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Echeveria subrigida is native to Mexico and its methanol extract (ME) shows relevant biological activities for human health, including the α-glucosidase inhibitory (αGI) activity that suggests its ...

ADP-glucose pyrophosphorylase (AGPase) is a key enzyme of starch synthesis in seeds, tubers and fruits. UDP-glucose pyrophosphorylase (UGPase) is an important enzyme of sucrose metabolism in the cytosol while alkaline phosphatase (ALP) is a marker enzyme of the amyloplast that keeps the production of ADPG by removing PPi. Unripe banana accumulates starch in the pulp during development, while ripe fruits are characterized by the accumulation of soluble sugars. The aim of the study was to compare starch granule structure, carbohydrate levels, subcellular location and activities of three enzymes: AGPase, UGPase and ALP. Protein extracts from the cytosolic and amyloplastidial fractions were obtained from the pulp of banana fruit at three developmental stages (11, 16 and 21 weeks after flowering) and analyzed by electrophoresis and immunodetection. Protein profiles were similar during ripening, showing a main electrophoretic band at 50-55 kDa. Higher protein content was found in the cytosolic than in the amyloplastidial fraction. Starch granules and ALP activity were enriched in the amyloplast, whereas AGPase showed a subcellular distribution similar to UGPase. Immunoblot analysis also confirmed the presence of AGPase in both cytosol and amyloplast. AGPase activity was higher in the cytosol than in the amyloplast. Both AGPase activity and western blot band intensity were highest at 16 weeks. UGPase activity was highest at 21 weeks. We conclude that cytosolic production of ADP-glucose is not an exclusive feature of cereal endosperms due to plant breeding, but it also occurs in fruits of non-domesticated plants such as tropical banana (Musa acuminata). This work increases our understanding about pyrophosphorylase activities in the pulp of banana fruit.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a major pest of the tomato (Solanum lycopersicum L.) crop worldwide, mostly because of resistance to insecticide that has been associated with endosymbionts. Although knowledge of biotypes in the B. tabaci complex and its endosymbionts is critical for developing pest management strategies, limited information is available on the pest in the Culiacan Province of Mexico. In total, 249 samples of whiteflies were collected from tomato plants of four growers in the Culiacan Valley to analyze the genetic identity of the B. tabaci complex and its endosymbionts. PCR and phylogenetic analyses on the mitochondrial cytochrome oxidase I gene (mtCO1) revealed MEAM1, MED, and indigenous genetic groups in 53, 19, and 28%, respectively, of B. tabaci on tomatoes. Individual whiteflies were examined for endosymbionts through specific primers and phylogenetic analyses on the 16S rRNA and the 23S rRNA genes. Incidence of Portiera, Hamiltonella, Rickettsia, Wolbachia, and Cardinium was 81, 46, 28, 9, and 13%, respectively, in the B. tabaci samples analyzed. A new bacterium-endosymbiont in the genus Orientia was found and described in 63% of the samples of B. tabaci. A mixture of the endosymbionts Rickettsia+UAS_511+Cardinium was observed in 10% of samples of B. tabaci MED, while in the MEAM1 biotype, a combination of Hamiltonella+UAS_511 was in 32% of the samples analyzed. The study highlights the diversity and distribution of B. tabaci and its endosymbionts throughout the Culiacan Province and provides evidence for control of the insect pest.

