Marco Fornaciari

Ozone and nitric oxide induce cGMP‐dependent and ‐independent transcription of defence genes in tobacco

Here, we analyse the temporal signatures of ozone (O3)-induced hydrogen peroxide(H2O2) and nitric oxide (NO) and the role of the second messenger guanosine3′,5′-cyclic monophosphate (cGMP) in transcriptional changes of genes diagnostic for biotic and abiotic stress responses. Within 90 min O3 induced H2O2 and NO peaks and we demonstrate that NO donors cause rapid H2O2 accumulation in tobacco (Nicotiana tabacum) leaf. Ozone also causes highly significant, late (> 2 h) and sustained cGMP increases, suggesting that the second messenger may not be required in all early (< 2 h) responses to O3,but is essential and sufficient for the induction of some O3-dependent pathways.This hypothesis was tested resolving the time course of O3-induced transcript accumulation of alternative oxidase (AOX1a), glutathione peroxidase (GPX),aminocyclopropancarboxylic acid synthase (ACS2) that is critical for the synthesis of ethylene, phenylalanine ammonia lyase (PALa) and the pathogenesis-related protein PR1a.The data show that early O3 and NO caused transcriptional activation of the scavenger encoding proteins AOX1a, GPX and the induction of ethylene production through ACS2 are cGMP independent. By contrast, the early response of PALa and the late response of PR1a show critical dependence on cGMP.

NO release by nitric oxide donors in vitro and in planta

Artificial nitric oxide (NO) donors are widely used as tools to study the role of NO in plants. However, reliable and reproducible characterisation of metabolic responses induced by different NO donors is complicated by the variability of their NO release characteristics. The latter are affected by different physical and biological factors including temperature and light. Here we critically evaluate NO release characteristics of the donors sodium nitroprusside (SNP), S-nitrosoglutathione (GSNO) and nitric oxide synthase (NOS), both in vitro and in planta (Nicotiana tabacum L. cv. BelW3) and assess their effects on NO dependent processes such as the transcriptional regulation of the mitochondrial alternative oxidase gene (AOX1a), accumulation of H(2)O(2) and induction of cell death. We demonstrate that, contrary to NOS and SNP, GSNO is not an efficient NO generator in leaf tissue. Furthermore, spectrophotometric measurement of NO with a haemoglobin assay, rather than diaminofluorescein (DAF-FM) based detection, is best suited for the quantification of tissue NO. In spite of the different NO release signatures by SNP and NOS in tissue, the NO dependent responses examined were similar, suggesting that there is a critical threshold for the NO response.

Aeropalynological and phenological study in two different Mediterranean olive areas: Cordoba (Spain) and Perugia (Italy)

ABSTRACT An aerobiological and phenological investigation on the olive tree was carried out during three years in two areas: Cordoba (Spain) and Perugia (Italy). In these countries, this species is economically important and those areas were chosen because of the long series of aeropalynological data (1982–1998) available, obtained by means of identical volumetric pollen traps. The aim of this study was to use phenological observations to prove the real contribution to the pollen curves in different cultivated areas. Results show that in Cordoba province (302.152 ha) the pollen curve is characterised by different peaks because of the pollination of different cultivated crops. In some cases, these crops are located far from the pollen trap (50 km) but pollen is transported thanks to favourable winds during the flowering period. In Perugia (750 ha) the pollen curve is characterised by only one peak; it is very concentrated because of the proximity of the investigated crops. The objective of this research was to obtain information on this species in order to elaborate statistical models aimed at forecasting the potential fruit production based on the amount of pollen released into the atmosphere.

A regression model for the start of the pollen season in Olea eropaea

The relationship between the timing of olive pollination and some parameters which can influence this phenomenon was investigated. A model is presented which shows a strong relationship between timing of pollination, selected meteorological variables (temperature and precipitation) and annual pollen quantity. The most important correlations are with mean temperature in February, and precipitation in May. The positive correlation with annual pollen quantity confirms the relationship of this variable to the time of pollination.

Reproductive biology of Olive (Olea europaea L.) DOP Umbria cultivars

Olive trees have a plentiful bloom but a low percentage of normal fruit set. To improve fruit set, numerous investigations have sought to identify the obstacles that prevent full production. In this work, flower development in five DOP Umbria cultivars (Leccino, Frantoio, Moraiolo, Dolce Agogia and San Felice) was studied throughout different developmental phases, from before microsporogenesis and megasporogenesis to post-anthesis, by morphological and cyto-histological observations. Dolce Agogia was the most precocious cultivar, while full flowering was simultaneous in Leccino, Frantoio, Moraiolo and San Felice. Frantoio and Leccino were also good pollen producers, having the highest percentage of pollen viability and germinability. Dolce Agogia can also be considered a good pollen producer in terms of the high quantity of released pollen, but it had the lowest levels of pollen viability and germinability and the highest percentage of aborted flowers and ovaries. Morphological and cyto-histological observations on the number of flowers per inflorescence and the number of aborted flowers and ovaries suggest that fruit set was not influenced by the number of flowers per inflorescence, but rather by the number of inflorescences, which depends on the global fruiting potential of the tree.

Olive flowering trends in a large Mediterranean area (Italy and Spain).

