Genome-wide analysis of G-type lectin genes in Fragaria vesca and functional characterization of FaMBL1 gene in defense response of F. × ananassa to fungal pathogens

Strawberry (Fragaria × ananassa) is an important soft fruit but easily to be infected by pathogens. Anthracnose and gray mold are two of the most destructive diseases of strawberry which lead to serious fruit rot. The first chapter introduced strawberry anthracnose caused by Colletotrichum acutatum. The infection strategy, disease cycle and management of C. acutatum on strawberry were reported. Likewise, the second chapter summarized the infection strategy of Botrytis cinerea and the defense responses of strawberry. As we already know white unripe strawberry fruits are more resistant to C. acutatum than red ripe fruits. During the interaction between strawberry white/red fruit and C. acutaum, a mannose binding lectin gene, FaMBL1, was found to be the most up-regulated gene and induced exclusively in white fruit. FaMBL1 belongs to the G-type lectin family which has important roles in plant development and defense process. To get insight into the role of FaMBL1, genome-wide identification was carried out on G-type lectin gene family in Fragaria vesca and the results were showed in chapter 3. G-type lectin genes make up a large family in F. vesca. Active expression upon biotic/abiotic stresses suggested a potential role of G-lectin genes in strawberry defenses. Hence, stable transgenic strawberry plants with FaMBL1 gene overexpressed were generated. Transformed strawberry plants were screened and identified. The results were showed in chapter 4, content of disease-related phytohormone, jasmonic acid, was found decreased in overexpressing lines compared with wild type (WT). Petioles inoculated by C. fioriniae of overexpressing lines had lower disease incidence than WT. Leaves of overexpressing lines challenged by B. cinerea showed remarkably smaller lesion diameters compared with WT. The chitinase 2-1 (FaChi2-1) showed higher expression in overexpressing lines than in WT during the interaction with B. cinerea, which could be related with the lower susceptibility of overexpressing lines.

I vini passiti: alcune tecniche di vinificazione a confronto

La pratica dell’appassimento fa parte della cultura mediterranea sin dall’era del Neolitico. Tale tecnica veniva praticata anche nell’Antica Grecia e successivamente nelle diverse regioni dell’Impero Romano. Il presente elaborato si propone, attraverso lo studio dello stato dell’arte relativamente ai vini passiti, di illustrare alcune tecniche di vinificazione, approfondire le caratteristiche salienti di questa tipologia di vino e delinearne il posizionamento sul mercato. In particolare, si sono prese in considerazione le tre tecniche di appassimento comunemente utilizzate: appassimento in campo, in fruttaio e in cella. Per quanto concerne l’appassimento in campo, si è fatto un approfondimento in merito agli effetti indotti dalla Botrytis cinerea, muffa nobile la cui presenza sulle uve permette di dar vita a vini botritizzati dalle caratteristiche distintive, come nel caso dell’Albana Passito di Romagna. I metodi di appassimento che prevedono l’utilizzo di un locale apposito (fruttaio e cella) si differenziano per il controllo della temperatura e dell’umidità e quindi il processo di disidratazione dell’acino avviene in tempi più rapidi e con minor rischio di sviluppo di patogeni. Lo sviluppo di queste tecniche come ha impattato sul consumo di vino passito? Negli ultimi anni il consumo di vino sta aumentando, e l’Italia è uno dei principali paesi produttori e consumatori. Il vino passito però, data la sua complessità e le particolari occasioni d’uso, occupa un piccolo spazio in questo scenario ed è caratterizzato da una concentrazione nel consumo delle produzioni locali, faticando a occupare un posto di rilievo sul mercato. Nonostante ciò, vi sono anche casi di successo, come quello dell’Amarone della Valpolicella DOCG, che lasciano ben sperare in uno sviluppo futuro.

Botrytis species: relentless necrotrophic thugs or endophytes gone rogue?

