Distinct cis-elements in the Asparagus officinalis asparagine synthetase promoter respond to carbohydrate and senescence signals

The Asparagus officinalis L. asparagine (Asn) synthetase (AS) promoter was analysed for elements responding to carbohydrate and senescence signals. Transgenic Arabidopsis thaliana L. plants containing deletion constructs of the –1958 bp AS promoter linked to the β-glucuronidase (GUS) reporter gene (AS::GUS) were analysed by measuring GUS specific activity. Inclusion of sucrose (Suc), glucose (Glc) or fructose (Fru) in plant media repressed levels of GUS activity in –1958AS::GUS plants, regardless of the light environment, with increases in GUS found 1 d after incubation on Suc-lacking media. Hexokinase is likely to be involved in the signal pathway, as Suc, Glc, Fru, 2-deoxy-d-glucose and mannose were more effective repressors than 3-O-methylglucose, and the hexokinase inhibitor mannoheptulose reduced repression. Plants containing AS::GUS constructs with deletions that reduced the promoter to less than –405 bp did not show low sugar induction. AS::GUS activity was significantly higher in excised leaves induced to senesce by dark storage for 24 h, compared to fresh leaves, for lines containing at least –640 bp of the AS promoter but not those with –523 bp or smaller promoter fragments. Fusion of the –640 to –523 bp region to a –381AS::GUS construct generated a promoter that retained senescence induction but lacked low sugar induction. Alignment of this region to the 33-bp senescence-related sequence of the Arabidopsis and Brassica napus L. SAG12 promoters identified the sequence TTGCACG as being conserved in all the promoters, and which may be an important senescence-responsive element.

Boron nutrition and chilling tolerance of warm climate crop species

Background Field observations and glasshouse studies have suggested links between boron (B)-deficiency and leaf damage induced by low temperature in crop plants, but causal relationships between these two stresses at physiological, biochemical and molecular levels have yet to be explored. Limited evidence at the whole-plant level suggests that chilling temperature in the root zone restricts B uptake capacity and/or B distribution/utilization efficiency in the shoot, but the nature of this interaction depends on chilling tolerance of species concerned, the mode of low temperature treatment (abrupt versus gradual temperature decline) and growth conditions (e.g. photon flux density and relative humidity) that may exacerbate chilling stress. Scope This review explores roles of B nutrition in chilling tolerance of continual root or transient shoot chills in crop species adapted to warm season conditions. It reviews current research on combined effects of chilling temperature (ranging from > 0 to 20 degrees C) and B deficiency on growth and B nutrition responses in crop species differing in chilling tolerance. Conclusion For subtropical/tropical species (e.g. cucumber, cassava, sunflower), root chilling at 10-17 degrees C decreases B uptake efficiency and B utilization in the shoot and increases the shoot : root ratio, but chilling-tolerant temperate species (e.g. oilseed rape, wheat) require much lower root chill temperatures (2-5 degrees C) to achieve the same responses. Boron deficiency exacerbates chilling injuries in leaf tissues, particularly under high photon flux density. Suggested mechanisms for B x chilling interactions in plants are: (a) chilling-induced reduction in plasmalemma hydraulic conductivity, membrane fluidity, water channel activity and root pressure, which contribute to the decrease in root hydraulic conductance, water uptake and associated B uptake; (b) chilling-induced stomatal dysfunction affecting B transport from root to shoot and B partitioning in the shoot; and (c) B deficiency induced sensitivity to photo-oxidative damage in leaf cells. However, specific evidence for each of the mechanisms is still lacking. Impacts of B status on chilling tolerance in crop species have important implications for the management of B supply during sensitive stages of growth, such as early growth after planting and early reproductive development, both of which can coincide with the occurrence of chilling temperatures in the field.

