An AP2/ERF Gene, HuERF1, from Pitaya (Hylocereus undatus) Positively Regulates Salt Tolerance.

Pitaya (Hylocereus undatus) is a high salt-tolerant fruit, and ethylene response factors (ERFs) play important roles in transcription-regulating abiotic tolerance. To clarify the function of HuERF1 in the salt tolerance of pitaya, HuERF1 was heterogeneously expressed in Arabidopsis. HuERF1 had nuclear localization when HuERF1::GFP was expressed in Arabidopsis protoplasts and had transactivation activity when HuERF1 was expressed in yeast. The expression of HuERF1 in pitaya seedlings was significantly induced after exposure to ethylene and high salinity. Overexpression of HuERF1 in Arabidopsis conferred enhanced tolerance to salt stress, reduced the accumulation of superoxide (O2∙) and hydrogen peroxide (H2O2), and improved antioxidant enzyme activities. These results indicate that HuERF1 is involved in ethylene-mediated salt stress tolerance, which may contribute to the salt tolerance of pitaya.

Transcriptomic analysis reveals hub genes and subnetworks related to ROS metabolism in Hylocereus undatus through novel superoxide scavenger trypsin treatment during storage.

BACKGROUND: It was demonstrated in our previous research that trypsin scavenges superoxide anions. In this study, the mechanisms of storage quality improvement by trypsin were evaluated in H. undatus. RESULTS: Trypsin significantly delayed the weight loss and decreased the levels of ROS and membrane lipid peroxidation. Transcriptome profiles of H. undatus treated with trypsin revealed the pathways and regulatory mechanisms of ROS genes that were up- or downregulated following trypsin treatment by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) enrichment analyses. The current results showed that through the regulation of the expression of hub redox enzymes, especially thioredoxin-related proteins, trypsin can maintain low levels of endogenous active oxygen species, reduce malondialdehyde content and delay fruit aging. In addition, the results of protein-protein interaction networks suggested that the downregulated NAD(P) H and lignin pathways might be the key regulatory mechanisms governed by trypsin. CONCLUSIONS: Trypsin significantly prolonged the storage life of H. undatus through regulatory on the endogenous ROS metabolism. As a new biopreservative, trypsin is highly efficient, safe and economical. Therefore, trypsin possesses technical feasibility for the quality control of fruit storage.

Molecular characterization and expression analysis of pitaya (Hylocereus polyrhizus) HpLRR genes in response to Neoscytalidium dimidiatum infection.

BACKGROUND: Canker disease caused by Neoscytalidium dimidiatum is a devastating disease resulting in a major loss to the pitaya industry. However, resistance proteins in plants play crucial roles to against pathogen infection. Among resistance proteins, the leucine-rich repeat (LRR) protein is a major family that plays crucial roles in plant growth, development, and biotic and abiotic stress responses, especially in disease defense. RESULTS: In the present study, a transcriptomics analysis identified a total of 272 LRR genes, 233 of which had coding sequences (CDSs), in the plant pitaya (Hylocereus polyrhizus) in response to fungal Neoscytalidium dimidiatum infection. These genes were divided into various subgroups based on specific domains and phylogenetic analysis. Molecular characterization, functional annotation of proteins, and an expression analysis of the LRR genes were conducted. Additionally, four LRR genes (CL445.Contig4_All, Unigene28_All, CL28.Contig2_All, and Unigene2712_All, which were selected because they had the four longest CDSs were further assessed using quantitative reverse transcription PCR (qRT-PCR) at different fungal infection stages in different pitaya species (Hylocereus polyrhizus and Hylocereus undatus), in different pitaya tissues, and after treatment with salicylic acid (SA), methyl jasmonate (MeJA), and abscisic acid (ABA) hormones. The associated protein functions and roles in signaling pathways were identified. CONCLUSIONS: This study provides a comprehensive overview of the HpLRR family genes at transcriptional level in pitaya in response to N. dimidiatum infection, it will be helpful to understand the molecular mechanism of pitaya canker disease, and lay a strong foundation for further research.

Cross-Linking Chitosan into Hydroxypropylmethylcellulose for the Preparation of Neem Oil Coating for Postharvest Storage of Pitaya (Stenocereus pruinosus).

