Environmental Stress and Plants 2.0

Following the success of our previous edition [1], this Special Issue ‘Environmental Stress and Plants 2.0’ includes 23 original articles and 3 reviews. The high level of participation and the large number of reports show that plant researchers are interested in this topic. We provide a brief overview of the papers, which include original articles (categorized by stress type) as well as reviews

Allergenic risk assessment of urban parks: Towards a standard index.

Allergenicity indices are a powerful tool to assess the health hazard posed by urban parks to pollen allergic subjects. Nonetheless, only few indices have been developed and applied to urban vegetation in the last decade, and they were never compared nor standardised over the same dataset. To address this issue, in this paper the two best-known allergenicity indices, the Urban Green Zones Allergenicity Index (IUGZA) and the Specific Allergenicity Index (SAI), have been calculated for the same park (the Botanical Garden of Bologna), collecting vegetation data through both systematic sampling and arboreal census. The results obtained with the two data collection methods were comparable for both indices, indicating systematic sampling as a reliable approximation of the total census. Besides, the allergenic risk resulted moderate to high according to SAI, and very low according to IUGZA. Since SAI does not consider the total volume of the vegetation, it was deemed less reliable than IUGZA in evaluating the allergenicity of an enclosed green space

Cytoskeleton, Transglutaminase and Gametophytic Self-Incompatibility in the Malinae (Rosaceae)

Self-incompatibility (SI) is a complex process, one out of several mechanisms that prevent plants from self-fertilizing to maintain and increase the genetic variability. This process leads to the rejection of the male gametophyte and requires the co-participation of numerous molecules. Plants have evolved two distinct SI systems, the sporophytic (SSI) and the gametophytic (GSI) systems. The two SI systems are markedly characterized by different genes and proteins and each single system can also be divided into distinct subgroups; whatever the mechanism, the purpose is the same, i.e., to prevent self-fertilization. In Malinae, a subtribe in the Rosaceae family, i.e., Pyrus communis and Malus domestica, the GSI requires the production of female determinants, known as S-RNases, which penetrate the pollen tube to interact with the male determinants. Beyond this, the penetration of S-RNase into the pollen tube triggers a series of responses involving membrane proteins, such as phospholipases, intracellular variations of cytoplasmic Ca2+, production of reactive oxygen species (ROS) and altered enzymatic activities, such as that of transglutaminase (TGase). TGases are widespread enzymes that catalyze the post-translational conjugation of polyamines (PAs) to different protein targets and/or the cross-linking of substrate proteins leading to the formation of cross-linked products with high molecular mass. When actin and tubulin are the substrates, this destabilizes the cytoskeleton and inhibits the pollen-tube's growth process. In this review, we will summarize the current knowledge of the relationship between S-RNase penetration, TGase activity and cytoskeleton function during GSI in the Malinae

Ancient Tomato (Solanum lycopersicum L.) Varieties of Tuscany Have High Contents of Bioactive Compounds

The Tuscan Region has a vast repertoire of ancient plants that have been recovered across the territory over the years. These plants thrive in an environment characterized by minimal human intervention and are thus the result of the process of adaptation to the territory of origin. In this work, we focused on the quantification of bioactive compounds in ancient tomato varieties. More specifically, we quantified polyphenols, flavonoids, carotenoids, and vitamin C in eight local Tuscan tomato varieties and found higher contents with respect to those in commercial tomatoes Polyphenol and antioxidant compounds in ancient varieties reported a two- and, in some instances, three-fold increase in concentration, compared to the commercial counterparts. Interestingly, the data relative to the carotenoids did not show any significant differences when comparing the ancient varieties with the commercial ones, a finding confirming the market selection criterion based on color. On a longer-term perspective, this study aims at drawing attention to the importance of preserving autochthonous natural plant biodiversity and towards promoting research on local varieties. We believe that this study will pave the way to the valorization of local plant biodiversity and promote an extended use of products in the nutraceutical sector derived from vegetables

