Fitorremediación con Brassicaceae y Apiaceae en suelos contaminados con metales pesados

Introducción: La cuenca media del río Bogotá viene sufriendo contaminación por metales pesados debido a los vertidos industriales. Esta fuente de agua ha sido utilizada para el riego de cultivos de ciclo corto, lo que ha provocado la contaminación por metales pesados de los suelos productivos. Objetivo: Evaluar un proceso de fitorremediación con las especies de Brócoli (Brassica oleracea var. italica Plenck) y Cilantro (Coriandrum sativum L.) en suelos contaminados con metales pesados en Mosquera, Colombia. Métodos: El muestreo se realizó con un diseño factorial al azar: a) proporciones de cilantro/brócoli (30 cilantro/70 brócoli, 50/50, 70 cilantro/30 brócoli y dos controles de tipo monocultivo de cada especie) y b) tiempo de fitorremediación (3, 6 y 9 meses), cada tratamiento con cuatro parcelas experimentales. El suelo y el tejido vegetal se analizó mediante espectrofotometría de absorción atómica para determinar la concentración de los metales pesados. Resultados: Las concentraciones de metales pesados en las muestras de tejido vegetal para brócoli y cilantro mostraron valores de Cd, Pb y Cr más altos que Hg y As, respecto al control. Las concentraciones de Cd, Pb y Cr presentaron valores significativamente más altos (P < 0.05) en suelos con cilantro y brócoli mezclados, respecto al sitio de control donde la concentración de Cd registró niveles significativamente más bajos que en los otros suelos. Las concentraciones de Cd, Pb y Cr fueron más altas en tejidos de cilantro y en suelos con cilantro en comparación con el control. Conclusiones: La fitorremediación por medio de las especies Brassica oleracea var. itálica Plenck y Coriandrum sativum L. en proporciones 70/30 o 30/70 a suelos con pH entre 5.5 y 6.0 permiten una buena recuperación de suelos con contenidos de metales pesados en términos de concentraciones altas a cortos plazos, iniciando la biorremediación a los tres meses y finalizando cargas de disponibilidad variable a los nueve meses.

Introduction: The middle basin of the Bogotá River has been suffering from heavy metal contamination due to industrial discharges. This water source has been used to irrigate short-cycle crops, which has resulted in heavy metal contamination of productive soils. Objective: To evaluate a phytoremediation process with the plant species Broccoli (Brassica oleracea var. Italica plenck) and Cilantro (Coriandrum sativum L.) in soils contaminated with heavy metals in Mosquera, Colombia. Methods: Sampling was performed with a randomized factorial design: a) cilantro/broccoli proportions (30 cilantro/70 broccoli, 50/50, 70 cilantro/30 broccoli and two monoculture type controls of each species) and b) phytoremediation time (3, 6 and 9 months), each treatment with four experimental plots. Soil and plant tissue were analyzed in the laboratory by atomic absorption spectrophotometry to determine the concentration of heavy metals. Results: The concentrations of heavy metals in plant tissue samples for broccoli and cilantro showed higher values of Cd, Pb and Cr than Hg and As, with respect to the control. The concentrations of Cd, Pb and Cr presented significantly higher values (P < 0.05) in soil where there was a mixed presence of cilantro and broccoli, with respect to the control site where the concentration of Cd registered significantly lower levels than in the other soils. Finally, Cd, Pb and Cr concentrations were higher in cilantro tissues and in soils with cilantro compared to the control. Conclusions: Phytoremediation by means of Brassica oleracea var. italica Plenck and Coriandrum sativum L. in 70/30 or 30/70 proportions in soils with pH between 5.5 and 6.0 allows a good recovery of soils with heavy metal contents in terms of high concentrations in short terms, starting bioremediation after three months and ending loads of variable availability after nine months.

Overexpression of BoLSU1 and BoLSU2 Confers Tolerance to Sulfur Deficiency in Arabidopsis by Manipulating Glucosinolate Metabolism.

