Small Amounts Of Plant Material Research Articles

Rayless goldenrod (Isocoma pluriflora) poisoning in chicks (Gallus gallusdomesticus)

Rayless goldenrod (Isocoma spp.) and white snakeroot (Ageratina altissima) poison livestock, wildlife and humans. The suggested toxin for both plants is tremetone, a mixture of benzofuran ketones. However, plant tremetone concentrations often do not correlate with poisoning, and they have not been identified in contaminated milk that poisons nursing neonates. This suggests there may be unidentified metabolites or toxins. Previous studies using various cell culture and large animal models have been inconsistent with varying animal response that often require large doses. The objective of this work is to document the toxicity of rayless goldenrod in California white leghorn chicks, a susceptible small animal model, that would require relatively small amounts of plant material or purified toxins. Four groups of 15 chicks were gavaged with finely ground I. pluriflora at rates of 0, 1%, 2% or 3% of their bodyweight per day for 7 days. After 7 exposure days the chicks were euthanized, necropsied and tissues were collected, fixed and examined microscopically. Myocyte damage was evaluated using clinical signs, weight gain, serum biochemical changes, and histologic lesions and scores. The 3% group had focally extensive myocyte degeneration and necrosis most severely affecting leg muscles (semitendinosus, iliotibialis, peroneus longus and gastrocnemius). This was supported by serum biochemical changes and reduced weight gains. These findings indicate young chicks are a sensitive model of toxicity that may be useful to better identify the rayless goldenrod and white snakeroot toxins, including those unidentified toxins of transmammary poisoning.

A new type of laboratory microdevice for distillation of lavender and herb raw materials

Abstract. Bulgaria is known for its essential oils and medicinal plants, which occupy increasing parts in the agricultural areas. In recent years, the boom in lavender production has made our country the largest producer of lavender oil in the world. This intensified the scientific work with it and led to the creation of new lavender varieties and lines that are used in production. All research and tests on an industrial scale go through the laboratory distillation of small amounts of plant material to determine the quantity and quality of essential oil. In laboratory practice, micro-apparatus for water distillation is widely used, which does not reflect the real results for the yield and quality of the product. This necessitated the creation of a new laboratory device for determining the content of essential oil in raw materials with labile (rich in esters) oil by the application of steam distillation. The design of this apparatus mirrors industrial installations, thus the results it produces accurately simulate those of industrial production. Tests with herbal raw materials, like Lavandula angustifolia Mill., Salva sclarea L. and Helichrysum italicum (Roth) G. Don showed that the model is effective and applicable. The oils obtained by the new apparatus retained an ester content of 40.2%, 75.0% and 4.8%, respectively; these figures represent a significant improvement over previous methods.

Bioprospecting of Essential Oil-Bearing Plants: Rapid Screening of Volatile Organic Compounds Using Headspace Bubble-in-Drop Single-Drop Microextraction for Gas Chromatography Analysis.

Essential oils are vital constituents of oil-bearing plants. However, their screening still demands harvesting of the plant for laboratory analysis. We report herein a simple, rapid and robust headspace bubble-in-drop microextraction screening technique (BID-SPME) requiring only small amounts of plant material. The optimised method uses 0.5 g of the crushed plant leaves sample obtained in a 2 mL capped chromatography vial, heated to 55 °C and sampled with 2 µL heptadecane in a Hamilton gastight syringe equilibrated for 15 min exposed to the headspace volume. The method was applied to three plants, Pinus radiata, Tagetes minuta and Artemisia afra, which are known for their essential oil content. The method was able to extract at least 80% of the oil constituents in such abundance that they could be easily annotated using the gas chromatography–mass spectrometry (GC–MS) mass spectral libraries. The major volatile organic compounds (VOCs) detected included tagetone, terpinen-4-ol, ocimenone, caryophyllene, dihydrotagetone, terpinolene and artemisia ketone, just to mention a few, at different concentrations in different plants. Importantly, these annotated VOCs were also reported in other studies in the same and even different plants, extracted using normal steam distillation and importantly those reported in the literature for different extraction techniques.

