Francesca Rapparini

Effect of arbuscular mycorrhizal (AM) colonization on terpene emission and content of Artemisia annua L.

Plant roots interact with a wide variety of rhizospheric microorganisms, including bacteria and the symbiontic arbuscular mycorrhizal (AM) fungi. The mycorrhizal symbiosis represents a series of complex feedbacks between plant and fungus regulated by their physiology and nutrition. Despite the widespread distribution and ecological significance of AM symbiosis, little is known about the potential of AM fungi to affect plant VOC metabolism. The purpose of this study was to investigate whether colonization of plant roots by AM fungi and associated soil microorganisms affects VOC emission and content of Artemisia annua L. plants (Asteraceae). Two inoculum types were evaluated: one consisted of only an arbuscular mycorrhizal (AM) fungus species (Glomus spp.), and the other was a mixture of different Glomus species and associated soil bacteria. Inoculated plants were compared with non-inoculated plants and with plants supplemented with extra phosphorus (P) to obtain plants of the same size as mycorrhizal plants, thus excluding potentially-confounding mycorrhizal effects on shoot growth. VOC emissions of Artemisia annua plants were analyzed by leaf cuvette sampling followed by off-line measurements with pre-concentration and gas chromatography mass spectrometry (GC-MS). Measurements of CO(2) and H(2)O exchanges were conducted simultaneously. Several volatile monoterpenes were identified and characterized from leaf emissions of Artemisia annua L. by GC-MS analysis. The main components identified belong to different monoterpene structures: alpha-pinene, beta-pinene, camphor, 1,8-cineole, limonene, and artemisia ketone. A good correlation between monoterpene leaf concentration and leaf emission was found. Leaf extracts included also several sesquiterpenes. Total terpene content and emission was not affected by AM inoculation with or without bacteria, while emission of limonene and artemisia ketone was stimulated by this treatment. No differences were found among treatments for single monoterpene content, while accumulation of specific sesquiterpenes in leaves was altered in mycorrhizal plants compared to control plants. Growth conditions seemed to have mainly contributed to the outcome of the symbiosis and influenced the magnitude of the plant response. These results highlight the importance of considering the below-ground interaction between plant and soil for estimating VOC emission rates and their ecological role at multitrophic levels.

Mycorrhizal Fungi to Alleviate Drought Stress on Plant Growth

Drought stress induces a range of metabolic responses in plants. Some of these responses are mediated by arbuscular mycorrhizae (AM), which occur almost ubiquitously in symbiotic associations. These changes are highly variable and depend on various factors related mainly to the diversity of plant and fungal species but are generally beneficial to the host plants. This chapter addresses the role of AM fungi in the amelioration and alleviation of drought stress in host plants and their positive effects on growth. We discuss the various biochemical, physiological, and molecular processes used by plants to alleviate drought stress. We provide an update of the recent progress in functional approaches for unraveling the mechanisms that promote resistance to drought stress and discuss their significance to the host plants. The positive aspects of AM are also discussed in the context of the ecosystem services provided by the symbiosis under environmental drought conditions.

Seasonal variation of monoterpene emission from Malus domestica and Prunus avium

Emission rates of monoterpenes released by apple (Malus domestica Borkh) and cherry (Prunus avium L.) were estimated at different phenological stages. These measurements employed a dynamic flow-through Teflon chamber, sample collection onto cartridges filled with graphitized carbon and thermal desorption gas chromatography-mass spectrometry (GC-MS) for identification and quantification of the emitted volatiles. At full bloom the release of monoterpene hydrocarbons from cherry flowers was 1213 ng g(-1) dry weight (DW) h(-1), exceeding by approximately three-fold the emission rate of apple flowers (366 ng g(-1) DW h(-1)). Observed seasonal variations in biogenic volatile organic compound (VOC) emissions ranged over several order of magnitudes. At fruit-set and ripening stages, in fact, the hydrocarbon emission dramatically decreased reaching the lowest values at harvest time when leaves were fully mature (3-9 ng g(-1) DW h(-1)). Wide diversity in the composition of compounds from the species studied was also recorded. At blooming, linalool contributed significantly to the monoterpene emission from apple (94% of the emitted carbon) while alpha-pinene and camphene represented on average more than 60% of the total emitted volatiles from cherry flowers. Among the monoterpenes identified in flowers, alpha-pinene, camphene and limonene were also found in the foliage emission of both species. Fruit trees are relevant monoterpene emitters only at blooming and thus for a short period of the vegetative cycle. When leaves are fully developed, the carbon loss due to monoterpene emissions related to the photosynthetically carbon gain is negligible.