El “permanente del tomate”, “manchado del tubérculo” o “zebra chip” en papa y “brotes cloróticos” del chile, son tres enfermedades descritas en México con signos coincidentes de aborto de flor, oscurecimiento de tejido vascular en la base del tallo y raíz de las plantas. Se ha mencionado la asociación entre estas enfermedades y la bacteria emergente Candidatus Liberibacter solanacearum (CLs) así como al psílido Bactericera cockerelli como su vector. Estas enfermedades, que en inicio se localizaban en tres estados de México se han diseminado a las principales regiones productoras de solanáceas, tanto en condiciones de campo como en invernadero. El objetivo del estudio fue conocer la presencia de CLs asociado a enfermedades que afectan los cultivos de tomate, para y chile en México. La bacteria se identificó por PCR del gen 16S de ADNr, clonación y secuenciación. La alineación de secuencias nucleotídicas se realizó con el método Clustal W y el árbol filogenético se construyó con el algoritmo de Neighbor-Joining a partir de distancias calculadas con el método de Tajima-Nei y un índice de Felsenstein de 1 000 réplicas, utilizando el software MEGA versión 5.05. En total se analizaron 167 muestras, de las cuales 86 resultaron positivas, provenientes de 14 estados de México. Se obtuvieron cinco secuencias nucleotídicas de Guanajuato, San Luis Potosí y Sinaloa correspondientes al tomate, papa y chile, adultos y huevecillos de B. cockerelli. El análisis de las secuencias mostró una identidad de 99.4% al comparase entre estas y hasta 99.8% con accesiones del GenBank descritas para CLs en EUA, Nueva Zelanda y Canadá.

The fruit of 18 Solanum lycopersicum var. cerasiforme accessions from Mexico were evaluated for total phenolics (TP) by the Folin-Ciocalteau assay, phenolic profiles by high performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS), antioxidant activity (AoxA) by 2,2´-azino-bis(3-ethylbenzothiazolin)-6-sulfonic acid (ABTS), 2,2-diphenil-1-pycrilhydrazyl (DPPH), and oxygen radical absorbance capacity (ORAC), and antimutagenic activity (AmuA) by the Ames assay. TP was measured as Gallic Acid Equivalents (GAE) and the AoxA as Trolox Equivalents (TE). TP varied from 37 to 86 mg GAE 100 g−1 fresh weight (fw). The AoxA by ABTS (568-1187 µmol TE 100 g−1 fw) and DPPH (157-350 µmol TE 100 g−1 fw) correlated with TP and the levels of caffeoylquinic acids and rutin. The AmuA did not correlate with the levels of phenolics. Some accessions had higher AoxA and AmuA than those reported for commercial cultivars and also showed high levels of caffeoylquinic acids and rutin; thus, their consumption could have good health promoting effects.

Anthracnose of papaya fruit caused by the fungus Colletotrichum gloeosporioides is one of the most economically important postharvest diseases. Hot water immersion (HW) and calcium chloride (Ca) treatments have been used to control papaya postharvest diseases; however, the effect of the combination HW-Ca on the pathogen growth and the development of the disease in infected papaya fruit has been scarcely studied. The aim of this study was to evaluate the effect of the HW-Ca treatment on the in vitro growth of C. gloesporioides conidia and the quality of infected papaya. In vitro, the HW-Ca treated conidia showed reduced mycelial growth and germination. In vivo, the HW-Ca treatment of infected papaya delayed for 5 days the onset of the anthracnose symptoms and improved the papaya postharvest quality. The combined treatment HW-Ca was better than any of the individual treatments to inhibit the in vitro development of C. gloeosporioides and to reduce the negative effects of papaya anthracnose.

A comparative proteomic approach was performed to analyze the differential accumulation of leaf proteins in response to the symbiosis between Solanum lycopersicum and the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis. Protein profiling was examined in leaves from tomato plants colonized with AMF (M), as well as non-colonized plants fertilized with low phosphate (20 μM P; NM-LP) and non-colonized plants fertilized with regular phosphate Hoagland’s solution (200 μM P; NM-RP). Comparisons were made between these groups, and 2D-SDS-PAGE revealed that 27 spots were differentially accumulated in M vs. NM-LP. Twenty-three out of the 27 spots were successfully identified by mass spectrometry. Two of these proteins, 2-methylene-furan-3-one reductase and auxin-binding protein ABP19a, were up-accumulated in M plants. The down-accumulated proteins in M plants were associated mainly with photosynthesis, redox, and other molecular functions. Superoxide dismutase, harpin binding protein, and thioredoxin peroxidase were down-accumulated in leaves of M tomato plants when compared to NM-LP and NM-RP, indicating that these proteins are responsive to AMF colonization independently of the phosphate regime under which they were grown. 14-3-3 protein was up-accumulated in NM-RP vs. NM-LP plants, whereas it was down-accumulated in M vs. NM-LP and M vs. NM-RP, regardless of their phosphate nutrition. This suggests a possible regulation by P nutrition and AMF colonization. Our results demonstrate AMF-induced systemic changes in the expression of tomato leaf proteins, including the down-accumulation of proteins related to photosynthesis and redox function.