Thermochemical characterisation of straws and high yielding perennial grasses

The research is concerned with thermochemical characterisation of straws and high yielding perennial grasses. Crops selected for this study include wheat straw (Triticum aestivum), rape straw (Brassica napus), reed canary grass (Phalaris arundinacea) and switch grass (Panicum virgatum). Thermogravimetric analysis (TGA) was used to examine the distribution of char and volatiles during pyrolysis up to 900 °C. Utilising multi-heating rate thermogravimetric data, the Friedman iso-conversional kinetic method was used to determine pyrolysis kinetic parameters. Light and medium volatile decomposition products were investigated using pyrolysis–gas chromatography–mass spectrometry (Py–GC–MS) up to 520 °C. The 22 highest yielding identifiable cellulose, hemicellulose and lignin biomass markers were semi-quantified taking into consideration peak areas from GC chromatograms. Notable differences can be seen in butanedioic acid, dimethyl ester (hemicelluloses decomposition products), 2-methoxy-4-vinylphenol (lignin marker) and levoglucosan (intermediate pyrolytic decomposition product of cellulose) content when comparing perennial grasses with straw. From results presented in this study, perennial grasses such as switch grass, have the most attractive properties for fast pyrolysis processing. This is because of the observed high volatile yield content of 82.23%, heating value of 19.64 MJ/kg and the relatively low inorganic content.

Genetic variation of temperature-regulated curd induction in cauliflower: elucidation of floral transition by genome-wide association mapping and gene expression analysis

Cauliflower (Brassica oleracea var. botrytis) is a vernalization-responsive crop. High ambient temperatures delay harvest time. The elucidation of the genetic regulation of floral transition is highly interesting for a precise harvest scheduling and to ensure stable market supply. This study aims at genetic dissection of temperature-dependent curd induction in cauliflower by genome-wide association studies and gene expression analysis. To assess temperature dependent curd induction, two greenhouse trials under distinct temperature regimes were conducted on a diversity panel consisting of 111 cauliflower commercial parent lines, genotyped with 14,385 SNPs. Broad phenotypic variation and high heritability (0.93) were observed for temperature-related curd induction within the cauliflower population. GWA mapping identified a total of 18 QTL localized on chromosomes O1, O2, O3, O4, O6, O8, and O9 for curding time under two distinct temperature regimes. Among those, several QTL are localized within regions of promising candidate flowering genes. Inferring population structure and genetic relatedness among the diversity set assigned three main genetic clusters. Linkage disequilibrium (LD) patterns estimated global LD extent of r(2) = 0.06 and a maximum physical distance of 400 kb for genetic linkage. Transcriptional profiling of flowering genes FLOWERING LOCUS C (BoFLC) and VERNALIZATION 2 (BoVRN2) was performed, showing increased expression levels of BoVRN2 in genotypes with faster curding. However, functional relevance of BoVRN2 and BoFLC2 could not consistently be supported, which probably suggests to act facultative and/or might evidence for BoVRN2/BoFLC-independent mechanisms in temperature regulated floral transition in cauliflower. Genetic insights in temperature-regulated curd induction can underpin genetically informed phenology models and benefit molecular breeding strategies toward the development of thermo-tolerant cultivars.

Posibles funciones de la familia génica de la glutamato descarboxilasa en Populus