AM fungi root colonization increases the production of essential isoprenoids vs. nonessential isoprenoids especially under drought stress conditions or after jasmonic acid application

Previous studies have shown that root colonization by arbuscular mycorrhiza (AM) fungi enhances plant resistance to abiotic and biotic stressors and finally plant growth. However, little is known about the effect of AM on isoprenoid foliar and root content. In this study we tested whether the AM symbiosis affects carbon resource allocation to different classes of isoprenoids such as the volatile nonessential isoprenoids (monoterpenes and sesquiterpenes) and the non-volatile essential isoprenoids (abscisic acid, chlorophylls and carotenoids). By subjecting the plants to stressors such as drought and to exogenous application of JA, we wanted to test their interaction with AM symbiosis in conditions where isoprenoids usually play a role in resistance to stress and in plant defence. Root colonization by AM fungi favoured the leaf production of essential isoprenoids rather than nonessential ones, especially under drought stress conditions or after JA application. The increased carbon demand brought on by AM fungi might thus influence not only the amount of carbon allocated to isoprenoids, but also the carbon partitioning between the different classes of isoprenoids, thus explaining the not previously shown decrease of root volatile isoprenoids in AM plants. We propose that since AM fungi are a nutrient source for the plant, other carbon sinks normally necessary to increase nutrient uptake can be avoided and therefore the plant can devote more resources to synthesize essential isoprenoids for plant growth.

Vesicular-arbuscular mycorrhizal inoculation of micropropagated fruit trees

Micropropagated plantlets of OH x F 51 and GF 677, respectively pear (Pyrus communis L.) and peach (Prunus persica x Prunus amygdalus) clonal rootstocks were inoculated during an early weaning stage of acclimatization with Glomus sp. Both rootstocks were well colonized, although the infection of OH x F 51 spread more slowly. At the end of initial vegetative growth, mycorrhizal plants of both rootstocks showed a three-fold increase in shoot length over control plants. Mycorrhizal plants also had longer internode and greater fresh mass. The root/shoot ratio was especially altered by arbuscular mycorrhizal inoculation in OH x F 51 plants, which showed a greater increase in shoot rather than in root biomass. The growth-promoting ability of endomycorrhizal fungus persisted throughout the experimental period: Glomus sp. induced a greater development of both rootstocks in the second growing year, after overwintering. Colonization of micropropagated plants by arbuscular mycorrhizal fungus appears to alter the car...

Ectopic expression of LEAFY COTYLEDON1-LIKE gene and localized auxin accumulation mark embryogenic competence in epiphyllous plants of Helianthus annuus × H. tuberosus

BACKGROUND AND AIMS The clone EMB-2 of the interspecific hybrid Helianthus annuus x H. tuberosus provides an interesting system to study molecular and physiological aspects of somatic embryogenesis. Namely, in addition to non-epiphyllous (NEP) leaves that expand normally, EMB-2 produces epiphyllous (EP) leaves bearing embryos on the adaxial surface. This clone was used to investigate if the ectopic expression of H. annuus LEAFY COTYLEDON1-LIKE (Ha-L1L) gene and auxin activity are correlated with the establishment of embryogenic competence. METHODS Ha-L1L expression was evaluated by semi-quantitative RT-PCR and in situ hybridization. The endogenous level and spatial distribution of free indole-3-acetic acid (IAA) were estimated by a capillary gas chromatography-mass spectrometry-selected ion monitoring method and an immuno-cytochemical approach. KEY RESULTS Ectopic expression of Ha-L1L was detected in specific cell domains of the adaxial epidermis of EP leaves prior to the development of ectopic embryos. Ha-L1L was expressed rapidly when NEP leaves were induced to regenerate somatic embryos by in vitro culture. Differences in auxin distribution pattern rather than in absolute level were observed between EP and A-2 leaves. More precisely, a strong IAA immuno-signal was detected in single cells or in small groups of cells along the epidermis of EP leaves and accompanied the early stages of embryo development. Changes in auxin level and distribution were observed in NEP leaves induced to regenerate by in vitro culture. Exogenous auxin treatments lightly influenced Ha-L1L transcript levels in spite of an enhancement of the regeneration frequency. CONCLUSIONS In EP leaves, Ha-L1L activity marks the putative founder cells of ectopic embryos. Although the ectopic expression of Ha-L1L seems to be not directly mediated by auxin levels per se, it was demonstrated that localized Ha-L1L expression and IAA accumulation in leaf epidermis domains represent early events of somatic embryogenesis displayed by the epiphyllous EMB-2 clone.