Plant pathology has a long-standing tradition of classifying microbes as pathogens, endophytes or saprophytes. Lifestyles of pathogens are categorized as biotrophic, necrotrophic or hemibiotrophic. Botrytis species are considered by many to be archetypal examples of necrotrophic fungi, with B. cinerea being the most extensively studied species because of its broad host range and economic impact. In this review, we discuss recent work which illustrates that B. cinerea is capable of colonizing plants internally, presumably as an endophyte, without causing any disease or stress symptoms. The extent of the facultative endophytic behaviour of B. cinerea and its relevance in the ecology and disease epidemiology may be vastly underestimated. Moreover, we discuss the recent discovery of a novel Botrytis species, B. deweyae, which normally grows as an endophyte in ornamental daylilies (Hemerocallis), but displays facultative pathogenic behaviour, and is increasingly causing economic damage. We propose that the emergence of endophytes ‘gone rogue’ as novel diseases may be related to increased inbreeding of hybrid lines and reduced genetic diversity. These observations lead us to argue that the sometimes inflexible classification of pathogenic microbes by their lifestyles requires serious reconsideration. There is much more variety to the interactions of Botrytis with its hosts than the eye (or the plant pathologist) can see, and this may be true for other microbes interacting with plants.

Analysis of cryptic, systemic Botrytis infections in symptomless hosts

Botrytis species are generally considered to be aggressive, necrotrophic plant pathogens. By contrast to this general perception, however, Botrytis species could frequently be isolated from the interior of multiple tissues in apparently healthy hosts of many species. Infection frequencies reached 50% of samples or more, but were commonly less, and cryptic infections were rare or absent in some plant species. Prevalence varied substantially from year to year and from tissue to tissue, but some host species routinely had high prevalence. The same genotype was found to occur throughout a host, representing mycelial spread. B. cinerea and B. pseudocinerea are the species that most commonly occur as cryptic infections, but phylogenetically distant isolates of Botrytis were also detected, one of which does not correspond to previously described species. Sporulation and visible damage occurred only when infected tissues were stressed, or became mature or senescent. There was no evidence of cryptic infection having a deleterious effect on growth of the host, and prevalence was probably greater in plants grown in high light conditions. Isolates from cryptic infections were often capable of causing disease (to varying extents) when spore suspensions were inoculated onto their own host as well as on distinct host species, arguing against co-adaptation between cryptic isolates and their hosts. These data collectively suggest that several Botrytis species, including the most notorious pathogenic species, exist frequently in cryptic form to an extent that has thus far largely been neglected, and do not need to cause disease on healthy hosts in order to complete their life-cycles.

Drying and storage of cauliflower (Brassica oleracea var. botrytis) hydroprimed seeds

Literature has documented beneficial effects of seed priming on speed, synchronization and uniformity of germination. often leading to improved stand establishment. However. doubts still persist about the possible reversal effects, after drying and during storage of primed seeds that could overcome, partial or totally, the improved performance. The objectives of this research were to identify drying and storage procedures that would maintain the physiological performance achieved after seed priming, without negative effects on storability. First. hydroprimed cauliflower Seeds cv. Sharon and cv. Teresopolis Gigante, each represented by three seed lots were submitted to fast drying, slow drying, and treatments of pre-drying incubation (exposure to 35 degrees C, to a polyethylene glycol 6000 solution or a heat shock) followed by fast drying. In the second phase of this study, hydroprimed seed samples were submitted to fast drying (30-35 degrees C and 40-50% R.H.) and stored under laboratory conditions or in a chamber at 20 degrees C and 50% relative humidity for six months. Seed physiological potential was evaluated after 60-day intervals for germination (speed and percentage), Seedling emergence and saturated salt accelerated aging tests. All drying treatments efficiently preserved the favourable priming effects, except for the incubation at 35 degrees C for 96-144 hours. The beneficial priming effects followed by fast drying persisted for four months under controlled conditions (20 degrees C and 50% relative humidity).