Sulfur is an essential element for plant growth, development and resistance to environmental stresses. Glucosinolates (GSLs), a group of sulfur rich secondary metabolites found in Brassicaceae plants, are known for their defensive properties against pathogens and herbivores. Due to their integration of a large proportion of total sulfur, their biosynthesis and degradation are closely linked to sulfur metabolism. It has been demonstrated that GSLs can be broken down to release sulfur and facilitate the production of other thio-metabolites when the plant is under stress. However, the regulation of this process is still not fully understood. In this study, we constructed two broccoli LSU (low sulfur responsive) gene overexpressing lines, 35S::BoLSU1 and 35S::BoLSU2, to detect changes in GSL metabolism after sulfur deficiency treatment. The results showed that BoLSU1 and BoLSU2 inhibit the biosynthesis of aliphatic GSLs, while also promoting their degradation and increasing the content of glutathione (GSH), leading to the reallocation of sulfur from the GSL pool to other thio-metabolites such as GSH. Furthermore, this regulation of GSL metabolism mediated by BoLSU1 and BoLSU2 is found to be dependent on myrosinases BGLU28 and BGLU30. Our study provides insight into the physiological role of LSU proteins and their regulation of sulfur metabolism.

Effect of Potassium Permanganate, Ultraviolet Radiation and Titanium Oxide as Ethylene Scavengers on Preservation of Postharvest Quality and Sensory Attributes of Broccoli Stored with Tomatoes.

This study introduces an effective solution to enhance the postharvest preservation of broccoli, a vegetable highly sensitive to ethylene, a hormone produced by climacteric fruits such as tomatoes. The proposed method involves a triple combination of ethylene elimination techniques: potassium permanganate (KMnO4) filters combined with ultraviolet radiation (UV-C) and titanium oxide (TiO2), along with a continuous airflow to facilitate contact between ethylene and these oxidizing agents. The effectiveness of this approach was evaluated using various analytical techniques, including measurements of weight, soluble solids content, total acidity, maturity index, color, chlorophyll, total phenolic compounds, and sensory analysis conducted by experts. The results demonstrated a significant improvement in the physicochemical quality of postharvest broccoli when treated with the complete system. Notably, broccoli subjected to this innovative method exhibited enhanced organoleptic quality, with heightened flavors and aromas associated with fresh green produce. The implementation of this novel technique holds great potential for the food industry as it reduces postharvest losses, extends the shelf life of broccoli, and ultimately enhances product quality while minimizing waste. The successful development and implementation of this new technique can significantly improve the sustainability of the food industry while ensuring the provision of high-quality food to consumers.

Anticancer activity of broccoli, its organosulfur and polyphenolic compounds.

The use of natural bioactive constituents from various food sources for anticancer purposes has become increasingly popular worldwide. Broccoli (Brassica oleracea var. italica) is on the top of the consumed vegetables by the masses. Its raw matrix contains a plethora of phytochemicals, such as glucosinolates and phenolic compounds, along with rich amounts of vitamins, and minerals. Consumption of broccoli-derived phytochemicals provides strong antioxidant effects, particularly due to its sulforaphane content, while modulating numerous molecules involved in cell cycle regulation, control of apoptosis, and tuning enzyme activity. Thus, the inclusion of broccoli in the daily diet lowers the susceptibility to developing cancers. Numerous studies have underlined the undisputable role of broccoli in the diet as a chemopreventive raw food, owing to the content in sulforaphane, an isothiocyanate produced as a result of hydrolysis of precursor glucosinolates called glucoraphanin. This review will provide evidence supporting the specific role of fresh florets and sprouts of broccoli and its key bioactive constituents in the prevention and treatment of different cancers; a number of studies carried out in the in vitro and in vivo conditions as well as clinical trials were analyzed.

Exogenous Postharvest Application of Calcium Chloride and Salicylic Acid to Maintain the Quality of Broccoli Florets.

The importance of broccoli (Brassica oleracea var. italica) consumption has increased in recent years due to its significant amount of anticarcinogenic and antioxidant compounds, as well as its many vitamins. However, broccoli florets are a highly perishable product which rapidly senesce and turn yellow after harvest, resulting in losses in nutritional and bioactive compounds. Thus, in this study, we evaluated the effect of postharvest exogenous of salicylic acid (SA) and calcium chloride (CaCl2) and their combination on the quality of broccoli florets stored at 5 °C for 28 days to minimize the rapid senescence of broccoli florets. Samples treated with 2 mM SA alone or in combination with 2% CaCl2 showed lower weight loss and lower losses of chlorophyll content, vitamin C, phenolic compounds, carotenoids, flavonoids, and glucosinolates compared with the control samples. Additionally, antioxidant activity was maintained by either SA or SA + CaCl2 treatments while peroxidase activity was decreased. For higher quality and lower losses in antioxidant compounds of broccoli florets during refrigerated storage at 5 °C, SA + CaCl2 treatment could be helpful for up to 21 days.