Red arils of Pithecellobium dulce fruit, commonly known as guamuchil, show high antioxidant (AOx) and α-glucosidase inhibitory (IαG) activities, which have been mainly associated with the content of unknown anthocyanins. In this study, the AOx (i.e., DPPH and ABTS as Trolox equivalents, μmol TE/g) and IαG (as half-maximal inhibitory concentration, IC50, mg/mL) activities of the anthocyanin-rich fraction (ARF) obtained from red arils were contrasted with those of the methanol extract (ME), and the main ARF anthocyanins were characterized by HPLC-DAD-ESI-MS, GC-MS and 1H-NMR. The AOx and IαG values of the ARF (DPPH = 597.8; ABTS = 884.01; IαG = 0.06) were better than those of the ME (DPPH = 41.5; ABTS = 142.3; IαG = 17.5); remarkably, the ARF IαG value was about 42 times lower than that of acarbose. The main anthocyanins in ARF were pelargonidin 3-O-glucoside and cyanidin 3-O-glucoside. Thus, the consumption of red P. dulce arils could provide health benefits for prevention/treatment of chronic degenerative diseases such as diabetes.

Context:Helicteres vegae Cristóbal (Sterculiaceae) (Hv) and Heliopsis sinaloensis B.L. Turner (Asteraceae) (Hs) are endangered and poorly studied plant species; related plants have been used against chronic-degenerative and infectious diseases. Therefore, Hv and Hs could be sources of bioactive compounds against these illnesses. Objective: To determine the chemical composition and biological activities (antioxidant, antimutagenic and antimicrobial) of Hv and Hs leaves (L) and stems (S). Materials and methods: Methanol extracts (ME) of each plant/tissue were evaluated for their phytochemicals; phenolics (HPLC-DAD-ESI-MS); antioxidant activity (AA) (0.125–4 mg/mL) (DPPH, ABTS, ORAC and β-carotene discoloration); antimutagenicity (0.5 and 1 mg/plate) (Ames assay, tester strain Salmonella enterica serovar Typhimurium YG1024, 1-nitropyrene as mutagen); activity against human pathogens (1 mg/mL); and toxicity (0.01–2 mg/mL) (Artemia salina assay). Results: All ME showed flavonoids and triterpenes/steroids. The ME-SHv had the highest content of total phenolics (TP) (2245.82 ± 21.45 mg GAE/100 g d.w.) and condensed tannins (603.71 ± 1.115 mg CE/100 g d.w.). The compounds identified were flavonoids (kaempferol 7-O-coumaroylhexoside, and two kaempferol 7-O-rhamnosylhexosides) and phenolics [rosmarinic acid, and 3′-O-(8″-Z-caffeoyl) rosmarinic acid]. The ME-LHs showed the highest content of flavonoids (357.88 mg RE/g d.w.) and phenolic acids (238.58 mg CAE/g d.w.) by HPLC. The ME-SHv showed the highest AA. All ME were strong antimutagens (63.3-85.7%). Only the Hs extracts were toxic (ME-LHs, LC50 = 94.9 ± 1.7 μg/mL; ME-SHs, LC50 = 89.03 ± 4.42 μg/mL). Discussion and conclusions: Both Hv and Hs are potential sources of preventive and therapeutic agents against chronic-degenerative diseases.