One prep to catch them all: “2 in 1”, an efficient method for the simultaneous extraction of DNA and RNA from Grapevine tissues

Recent advances in our understanding of plant physiology and adaptation to the environment are tightly related to the development of ‘omics’ technologies such as metabolomics, transcriptomics, genomics and epigenomics that allow a more comprehensive view of the plant functioning. In this context, the ability to extract DNA and RNA from small amounts of plant material can be a limiting factor, worse in the case of non-model plants for which efficient nucleic extraction procedures are lacking. In the case of grapevine, extraction of high-quality DNA is typically limited by the high polyphenolic and polysaccharide contents of the different tissues. Here, we propose an adaptation of the method of Reid et al. (2006) that allows the simultaneous and efficient extraction of DNA and RNA from grapevine vegetative and berry tissues from in vitro grown grapevine plants and cells and from other plants. The protocol allows the extraction of high-quality RNA and DNA for standard molecular biology methods as well as for Next Generation Sequencing (NGS). It also works with a limited amount of plant material, such as young developing buds, and provides the means to analyse “omics” data from a single plant sample.

Rapid Affinity Purification of Tagged Plant Mitochondria (Mito-AP).

This protocol describes the isolation of mitochondria by affinity chromatography using magnetic beads coated with Strep-Tactin in a timeframe of ca. 30min. Compared to a classic differential and density gradient centrifugation this protocol enables a more rapid and efficient isolation of mitochondria even with small amounts of plant material. Transgenic plants with mitochondria that are decorated with a protein that is integrated into the outer mitochondrial membrane and fused to a green fluorescent protein (GFP) and a TwinStrep-tag facing the cytosol. This tag can bind to Strep-Tactin coated magnetic beads. Isolated mitochondria still bound to magnetic beads are uniquely suited for measuring oxygen consumption rates since this measurement needs mitochondria to be immobilized on the bottom of the measuring well. Furthermore, the isolated mitochondria can be used for downstream applications such as proteomics and metabolomics. This technique also allows for the isolation of mitochondria from specific cell types and tissues by altering the expression of the protein decorating the mitochondria.

A Rapid Method for Quantifying RNA and Phytohormones From a Small Amount of Plant Tissue.

Phytohormones are involved in most plant physiological processes and the quantification of endogenous phytohormone levels and related gene expressions is an important approach to studying phytohormone functions. However, the quantification of phytohormones is still challenging due to their extremely low endogenous level in plant tissues and their high chemical diversity. Therefore, developing a method to simultaneously quantify phytohormone levels and RNA would strongly facilitate comparative analyses of phytohormones and gene expression. The present work reports a convenient extraction protocol enabling multivariate analysis of phytohormones and RNA from small amounts of plant material (around 10 mg). This high-throughput ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method demonstrates quantification of phytohormones and their related metabolites from four plant hormone classes: cytokinin, auxin, abscisic acid, and gibberellin. The UPLC-MS/MS method can quantify thirteen phytohormones and their metabolites simultaneously in 14 min. To validate the developed method, we determined the dynamic profiles of phytohormones and gene expressions in small axillary shoot buds in garden pea. This new method is applicable to quantification analysis of gene expression and multiple phytohormone classes in small amounts of plant materials. The results obtained using this method in axillary buds provide a basis for understanding the phytohormone functions in shoot branching regulation.

Quantification and Localization of Formylated Phloroglucinol Compounds (FPCs) in Eucalyptus Species.

The Eucalyptus genus is a hyper-diverse group of long-lived trees from the Myrtaceae family, consisting of more than 700 species. Eucalyptus are widely distributed across their native Australian landscape and are the most widely planted hardwood forest trees in the world. The ecological and economic success of Eucalyptus trees is due, in part, to their ability to produce a plethora of specialized metabolites, which moderate abiotic and biotic interactions. Formylated phloroglucinol compounds (FPCs) are an important class of specialized metabolites in the Myrtaceae family, particularly abundant in Eucalyptus. FPCs are mono- to tetra-formylated phloroglucinol based derivatives, often with an attached terpene moiety. These compounds provide chemical defense against herbivory and display various bioactivities of pharmaceutical relevance. Despite their ecological and economic importance, and continued improvements into analytical techniques, FPCs have proved challenging to study. Here we present a simple and reliable method for FPCs extraction, identification and quantification by UHPLC-DAD-ESI-Q-TOF-MS/MS. The method was applied to leaf, flower bud, and flower samples of nine different eucalypt species, using a small amount of plant material. Authentic analytical standards were used to provide high resolution mass spectra and fragmentation patterns. A robust method provides opportunities for future investigations into the identification and quantification of FPCs in complex biological samples with high confidence. Furthermore, we present for the first time the tissue-based localization of FPCs in stem, leaf, and flower bud of Eucalyptus species measured by mass spectrometry imaging, providing important information for biosynthetic pathway discovery studies and for understanding the role of those compounds in planta.