Enhancement of Glucosinolate Formation in Broccoli Sprouts by Hydrogen Peroxide Treatment.

Broccoli sprouts are known as a rich source of health-beneficial phytonutrients: glucosinolates and phenolic compounds. The production of phytonutrients can be stimulated by elicitors that activate the plant stress response. The aim of this study was enhancing the nutritional value of broccoli sprouts using hydrogen peroxide (H2O2) as an elicitor. Daily spraying with H2O2 (500-1000 mM) enhanced the accumulation of glucosinolates, doubling their content in the cotyledons of 16/8 h photoperiod-grown 7-day sprouts compared to the water-treated controls. The application of H2O2 on dark-grown sprouts showed a smaller extent of glucosinolate stimulation than with light exposure. The treatment affected sprout morphology without reducing their yield. The H2O2-treated sprouts had shorter hypocotyls and roots, negative root tropism and enhanced root branching. The activated glucosinolate production became evident 24 h after the first H2O2 application and continued steadily until harvest. Applying the same treatment to greenhouse-grown wild rocket plants caused scattered leaf bleaching, a certain increase in glucosinolates but decline in phenolics content. The H2O2 treatment of broccoli sprouts caused a 3.5-fold upregulation of APK1, a gene related to sulfur mobilization for glucosinolate synthesis. Comparing the APK1 expression with the competing gene GSH1 using sulfur for antioxidant glutathione production indicated that glutathione synthesis prevailed in the sprouts over the formation of glucosinolates.

Combination of Selenium and UVA Radiation Affects Growth and Phytochemicals of Broccoli Microgreens.

Addition of selenium or application of ultraviolet A (UVA) radiation for crop production could be an effective way of producing phytochemical-rich food. This study was conducted to investigate the effects of selenium and UVA radiation, as well as their combination on growth and phytochemical contents in broccoli microgreens. There were three treatments: Se (100 µmol/L Na2SeO3), UVA (40 µmol/m2/s) and Se + UVA (with application of Se and UVA). The control (CK) was Se spraying-free and UVA radiation-free. Although treatment with Se or/and UVA inhibited plant growth of broccoli microgreens, results showed that phytochemical contents increased. Broccoli microgreens under the Se treatment had higher contents of total soluble sugars, total phenolic compounds, total flavonoids, ascorbic acid, Fe, and organic Se and had lower Zn content. The UVA treatment increased the contents of total chlorophylls, total soluble proteins, total phenolic compounds, and FRAP. However, the Se + UVA treatment displayed the most remarkable effect on the contents of total anthocyanins, glucoraphanin, total aliphatic glucosinolates, and total glucosinolates; here, significant interactions between Se and UVA were observed. This study provides valuable insights into the combinational selenium and UVA for improving the phytochemicals of microgreens grown in an artificial lighting plant factory.

Preharvest UVB Application Increases Glucosinolate Contents and Enhances Postharvest Quality of Broccoli Microgreens.

Broccoli microgreens have shown potential health benefits due to their high glucosinolate (GL) levels. Previously, we observed that postharvest UVB treatment did not have much effect on increasing GLs in broccoli microgreens. In this study, we investigated the influence of preharvest UVB irradiation on GL levels in broccoli microgreens. UHPLC-ESI/ITMS analysis showed that preharvest UVB treatments with UVB 0.09 and 0.27 Wh/m2 significantly increased the glucoraphanin (GLR), glucoerucin (GLE), and total aliphatic GL levels by 13.7 and 16.9%, respectively, in broccoli microgreens when measured on harvest day. The nutritional qualities of UVB-treated microgreens were stable during 21-day storage, with only small changes in their GL levels. Broccoli microgreens treated before harvest with UVB 0.27 Wh/m2 and 10 mM CaCl2 spray maintained their overall quality, and had the lowest tissue electrolyte leakage and off-odor values during the storage. Furthermore, preharvest UVB 0.27 Wh/m2 treatment significantly increased GL biosynthesis genes when evaluated before harvest, and reduced the expression level of myrosinase, a gene responsible for GL breakdown during postharvest storage. Overall, preharvest UVB treatment, together with calcium chloride spray, can increase and maintain health-beneficial compound levels such as GLs and prolong the postharvest quality of broccoli microgreens.