Small extracellular vesicles released from germinated kiwi pollen (pollensomes) present characteristics similar to mammalian exosomes and carry a plant homolog of ALIX

Introduction: In the last decade, it has been discovered that allergen-bearing extracellular nanovesicles, termed “pollensomes”, are released by pollen during germination. These extracellular vesicles (EVs) may play an important role in pollen-pistil interaction during fertilization, stabilizing the secreted bioactive molecules and allowing long-distance signaling. However, the molecular composition and the biological role of these EVs are still unclear. The present study had two main aims: (I) to clarify whether pollen germination is needed to release pollensomes, or if they can be secreted also in high humidity conditions; and (II) to investigate the molecular features of pollensomes following the most recent guidelines for EVs isolation and identification. Methods: To do so, pollensomes were isolated from hydrated and germinated kiwi (Actinidia chinensis Planch.) pollen, and characterized using imaging techniques, immunoblotting, and proteomics. Results: These analyses revealed that only germinated kiwi pollen released detectable concentrations of nanoparticles compatible with small EVs for shape and protein content. Moreover, a plant homolog of ALIX, which is a well-recognized and accepted marker of small EVs and exosomes in mammals, was found in pollensomes. Discussion: The presence of this protein, along with other proteins involved in endocytosis, is consistent with the hypothesis that pollensomes could comprehend a prominent subpopulation of plant exosome-like vesicles

Small extracellular vesicles released from germinated kiwi pollen (pollensomes) present characteristics similar to mammalian exosomes and carry a plant homolog of ALIX

IntroductionIn the last decade, it has been discovered that allergen-bearing extracellular nanovesicles, termed “pollensomes”, are released by pollen during germination. These extracellular vesicles (EVs) may play an important role in pollen-pistil interaction during fertilization, stabilizing the secreted bioactive molecules and allowing long-distance signaling. However, the molecular composition and the biological role of these EVs are still unclear. The present study had two main aims: (I) to clarify whether pollen germination is needed to release pollensomes, or if they can be secreted also in high humidity conditions; and (II) to investigate the molecular features of pollensomes following the most recent guidelines for EVs isolation and identification.MethodsTo do so, pollensomes were isolated from hydrated and germinated kiwi (Actinidia chinensis Planch.) pollen, and characterized using imaging techniques, immunoblotting, and proteomics.ResultsThese analyses revealed that only germinated kiwi pollen released detectable concentrations of nanoparticles compatible with small EVs for shape and protein content. Moreover, a plant homolog of ALIX, which is a well-recognized and accepted marker of small EVs and exosomes in mammals, was found in pollensomes.DiscussionThe presence of this protein, along with other proteins involved in endocytosis, is consistent with the hypothesis that pollensomes could comprehend a prominent subpopulation of plant exosome-like vesicles

New insight into quinoa seed quality under salinity: Changes in proteomic and amino acid profiles, phenolic content, and antioxidant activity of protein extracts

Quinoa (Chenopodium quinoa Willd) is an ancient Andean seed-producing crop well known for its exceptional nutritional properties and resistance to adverse environmental conditions, such as salinity and drought. Seed storage proteins, amino acid composition, and bioactive compounds play a crucial role in determining the nutritional value of quinoa. Seeds harvested from three Chilean landraces of quinoa, one belonging to the salares ecotype (R49) and two to the coastal-lowlands ecotype, VI-1 and Villarrica (VR), exposed to two levels of salinity (100 and 300 mM NaCl) were used to conduct a sequential extraction of storage proteins in order to obtain fractions enriched in albumins/globulins, 11S globulin and in prolamin-like proteins. The composition of the resulting protein fractions was analyzed by one- and two-dimensional polyacrylamide gel electrophoresis. Results confirmed a high polymorphism in seed storage proteins; the two most representative genotype-specific bands of the albumin/globulin fraction were the 30- and 32-kDa bands, while the 11S globulin showed genotype-specific polymorphism for the 40- and 42-kDa bands. Spot analysis by mass spectrometry followed by in silico analyses were conducted to identify the proteins whose expression changed most significantly in response to salinity in VR. Proteins belonging to several functional categories (i.e., stress protein, metabolism, and storage) were affected by salinity. Other nutritional and functional properties, namely amino acid profiles, total polyphenol (TPC) and flavonoid (TFC) contents, and antioxidant activity (AA) of protein extracts were also analyzed. With the exception of Ala and Met in R49, all amino acids derived from protein hydrolysis were diminished in seeds from salt-treated plants, especially in landrace VI-1. By contrast, several free amino acids were unchanged or increased by salinity in R49 as compared with VR and VI-1, suggesting a greater tolerance in the salares landrace. VR had the highest TPC and AA under non-saline conditions. Salinity increased TPC in all three landraces, with the strongest increase occurring in R49, and enhanced radical scavenging capacity in R49 and VR. Overall, results show that salinity deeply altered the seed proteome and amino acid profiles and, in general, increased the concentration of bioactive molecules and AA of protein extracts in a genotype-dependent manner

