locations were set up on along a transect from the water inlet into the field. (Water warms up as it moves into the field.) T_a averaged 22.7 °C across sampling locations within each field, but average seasonal T_w increased from 22 °C near the inlet to 23.4 °C furthest away from the inlet. Relative to T_w furthest from the inlet, low T_w near the inlet delayed time to panicle initiation (PI 5 days) and heading (HD 8 days) and the appearance of one yellow hull on the main stem panicle (R7 9 days). Using T_w instead of T_a when the active growing point is under water until booting (midway between PI and HD) in a thermal time model improved accuracy (root-mean-square error, RMSE) for predicting time to PI by 2.5 days and HD by 1.6 days and R7 by 1.8 days. This model was further validated under more typical field conditions (i.e., not close to cold water inlets) in six locations in California. Under these conditions, average T_w was 2.6 °C higher than T_a between planting and booting, primarily due to higher daily maximum T_w values. Using T_w in the model until booting improved RMSE by 1.2 days in predicting time to HD. Using T_w instead of T_a during this period could improve the accuracy of rice phenology models.     

<Emphasis Type="Italic">t</Emphasis>-Resveratrol Protects against Acute High Glucose Damage in Endothelial Cells

Postprandial hyperglycemia in diabetic and nondiabetic subjects is associated with endothelial dysfunction. Evidence shows that high glucose generates oxidative stress and a pro-inflammatory state promoting the development of cardiovascular diseases. trans-Resveratrol (t-RV) has been shown to reduce cardiovascular risk. To determine whether t-RV acts as a protector against acute high glucose (AHG)-induced damage, two in vitro models, rat aortic rings (RAR) and human umbilical vein endothelial cells (HUVEC) were used. RAR pretreated with AHG (25 mM D-glucose) for 3 h dramatically decreased the endothelium-dependent relaxation (EDR) induced by acetylcholine in phenylephrine (PE)-precontracted vessels. However, coincubation with t-RV significantly mitigated the damage induced by AHG on EDR. Pretreatment with AHG did not affect the vasodilation induced by sodium nitroprusside. HUVEC treated with t-RV decreased cytotoxicity and reduced radical oxygen species production induced by AHG. Taken together, these results suggest that t-RV can mitigate the AHG-induced EDR damage through a mechanism involving ROS scavenging and probably an increase in the bioavailability of NO.     

Identification and fine mapping of <Emphasis Type="Italic">Bph33</Emphasis>, a new brown planthopper resistance gene in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Background

Host-plant resistance is the most desirable and economic way to overcome BPH damage to rice. As single-gene resistance is easily lost due to the evolution of new BPH biotypes, it is urgent to explore and identify new BPH resistance genes.

Results

In this study, using F_2:3 populations and near-isogenic lines (NILs) derived from crosses between two BPH-resistant Sri Lankan rice cultivars (KOLAYAL and POLIYAL) and a BPH-susceptible cultivar 9311, a new resistance gene Bph33 was fine mapped to a 60-kb region ranging 0.91–0.97 Mb on the short arm of chromosome 4 (4S), which was at least 4 Mb distant from those genes/QTLs (Bph12, Bph15, Bph3, Bph20, QBph4 and QBph4.2) reported before. Seven genes were predicted in this region. Based on sequence and expression analyses, a Leucine Rich Repeat (LRR) family gene (LOC_Os04g02520) was identified as the most possible candidate of Bph33. The gene exhibited continuous and stable resistance from seedling stage to tillering stage, showing both antixenosis and antibiosis effects on BPH.

Conclusion

The results of this study will facilitate map-based cloning and marker-assisted selection of the gene.

     

Variation in the Carotenoid Composition of the Lycopene-Rich Brazilian Fruit <Emphasis Type="Italic">Eugenia uniflora</Emphasis> L.

The indigenous pitanga (Eugenia uniflora L.) is now marketed and commercially processed in Brazil. In the present work, the carotenoids of the pitanga fruit from two states and at two stages of ripening, as well as of processed pitanga products (frozen pulp and juice, the brands being designated as A, B and C) commercialized in Campinas, São Paulo, were determined by HPLC. As compared to ripe pitanga from Medianeira, Paraná, those from Campinas had significantly higher (all-E)-lycopene (14.0 vs. 71.1 μg/g), (13Z)-lycopene (1.1 vs. 5.0 μg/g) and (all-E)-γ-carotene (1.6 vs. 3.8 μg/g) levels. Significant increases in most of the carotenoids occurred from the partially ripe to the ripe fruits, with (all-E)-lycopene doubling its concentration in fruits from both states. Pitanga was found to be one of the richest fruit sources of carotenoids, particularly lycopene, but the processed products had much lower lycopene content. The mean (all-E)-lycopene concentration was 16.6 μg/g for frozen pulp brand A, 23.0 μg/g for bottled juice brand B and 25.6 μg/g for bottled juice brand C. Optimization of processing is therefore needed to guarantee better retention of this important carotenoid.     