Postharvest LED lighting: effect of red, blue and far red on quality of minimally processed broccoli sprouts.

BACKGROUND: The main objective of this study was to evaluate physiological and quality changes of minimally processed broccoli sprouts illuminated during postharvest storage under blue, red and far-red LED lighting as compared to darkness or illumination with fluorescent light, as control treatments. RESULTS: Morphological and microbiological changes were determined during 15 days at 5 °C. In addition, total antioxidant activity and bioactive compound changes throughout the shelf life were also monitored. Results showed that far-red LED lighting increased hypocotyl and sprout length, decreased microbial growth and improved the total antioxidant and scavenging activities, compared to darkness and fluorescent lighting treatments. However, it did not stimulate the biosynthesis of phenolic acids. In contrast, blue LED light reduced by 50% the total antioxidant capacity of broccoli sprouts compared to far-red treatment, as well as morphological development. In addition, total scavenging activity was increased under far-red LED light compared with the other treatments by 12-10% (darkness and fluorescence) and 33-31% (blue and red LEDs). CONCLUSIONS: Our results suggest that minimally processed sprouts may benefit from LED lighting during shelf life in terms of quality, although further experiments should be conducted to optimize a proper exposure cycle and intensity aiming for use in the distribution chain. The results also open the way for further development towards the integration of this technology in the food distribution chain.

Revalorization of Broccoli By-Products for Cosmetic Uses Using Supercritical Fluid Extraction.

The agri-food industry is currently one of the main engines of economic development worldwide. The region of Murcia is a reference area in Europe for the cultivation of fruits and vegetables and produces the bulk of Spanish exports of broccoli (Brassica oleracea var. italica). The processing of fresh produce generates a huge number of by-products that represent an important economic and environmental problem when discarded. In this work, an advanced extraction technique using environmentally friendly solvents was applied to assess the revalorization of broccoli by-products, by performing a comparative analysis with conventional extraction. To achieve this goal, supercritical fluid extraction based on response surface methodology was performed using CO2 and ethanol as solvents. The results obtained showed that the supercritical fluid extracts were rich in ß-carotene, phenolic compounds, chlorophylls and phytosterols. Moreover, in bioactivity assays, the supercritical fluid extracts exhibited a high antioxidant activity and a cytoprotective effect in a non-tumorigenic keratinocyte cell line exposed to ultraviolet B light. The results indicate that supercritical fluid extracts from broccoli by-products could potentially serve as an ingredient for cosmetic purposes.

Pseudomonas brassicae sp. nov., a pathogen causing head rot of broccoli in Japan.

Phytopathogenic bacteria, MAFF 212426, MAFF 212427T, MAFF 212428 and MAFF 212429, were isolated from head rot lesions of broccoli (Brassica oleracea L. var. italica Plenck) in Hokkaido, Japan, and subjected to polyphasic taxonomic characterization. The cells were Gram-reaction-negative, aerobic, non-spore-forming, motile with one or two polar flagella, rod-shaped and formed pale yellow colonies. Results of 16S rRNA gene sequence analysis showed that they belong to the genus Pseudomonas with the highest similarity to 'Pseudomonas qingdaonensis' JJ3T (99.86 %), Pseudomonas laurentiana GSL-010T (99.22 %), Pseudomonas huaxiensis WCHPs060044T (99.01 %), Pseudomonas japonica NBRC 103040T (98.87 %) and Pseudomonas alkylphenolica KL28T (98.73 %). The genomic DNA G+C content was 63.4 mol% and the major fatty acids (>5 % of the total fatty acids) were summed feature 3 (C16 : 1 ω7c / C16 : 1 ω6c), C16 : 0, summed feature 8 (C18 : 1 ω7c / C18 : 1 ω6c) and C17 : 0 cyclo. Multilocus sequence analysis using the partial rpoD, gyrB and rpoB gene sequences and phylogenomic analyses based on the whole genome sequences demonstrated that the strains are members of the Pseudomonas putida group, but form a monophyletic, robust clade separated from their closest relatives. Average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values corroborated their novel species status, with 88.39 % (ANI) and 35.8 % (dDDH) as the highest scores with 'P. qingdaonensis' JJ3T. The strains were differentiated from their closest relatives by phenotypic characteristics, pathogenicity on broccoli, and whole-cell MALDI-TOF mass spectrometry profiles. The phenotypic, chemotaxonomic and genotypic data showed that the strains represent a novel Pseudomonas species, for which the name Pseudomonas brassicae sp. nov. is proposed. The type strain is MAFF 212427T (=ICMP 23635T).