The market trend for pitaya is increasing, although the preservation of the quality of this fruit after the harvest is challenging due to microbial decay, dehydration, and oxidation. In this work, the application of antimicrobial chitosan-based coatings achieved successful postharvest preservation of pitaya (Stenocereus pruinosus) during storage at 10 ± 2 °C with a relative humidity of 80 ± 5%. The solution of cross-linked chitosan with hydroxypropylmethylcellulose with entrapped Neem oil (16 g·L-1) displayed the best postharvest fruit characteristics. The reduction of physiological weight loss and fungal contamination, with an increased redness index and release of azadirachtin from the microencapsulated oil, resulted in up to a 15 day shelf life for this fruit. This postharvest procedure has the potential to increase commercial exploitation of fresh pitaya, owing to its good taste and high content of antioxidants.

Comparative volatile compounds and primary metabolites profiling of pitaya fruit peel after ozone treatment.

BACKGROUND: Ozone treatment can effectively inhibit fruit decay in many fruits during postharvest storage. However, little information is available for pitaya fruit. RESULTS: Ozone treatment significantly reduced the decay rate and induced the enzyme activities of peroxidase and polyphenol oxidase, and also reduced the levels of reactive oxygen species. In total, 103 metabolites were detected and changed the content after ozone treatment, including 54 primary metabolites and 49 aromatic compounds. After significance and importance analysis, 37 metabolites were important. Some metabolites were induced by peel senescence to respond to senescence stress, including d-fructose, d-glucose, mannose, inositol, galactonic acid, ethanedioic acid and stearic acid. Some metabolic products of peel senescence were reduced by ozone treatment, including d-arabinose, glucaric acid, galacturonic acid, 1-hexanol, 4-ethylcyclohexanol, ß-linalool, palmitoleic acid and 2-hydroxy-cyclopentadecanone. Some metabolites induced by ozone treatment might play a vital role in delaying the senescence and decay, including malic acid, succinic acid, pentenoic acid, eicosanoic acid, 2-hexenal, hexanal, 2-heptenal, 4-heptenal, 2-octenal and nitro m-xylene. CONCLUSION: Ozone treatment significantly reduced decay and prolonged shelf-life without reducing fruit quality. In total, 37 metabolites might play an important role in ozone delayed fruit decay. © 2018 Society of Chemical Industry.

Effect of postharvest spray of apple polyphenols on the quality of fresh-cut red pitaya fruit during shelf life.

Fresh-cut (FC) red pitaya fruit were treated with 5ga.i.l-1 apple polyphenols (APP) and then stored at 20°C for up to 4days to evaluate the effects on attributes. Results showed that FC pitaya fruit with APP treatment showed greater colour retention, delayed softening, reduced loss of soluble solids content, titratable acidity, betacyanin and total phenolics compared with untreated FC fruit. APP treatment also maintained antioxidant activity, as indicated by higher DPPH radical-scavenging activity and reducing power compared with untreated FC pitaya fruit. APP treatment strongly suppressed microbial growth, contributing to improvement of product safety. Because APP is a natural product, we propose that application of APP could be a convenient, safe and low-cost approach to maintain the quality and extend the shelf life of FC red pitaya fruit.

Morphological response of a cactus to cement dust pollution.

Cement dust from cement plants around the world has multiple negative effects on organisms and their environment. Cement's effects come from its strongly alkaline nature and high content of heavy metals. Previous studies on plants have documented that cement dust deposition can influence plant vegetative growth, the lipid and ionic composition of tissues, and foliar temperature. Here we evaluate the effects of cement dust coming from a plant in western Argentina on the morphology of the cactus Tephrocactus aoracanthus. In sites located at 0.15km, 2km and 6km from the cement plant, we recorded five morphological attributes of the cactus: length and number of spines, cladode (stem) diameter, and fresh and dry weight. We also transplanted plants in situ to evaluate the effect of distance from the cement plant. In addition, we set an experiment spreading cement dust weekly on the aerial and ground parts of the cactus. Results of our field observational and experimental studies indicate that cement dust deposition on aerial parts of the plant leads to increased spine length, number of spines, and wet and dry weights of cladodes.

Callus Induction from Various Organs of Dragon Fruit, Apple and Tomato on some Mediums.