Aunque hace más de 50 años que se describió que la glutamato descarboxilasa (GAD) lleva a cabo la descarboxilación del glutamato para producir GABA, y en animales ha sido muy estudiada debido al papel del GABA como neurotransmisor, la información disponible sobre las GADs de plantas es aún limitada, conociéndose sólo algunos aspectos de la regulación por calcio de su actividad enzimática o de expresión de algunos de los genes de su familia génica. El GABA es un metabolito que tradicionalmente se ha asociado a estrés, pero su papel en plantas todavía no está claro. En las últimas dos décadas los resultados experimentales obtenidos sobre la GAD y el GABA, destacando las alteraciones fenotípicas mostradas por plantas tratadas con GABA y por plantas transgénicas para GAD, han generado preguntas interesantes sobre el posible papel de este metabolito y la enzima en señalización en plantas. En plantas, son varios los papeles que se han propuesto para el metabolismo del GABA tales como su participación como componente del metabolismo del carbono y del nitrógeno (Fait y col., 2008), protección frente especies reactivas de oxigeno (Liu y col., 2011), regulación de la expresión génica incluyendo la regulación de genes implicados en la síntesis de hormonas (Khatiresan y col., 1997; Shi y col., 2010; Lancien y Roberts, 2006) y señalización a larga distancia (Beuve y col., 2004) y en gradiente guiando el crecimiento del tubo polínico (Palanivelu y col., 2013). Nuestro grupo de investigación ha sugerido un papel novedoso para la producción de GABA durante la xilogénesis en pino (Molina-Rueda y col., 2010, 2015). En base a estos antecedentes, los objetivos planteados para este trabajo han sido: la asignación de posibles funciones a las GADs de Populus en condiciones normales de crecimiento y en estrés abióticos, estudiar la adquisición del dominio de unión a calmodulina (CaMBD) de las GADs de plantas vasculares y analizar el efecto del GABA y del glutamato en las raíces de Populus. Las conclusiones que se derivan de los resultados de este trabajo se detallan a continuación. El dominio de unión a calmodulina de la GAD de plantas esta conservado en GADs de plantas consideradas ancestros de plantas vasculares y ausente en plantas no vasculares, lo que sitúa juntos en la evolución los eventos de adquisición del dominio de unión a CaM y el desarrollo del tejido vascular de plantas. Los resultados similares de la localización de GABA en xilema y una expresión GAD asociada a la formación de madera de reacción tanto en pino como en chopo apuntan a un papel relevante de la producción de GABA durante la xilogénesis en leñosas. La familia génica GAD posee seis genes codificando todos ellos para proteínas aparentemente funcionales y susceptibles de ser reguladas por calcio. Esta familia génica ha sufrido duplicaciones y eventos de especialización durante la evolución de Populus. Este trabajo ha posibilitado la asociación entre papeles específicos y los diferentes genes de esta familia. Beuvé N, Rispail N, Laine P, Cliquet J-B, Ourry A, Deunff F (2004) Putative role of Υ-aminobutyric acid as a long-distance signal in up-regulation of nitrate uptake in Brassica napus L. Plant Cell Environ. 27: 1035-1046 Fait A, Fromm H, Walter D, Galili G, Fernie AR (2008) Highway or byway: the metabolic role of the GABA shunt in plants. Trends in plant science 13: 14-19 Kathiresan A, Tung P, Chinnappa CC, Reid DM (1997) gamma-Aminobutyric acid stimulates ethylene biosynthesis in sunflower. Plant Physiol. 115: 129-135 Lancien M, Roberts MR (2006) Regulation of Arabidopsis thaliana 14-3-3 gene expression by ϒ-aminobutyric acid. Plant Cell Environ. 29: 1430-1436 Liu C, Zhao L, Yu G (2011) The dominant glutamic acid metabolic flux to produce gamma-amino butyric acid over proline in Nicotiana tabacum leaves under water stress relates to its significant role in antioxidant activity. Journal of integrative plant biology 53: 608-618 Molina-Rueda JJ, Pascual MB, Canovas FM, Gallardo F (2010) Characterization and developmental expression of a glutamate decarboxylase from maritime pine. Planta 232: 1471-1483 Molina-Rueda, J.J. y col., 2015. A putative role for γ-aminobutyric acid (GABA) in vascular development in pine seedlings. Planta 241: 257-267 Palanivelu R, Brass L, Edlund AF, D P (2003) Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels. Cell 114: 47-59 Shi SQ, Shi Z, Jiang ZP, Qi LW, Sun XM, Li CX, Liu JF, Xiao WF, Zhang SG (2010) Effects of exogenous GABA on gene expression of Caragana intermedia roots under NaCl stress: regulatory roles for H2O2 and ethylene production. Plant, cell & environment 33: 149-162

Effects of population density on cauliflower crop (Brassica oleracea L. var. botrytis)

An experiment was carried out to study the effects of the following population densities cauliflowers (plants per ha): 20,833 (0.60 m x 0.80 m), 25,641 (0.60 m x 0.65 m), ....37.037 (0.60 m x 0.45 m) , 55.555 (.0.60 m x 0.30 m), and 111,111 (0,60 m x 0,15 m) ; variety Snow ball. It was concluded that the effects of plant population density are greater on curd quality (weight and size) than on production per ha. The best plant population density to produce cauliflowers curd for Brazil market is from 20,000 to 25,000 plants/ha while for mini-curd is above 55,000 plants/ha.