Digitization of Broccoli Freshness Integrating External Color and Mass Loss.

Yellowing of green vegetables due to chlorophyll decomposition is a phenomenon indicating serious deterioration of freshness, and it is evaluated by measuring color space values. In contrast, mass reduction due to water loss is a deterioration of freshness observed in all horticultural crops. Therefore, in this study, we propose a novel freshness evaluation index for green vegetables that combines the degree of greenness and mass loss. The green color retention rate was measured using a computer vision system, and the mass retention rate was measured by weighing. Linear discriminant analysis (LDA) was performed using both variables (greenness and mass) as covariates to obtain a single freshness evaluation value (first canonical variable). The correct classification of storage period length by LDA was 96%. Green color retention alone allowed for classification of storage durations between 0 day and 10 days, whereas LDA could classify storage durations between 0 day and 12 days. The novel freshness evaluation index proposed by this research, which integrates greenness and mass, has been shown to be more accurate than the conventional evaluation index that uses only greenness.

NMR-Based Metabolomic Comparison of Brassica oleracea (Var. italica): Organic and Conventional Farming.

Brassicaceae family provides several crops which are worldwide known for their interesting phytochemical profiles, especially in terms of content of glucosinolates. These secondary metabolites show several beneficial effects toward consumers' health, and several studies have been conducted to identify cultivation factors affecting their content in crops. One of the agronomic practices which is attracting growing interest is the organic one, which consists in avoiding the use of mineral fertilizers as well as pesticides. The aim of this study is to define the metabolic profile of Brassica oleracea (var. italica) and to compare the samples grown using organic and conventional fertilization methods. The hydroalcoholic and organic extracts of the samples have been analyzed by NMR spectroscopy. Forty-seven metabolites belonging to the categories of organic acids, amino acids, carbohydrates, fatty acids, sterols, and other molecules have been identified. Thirty-seven metabolites have been quantified. Univariate and multivariate PCA analyses allowed to observe that the organic practice influenced the nitrogen transport, the carbohydrate metabolism, the glucosinolate content and the phenylpropanoid pathway in B. oleracea (var. italica).

Pectin from Brassica oleracea var. italica triggers immunomodulating effects in vivo.

The immunomodulatory ability of pectins is related with structural features such as the degree of methyl-esterification and branching, as well as the molecular mass. The pectin FB, extracted from broccoli stalks (Brassica oleracea var. italica) had low molecular mass, 56% methyl-esterification and galactose as the main neutral sugar, sharing some characteristics with the modified citrus pectin (MCP), which has been extensively studied in vivo and in vitro due its immunomodulator potential. Considering that broccoli has an important role in the diet, the main objective of this study was to investigate the ability of FB in modulating the immune system in vivo. At concentrations 100-500 µg/mL, FB did not affect the viability of macrophages. Evaluations on morphology and phagocytic activity showed that FB (500 µg/mL) increased the number of activated macrophages by 39% and phagocytic activity by 30% within 48 h. FB (200 mg/kg) administered intraperitoneally increased the number of peritoneal macrophages in mice by 490% after 24 h and modulated these cells for an activated phenotype. In mice, oral administration of FB (200 mg/kg) stimulated lymphocytes from spleen and bone marrow proliferation. FB did not induce nitric oxide (NO) production by macrophages and also did not affect the levels of pro-inflammatory interleukins IL-1ß and IL-12 by peritoneal macrophages, but induced the production of the anti-inflammatory interleukin IL-10. The results could suggest that the anti-inflammatory effects triggered by FB could be related to its degree of esterification and pointed this polysaccharide as a target for the development of new immunomodulatory drugs.

