Catechin promotes the germination of Pistacia chinensis seeds via GA biosynthesis.

BACKGROUND AND AIMS: Chinese pistachio (Pistacia chinensis), an important horticultural plant species, holds great ornamental value with beautiful leaves and fruits. Seedling propagation of this tree species is restricted by its erratic seed germination; however, the germination mechanism is ambiguous. The aim of this study was to determine the germination mechanism from a novel perspective based on the multi-omics data. METHODS: The multi-omics technique combined with hormone content measurement was applied to seed germination of Chinese pistachio. KEY RESULTS: Due to its great accumulation during seed germination, catechin stood out from the identified metabolites in a broadly targeted metabolomic analysis. Exogenous catechin at 10 mg L-1 significantly improved the germination of Chinese pistachio seeds. An interesting result of hormone analysis was that the improving effect of catechin could be attributed to an increase in gibberellic acid 3 (GA3) content rather than a decrease in abscisic acid (ABA) content before germination. Treatments with paclobutrazol (PAC, a GA biosynthesis inhibitor) and PAC + catechin also showed that the promoting effect of catechin on seed germination depends on GA biosynthesis. Transcriptome analysis and qRT‒PCR further revealed that catechin induced the expression of PcGA20ox5 to activate GA biosynthesis. Several transcription factors were induced by catechin and GA treatments, such as TCP, bZIP and C3H, which may play an important regulatory role in GA biosynthesis in a catechin-mediated way. CONCLUSIONS: Catechin promotes seed germination via GA biosynthesis in Chinese pistachios. This study proposes a novel mechanism by which catechin promotes seed germination via the GA pathway, which provides new insight into a comprehensive understanding of seed dormancy and germination.

Exogenous ß-aminobutyric acid application attenuates Aspergillus decay, minimizes aflatoxin B1 accumulation, and maintains nutritional quality in fresh-in-hull pistachio kernels.

BACKGROUND: Pistachio fruits suffer from postharvest decay, caused by Aspergillus flavus. This results in aflatoxin B1 (AFB1 ) accumulation in kernels, which is hazardous for human health due to its carcinogenic activity. In this study, the mechanism used by exogenous ß-aminobutyric acid (BABA) treatment for attenuating Aspergillus decay, minimizing aflatoxin B1 (AFB1 ) accumulation, and maintaining nutritional quality in fresh-in-hull pistachio kernels, infected by A. flavus during storage at 25 °C for 18 days, was investigated. RESULT: Results of an in vivo assay showed that the spore germination and germ tube elongation of A. flavus was repressed by BABA treatment at 7.5 mM. Aspergillus decay accompanied by AFB1 accumulation was also minimized in fresh-in-hull pistachio kernels treated with BABA at 7.5 mM and infected by A. flavus. Fresh-in-hull pistachio kernels, infected by A. flavus, treated with BABA at 7.5 mM, also exhibited higher phenol and flavonoid accumulation and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging capacity accompanied by higher phenylalanine ammonia lyase (PAL) enzyme activity. CONCLUSION: Promoting phenylpropanoid pathway activity with higher PAL enzyme activity in fresh-in-hull pistachio kernels treated with BABA may not only reduce Aspergillus decay in kernels by cell wall fortification but also may be favorable for maintaining the kernels' nutritional quality through its effects on ROS scavenging capacity. As oxidative stress, represented by ROS accumulation, is responsible for A. flavus growth and AFB1 accumulation, higher phenol and flavonoid accumulation in fresh-in-hull pistachio kernels treated with BABA may be beneficial for attenuating Aspergillus decay and minimizing AFB1 accumulation. © 2019 Society of Chemical Industry.

Effect of phosphorus concentration on the photochemical stability of PSII and CO2 assimilation in Pistacia vera L. and Pistacia atlantica Desf.