Efeito inibitório na germinação, induzido pelo extrato de couve (Brassica oleracea L. var. acephala DC.)

Com a finalidade de se estudar as possíveis propriedades alelopáticas da couve (Brassica oleracea L. var. acephala DC), foi conduzido um bioensaio, em condições de laboratório, na E.S.A."Luiz de Queiroz", em Piracicaba, SP. Foram utilizadas sementes de tomate (Lycopersicon esculentum Mill. cv.Santa Cruz) colocadas para germinar em soluções com diferentes concentrações (0; 25; 50 e 100%) do extrato obtido da parte aérea (folhas) da couve. Constataram-se efeitos pronunciadamente inibitórios de germinação das sementes de tomate nas maiores concentrações do extrato de couve. As plântulas que germinaram, nas parcelas tratadas com o extrato, mostraram-se com o crescimento reduzido, morfo-fisiologicamente anormais, e com maior tempo para o início da germinação. Todas essas características foram mais pronunciadas nas parcelas com maiores concentrações do extrato.

Simultaneous Evaluation of Intact Glucosinolates and Phenolic Compounds by UPLC-DAD-MS/MS in Brassica oleracea L. var. botrytis

A method for the simultaneous determination of intact glucosinolates and main phenolic compounds (flavonoids and sinapic acid derivatives) in Brassica oleracea L. var. botrytis was proposed. A simplified sample extraction procedure and a UPLC separation were carried out to reduce the total time of analysis. Brassica oleracea samples were added with internal standards (glucotropaeolin and rutin), and extracted with boiling methanol. Crude extracts were evaporated under nitrogen, redissolved in mobile phase and analyzed by UPLC with double detection (ESI--MRM for glucosinolates and flavonoids, and DAD for main sinapic acid derivatives). The proposed method allowed a satisfactory quantification of main native sinapic acid derivatives, flavonoids and glucosinolates with a reduced time of analysis.

Assessing the impact of high-pressure processing on selected physical and biochemical attributes of white cabbage (Brassica oleracea L. var. capitata alba)

The aim of this study was to investigate the effect of combined pressure/temperature treatments (200, 400 and 600 MPa, at 20 and 40 °C) on key physical and chemical characteristics of white cabbage (Brassica oleracea L. var. capitata alba). Thermal treatment (blanching) was also investigated and compared with high-pressure processing (HPP). HPP at 400 MPa and 20–40 °C caused significantly larger colour changes compared to any other pressure or thermal treatment. All pressure treatments induced a softening effect, whereas blanching did not significantly alter texture. Both blanching and pressure treatments resulted in a reduction in the levels of ascorbic acid, effect that was less pronounced for blanching and HPP at 600 MPa and 20–40 °C. HPP at 600 MPa resulted in significantly higher total phenol content, total antioxidant capacity and total isothiocyanate content compared to blanching. In summary, the colour and texture of white cabbage were better preserved by blanching. However, HPP at 600 MPa resulted in significantly higher levels of phytochemical compounds. The results of this study suggest that HPP may represent an attractive technology to process vegetable-based food products that better maintains important aspects related to the content of health-promoting compounds. This may be of particular relevance to the food industry sector involved in the development of convenient novel food products with excellent functional properties

Forced-air, vacuum, and hydro precooling of cauliflower (Brassica oleracea L. var. botrytis cv. Freemont): part I. determination of precooling parameters