Efeito da densidade de população nos teores de ácido ascórbico e carboidratos solúveis de couve-flor (Brassica oleracea var. botrytis)

Os teores de carboidratos solúveis, glucose, frutose, sacarose e ácido ascórbico foram determinados em amostras de couve-flor (Brassica oleracea var. botrytis), obtidas de tratamentos apresentando diversas densidades de plantais. O efeito do espaçamento não foi significativo para os dviersos constituintes analisados, embora o tratamento com 20.833 plantas/ha apresentasse maior teor de ácido ascórbico. O peso seco das amostras variou de 8,24 a 10,13% e ácido ascórbico de 93,1 a 113,5 mg/100 g de peso fresco. Os teores de carboidratos solúveis, expressos em g/100 g de peso fresco foram: totais (3,17-4,54) sacarose (0,26-0,66), glucose (0,92-1,50) e frutose (0,91-145).

Aplicação de alguns herbicidas na cultura da couve-flor (Brassica oleracea var. botrytis)

Foi conduzido na Área Experimental do Setor de Horticultura, DAH-ESALQ, Piracicaba, um experimento sobre o uso de alguns herbicidas na cultura da couve-flor (Brassica oleracea var. botrytis). Os herbicidas foram Dacthal (DCPA), Tenoran (cloroxuron), Afalon (linuron) e Ramrod (propacloro). O Dacthal, Ramrod, e o Tenoran não foram fitotóxicos à couve-flor, enquanto que o Afalon foi muito fitotóxico. O herbicida que melhor resultado apresentou para a couve-flor foi o Ramrod.

Teor de ácido ascórbico de couve-flor (Brassica oleracea var. botrytis) conservada à temperatura ambiente, em geladeira e em congelador

Amostras de couve-flor (Brassica oleracea var. botrytis) foram armazenadas à temperatura ambiente, a 5°C (geladeira) e a - 20°C (congelador) e foram feitas as análises para ácido ascórbico. O teor de ácido ascórbico não variou durante o período de resfriamento e as amostras de couve-flor congelada mostrou uma redução de 50% durante 12 dias de armazenamento. Amostras colocadas à temperatura ambiente apresentaram um pequeno aumento até os 3 dias. Não foram observadas diferenças significativas entre os tipos de embalagens usadas (garrafa de vidro, sacos plásticos e folhas de alumínio) quanto ao teor de ácido ascórbico.

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.

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.

Highly variable social organisation of colonies in the ant Formica cinerea.

Social organisation of colonies was examined in the ant Formica cinerea by estimating the coefficient of genetic relatedness among worker nest mates. The estimates based on microsatellite genotypes at three loci ranged from values close to zero to 0.61 across the populations studied in Finland. These results showed that a fundamental feature of colonies, the number of reproductive queens, varied greatly among the populations. Colonies in some populations had a single queen, whereas the nests could have a high number number of queens in other populations. There was a weak but non-significant correlation between the genetic and metric distance of nests within two populations with intermediate level of relatedness. Differentiation among nearby populations (within the dispersal distance of individuals) in one locality indicated limited dispersal or founder effects. This could occur when females are philopatric and stay in the natal polygynous colony which expands by building a network of nest galleries within a single habitat patch.

Isolamento e avaliação da atividade nematicida de constituintes químicos de Mucuna cinerea contra Meloidogyne incognita e Heterodera glycines

Phytochemical investigation of the aerial parts and roots of Mucuna cinerea led to the isolation of a mixture of fatty acids, triacylglicerols, beta-sitosterol, stigmasterol, stigmasterol glucoside, daucosterol, asperglaucide (4) and the isoflavonoids prunetin (1), genistein (2), medicarpin (3), daidzein (5), 7-O-alpha-glycopiranosyl daidzein (6). An in vitro bioassay was carried out with compounds 1-4, at the concentration of 50 and 5 mug mL-1 against the phytonematodes M. incognita and H. glycines. Although the four compounds showed some nematocidal property, the most active was (1), causing 70% mortality of M. incognita at the concentration of 50 mug mL-1.