A greenhouse pot experiment was conducted at the faculty of sciences of Gafsa to evaluate the effect of phosphorus treatment on two pistachio species. The seedlings of Pistacia vera and Pistacia atlantica were subjected to six levels of phosphoric acid (P2O5) (0, 5, 15, 30, 60 and 120 ppm). Stomatal conductance, net photosynthesis, chlorophyll fluorescence (OJIP) and total chlorophyll content were measured after 1, 2, 3, 6, 8, 9 and 12 weeks of treatment. During the experiment, phosphorus application at 5 ppm increased photosynthesis and stomatal conductance, relative to the treatment 0 ppm only in P. atlantica. However, phosphorus supply at 60 and 120 ppm induced toxicity leading to an inhibition of CO2 photo-assimilation rate, an alteration of photosystem II (PSII) structure and function and reduction in leaf chlorophyll content in both species. The (OJIP) transient showed complex changes in O-J, J-I and I-P phases of fluorescence. Due to phosphorus toxicity, both donor and acceptor sides of PSII were damaged, electron transport perturbed and chlorophyll pigment reduced which resulted in the fall of CO2 photo-assimilation rate, followed by mortality in both species.

Computer-based tools provide new insight into the key factors that cause physiological disorders of pistachio rootstocks cultured in vitro.

During the in vitro culture of plants some physiological disorders caused major problems that have been associated with culture media composition. The objective of this study was to better understand the abnormal physiological response of two pistachio rootstocks to changes in culture media ingredients. On this purpose, two computer-based tools were employed: design of experiment (DOE) and neurofuzzy logic. DOE was employed to generate a five-dimensional IV-design spaces allowing to reduce the number of treatments from 6,250 to 61. The second one, an artificial intelligence (AI) tool, neurofuzzy logic, was used to understand the cause-effect relationships between the factors studied (25) and seven physiological disorders including shoot-tip necrosis (STN), leaf necrosis (LN), leaf color (LC), basal callus (BC) formation, shoot fasciation (SF), hyperhydricity and epinasty, typically described during pistachio in vitro culture. Four out of the seven disorders were successfully modeled, being significantly affected by a limited number of factors. STN and BC were significantly affected by the concentration of EDTA-. However, while a low concentration of EDTA- reduces the STN, promotes BC. LN and LC were strongly alleviated by high amounts of thiamine-HCl. Undoubtedly, the results demonstrate the importance of recording and using data related to physiological disorders along with growth parameters when developing suitable culture media for plant tissues. The computer-based tools have been useful to: i) well sample experimental design; ii) reduce the final number of treatments and the experimental work; iii) identify the key factors affecting each disorder; iv) get insight about the causes that promote the appearance of physiological disorders. Our findings demonstrate that the recently AI designed POM media, although not optimal, is the most suitable (favouring growth and limiting physiological abnormalities) media for in vitro culture of pistachio compared to those media, currently used.

Ion homeostasis, osmoregulation, and physiological changes in the roots and leaves of pistachio rootstocks in response to salinity.

Pistachio, one of the important tree nuts, is cultivated in arid and semi-arid regions where salinity is the most common abiotic stress encountered by this tree. However, the mechanisms underlying salinity tolerance in this plant are not well understood. In the present study, five 1-year-old pistachio rootstocks (namely Akbari, Badami, Ghazvini, Kale-Ghouchi, and UCB-1) were treated with four saline water regimes (control, 8, 12, and 16 dS m-1) for 100 days. At high salinity level, all rootstocks showed decreased relative water content (RWC), total chlorophyll content (TCHC), and carotenoids in the leaf, while ascorbic acid (AsA) and total soluble proteins (TSP) were reduced in both leaf and root organs. In addition, the total phenolic compounds (TPC), proline, glycine betaine, total soluble carbohydrate (TSC), and H2O2 content increased under salinity stress in all studied rootstocks. Three different ion exclusion strategies were observed in the studied rootstocks: (i) Na+ exclusion in UCB-1, because most of its Na+ is retained in the roots; (ii) Cl- exclusion in Badami, in which most of its Cl- remained in the roots; and (iii) similar concentrations of Na+ and Cl- were observed in the leaves and roots of Ghazvini, Akbari, and Kale-Ghouchi. Transport capacity (ST value) of K+ over Na+ from the roots to the leaves was more observable in UCB-1 and Ghazvini. Overall, the root system cooperated more effectively in UCB-1 and Badami for retaining and detoxifying an excessive amount of Na+ and Cl-. The results presented here provide important inputs to better understand the salt tolerance mechanism in a tree species for developing more salt-tolerant genotypes. Based on the results obtained here, the studied rootstocks from tolerant to susceptible are arranged as follows: UCB-1 > Badami > Ghazvini > Kale-Ghouchi > Akbari.