BACKGROUND AND OBJECTIVE: Dragon fruit (Hylocereus spp.), apple (Malus sylvestris Mill.) and tomato (Solanum lycopersicum L.) are high potential sources of antioxidant compounds such as phenolics. The compounds have the capability of protecting cells and tissues against free radicals. Secondary metabolite produced by callus cell culture from plant organs also acts as a source of antioxidants. This study aimed to determine the optimal ratio of sucrose and 2,4-D in Murashige and Skoog (MS) medium for callus induction from different plant organ explants. With all of characteristic, callus can be used further for the development of natural cell regeneration agent. METHODOLOGY: This study was conducted using analytical technique. Suitable explants were obtained. They were developed in various concentrations of combination between MS medium and 2,4-D. Callus growth, including their weight and surface was then measured and analyzed by using one-way analysis of variance (ANOVA). RESULTS: Callus was able to grow from its explants in 5-7 days after induction process. They were clear in color and had friable texture. The highest value of fresh weight of dragon fruit callus was obtained through MS supplemented with 1 µL L-1 2,4-D and 30 g sucrose. However, apple and tomato callus induction and growth maintenance reached optimal medium on MS supplemented with 30 g sucrose and 2 µL L-1 2,4-D. CONCLUSION: Callus of apple, dragon fruit and tomato was maintained upon MS supplemented with 30-40 g sucrose and 1-2 µL L-1 2,4-D for optimum induction and growth. The optimization of growth medium will give advantages for further development of natural cell regeneration agent.

Isolation and characterization of a catalase gene "HuCAT3" from pitaya (Hylocereus undatus) and its expression under abiotic stress.

Abiotic stresses usually cause H2O2 accumulation, with harmful effects, in plants. Catalase may play a key protective role in plant cells by detoxifying this excess H2O2. Pitaya (Hylocereus undatus) shows broad ecological adaptation due to its high tolerance to abiotic stresses, e.g. drought, heat and poor soil. However, involvement of the pitaya catalase gene (HuCAT) in tolerance to abiotic stresses is unknown. In the present study, a full-length HuCAT3 cDNA (1870 bp) was isolated from pitaya based on our previous microarray data and RACE method. The cDNA sequence and deduced amino acid sequence shared 73-77% and 75-80% identity with other plant catalases, respectively. HuCAT3 contains conserved catalase family domain and catalytic sites. Pairwise comparison and phylogenetic analysis indicated that HuCAT3 is most similar to Eriobotrya japonica CAT, followed by Dimocarpus longan CAT and Nicotiana tabacum CAT1. Expression profile analysis demonstrated that HuCAT3 is mainly expressed in green cotyledons and mature stems, and was regulated by H2O2, drought, cold and salt stress, whereas, its expression patterns and maximum expression levels varied with stress types. HuCAT activity increased as exposure to the tested stresses, and the fluctuation of HuCAT activity was consistent with HuCAT3 mRNA abundance (except for 0.5 days upon drought stress). HuCAT3 mRNA elevations and HuCAT activities changes under cold stress were also in conformity with the cold tolerances among the four genotypes. The obtained results confirmed a major role of HuCAT3 in abiotic stress response of pitaya. This may prove useful in understanding pitaya's high tolerance to abiotic stresses at molecular level.

Influence of abscisic acid and sucrose on somatic embryogenesis in Cactus Copiapoa tenuissima Ritt. forma mostruosa.

Having produced the embryos of cactus Copiapoa tenuissima Ritt. forma monstruosa at the globular stage and callus, we investigated the effect of abscisic acid (ABA) in the following concentrations: 0, 0.1, 1, 10, and 100 µ M on successive stages of direct (DSE) and indirect somatic embryogenesis (ISE). In the indirect somatic embryogenesis process we also investigated a combined effect of ABA (0, 0.1, 1 µ M) and sucrose (1, 3, 5%). The results showed that a low concentration of ABA (0-1 µ M) stimulates the elongation of embryos at the globular stage and the number of correct embryos in direct somatic embryogenesis, while a high ABA concentration (10-100 µ M) results in growth inhibition and turgor pressure loss of somatic embryos. The indirect somatic embryogenesis study in this cactus suggests that lower ABA concentrations enhance the increase in calli fresh weight, while a high concentration of 10 µ M ABA or more changes calli color and decreases its proliferation rate. However, in the case of indirect somatic embryogenesis, ABA had no effect on the number of somatic embryos and their maturation. Nevertheless, we found a positive effect of sucrose concentration for both the number of somatic embryos and the increase in calli fresh weight.