The aim of the present study was to precool cauliflower using forced-air, vacuum and high and low flow hydro cooling methods. The weight of the precooled cauliflower heads (5000±5 g) was measured before they were placed in standard plastic crates. Cauliflower heads, whose initial temperature was 23.5 ± 0.5 ºC, were cooled until the temperature reached at 1 ºC. During the precooling process, time-dependent temperature and energy consumption were measured, and during vacuum precooling, the decreasing pressure values were recorded, and a curve of time-dependent pressure decrease (vacuum) was built. The most suitable cooling method to precool cauliflower in terms of cooling time and energy consumption was vacuum, followed by the high and low flow hydro and forced-air precooling methods, respectively. The highest weight loss was observed in the vacuum precooling method, followed by the forced-air method. However, there was an increase in the weight of the cauliflower heads in the high and low flow hydro precooling method. The best colour and hardness values were found in the vacuum precooling method. Among all methods tested, the most suitable method to precool cauliflower in terms of cooling and quality parameters was the vacuum precooling method.

Forced-air, vacuum, and hydro precooling of cauliflower (Brassica oleracea L. var. botrytis cv. Freemont): Part II. Determination of quality parameters during storage

Cauliflower heads, which were precooled using four different methods including vacuum, forced-air, and high and low flow hydro precooling, were stored under controlled atmosphere and room conditions. Controlled atmosphere conditions (CA) were as follows: 1°C temperature, 90 ± 5% relative humidity, and 0:21 [(%CO2:%O2) – (0:21) control] atmosphere composition. Room conditions (RC) were: 22±1°C temperature and 55-60% humidity. Various quality parameters of the cauliflower heads were assessed during storage (days 0, 7, 14, 21, 28, and 35) under controlled atmosphere and room conditions (days 0, 5, and 10). During storage, weight loss, deterioration rate, overall sensory quality score, hardness, and colour (L, a, b, C and α) were evaluated. In the present study, the strength and quality parameters of cauliflower under CA and RC conditions were obtained. Vacuum precooling was found to be most suitable method before cauliflower was submitted to cold storage and sent to market. Furthermore, the storage of cauliflower without precooling resulted in a significant decrease in quality parameters.

Comparação entre métodos para a avaliação do vigor de lotes de sementes de couve-brócolos (Brassica oleracea L. var. italica Plenk)

Os testes de vigor e o teste de germinação são componentes essenciais no controle de qualidade das empresas de produção de sementes. Com o objetivo de verificar a eficiência de diferentes testes de vigor e de variações de suas metodologias na avaliação da qualidade de sementes de couve-brócolos visando diferenciação de lotes e previsão de emergência em bandeja, cinco lotes de sementes do híbrido Flórida foram submetidos aos seguintes testes: germinação; primeira contagem de germinação; emissão de raiz primária (após 48, 56, 72, 80 e 96 h após a instalação do teste de germinação); emergência de plântulas em substrato; envelhecimento acelerado com água (1g de sementes mantidas a 41ºC por 48 e 72 h a 100%UR); envelhecimento acelerado com solução saturada de sal (mesmo procedimento do item anterior, porém usando solução de NaCl, 40% e 76%UR); condutividade elétrica (50 sementes em 25 mL de água destilada a 25ºC e leituras após 2, 4, 6, 8 e 24 h). Todos os testes apresentaram correlação significativa com a porcentagem de emergência de plântulas em substrato, a 1% de probabilidade. Os testes de envelhecimento acelerado com solução saturada de sal por 48 h e de condutividade elétrica após 8 e 24 h de embebição foram eficientes e tiveram resultados semelhantes aos da emergência em substrato. Os testes da primeira contagem de germinação, emissão da raiz primária após 56 h e envelhecimento acelerado com solução saturada de sal por 72 h, apresentaram-se mais eficientes que a emergência de plântulas em substrato na diferenciação do vigor dos lotes.