Identification of Reference Genes for Quantitative Gene Expression Studies in a Non-Model Tree Pistachio (Pistacia vera L.).

The tree species, Pistacia vera (P. vera) is an important commercial product that is salt-tolerant and long-lived, with a possible lifespan of over one thousand years. Gene expression analysis is an efficient method to explore the possible regulatory mechanisms underlying these characteristics. Therefore, having the most suitable set of reference genes is required for transcript level normalization under different conditions in P. vera. In the present study, we selected eight widely used reference genes, ACT, EF1α, α-TUB, ß-TUB, GAPDH, CYP2, UBQ10, and 18S rRNA. Using qRT-PCR their expression was assessed in 54 different samples of three cultivars of P. vera. The samples were collected from different organs under various abiotic treatments (cold, drought, and salt) across three time points. Several statistical programs (geNorm, NormFinder, and BestKeeper) were applied to estimate the expression stability of candidate reference genes. Results obtained from the statistical analysis were then exposed to Rank aggregation package to generate a consensus gene rank. Based on our results, EF1α was found to be the superior reference gene in all samples under all abiotic treatments. In addition to EF1α, ACT and ß-TUB were the second best reference genes for gene expression analysis in leaf and root. We recommended ß-TUB as the second most stable gene for samples under the cold and drought treatments, while ACT holds the same position in samples analyzed under salt treatment. This report will benefit future research on the expression profiling of P. vera and other members of the Anacardiaceae family.

Seasonal variation and gender pattern of phenolic and flavonoid contents in Pistacia chinensis Bunge inflorescences and leaves.

Pistacia chinensis Bunge (P. chinensis) is a deciduous and dioecious perennial arbor of the family Anacardiaceae that flowers from March to April and bears fruit from September to October. There are three rapidly growing stages in the annual growth process of P. chinensis. However, the knowledge of the secondary metabolites related to P. chinensis gender and growth season remains scant. In this study, HPLC was used to qualitatively and quantitatively determine the content of the catechin hydrate, rutin, quercetin, and kaempferol contents in male and female tree inflorescences and leaves. Total phenolics and flavonoids were also detected using a spectrophotometer. The results indicated that the contents of these compounds fluctuated with seasons and they reached the highest levels in nascent leaves. The fluctuations of these compounds followed different pathways of evolution, by increasing or decreasing in male and female trees throughout the whole growth process because they had their own biological functions. Moreover, the extracts exhibited DPPH radical scavenging bioactivity and showed no significant cytotoxicity towards 3T3-L1 preadipocytes. Together, these results demonstrated that P. chinensis has great potential as an antioxidant medicine, and the best harvest time is in the spring.

CO2 gasification reactivity of biomass char: catalytic influence of alkali, alkaline earth and transition metal salts.