Tomato apex necrosis virus (ToANV) is a new virus that causes important damage in tomato crops from the Culiacan Valley, Sinaloa, Mexico. To understand the relationship between ToANV and its vector Bermisia tabaci (Hemiptera: Aleyrodidae) (Gennadius) biotype B, laboratory and greenhouse trials were completed to: 1) determine the acquisition and inoculation access periods of ToANV by B. tabaci from tomato to tomato, 2) understand the transmission efficiency at different B. tabaci population densities, 3) estimate the time from inoculation of the virus at different B. tabaci densities to manifestation of symptoms in the plants, and 4) determine the retention time of the virus by the insect vector. The presence of the virus in plants was determined by reverse transcription-polymerase chain reaction amplification ofa 795-bp fragment (GenBank JN704068), which is phylogenetically related to ToANV (GenBank EF063242). The results showed that B. tabaci is an effective vector for ToANV with relatively long acquisition (12 h) and inoculation (9 h) access periods; a single adult is capable of transmitting and retaining the virus for up to 7d, suggesting a persistent mode of transmission. These results will help in the development of management strategies for controlling the vector and the disease.

Salmonellosis is an important zoonotic disease but little is known about the role that free-living animals play as carriers of this pathogen. Moreover, the primary route of infection in the wild needs to be elucidated. The aim of this study was to determine the source and the route of transmission of Salmonella enterica serovar Albany (S. Albany) infection in captive zoo wild animals in the Culiacán Zoo. A total of 267 samples were analyzed including 220 fecal samples from zoo animals, 15 fecal samples from rodents, 5 pooled samples each of two insects (Musca domestica and Periplaneta americana), and 22 samples of animal feed. We detected S. Albany in 28 (10.5%) of the samples analyzed, including in samples from raw chicken meat. Characterization of isolates was performed by serotyping and pulsed-field gel electrophoresis. All isolates shared a single pulsed-field gel electrophoresis profile, indicating a possible common origin. These data suggest that the infected meat consumed by the wild felines was the primary source of infection in this zoo. It is likely that the pathogen was shed in the feces and disseminated by insects and rats to other locations in the zoo.

Changes in phenylalanine ammonia-lyase (PAL) activity and chilling injury (CI) index were evaluated in tomato (Lycopersicon esculentum M.) during storage and ripening. Fruits were stored at 12C, 4C and from 12 to 4C (gradual cooling, decreasing 2C from 12 to 8C at 3 days intervals and then at 4C) and 85–90% relative humidity for up to 21 days followed by a ripening period of 8 days at 21C. PAL activity showed an increase during the storage period, showing a maximum value after 21 days of storage at 12–4C and 4C. Development of CI symptoms was coincident, and closely correlated with the increase in PAL activity. Gradual cooling reduced 1.5-fold the CI symptoms. It appears that low temperatures induced the tomato fruit defense response, as suggested by the behavior of PAL activity, and gradual cooling increased chilling tolerance. PRACTICAL APPLICATIONS The storage of tomatoes at temperatures below 12C could lead to chilling injury, a physiological disorder that affects the quality of tropical and subtropical fruits resulting in commercial losses or reduced consumer acceptability. In this study, a gradual cooling storage treatment was used to induce tolerance of tomato against this disorder. The results of this research showed that gradual cooling could be an alternative for using temperatures below 12C for the storage of tomato, increasing its shelf life, maintaining the quality and benefiting the distribution chain.

Tomato (Solanum lycopersicum) fruit is susceptible to chilling injury (CI), a physiological disorder caused by low, non-freezing temperatures that affects fruit postharvest quality. Little is known about the biochemical basis of CI, and the aim of this study was to identify proteins related to this disorder in ‘Imperial’ tomato fruit. CI and protein expression changes were analyzed during fruit ripening (0, 4, 8, and 12 days at 21 °C) after storage under chilling (5 °C) and non-chilling conditions (21 °C) for 5, 15, and 25 days. The main CI symptoms observed were uneven fruit ripening and color development, pitting, and decay. Protein analysis of two-dimensional gels showed that 6% of the detected spots (≈300) changed their expression in response to cold. The identified proteins are involved in carbon metabolism, oxidative stress, photosynthesis, and protein processing and degradation; two were related to cold stress, showing higher accumulation in non-damaged tissue of chilled fruit: thioredoxin peroxidase (TPxI) and glycine-rich RNA-binding protein (GR-RBP). This is the first report suggesting an important role for TPxI and GR-RBP in cold response during tomato fruit ripening, and they may be acting through redox sensing and regulation of gene expression at low temperature. These enzymes and the other chilling-related proteins might be working together to maintain the cellular homeostasis under cold stress conditions.