Nopalea cochenillifera, a potential chromium (VI) hyperaccumulator plant.

Hexavalant chromium [Cr(VI)] tolerance and accumulation in in vitro grown Nopalea cochenillifera Salm. Dyck. plants was investigated. A micropropagation protocol was establish for a rapid multiplication of N. cochenillifera and [Cr(VI)] tolerance and accumulation was studied in in vitro grown cultures. Cr concentration was estimated by atomic absorption spectroscopy in roots and shoots to confirm plant's hyperaccumulation capacity. Plants showed tolerance up to 100 µM K(2)Cr(2)O(7) without any significant changes in root growth after 16 days treatment; whereas, chlorophyll content in plants treated with 1 and 10 µM K(2)Cr(2)O(7) were not so different than the control plant. The levels of lipid peroxidation and protein oxidation increased significantly (p < 0.01) with increasing concentration of chromium. Exposures of N. cochenillifera to lower concentrations of K(2)Cr(2)O(7) (≤ 10 µM) induced catalase (CAT) and superoxide dismutase (SOD) significantly (p < 0.001) but higher concentrations of K(2)Cr(2)O(7) (>100 µM) inhibited the activities of CAT and SOD. Roots accumulated a maximum of 25,263.396 ± 1,722.672 mg Cr Kg(-1) dry weight (DW); while the highest concentration of Cr in N. cochenillifera shoots was 705.714 ± 32.324 mg Cr Kg(-1) DW. N. cochenillifera could be a prospective hyperaccumulator plant of Cr(VI) and a promising candidate for phytoremediation purposes.

Clumping and dispersal of chloroplasts in succulent plants.

Plants have evolved various photoprotective mechanisms to mitigate photodamage. Here we report the diurnal movement of chloroplasts in the leaves of succulent crassulacean acid metabolism (CAM) plants under combined light and water stress. In leaves of water-stressed plants, the chloroplasts became densely clumped in one or sometimes two areas in the cytoplasm under light and dispersed during darkness. The chloroplast clumping resulted in leaf optical changes, with a decrease in absorptance and an increase in transmittance. The plant stress hormone abscisic acid induced chloroplast clumping in the leaf cells under light. We suggest that the marked chloroplast movement in these CAM plants is a photoprotective strategy used by the plants subjected to severe water stress.

Effects of benomyl and drought on the mycorrhizal development and daily net CO2 uptake of a wild platyopuntia in a rocky semi-arid environment.

The effects of drought and the fungicide benomyl on a wild platyopuntia, Opuntia robusta Wendl., growing in a rocky semi-arid environment were assessed. Cladode phosphorus content, cladode water potential and daily net CO2 uptake were measured monthly in 2000 and 2001 before, during and after the summer rainy period. During 2000, the formation of new roots and new cladodes was severely suppressed in response to a prolonged drought, impairing the development of the symbiotic relationship between the arbuscular mycorrhizal (AM) fungi and the roots. Hence no effect of benomyl application was observed on daily carbon assimilation by this Crassulacean acid metabolism plant. During 2001, drought was interrupted, and new cladodes and roots were formed in response to rainfall. Benomyl was highly effective in suppressing root colonization by AM-fungi; however, daily C assimilation was reduced by benomyl application only in October. Thus, the inhibition of AM-fungal colonization by benomyl did not affect photosynthesis, water uptake and P uptake under prolonged drought.

Alkaloid production by callous tissue cultures of Cereus peruvianus (Cactaceae).

The morphologically undifferentiated cells of nonregenerant callous tissue of Cereus peruvianus cultured in the original medium and in medium supplemented with tyrosine were used as an alkaloid source. Comparison of alkaloid production by C. peruvianus plants and by callous tissues indicated that alkaloid levels were almost twice as high in callous tissues as in shoots of C. peruvianus plants. The ratio of alkaloid concentration between mature plant and morphologically undifferentiated cells of callous tissue was 1:1.7. A relationship between culture medium containing tyrosine and alkaloid production was also observed in the callous tissues of C. peruvianus. Since increased alkaloid production may be induced by additional factors such as tyrosine, increasing levels of tyrosine or other conditions of the culture medium may be considered factors for inducing higher alkaloid production by C. peruvianus callous tissues.