This study investigates the influence of alkali (Na, K), alkaline earth (Ca, Mg) and transition (Fe) metal nitrates on CO2 gasification reactivity of pistachio nut shell (PNS) char. The preliminary gasification experiments were performed in thermogravimetric analyzer (TGA) and the results showed considerable improvement in carbon conversion; Na-char>Ca-char>Fe-char>K-char>Mg-char>raw char. Based on TGA studies, NaNO3 (with loadings of 3-7 wt%) was selected as the superior catalyst for further gasification studies in bench-scale reactor; the highest reactivity was devoted to 5 wt% Na loaded char. The data acquired for gasification rate of catalyzed char were fitted with several kinetic models, among which, random pore model was adopted as the best model. Based on obtained gasification rate constant and using the Arrhenius plot, activation energy of 5 wt% Na loaded char was calculated as 151.46 kJ/mol which was 53 kJ/mol lower than that of un-catalyzed char.

Response of Holm oak (Quercus ilex subsp. ballota) and mastic shrub (Pistacia lentiscus L.) seedlings to high concentrations of Cd and Tl in the rhizosphere.

The impairment of root growth and photosynthetical functioning are the main impacts of trace elements on woody plant seedlings. In this work, we assessed the response of Holm oak (Quercusilex subsp. ballota) and mastic shrub (Pistacia lentiscus) seedlings to high concentrations of Cd and Tl in the rhizosphere. These are non-essential trace elements, with a potential high mobility in the soil-plant system. Seedlings of these species are frequently used in the afforestation of degraded soils in mining areas. Plants were exposed to different levels of Cd (20, 80 and 200 mg L(-1)) and Tl (2, 10 and 20 mg L(-1)) in a sand culture. Biomass allocation, growth rates, chlorophyll fluorescence and gas exchange were studied. Both metals affected root biomass. Cadmium produced an increase in the root mass ratio and a decrease in the specific leaf area of the plants in oak seedlings, while Tl did not provoke such response. Mastic plants were more sensitive to Tl and Cd than oak plants. Between elements, Tl provoked more severe toxic effects in the plants, affecting the antennae complexes and reaction centers of the photosystem II. Both elements decreased net assimilation rates (down to a 20% of the control plants) and stomatal conductance (5-10% of the values for the control plants). Cadmium was highly retained in the roots of both species, while Tl was highly translocated into the leaves. In general, Holm oak showed a higher tolerance for Cd than for Tl, and a higher resistance to both metals than mastic shrub, due to a high capacity for Cd retention at the root level. However, such accumulation in roots may induce water stress in the seedling exposed to Cd.

Salt-driven interactions between Pistacia lentiscus and Salsola inermis.

BACKGROUND, AIM, AND SCOPE: It can be learned from the Pistacia spp. germplasm collection (http://www.bgu.ac.il/pistacia) that the growth of Salsola inermis is inhibited in the vicinity of the evergreen Pistacia lentiscus, but not in the surroundings of the deciduous Pistacia atlantica and Pistacia chinensis. Irrigation of trees during the summer months increases soil salinity around the trees. It was therefore hypothesized that inhibition of S. inermis around P. lentiscus is related to depletion of salt in the vicinity of the latter species. MATERIALS AND METHODS: A multi-approach experimental scheme was carried out which included soil edaphic characterization and germination tests. To test salt tolerance of P. lentiscus, plants were grown in a hydroponic system for a month in medium containing NaCl, while physiological and growth parameters were measured. RESULTS: Conductivity measurements in summer, during the growth season of S. inermis, indicated that soil salinity beneath deciduous Pistacia trees was significantly higher than that below P. lentiscus. Germination of S. inermis seeds on filter paper moistened with P. lentiscus low-conductivity soil filtrate was twice as high as that of the deciduous trees high-conductivity soil filtrates. Nevertheless, fresh and dry weights of mature S. inermis growing next to P. atlantica and P. chinensis were 2.9 to 4.8 times higher than those of plants growing in the vicinity of P. lentiscus. In a hydroponic system, no significant differences were found in growth parameters and stomatal conductance between P. lentiscus growing in control and salt treatments. It was therefore proposed that salt depletion in the vicinity of P. lentiscus inhibits the growth, but not germination, of S. inermis thus confirming the halophylic characteristics of this plant. DISCUSSION: The nature of Salsola-Pistacia interactions cannot be explained by allelopathic effects; hence, plausible salt-driven interactions were considered. Our data showed that S. inermis accumulated salt and has halophytic characteristics. Interestingly, germination of S. inermis was inhibited in medium containing salt, but the salt was obligatory for further growth, development, and fast biomass production. These results explained the observation of large biomass accumulation in the more saline soil around the deciduous P. atlantica and P. chinensis and the lack of development in the salt-depleted soil around the salt-tolerant accumulator P. lentiscus. CONCLUSIONS: Soil salinity around Pistacia trees critically affects the growth of S. inermis. Inhibition of S. inermis growth, but not germination, around the evergreen P. lentiscus, stems from the latter's ability to deplete salt from its surroundings. The results indicated that P. lentiscus is able to tolerate and accumulate salt, which we assume contributes to its wide distribution along the Mediterranean coast in Israel. Recommendations and perspectives While the phytoremediation potential of Salsola spp. has been explored to some extent, this of P. lentiscus has not been tested and proven before. The results suggest that the evergreen perennial salt-tolerant P. lentiscus can be recommended for horticulture purposes and soil stabilization in relatively saline environments.