The use of micro-energy dispersive X-ray fluorescence spectrometry combined with a multivariate approach to determine element variation and distribution in tobacco seedlings exposed to arsenate

Here, we present a new scheme of analysis combining micro-energy dispersive X-ray fluorescence spectrometry (μ-XRF) with a multivariate approach that allows to establish the inter-correlation of multiple elements and their elemental map in plants. The main advantage of this procedure is that XRF spectral profiles can be analysed directly, by means of principal component analysis (PCA), allowing a quick interpretation of the results. Furthermore, this analysis requires small amounts of plant material and can be performed in whole individual seedlings in the hydrated state without chemical extraction. With this technology, we determined the distribution of arsenic (As) and the variation and spatial distribution of multiple elements in whole tobacco seedlings grown in the presence of different arsenate concentrations. We observed that As is detectable mainly in roots, primarily in the basal part, but also in the root apex in seedlings grown in the highest arsenate concentration. The low rate of As translocation from roots to shoots and the significant increase in S are consistent with previous evidence showing that As is retained in roots by forming complexes with thiol peptides. We also found a significant non-linear increase in P, as arsenate is taken up by phosphate transporters and induces the expression of genes encoding them. A decrease in Mn, Fe and Zn proportional to the accumulation of As and in the same tissues, suggests a competition of these elements with As for cellular transporters.

Lily poisoning

Lilies, that is, species of Lilium (Liliaceae) (true lily) and Hemerocallis (Hemerocallidaceae) (day lily), cause acute kidney injury in cats. This does not occur in dogs, rats or rabbits. All parts of the plant are toxic to cats and a small amount of plant material can have serious consequences if ingested. The mechanism of toxicity remains unknown. Signs develop rapidly, with gastrointestinal irritation followed by polyuria, dehydration and then renal failure. Seizures can occur in severe cases. Treatment is aimed at reducing absorption using emesis and/or activated charcoal; preventing renal shutdown; and enhancing renal perfusion with intravenous fluid diuresis for at least 48 hours. Once renal failure has occurred treatment options are limited. Prognosis is good in cats treated promptly and before the onset of acute kidney injury. Cats with renal failure have a more guarded prognosis.

Outstanding Anti-inflammatory Potential of Selected Asteraceae Species through the Potent Dual Inhibition of Cyclooxygenase-1 and 5-Lipoxygenase.

Cyclooxygenase and 5-lipoxygenase are enzymes that catalyze important inflammatory pathways, suggesting that dual cyclooxygenase/lipoxygenase inhibitors should be more efficacious as anti-inflammatory medicines with lower side effects than the currently available nonsteroidal anti-inflammatory drugs. Many plants from the family Asteraceae have anti-inflammatory activities, which could be exerted by inhibiting the cyclooxygenase-1 and 5-lipoxygenase enzymes. Nevertheless, only a small number of compounds from this family have been directly evaluated for their ability to inhibit the enzymes in cell-free assays. Therefore, this study systematically evaluated 57 Asteraceae extracts in vitro in enzyme activity experiments to determine whether any of these extracts exhibit dual inhibition of cyclooxygenase-1 and 5-lipoxygenase. The chemical profiles of the extracts were obtained by the high-performance liquid chromatography-ultraviolet-diode array detector method, and their major constituents were dereplicated. Of the 57 tested extracts, 13 (26.6 %, IC50 range from 0.03-36.2 µg/mL) of them displayed dual inhibition. Extracts from known anti-inflammatory herbs, food plants, and previously uninvestigated species are among the most active. Additionally, the extract action was found to be specific with IC50 values close to or below those of the standard inhibitors. Thus, the active extracts and active substances of these species are potent inhibitors acting through the mechanism of dual inhibition of cyclooxygenase-1 and 5-lipoxygenase. The extracts were prepared for this study using nontoxic extraction solvents (EtOH-H2O), requiring only a small amount of plant material to carry out the bioassays and the phytochemical analyses. In summary, this study demonstrated the potential of the investigated species as dual inhibitors, revealing their potential as pharmaceuticals or nutraceuticals.