The aim of this study was to investigate the main climatic and biological trends related to olive flowering in central-southern Italy compared to those in Andalusia, Spain. Results since 1982 were compared for the two long-series monitoring areas of Cordoba and Perugia, and since 1992–1999 for the short-series areas. The relationship between climatic trends and the biological response of the olive, a widespread culture in the Mediterranean basin, were investigated. An aerobiological method involving capturing pollen released into the atmosphere was utilised as a bioindicator of flowering phenology. The study results confirm the strong relationship between flowering periods and spring temperature trends for the olive. Temperature during March, April and May was the parameter most related to flowering date in the study areas, particularly in Italy. In some cases we found a significant correlation between flowering and past autumn temperatures, probably due to their effect on floral bud dormancy induction, but this phenomenon appeared to be of minor importance in the studied areas. The phenological trend results show the continuous advance of flowering dates to the late 1990s, followed by a relatively stationary time series related to a short-term temperature fluctuation in the Mediterranean area. This latter period probably represents a mesoscale event forced by a macroscale event—the North Atlantic Oscillation. The results reveal that the trend towards increased temperatures, and the consequent flowering advance of some species, indicated some years ago is nowadays not as clear as was expected and should be confirmed over the next few years in the Mediterranean areas under investigation.

Morphological and cytological development and starch accumulation in hermaphrodite and staminate flowers of olive (Olea europaea L.)

In olive (Olea europaea L.), the formation of functionally staminate flowers rather than fully functional hermaphrodites is one of the major factors limiting fruit set, as flowers with aborted pistils are incapable of producing fruit. Studies conducted on various angiosperm species have shown a correlation between flower abortion and starch content. Thus, it is important to know if starch content plays a role in regulating pistil development in olive and if so, what mechanism regulates starch distribution. Cyto-histological observations of staminate and hermaphrodite olive flowers show that pistil development in staminate flowers is interrupted after the differentiation of the megaspore mother cell. At that stage, starch grains were only detected in the ovary, style and stigma of the hermaphrodite flowers. No starch was observed in the pistils of the staminate flowers. This finding suggests a tight correlation between starch content and pistil development. The secondary origin of starch within the flower is indicated by low chlorophyll content in the gynoecium, undetectable Rubisco activity in the pistils of these two kinds of flowers and by the ultrastructure of the plastids observed by transmission electron microscope analysis. The plastids have few thylakoid membranes and grana and in the staminate flowers appeared very similar to proplastids. Considering differences in starch content between staminate and hermaphrodite flowers and the secondary origin of the starch, differences in pistil development in the staminate and hermaphrodite flowers could be related to differences in the sink strength of these two types of flowers.

A New Approach to Consider the Pollen Variable in Forecasting Yield Models1

Methods for forecasting harvest yields have been improved considerably in the last 20 years with the development of new data survey (remote sensing) and statistical techniques. One of these methods, based on pollen release in the atmosphere, is especially important for anemophilous species such as olive. The aim of the present work is to use a different approach to forecast the olive harvest by considering the pollen variable as “endogenous” because it is involved in the consequential processes from the formation of pollen to fruiting, the complex of which determines, more or less, the final production. Unlike models built upon a single equation (multiple linear regression analysis), the proposed estimate, based on an incomplete system of equations, recovers the consistency associated with the inference of parameters while avoiding the errors of “over-estimation.” The study, based on 17 years of data considers the quantity of olive pollen monitored and the relative annual olive production in addition to climatic, agronomic, and pathological variables associated with production. The harvest forecast provides the possibility for planning and optimizing the various stages of olive production from cultivation to distribution, including sound management of the olive supply.RésuméPendant les vingt dernières années les méthodes de prévision des rendements de récolte ont été considérablement améliorées grâce au développement de nouvelles techniques statistiques et d’ enquête des données (télédétection). Parmi ces méthodes celle basée sur l’émission du pollen dans l’atmosphère se révèle particulièrement importante pour les espèces anémophiles comme l’olivier. Le but de ce travail est celui d’arriver à employer une méthode différente dans la prévision des rendements de récolte de l’olivier, le tout en considérant comme “endogène” la variable pollen. Ce dernier est, en effect, impliqué dans les processus d’évolution qui vont de sa formation à la fructification, de manière à déterminer la production finale. Contrairement aux modèles établis sur une équation simple (analyse multiple de régression linéaire), l’évaluation proposée, basée sur un système inachevé d’équations, récupère la consistance connexe à l’ inférence des paramètres tout en évitant les erreurs de “surestimation”. L’étude, basée sur dix-sept ans de données, considère la quantité de pollen d’olivier détectée et conséquemment la production oléicole annuelle, outre aux variables climatiques, agronomiques et pathologiques liées à la production. Les modèles de prévision offrent la possibilité de rationaliser les différentes phases de la filière oléicole en optimisant les procédés, de la production à la distribution, y compris la gestion rationnelle des stocks.

Heat accumulation period in the Mediterranean region: phenological response of the olive in different climate areas (Spain, Italy and Tunisia)

The main characteristics of the heat accumulation period and the possible existence of different types of biological response to the environment in different populations of olive through the Mediterranean region have been evaluated. Chilling curves to determine the start date of the heat accumulation period were constructed and evaluated. The results allow us to conclude that the northern olive populations have the greatest heat requirements for the development of their floral buds, and they need a period of time longer than olives in others areas to completely satisfy their biothermic requirements. The olive trees located in the warmest winter areas have a faster transition from endogenous to exogenous inhibition once the peak of chilling is met, and they show more rapid floral development. The lower heat requirements are due to better adaptation to warmer regions. Both the threshold temperature and the peak of flowering date are closely related to latitude. Different types of biological responses of olives to the environment were found. The adaptive capacity shown by the olive tree should be considered as a useful tool with which to study the effects of global climatic change on agro-ecosystems.

Time linkages between pollination onsets of different taxa over an 11-year period in Perugia, Central Italy

Times of pollination of different taxa in the atmosphere of Perugia (Central Italy) over an 11-year period (1982–1992) were recorded and analysed by means of a 7-day volumetric Hirst-type pollen trap. For some taxa, the pollination period varied from year to year from a chronological and/or quantitative point of view. Several taxa showed a linkage in their starting dates of pollination. Knowledge of this kind of linkage allows us to build a forecasting model.