[Effects of salt stress on germination and in vitro growth of pistachio (Pistacia vera L.)]. / Effet du stress salin sur la germination et la croissance in vitro du pistachier (Pistacia vera L.).

In order to study the salinity tolerance of pistachio (Pistacia vera L.), embryos developed from mature seeds were isolated and cultured in vitro and subjected to different NaCl concentrations (0, 42.8, 85.5, 171.1 and 256.6 mM) for 30 days. The results showed that in vitro germination of embryonic axes was not affected by the salt concentration. However, the germinated embryo survival rates decreased from 100% for the control to 62.9% for the highest salt concentration (256.6 mM). In addition, the plantlet growth (length of aerial and root parts, number of leaf produced per embryo, as well as the production of total fresh and dry matter for both aerial parts and roots) showed significant differences according the various salt concentrations.

Arsenic- and mercury-induced phytotoxicity in the Mediterranean shrubs Pistacia lentiscus and Tamarix gallica grown in hydroponic culture.

Hg and As resistance and bioaccumulation were studied in hydroponically grown Pistacia lentiscus and Tamarix gallica plants. Both elements caused growth inhibition in roots and shoots, with mercury showing greater phytotoxicity than arsenic. Accumulation of both elements by plants increased in response to element supply, with the greatest uptake found in T. gallica. Both elements affected P and Mn status in plants, reduced chlorophyll a concentration and increased MDA and thiol levels. These stress indices showed good correlations with As and Hg concentration in plant tissues, especially in the roots. Toxic responses to mercury were more evident than for arsenic, especially in shoot tissues. T. gallica showed higher resistance to both Hg and As than P. lentiscus, as well accumulating more As and Hg.

Responses to changes in Ca2+ supply in two Mediterranean evergreens, Phillyrea latifolia and Pistacia lentiscus, during salinity stress and subsequent relief.