Simultaneous determination of plant hormones in peach based on dispersive liquid-liquid microextraction coupled with liquid chromatography-ion trap mass spectrometry.

Fruit development is influenced greatly by endogenous hormones including salicylic acid (SA) and abscisic acid (ABA). Mass spectrometry with high sensitivity has become a routine technology to analyze hormones. However, pretreatment of plant samples remains a difficult problem. Thus, dispersive liquid-liquid microextraction (DLLME) was used to concentrate trace plant hormones before liquid chromatography-ion trap mass spectrometry (LC-ITMS) analysis. Standard curves were linear within the ranges of 0.5-50, 0.2-20ng/mL for SA and ABA, respectively. The correlation coefficients were greater than 0.9995 with recoveries above 87.5%. The limits of detection were 0.2ng/mL for SA and 0.1ng/mL for ABA in spiked water solution, respectively (injection 20μL). The successful analysis of SA and ABA in fruit samples indicated our DLLME-LC-ITMS approach was efficient, allowing reliable quantification of both two compounds from very small amounts of plant material. Moreover, this research revealed the relationship between SA and ABA content and development of peach fruit at different growth stages.

A new method to isolate total dsRNA.

When a diseased plant is suspected to be infected with unknown viruses, the approach of isolating double-stranded RNA (dsRNA) from diseased tissues and analyzing the sequence has been useful for detecting the viruses. This procedure owes its success to the majority of plant pathogenic viruses being RNA viruses, which accumulate dsRNAs as copies of their genome or as a replicative intermediate in infected cells. Conventional dsRNA isolation methods (e.g., chromatography using CF-11 cellulose) require a significant amount of plant material and are laborious and time consuming. Therefore, it has been impractical to isolate dsRNA from many samples at the same time. To overcome these problems, we developed a novel dsRNA isolation method involving a recombinant dsRNA-binding protein. Using this method, we can readily isolate viral dsRNA from a small amount of plant material, and can process numerous samples simultaneously. Purified dsRNA can be used as a template for cDNA synthesis and sequencing, enabling detection of both known and unknown viruses.

Simplification of a haemolytic micromethod for toxic saponin quantification in alfalfa

A simplification of a haemolytic micromethod is presented. In the orginal method, alfalfa (<i>Medicago media</i> Pers.) leaf sap is spotted on the plate covered with a blood-gelatine suspension. In the presented method, the mashed alfalfa pulp is used instead of sap. Due to saponin diffusion and the reaction with erythrocytes, a haemolytic ring appears, which has a width proportional to the concentration of toxic saponins. It is shown that the width of haemolytic ring does not depend on the sample weight ranging from 20 to 100 mg. This allows for the omission of laborious sap pressing and sample quantification. Individual alfalfa plants with different saponin contents were tested using leaf sap and leaf pulp for analyses. Good agreement was found with sap and leaf pulp methods. The correlation obtained by both methods was high, <i>r</i> = 0.87. The modified method requires only a small amount of plant material and makes the analyses of large numbers of individual plants per day possible. The method is especially recommended for breeding purposes.

Feasibility of using atmospheric pressure matrix-assisted laser desorption/ionization with ion trap mass spectrometry in the analysis of acetylated xylooligosaccharides derived from hardwoods and Arabidopsis thaliana

The atmospheric pressure matrix-assisted laser desorption/ionization with ion trap mass spectrometry (AP-MALDI-ITMS) was investigated for its ability to analyse plant-derived oligosaccharides. The AP-MALDI-ITMS was able to detect xylooligosaccharides (XOS) with chain length of up to ten xylopyranosyl residues. Though the conventional MALDI-time-of-flight/mass spectrometry (TOF/MS) showed better sensitivity at higher mass range (>m/z 2,000), the AP-MALDI-ITMS seems to be more suitable for detection of acetylated XOS, and the measurement also corresponded better than the MALDI-TOF/MS analysis to the actual compositions of the pentose- and hexose-derived oligosaccharides in a complex sample. The structures of two isomeric aldotetrauronic acids and a mixture of acidic XOS were elucidated by AP-MALDI-ITMS using multi-stages mass fragmentation up to MS(3). Thus, the AP-MALDI-ITMS demonstrated an advantage in determining both mass and structures of plant-derived oligosaccharides. In addition, the method of combining the direct endo-1,4-β-D-xylanase hydrolysis of plant material, and then followed by AP-MALDI-ITMS detection, was shown to recognize the substitution variations of glucuronoxylans in hardwood species and in Arabidopsis thaliana. To our knowledge, this is the first report to demonstrate the acetylation of glucuronoxylan in A. thaliana. The method, which requires only a small amount of plant material, such as 1 to 5 mg for the A. thaliana stem material, can be applied as a high throughput fingerprinting tool for the fast comparison of glucuronoxylan structures among plant species or transformants that result from in vivo cell wall modification.