Adequação da metodologia do teste de deterioração controlada para sementes de brócolis (Brassica oleracea L. - var. itálica)

A eficiência dos testes de vigor na avaliação da qualidade de sementes, depende da sua padronização para as diferentes espécies. Neste contexto, os objetivos deste trabalho foram elaborar as curvas de absorção de água para determinar o período necessário ao ajuste do teor de água das sementes e aprimorar o teste de deterioração controlada para sementes de brócolis. Foram utilizadas sementes de três variedades: dois lotes de Piracicaba precoce, dois lotes de Flórida e quatro lotes de Ramoso. Os lotes foram caracterizados mediante a determinação do teor de água e germinação; o vigor foi determinado pelos testes de primeira contagem, comprimento de plântula, emergência e índice de velocidade de emergência. Foram determinadas as curvas de absorção de água por 84 horas. Para averiguar a eficiência do teste de deterioração controlada para sementes de brócolis, foi feito o ajuste do conteúdo de água das sementes para: 18, 20, 21, 23 e 24%, sendo estas expostas a 45ºC em banho-maria, por 24 horas. O delineamento utilizado foi o inteiramente casualizado. As médias foram comparadas pelo teste de Tukey a 5% de probabilidade. As curvas de absorção permitiram estimar os períodos de 15, 20, 22, 32 e 48 horas para elevação do teor de água a 18, 20, 21, 23 e 24% respectivamente. O teste de deterioração controlada, para sementes de brócolis, propiciou resultados promissores, tendo sido eficiente na separação de lotes, em todos os teores de água estudados, tanto quanto os testes de emergência de plântula e índice de velocidade de emergência.

Thermal and pressure stability of myrosinase enzymes from black mustard (Brassica nigra L. W.D.J Koch. var. nigra), brown mustard (Brassica juncea L. Czern. var. juncea) and yellow mustard (Sinapsis alba L. Subsp Maire) seeds

This study investigates the effects of temperature and pressure on inactivation of myrosinase extracted from black, brown and yellow mustard seeds. Brown mustard had higher myrosinase activity (2.75 un/mL) than black (1.50 un/mL) and yellow mustard (0.63 un/mL). The extent of enzyme inactivation increased with pressure (600-800 MPa) and temperature (30-70 °C) for all the mustard seeds. However, at combinations of lower pressures (200-400 MPa) and high temperatures (60-80 °C), there was less inactivation. For example, application of 300 MPa and 70 °C for 10 minutes retained 20%, 80% and 65% activity in yellow, black and brown mustard, respectively, whereas the corresponding activity retentions when applying only heat (70 °C, 10min) were 0%, 59% and 35%. Thus, application of moderate pressures (200-400 MPa) can potentially be used to retain myrosinase activity needed for subsequent glucosinolate hydrolysis.

Influência de genótipos de couve (Brassica oleracea L. var. acephala DC.) na biologia de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae)

Objetivou-se, com este trabalho, avaliar a influência de alguns genótipos de couve (Brassica oleracea L. var. acephala DC.) no desenvolvimento de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae). Os genótipos avaliados foram: Manteiga de Ribeirão Pires I-2620, Roxa I-919, Manteiga de São José, Manteiga de Monte Alegre, Pires 2 de Campinas, Couve Comum, Couve de Arthur Nogueira 2, Couve de Arthur Nogueira 1. Lagartas recém-eclodidas foram mantida em discos foliares de 8 cm de diâmetro para cada genótipo. Foram analisados os seguintes parâmetros: duração e viabilidade das fases larval e pupal, longevidade e fecundidade de adultos, utilizando análises paramétricas e de agrupamentos para interpretação dos dados. Observou-se um prolongamento em dias no ciclo de P. xylostella, aumento no peso de pupa e maiores valores de viabilidade e fecundidade, durante a segunda geração. O genótipo Couve de Arthur Nogueira 2 foi menos favorável ao desenvolvimento de P. xylostella nas duas gerações, e Couve Comum demonstrou maior influência negativa ao inseto na segunda geração. Manteiga de Ribeirão Pires I-2620 foi o mais suscetível nas duas gerações, agrupando com este na segunda geração Pires 2 de Campinas e Manteiga de São José.