Expression of clementine asp-rich proteins (CcASP-RICH) in tobacco plants interferes with the mechanism of pollen tube growth

Low-molecular-weight, aspartic-acid-rich proteins (ASP-RICH) have been assumed to be involved in the self-incompatibility process of clementine. The role of ASP-RICH is not known, but hypothetically they could sequester calcium ions (Ca2+) and affect Ca2+-dependent mechanisms. In this article, we analyzed the effects induced by clementine ASP-RICH proteins (CcASP-RICH) when expressed in the tobacco heterologous system, focusing on the male gametophyte. The aim was to gain insight into the mechanism of action of ASP-RICH in a well-known cellular system, i.e., the pollen tube. Pollen tubes of tobacco transgenic lines expressing CcASP-RICH were analyzed for Ca2+ distribution, ROS, proton gradient, as well as cytoskeleton and cell wall. CcASP-RICH modulated Ca2+ content and consequently affected cytoskeleton organization and the deposition of cell wall components. In turn, this affected the growth pattern of pollen tubes. Although the expression of CcASP-RICH did not exert a remarkable effect on the growth rate of pollen tubes, effects at the level of growth pattern suggest that the expression of ASP-RICH may exert a regulatory action on the mechanism of plant cell growth

Changes in the accumulation of alfa- and beta-tubulin during bud development in Vitis vinifera L.

3Microtubules play important roles duringgrowth and morphogenesis of plant cells. Multiple isoformsof - and -tubulin accumulate in higher plant cells andoriginate either by transcription of diVerent genes or bypost-translational modiWcations. The use of diVerenttubulin isoforms involves the binding of microtubules todiVerent associated proteins and therefore generates microtubuleswith diVerent organizations and functions. Tubulinisoforms are diVerentially expressed in vegetative andreproductive structures according to the developmental programof plants. In grapevine (Vitis vinifera L.), vegetativeand reproductive structures appear on the same stem, makingthis plant species an excellent model to study the accumulationof tubulin isoforms. Proteins were extracted fromgrapevine samples (buds, leaves, Xowers and tendrils)using an optimized extraction protocol, separated by twodimensionalelectrophoresis and analyzed by immunoblotwith anti-tubulin antibodies. We identiWed eight -tubulinand seven -tubulin isoforms with pI around 4.8–5 thatgroup into separate clusters. More acidic -tubulin isoformswere detected in buds, while more basic -isoforms wereprevalently found in tendrils and Xowers. Similarly, moreacidic -tubulin isoforms were used in the bud stage whilea basic -tubulin isoform was essentially used in leaves andtwo central -tubulin isoforms were characteristically usedin tendrils and Xowers. Acetylated -tubulin was notdetected in any sample while tyrosinated -tubulin wasessentially found in large latent buds and in bursting budsin association with a distinct subset of tubulin isoformsreservedmixedParrotta, Luigi; Cai, Giampiero; Cresti, MauroParrotta, Luigi; Cai, Giampiero; Cresti, Maur