Prediction of Degreening Velocity of Broccoli Buds Using Hyperspectral Camera Combined with Artificial Neural Networks.

Developing a noninvasive technique to estimate the degreening (loss of green color) velocity of harvested broccoli was attempted. Loss of green color on a harvested broccoli head occurs heterogeneously. Therefore, hyperspectral imaging technique that stores spectral reflectance with spatial information was used in the present research. Using artificial neural networks (ANNs), we demonstrated that the reduction velocity of chlorophyll at a site on a broccoli head was related to the second derivative of spectral reflectance data at 15 wavelengths from 405 to 960 nm. The reduction velocity was predicted using the ANNs model with a correlative coefficient of 0.995 and a standard error of prediction of 5.37 × 10-5 mg·g-1·d-1. The estimated reduction velocity was effective for predicting the chlorophyll concentration of broccoli buds until 7 d of storage, which was established as the maximum time for maintaining marketability. This technique may be useful for nondestructive prediction of the shelf life of broccoli heads.

Induction of Terminal Oxidases of Electron Transport Chain in Broccoli Heads under Controlled Atmosphere Storage.

Controlled atmosphere (CA) storage, that is, at low O2 and high CO2 concentrations, effectively extends the shelf life of horticultural products. The influence of CA storage (O2/CO2: 2.5%/6.0% or 2.5%/0.0%) and in normal air (both at 1 °C for 21 d) on the physicochemical (O2 uptake, mass loss and L-ascorbate) and biological properties of broccoli (Brassica oleracea var. italica, Plenck, 1794) via amounts and activities of terminal oxidases of the electron transport chain was investigated. Mass loss, a sensitive index of freshness for broccoli heads under CA, was significantly lower under CA than under normoxia (p < 0.05). Mass loss was depressed 7 d earlier under CA, including 6.0% CO2 than under CA without CO2. High CO2 effectively depressed the degradation of L-ascorbate. During storage, the activity of the alternative oxidase (AOX) was lower under CA than in normal air (p < 0.05), while the amount of cytochrome c oxidase (COX), and the AOX/COX activity ratio (based on oxygen isotope discrimination), were not affected during storage. Our results indicate that CA storage effectively retained the freshness of broccoli heads by depressing the induction of AOX. However, depression of AOX amount was not associated with CO2 around broccoli heads.

Class I defensins (BraDef) from broccoli (Brassica oleracea var. italica) seeds and their antimicrobial activity.

The objective of this study was to determine whether seeds of Brassica oleracea var. italica (i.e. broccoli, an edible plant) produce defensins that inhibit phytopathogenic fungi and pathogenic bacteria of clinical significance. Crude extracts obtained from broccoli seeds were fractioned by molecular exclusion techniques and analyzed by liquid chromatography-high-resolution mass spectrometry. Two peptides were identified, BraDef1 (10.68 kDa) and BraDef2 (9.9 kDa), which were categorized as Class I defensins based on (a) their primary structure, (b) the presence of four putative cysteine disulfide bridges, and (c) molecular modeling predictions. BraDef1 and BraDef2 show identities of, respectively, 98 and 71%, and 67 and 85%, with defensins from Brassica napus and Arabidopsis thaliana. BraDef (BraDef1 + BraDef2) disrupted membranes of Colletotrichum gloeosporioides and Alternaria alternata and also reduced hyphal growth of C. gloeosporioides by ~ 56% after 120 h of incubation. Pathogenic bacteria (Bacillus cereus 183, Listeria monocytogenes, Salmonella typhimurium, Pseudomonas aeruginosa, and Vibrio parahaemolitycus) were susceptible to BraDef, but probiotic bacteria such as Bifidobacterium animalis, Lactobacillus acidophilus, and Lactobacillus casei were not inhibited. To our knowledge, this is the first report of defensins present in seeds of B. oleracea var. italica (i.e. edible broccoli). Our findings suggest an applied value for BraDef1/BraDef2 in controlling phytopathogenic fungi and pathogenic bacteria of clinical significance.

Genetics and Expression Analysis of Anthocyanin Accumulation in Curd Portion of Sicilian Purple to Facilitate Biofortification of Indian Cauliflower.