Development of a weather-based model for Botrytis leaf blight of onion

ABSTRACT In the present study, onion plants were tested under controlled conditions for the development of a climate model based on the influence of temperature (10, 15, 20 and 25°C) and leaf wetness duration (6, 12, 24 and 48 hours) on the severity of Botrytis leaf blight of onion caused by Botrytis squamosa. The relative lesion density was influenced by temperature and leaf wetness duration (P <0.05). The disease was most severe at 20°C. Data were subjected to nonlinear regression analysis. Beta generalized function was used to adjust severity and temperature data, while a logistic function was chosen to represent the effect of leaf wetness on the severity of Botrytis leaf blight. The response surface obtained by the product of two functions was expressed as ES = 0.008192 * (((x-5)1.01089) * ((30-x)1.19052)) * (0.33859/(1+3.77989 * exp (-0.10923*y))), where ES represents the estimated severity value (0.1); x, the temperature (°C); and y, the leaf wetness (in hours). This climate model should be validated under field conditions to verify its use as a computational system for the forecasting of Botrytis leaf blight in onion.

Superação da profundidade de semeadura e densidades de palha para Mucuna aterrima, Mucuna deeringiana e Mucuna cinerea

O experimento foi desenvolvido em casa de vegetação, com o objetivo de verificar a capacidade das sementes de Mucuna aterrima, Mucuna cinerea e Mucuna deeringiana para superar diferentes profundidades do solo acrescido de densidades de palha de cana-de-açúcar. O delineamento utilizado foi o inteiramente casualizado; os tratamentos foram distribuídos em esquema fatorial 3 x 3 x 3, com quatro repetições. Os fatores foram constituídos por camada de palha (0, 5 e 10 cm), espécies de Mucuna (M. aterrima, M. cinerea e M. deeringiana) e profundidade de semeadura (1, 4 e 8 cm). As espécies de Mucuna superaram até 23 t ha-1 de palha (10 cm) e até 8 cm de profundidade de semeadura, expondo as partes aéreas à superfície a partir dos 10 dias após emergência (DAS); elas foram capazes também de desenvolver a altura (>96 cm por planta), a área foliar (>723 cm-2 por planta) e o acúmulo de massa seca (>2,09 g por planta) até os 35 DAS.

Sabicea cinerea Aubl. (Rubiaceae): distilia e polinização em um fragmento de floresta Atlântica em Pernambuco, Nordeste do Brasil

O estudo foi desenvolvido no Parque Estadual de Dois Irmãos, um fragmento de Floresta Atlântica, em Recife - PE, no período de maio/1998 a dezembro/2000. Sabicea cinerea é uma espécie lianescente encontrada na borda da mata, que produz flores por todo ano, com pico de floração na estação seca (setembro a fevereiro). Apresenta flores distílicas, actinomorfas, tubulosas, com pétalas brancas e antese iniciando por volta das 5:00 h, quando o néctar já está disponível; a produção de néctar estende-se até às 16:00 h. O volume médio diário de néctar foi de 8,0 µL e a concentração média de açúcares, de 24%, em ambos os morfos florais. As flores brevistilas possuem a corola e os grãos de pólen maiores em relação às flores longistilas, não havendo, entretanto, diferenças na quantidade dos grãos produzidos entre os dois morfos. Como visitantes florais, foram observados Phaethornis ruber, um dos principais polinizadores, e Amazilia sp., um pilhador. Além desses beija-flores, várias espécies de abelhas Apidae, Anthophoridae e Halictidae, borboletas Hesperiidae e Nymphalidae e moscas Syrphidae, visitavam as flores. A auto-incompatibilidade foi constatada através dos testes de polinização controlada, que resultaram na formação de frutos apenas nos cruzamentos intermorfo. A análise do crescimento dos tubos polínicos confirmou a ocorrência da auto-incompatibilidade esporofítica. A eficiência no transporte de pólen entre os morfos florais é a principal e mais importante função desempenhada pelos polinizadores para que ocorra a manutenção dos agrupamentos de indivíduos de S. cinerea, principalmente por esta espécie apresentar um sistema reprodutivo do tipo xenógamo obrigatório.