Rapid and sensitive hormonal profiling of complex plant samples by liquid chromatography coupled to electrospray ionization tandem mass spectrometry.

BackgroundPlant hormones play a pivotal role in several physiological processes during a plant's life cycle, from germination to senescence, and the determination of endogenous concentrations of hormones is essential to elucidate the role of a particular hormone in any physiological process. Availability of a sensitive and rapid method to quantify multiple classes of hormones simultaneously will greatly facilitate the investigation of signaling networks in controlling specific developmental pathways and physiological responses. Due to the presence of hormones at very low concentrations in plant tissues (10-9 M to 10-6 M) and their different chemistries, the development of a high-throughput and comprehensive method for the determination of hormones is challenging.ResultsThe present work reports a rapid, specific and sensitive method using ultrahigh-performance liquid chromatography coupled to electrospray ionization tandem spectrometry (UPLC/ESI-MS/MS) to analyze quantitatively the major hormones found in plant tissues within six minutes, including auxins, cytokinins, gibberellins, abscisic acid, 1-amino-cyclopropane-1-carboxyic acid (the ethylene precursor), jasmonic acid and salicylic acid. Sample preparation, extraction procedures and UPLC-MS/MS conditions were optimized for the determination of all plant hormones and are summarized in a schematic extraction diagram for the analysis of small amounts of plant material without time-consuming additional steps such as purification, sample drying or re-suspension.ConclusionsThis new method is applicable to the analysis of dynamic changes in endogenous concentrations of hormones to study plant developmental processes or plant responses to biotic and abiotic stresses in complex tissues. An example is shown in which a hormone profiling is obtained from leaves of plants exposed to salt stress in the aromatic plant, Rosmarinus officinalis.

Product quality and energy consumption when drying peppermint by rotary drum dryer

Peppermint is one of the most important aromatic plants because it has wide range of industrial uses and is widely accepted by the public for its flavoring and pharmaceutical properties. Hot air or natural drying of medicinal and aromatic plants, including peppermint, is a common postharvest operation. Drying the product reduces biochemical and microbiological degradation. However, many studies in the literature report the results of drying only a small amount of plant material (less than 1 kg) and do not consider the performances of industrial scale dryers in terms of their effect on product quality, energy consumption and throughput. A rotary drum dryer was used to dry 15 kg of fresh chopped peppermint plant. The rotary drum dryer was programmed to operate under two different drying schemes: constant air temperature drying and rectangular wave-shaped drying air temperature profiles. The drying durations were 15–18 h and 12–15 h for constant temperature profile and rectangular wave-shaped temperature profiles, respectively. Hot air drying caused considerable darkening of the dried peppermint leaves. Essential oil contents of peppermint samples dried by the two different drying procedures were relatively unaffected by drying scheme (2.08–2.7 mL 100 g dry matter −1). The menthol content of the leaves increased from 32.52% (fresh) up to 44.52% (dried), their menthone content decreased from 24.85% (fresh) down to 9.07% (dried). However, the variations within replications were relatively high in terms of the menthol (25.39–44.52%) and menthone (9.07–18.49%) contents of the dried leaves. These variations might have been caused by the uncontrolled morphological and physiological changes of the fresh peppermint materials associated with their maturity. These variations are the main obstacle to obtaining reproducible results and therefore dried peppermint products with consistent quality. The specific energy consumption values ranged from 7.88 to 15.08 MJ (kg water removed) −1. Daily fluctuations in ambient air conditions directly affected the specific energy consumption of rotary drum dryer.