The Mexican fruit fly, Anastrepha ludens Loew (Diptera: Tephritidae), is one of the most harmful pests of mango causing direct damage by oviposition on the fruit pulp. Mango for export is subjected to hydrothermal treatment as a quarantine method for the control of this pest, but exposure to heat for long periods of time reduces considerably the quality and shelf-life of treated fruit. The aim of this work was to study morphological changes of third-instar larvae and adults of A. ludens after in vitro exposure to high temperature at sublethal times. A heating block system was used to expose larvae at 46.1°C for 19.6 and 12.9 min, producing 94.6 and 70% mortality, respectively. Treated larvae were processed for optical microscopy. A fraction of surviving treated larvae was separated into containers with artificial diet to allow development into adults. Adult sexual organs were dissected and processed for transmission electron microscopy analysis. Results showed that 94.6% of the treated larvae died at 46.1°C for 19.6 min and none of the surviving larvae eclosed to adulthood, as they developed as malformed puparia. For the in vitro treatment at 46.1°C during 12.9 min, 70% of the treated larvae died and only 3.75% reached the adult stage, but ultrastructural damage in the male testes and in the female ovaries was observed. Additionally, 11.1% of the adult flies from the in vitro treatment also showed wing malformation and were incapable of flying. The analysis showed that surviving flies were unable to reproduce.

The pulp of the Vitex mollis fruit is edible and traditionally used to treat diarrhoea. The antibacterial activity of this fruit is reported here for the first time. The fruit pulp was extracted with methanol (ME) and the extract was fractionated with solvents. ME and their fractions [hexanic (HF), chloroformic (CF), ethyl acetate (EAF) and aqueous (AqF)] were assayed against human pathogenic bacteria (microdilution test) and their phytochemicals determined (qualitative chemical determinations). The samples (i.e., ME, HE, CF, EAF and AqF) showed antibacterial activity; EAF was the most active, showing such activity against Shigella dysenteriae [minimal inhibitory concentration (MIC)=2 mg/ml]. Phenolics were mainly found in ME and EAF; compounds of this chemical family are well known for their antidiarrhoeal and antimicrobial activities. The reported antibacterial activity and phenolics content of V. mollis fruit could be associated with its use in the treatment of diarrhoea. Key words: Antibacterial activity, diarrhoea, phytochemical analysis, Vitex mollis

A simplified protocol to obtain transgenic tomato plants was established. The effects of culture media composition and Agrobacterium concentration were evaluated. The highest shoot-forming capacity index (5.6) was observed when leaf explants were cultured for 6 weeks with 2 mg·L−1 zeatin, 0.1 mg·L−1 indoleacetic acid, and 300 mg·L−1 timentin. Shoot elongation and root formation were performed in one step on growth regulator-free media. The highest percentage (82%) of fully developed plantlets was obtained when shoots were cultured for 4 weeks with 0.5× Murashige and Skoog (MS) media and 15 g·L−1 sucrose. A 100% of plant survival rate was observed after 4 weeks of being transplanted to ex vitro conditions followed by fruit production (15 fruits/plant) after 2 more weeks. Transient expression of β-glucuronidase was visualized in 100% of the leaf explants infected with Agrobacterium at an OD600 = 0.5 and cocultured for 48 h with 2 mg·L−1 benzylaminopurine, 0.1 mg·L−1 naphthaleneacetic acid, and 100 μM acetosyringone. Stable transformation was confirmed by histochemical glucuronidase assay and polymerase chain reaction (PCR) analysis with a total efficiency of 19.1%. The complete protocol, from shoot induction to fruit production of soil-adapted transgenic plants can be accomplished in only 4 months, and it seems to be very useful for both micropropagation and genetic transformation purposes.