The application of chitosan and benzothiadiazole in vineyard (Vitis vinifera L. cv Groppello Gentile) changes the aromatic profile and sensory attributes of wine.

This work reports the effects of resistance inducers on wine aroma compounds and sensory attributes. Resistance inducers are a class of products able to elicit the plant defence mechanisms against pathogens, incurring lower toxicological risks than conventional agrochemicals. Among them, chitosan (CHT) and benzothiadiazole (BTH) are particularly effective in stimulating the biosynthesis of bioactive phytochemicals. They were used in a two-year survey conducted to assess experimental wines obtained from elicitor-treated grapes. Compared with conventional fungicides (penconazole and methyldinocap), in 2009, BTH increased total acetals and esters, while CHT raised the levels of total acetals and alcohols. Sensory analysis revealed that overall acceptance was higher in CHT than in BTH. In 2010, differences were not significant. Therefore, plant activators deserve attention beyond their efficacy in crop protection. In particular, in our experimental conditions, CHT improved the volatile profile, flavour and taste of Groppello wine.

Emission of isoprenoids from natural vegetation in the Beijing region (Northern China)

ABSTRACT A survey was conducted to identify plants emitting isoprenoids in the Beijing area which could potentially contribute to smog episodes when combined with anthropogenic pollutants. The emission pattern was similar to that observed in the previously surveyed boreal ecosystems (Europe, North America). Most deciduous oaks are strong isoprene emitters; however, some of them do not emit isoprenoids and are therefore more suitable for the urban environment of Beijing. No emission of monoterpenes was found in Chinese oaks, and this trait seems therefore confined to the Mediterranean environment. The emission of isoprene was found in poplars and in some of the bamboo widespread in city parks and in the riparial vegetation surrounding the city. Chinese pine species emit monoterpenes when wounded, and the emission is not qualitatively different among species. Pinus tabulaeformis, one of the most important trees in China, is a low emitter compared to other pine species.

Blue light regulation of the growth of Prunus persica plants in a long term experiment: morphological and histological observations

Abstract Prunus persica plants were grown under prolonged exposure to different light treatments to determine the interaction between the blue light (BL) receptor and phytochrome and/or an independent BL response in the photoregulation of shoot and leaf development. Different light conditions were established in growth chambers by changing both the state of phytochrome and the BL photon flux density (PFD) at constant photosynthetically active radiation (PAR). Furthermore, to evaluate the independent action of the BL photoreceptor, increasing amounts of BL photons were added to the light emitted by low-pressure sodium (LPS) lamps without altering irradiance and phytochrome photoequilibrium. Applying the principle of equivalent light action, the observed blue inhibition of shoot elongation, leaf expansion and thickness were clearly related to a specific BL receptor because the state of phytochrome for each treatment was nearly identical. Increasing amounts of blue photons to light emitted from LPS lamps decreased shoot elongation, whereas leaf expansion was negatively affected only at the highest blue level, suggesting a specific fluence dependence response to BL for each organ and tissue. The BL effect was evident in reducing the thickness of all the leaf tissues except for the upper epidermis, which became thicker. This could be the result of an adaptation to protect the underlying photosynthetic apparatus. Other morphological and anatomical responses to the action of the BL receptor were greatly altered when the state of phytochrome changed in the plant tissues.

Effects of simulated light environments on growth and leaf morphology of peach plants

SummaryThe role of light quality and quantity in regulating growth and differentiation of Prunus persica plants was evaluated using different coloured filters in an outdoor experiment. Examination of total growth showed that neutral shading (80% of transparent control) did not affect shoot length, internode elongation, leaf number, and branching, but strongly influenced the anatomical characteristics of leaves. The combination blue + far red (B + FR) acted quite differently from the combination red + far red (R + FR) and caused a general inhibition of growth phenomena. Shorter and more compact plants were produced under B + FR tunnels where the reduction in shoot elongation was consistent with the reduction in internode length. Furthermore, light transmitted through the B + FR filter resulted in lower branching and in smaller and thinner leaves compared with those grown under the other light treatments. The number of flowers was not affected by either light quality or quantity. The negative effect on grow...