A New Approach to Reconstruct Paleogene Atmospheric Hydrology at High Latitudes

Reconstruction of ancient atmospheric conditions through the analysis of precipitation patterns is a novel endeavor in the study of paleoclimate. A new approach is now available for a quantitative reconstruction of Paleogene atmospheric hydrological conditions in High Arctic. It is based on 1) the discovery of exceptionally-preserved Paleogene plant fossils from the Canadian Arctic which yielded in situ labile biomolecules; 2) the development of compound-specific hydrogen isotope analysis which can be applied to small amount of plant material; and 3) taxon-specific apparent hydrogen isotope fractionation factors obtained from empirical measurement. A new moisture recycling model is established to explain the reconstructed paleohydrologic pattern in the High Arctic during Paleogene.

Maximum yields of microsomal-type membranes from small amounts of plant material without requiring ultracentrifugation

Isolation of a microsomal membrane fraction is a common procedure in studies involving membrane proteins. By conventional definition, microsomal membranes are collected by centrifugation of a postmitochondrial fraction at 100,000g in an ultracentrifuge, a method originally developed for large amounts of mammalian tissue. We present a method for isolating microsomal-type membranes from small amounts of Arabidopsis thaliana plant material that does not rely on ultracentrifugation but instead uses the lower relative centrifugal force (21,000g) of a microcentrifuge. We show that the 21,000g pellet is equivalent to that obtained at 100,000g and that it contains all of the membrane fractions expected in a conventional microsomal fraction. Our method incorporates specific manipulation of sample density throughout the procedure, with minimal preclearance, minimal volumes of extraction buffer, and minimal sedimentation pathlength. These features allow maximal membrane yields, enabling membrane isolation from limited amounts of material. We further demonstrate that conventional ultracentrifuge-based protocols give submaximal yields due to losses during early stages of the procedure; that is, extensive amounts of microsomal-type membranes can sediment prematurely during the typical preclearance steps. Our protocol avoids such losses, thereby ensuring maximal yield and a representative total membrane fraction. The principles of our method can be adapted for nonplant material.

Analysis of metabolic variation and galanthamine content in Narcissus bulbs by 1H NMR

Galanthamine is a benzazepine alkaloid used as a drug to relieve symptoms of Alzheimer's disease. For pharmaceutical use this natural product has been extracted from the plant Leucojum aestivum (Amaryllidaceae) or produced synthetically. Limited supply of the natural source and high cost of synthetic production has led to a search for alternative sources of galanthamine. The bulbs of Narcissus pseudonarcissus (Amaryllidaceae) have been identified as a potential source of raw material for galanthamine extraction. Since inconsistent chemical composition can be an issue with medicinal plant material, it is of interest to know whether large variations occur between Narcissus bulbs grown in different geographical locations. To evaluate whether large differences exist in the overall metabolic profiles of Narcissus bulbs grown in the two most important cultivation regions. (1)H NMR and principal component analysis were used for an unbiased comparison of the bulb samples. Overall metabolite profiles were quite similar, but galanthamine levels could slightly discriminate samples by geographical region. (1)H NMR was used for quantitation of galanthamine, and was found to be comparable to quantitation by HPLC. Compared with conventional chromatographic methods, sample preparation for (1)H NMR analysis is simple and rapid, and only a small amount of plant material is required. Since useful qualitative and quantitative information about the metabolic state of Narcissus bulbs can be obtained by (1)H NMR, this method is useful for agricultural applications, and for quality control of raw material used in the pharmaceutical industry.

Is xylem cavitation resistance a relevant criterion for screening drought resistance among Prunus species?

Fruit trees are likely to suffer from the effects of severe drought in the future; however, sound criteria for evaluating the species' ability to survive these extreme conditions are largely missing. Here, we evaluated the feasibility of using xylem cavitation resistance as a tool for screening Prunus species for drought resistance. Ten different Prunus species were selected to cover a large range of water requirements, from hydrophilic to xerophilic types. Shoot cavitation resistance was evaluated with the new Cavitron technique. At this inter-specific level, cavitation resistance was related to species drought resistance, with xerophilic species being less vulnerable to cavitation. The Cavitron technique enabled species characterization that required a short time and small amounts of plant material. This technique could be used to evaluate the drought resistance of a limited number of fruit tree genotypes. Genotype screening on a larger scale, however, would likely require another approach. Out of a number of anatomical traits tested, a significant correlation was found between cavitation resistance and inter-vessel wall thickness across species. This anatomical trait is, therefore, suggested as a possible alternative to direct cavitation estimates and could be included as a screening criterion in breeding programs for drought resistance of Prunus genotypes.