BACKGROUND AND AIMS: Changes in root-zone Ca(2+) concentration affect a plant's performance under high salinity, an issue poorly investigated for Mediterranean xerophytes, which may suffer from transient root-zone salinity stress in calcareous soils. It was hypothesized that high-Ca(2+) supply may affect differentially the response to salinity stress of species differing in their strategy of Na(+) allocation at organ level. Phillyrea latifolia and Pistacia lentiscus, which have been reported to greatly differ for Na(+) uptake and transport rates to the leaves, were studied. Methods In plants exposed to 0 mM or 200 mM NaCl and supplied with 2.0 mM or 8.0 mM Ca(2+), under 100 % solar irradiance, measurements were conducted of (a) gas exchange, PSII photochemistry and plant growth; (b) water and ionic relations; (c) the activity of superoxide dismutase and the lipid peroxidation; and (d) the concentration of individual polyphenols. Gas exchange and plant growth were also estimated during a period of relief from salinity stress. Key Results The performance of Pistacia lentiscus decreased to a significantly smaller degree than that of Phillyrea latifolia because of high salinity. Ameliorative effects of high-Ca(2+) supply were more evident in Phillyrea latifolia than in Pistacia lentiscus. High-Ca(2+) reduced steeply the Na(+) transport to the leaves in salt-treated Phillyrea latifolia, and allowed a faster recovery of gas exchange and growth rates as compared with low-Ca(2+) plants, during the period of relief from salinity. Salt-induced biochemical adjustments, mostly devoted to counter salt-induced oxidative damage, were greater in Phillyrea latifolia than in Pistacia lentiscus. CONCLUSIONS: An increased Ca(2+) : Na(+) ratio may be of greater benefit for Phillyrea latifolia than for Pistacia lentiscus, as in the former, adaptive mechanisms to high root-zone salinity are primarily devoted to restrict the accumulation of potentially toxic ions in sensitive shoot organs.

Effects of ozone on the foliar histology of the mastic plant (Pistacia lentiscus L.).

An open-top chamber study was conducted to investigate the tissue and cellular-level foliar effects of ozone (O3) on a Mediterranean evergreen species, the mastic plant (Pistacia lentiscus L.). Plants were exposed at three different O3 levels, and leaf samples were collected periodically from the beginning of the exposure. Although no visible foliar injury was evident, alterations of the plastids and vacuoles in the mesophyll were observed. Senescence processes were accelerated with an anomalous stacking of tannin vacuoles, and a reduction in the size and number of the chloroplasts. Overall, most of the modifications induced by O3 were consistent with previously reported observations on deciduous broadleaf species, with the exception of alterations in the cells covering the secretory channels, reported here as a new finding. Comments on the feasibility of using microscopy to validate O3 related field observations and subtle foliar injury are also given.

Water sources and water-use efficiency in mediterranean coastal dune vegetation.

In coastal environments plants have to cope with various water sources: rainwater, water table, seawater, and mixtures. These are usually characterized by different isotopic signatures ( (18)O/ (16)O and D/H ratios). Xylem water reflects the isotopic compositions of the water sources. Additionally, water-use efficiency (WUE) can be assessed with carbon isotope discrimination (Delta) analyses. Gas exchange, Delta of leaf dry matter, and isotopic composition (delta (18)O) of xylem water were measured from June to August 2001 in herbaceous perennials of mobile dunes (Ammophila littoralis, Elymus farctus) and sclerophyllous shrubs and climbers (Arbutus unedo, Pistacia lentiscus, Phillyrea angustifolia, Qercus ilex, Juniperus oxycedrus, Smilax aspera) of consolidated dunes. Assimilation rates were rather low and did not show clear seasonal patterns, possibly due to limited precipitation and generally low values of stomatal conductance. The lowest values were shown in S. aspera. Different physiological patterns were found, on the basis of delta (18)O and Delta analyses. Values of delta (18)O of xylem water of phanerophytes were remarkably constant and matched those of the water table, indicating dependence on a reliable water source; values of Delta were relatively high, indicating low intrinsic WUE, with the exception of J. oxycedrus. Surprisingly, very high delta (18)O values were found for the xylem water from S. aspera in August. This suggests retrodiffusion of leaf water to xylem sap in the stem or direct uptake of water by leaves or stems, owing to dew or fog occurrence. Low Delta values indicated high WUE in S. aspera. Contrasting strategies were shown by the species of mobile dunes: E. farctus relied on superficial water and exhibited low WUE, accordingly to its therophyte-like vegetative cycle; on the contrary, A. littoralis used deeper water sources, showing higher WUE in relation to its long-lasting vegetative habit.