The present study was undertaken to know the genetics of purple color of cauliflower curds using a Sicilian purple 'PC-1' and a white curding mid-late group genotype of Indian cauliflower. For this, a cross was attempted between 'DC-466' (white curd) and 'PC-1' (purple curd) and observed intermediate level of purple pigmentation on curds in F1 plants. Segregation of F2 population (173) revealed that the purple color of the curd was governed by a single gene dominant over white, but the expression of trait was incomplete. It was substantiated by segregation of plants of BC1 and F2:3(intermediate) generations into 1(white):1(intermediate) and 1(white):2(intermediate):1(intense), respectively. The F2, B1, and B2 generations segregated into purple(intermediate to intense): white curding plants in the ratio of 126: 47, 26:24, and 40:0, respectively fitting well with the Mendelian ratio of single gene for purple curds. However, purple pigmentation on curds ranged from very light to intense, which corroborated with the wide range of anthocyanin content in F2 (3.81-48.21 mg/100 g fw). Out of three molecular markers from high resolution map of Pr gene in purple color cauliflower 'Graffiti', only BoMYB3 marker could distinguish purple and white curding parents but did not show co-segregation while investigated in F2 population. Expression of BoMYB1 gene was up regulated in both the purple curd genotypes 'PC-1' and 'Graffiti' in comparison to white curded 'DC-466', while BoMYB2 gene was slightly upregulated in 'PC-1' but down regulated in 'Graffiti'. Occurrence of 'broccoli type' F2 individuals and their genetic stability in F2:3 support the intermediate position of 'Sicilian purple' between broccoli (Calabrese) and cauliflower. There was not any correlation between curd coloration and pigmentation on apical leaf and stem portion, indicating difference of expression in 'PC-1' than 'Graffiti'. The information obtained is useful for breeding anthocyanin rich attractive purple curding 'specialty cauliflower' for better consumer health and growers' earnings.

Influence of a Modified Atmosphere on the Induction and Activity of Respiratory Enzymes in Broccoli Florets during the Early Stage of Postharvest Storage.

Modified atmosphere packaging and controlled atmosphere storage (hypoxia conditions) extend shelf lives of horticultural products by depressing the O2 uptake rate. We investigated the relationship between atmospheres and alternative oxidase (AOX) to cytochrome c oxidase (COX) activities (on the basis of oxygen isotope discrimination) and the relative amounts of two respiratory enzymes, AOX and COX, during the early stage of storage. Broccoli florets, with high O2 uptake rates, were stored under hypoxia and normoxia at 25 °C. O2 uptake rates, weight loss, and yellowing of broccoli florets were significantly lower when stored under hypoxia than when stored under normoxia. Significantly more AOX proteins were produced during storage under normoxia, but COX proteins were more consistent than those of AOX proteins. Hypoxia may depress the expression of AOX and prolong the shelf life. Oxygen isotope discrimination was elevated under hypoxia after 50.5 h. AOX production in broccoli was controlled more by changing atmospheres than by COX.

Small RNA Sequencing Reveals Differential miRNA Expression in the Early Development of Broccoli (Brassica oleracea var. italica) Pollen.

Pollen development is an important and complex biological process in the sexual reproduction of flowering plants. Although the cytological characteristics of pollen development are well defined, the regulation of its early stages remains largely unknown. In the present study, miRNAs were explored in the early development of broccoli (Brassica oleracea var. italica) pollen. A total of 333 known miRNAs that originated from 235 miRNA families were detected. Fifty-five novel miRNA candidates were identified. Sixty of the 333 known miRNAs and 49 of the 55 predicted novel miRNAs exhibited significantly differential expression profiling in the three distinct developmental stages of broccoli pollen. Among these differentially expressed miRNAs, miRNAs that would be involved in the developmental phase transition from uninucleate microspores to binucleate pollen grains or from binucleate to trinucleate pollen grains were identified. miRNAs that showed significantly enriched expression in a specific early stage of broccoli pollen development were also observed. In addition, 552 targets for 127 known miRNAs and 69 targets for 40 predicted novel miRNAs were bioinformatically identified. Functional annotation and GO (Gene Ontology) analysis indicated that the putative miRNA targets showed significant enrichment in GO terms that were related to plant organ formation and morphogenesis. Some of enriched GO terms were detected for the targets directly involved in plant male reproduction development. These findings provided new insights into the functions of miRNA-mediated regulatory networks in broccoli pollen development.