An <Emphasis Type="Italic">In Vitro</Emphasis> Assay Method for Resistance to Bacterial Wilt (<Emphasis Type="Italic">Ralstonia solanacearum</Emphasis>) in Potato

To develop an in vitro assay method for bacterial wilt resistance in potato, resistant and susceptible standard genotypes were grown in vitro, and different conditions of inoculation with Ralstonia solanacearum phylotype I/biovar 4 were examined. The optimal condition was the inoculation of 6–8 leaf stage plants with a bacterial concentration of 10² CFU ml^?1 and an incubation temperature of 28 °C. Evaluation of stem wilting was more reliable than that of leaf wilting. Using this method, nine genotypes with different resistance levels in the field were evaluated. Lower disease indices were obtained for genotypes with high resistance levels in the field, suggesting that this assay is useful for evaluating bacterial wilt resistance in a controlled environment.     

Divergent evolution of rice blast resistance <Emphasis Type="Italic">Pi54</Emphasis> locus in the genus <Emphasis Type="Italic">Oryza</Emphasis>

Background

The rice blast resistance gene Pi54 was cloned from Oryza sativa ssp. indica cv. Tetep, which conferred broad-spectrum resistance against Magnaporthe oryzae. Pi54 allelic variants have been identified in not only domesticates but also wild rice species, but the majority of japonica and some indica cultivars lost the function.

Results

We here found that Pi54 (Os11g0639100) and its homolog Os11g0640600 (named as #11) were closely located on a 25 kbp region in japonica cv. Sasanishiki compared to a 99 kbp region in japonica cv. Nipponbare. Sasanishiki lost at least six genes containing one other R-gene cluster (Os11g0639600, Os11g0640000, and Os11g0640300). Eight AA-genome species including five wild rice species were classified into either Nipponbare or Sasanishiki type. The BB-genome wild rice species O. punctata was Sasanishiki type. The FF-genome wild rice species O. brachyantha (the basal lineage of Oryza) was neither, because Pi54 was absent and the orientation of the R-gene cluster was reversed in comparison with Nipponbare-type species. The phylogenetic analysis showed that #11gene of O. brachyantha was on the root of both Pi54 and #11 alleles. All Nipponbare-type Pi54 alleles were specifically disrupted by 143 and 37/44?bp insertions compared to Tetep and Sasanishiki type. In addition, Pi54 of japonica cv. Sasanishiki lost nucleotide-binding site and leucine-rich repeat (NBS–LRR) domains owing to additional mutations.

Conclusions

These results suggest that Pi54 might be derived from a tandem duplication of the ancestor #11 gene in progenitor FF-genome species. Two divergent structures of Pi54 locus caused by a mobile unit containing the nearby R-gene cluster could be developed before domestication. This study provides a potential genetic resource of rice breeding for blast resistance in modern cultivars sustainability.

     

Effect of Inulin on the Viability of <Emphasis Type="Italic">L. plantarum</Emphasis> during Storage and <Emphasis Type="Italic">In Vitro</Emphasis> Digestion and on Composition Parameters of Vegetable Fermented Juices

The prebiotic effect of different concentrations of inulin (0, 1 and 2%) on the growth and survival of Lactobacillus plantarum (LP) CECT 220 in blended carrot and orange juices was investigated after 24 h of fermentation, during 30 days of storage at 4 °C and through the phases of gastrointestinal digestion after different storage periods. Microbiological and chemical determinations were also carried out in all juices. The lactic fermentation increased the shelf life of the fermented juices with inulin. The hygienic-sanitary quality in fermented juices was better than the control juices. During storage, the inulin improved the viability of LP and the monosaccharide concentration remained higher with respect to the juice without inulin (40% lower). At 30 days, the fermented juices with 2% inulin after in vitro digestion presented the highest survival of L. plantarum.     

Effects of Different Soil Treatments on the Development of <Emphasis Type="Italic">Spongospora subterranea</Emphasis> f. sp. <Emphasis Type="Italic">subterranea</Emphasis> in Potato Roots and Tubers in the Greenhouse

Powdery scab caused by Spongospora subterranea f. sp. subterranea (Sss) causes extensive losses in potato production systems globally. Two pot experiments were established in the greenhouse in summer 2013 and winter 2014 to evaluate the effectiveness of different soil chemicals, fumigant, amendments and biological control agents (BCAs) against Sss in the rhizospheric soil, potato roots and tubers. The study used visual assessment methods to assess the effect of treatments on root galling and zoosporangia production, and qPCR to measure Sss concentration in the soil and in the potato roots and tubers. All six soil treatments, namely metam sodium, fluazinam, ZincMax, calcium cyanamide, Biocult and a combination of Bacillus subtilis and Trichoderma asperellum recorded significantly (P < 0.05) lower numbers of zoosporangia in the roots compared to the untreated control. The same effect was observed on the concentration of Sss DNA in the roots at tuber initiation. A more diverse picture was obtained when root gall scores at tuber initiation and Sss DNA in the rhizospheric soil at tuber initiation and harvesting were compared. Significant differences (P < 0.05) were also noted in disease severity, disease incidence, and tuber yield between metam sodium, fluazinam, ZincMax, calcium cyanamide and the untreated control. Calcium cyanamide gave the highest tuber yield. The study demonstrated the potential of soil treatments such as metam sodium, fluazinam, ZincMax and calcium cyanamide in managing Sss in potatoes by reducing the pathogen both in the rhizospheric soil and the roots of the potato plant.     

Functional Teas from the Leaves of <Emphasis Type="Italic">Arbutus unedo</Emphasis>: Phenolic Content,Antioxidant Activity,and Detection of Efficient Radical Scavengers

The phenolic content/composition and antioxidant activity of hot/cold infusion and decoction from the leaves of Arbutus unedo were studied for the first time. 1,1-diphenyl-2-picrylhydrazyl (DPPH^●), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical cation (ABTS^●+), crocin-bleaching, copper-reducing, and liposome accelerated oxidation assays were used for the evaluation of the activity in vitro. In vivo, the extracts were examined for their ability to protect S. cerevisiae cells from H₂O₂ induced oxidative stress. An on-line high-performance liquid chromatography-DPPH^● assay was applied to identify potent radical scavengers and comment on their contribution to the total activity. The addition of leaves to boiling water (decoction) was the most appropriate practice to apply since the highest phenol intake (220.2 mg gallic acid/cup served) was obtained. Additionally, its antioxidant activity was equal or superior to that of the other extracts. Flavonols (~51–61 mg/g dry extract) were the main phenols in all the extracts, with quercitrin accounting for ~20% of the total phenol amount. The on-line DPPH^● method verified the high potency of the decoction and indicated as the most active radical scavengers, two galloylquinic acid derivatives and myricitrin, accounting for ~28–45% and ~11–13% of the total scavenging, respectively. Present data may contribute to the future exploitation of A. unedo leaves by the food industry for health-promoting herbal tea preparations and dietary supplements.     

Proteomic analysis of the defense response to <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis> in rice harboring the blast resistance gene <Emphasis Type="Italic">Piz-t</Emphasis>

Background

Rice blast (caused by Magnaporthe oryzae) is one of the most destructive diseases of rice. While many blast resistance (R) genes have been identified and deployed in rice cultivars, little is known about the R gene-mediated defense mechanism. We used a rice transgenic line harboring the resistance gene Piz-t to investigate the R gene-mediated resistance response to infection.

Results

We conducted comparative proteome profiling of the Piz-t transgenic Nipponbare line (NPB-Piz-t) and wild-type Nipponbare (NPB) inoculated with M. oryzae at 24, 48, 72 h post-inoculation (hpi) using isobaric tags for relative and absolute quantification (iTRAQ) analysis. Comparative analysis of the response of NPB-Piz-t to the avirulent isolate KJ201 and the virulent isolate RB22 identified 114 differentially expressed proteins (DEPs) between KJ201-inoculated NPB-Piz-t (KJ201-Piz-t) and mock-treated NPB-Piz-t (Mock-Piz-t), and 118 DEPs between RB22-inoculated NPB-Piz-t (RB22-Piz-t) and Mock-Piz-t. Among the DEPs, 56 occurred commonly in comparisons KJ201-Piz-t/Mock-Piz-t and RB22-Piz-t/Mock-Piz-t. In a comparison of the responses of NPB and NPB-Piz-t to isolate KJ201, 93 DEPs between KJ201-Piz-t and KJ201-NPB were identified. DEPs in comparisons KJ201-Piz-t/Mock-Piz-t, RB22-Piz-t/Mock-Piz-t and KJ201-Piz-t/KJ201-NPB contained a number of proteins that may be involved in rice response to pathogens, including pathogenesis-related (PR) proteins, hormonal regulation-related proteins, defense and stress response-related proteins, receptor-like kinase, and cytochrome P450. Comparative analysis further identified 7 common DEPs between the comparisons KJ201-Piz-t/KJ201-NPB and KJ201-Piz-t/RB22-Piz-t, including alcohol dehydrogenase I, receptor-like protein kinase, endochitinase, similar to rubisco large subunit, NADP-dependent malic enzyme, and two hypothetical proteins.

Conclusions

Our results provide a valuable resource for discovery of complex protein networks involved in the resistance response of rice to blast fungus.