Pecan Research Articles

Evaluation of Natural Spring Freeze Tolerance of Five Pecan Scion–Rootstock Combinations

Pecan [Carya illinoinensis (Wangenh.) K. Koch] is a member of the Juglandaceae family. During spring, pecan trees break their bud dormancy and produce new leaves and flowers. Carbohydrates stored in roots and shoots are thought to support the bloom and early vegetative growth during this time until new leaves start the full photosynthetic activity. Spring freeze is known for its damaging effects on pecan bud and flower growth and development. Pecan shoots with leaves and flowers from five scion–rootstock combinations were collected hours before and after a recent spring freeze (below 0 °C for 6 hours, 21 Apr 2021, Perkins, OK, USA). Morphologies of the leaf, bud, and catkin were visually observed, and the morphologies of the anther and pollen in paraffin sections were investigated by light microscopy. Soluble sugar and starch from bark and wood were analyzed using the anthrone reagent method. The Kanza–Mount showed the maximum damage to terminal leaves, buds, and catkins, whereas Maramec–Colby had the minimum damage only to leaves. Pollen grains were shrunk and reduced in number in the anthers in the protandrous Pawnee scions, whereas no pollen damage was observed in the protogynous Kanza scion. Furthermore, bark soluble sugar levels increased in all the scion–rootstock combinations after the freeze, which may indicate a physiological response to the cold stress. Overall, the extent of spring freeze damage of pecans is affected by the growth stage, types of scion and rootstock, and the scion–rootstock interactions. Furthermore, in addition to low temperature, scion–rootstock interactions also affected the starch and soluble sugar contents in wood and bark tissues.

Development and validation of sensitive and rapid immunoassays to detect minute amounts of hazelnut in processed food and working surfaces

Hazelnut (Corylus avellana L.) represents one of the most allergenic nuts and it can be found as a hidden allergen in processed food due to cross contamination. Therefore, sensitive and specific analytical techniques are in high demand to be used in allergen risk management plans at food industry. In this study, sandwich ELISA and Lateral Flow Immunoassay (LFIA) to detect hazelnut have been developed based on the determination of Cor a 9, one of the most abundant and allergenic proteins of hazelnut. Results showed that cross-reactivity was only found with walnut and Pecan nut, which was lower than 0.1%. When analyzing food spiked with a hazelnut extract or blended with hazelnut flour, ELISA and LFIA were able to detect 0.1 ppm and 0.5 ppm of hazelnut protein with a recovery from 82 to 110%. ELISA and LFIA could also detect 0.15 and 0.6 ppm of hazelnut protein in baked cookies incurred with ground hazelnut, respectively. Furthermore, LFIA could detect 1.25 μg of hazelnut protein in working surfaces of stainless steel and melamine. The sandwich ELISA was in-house validated, showing acceptable results of precision. Likewise, ELISA and LFIA showed to be robust tests. The combined use of both assays could improve the allergen risk management plans in food industry to monitor the presence of hazelnut traces in raw ingredients, processed food and working surfaces.

Targeted metabolomics reveals key phenolic changes in pecan nut quality deterioration under different storage conditions.

Pecan nuts are highly enriched in phenolic compounds, which contribute to the health benefits of pecans. Phenolic compounds represent the main oxidation reaction substrates, thus leading to quality deterioration, namely pellicle browning or a decrease in beneficial effects during pecan storage. Hence, four different storage conditions were performed for 180 d to simulate real production situations. Targeted metabolomics was chosen to identify the specific phenolic compounds involved in quality deterioration under different storage conditions in 0, 90, and 180 d samples. A total of 118 phenolic compounds were detected, nine of which were identified for the first time in pecan. The total phenolic content (TPC) and antioxidant capacities initially demonstrated high scores, after which they tended to decrease during the storage process. The significantly modified phenolic compounds during storage were selected as the metabolite markers of pecan quality deterioration, including catechin, procyanidin (PA) trimer, PA tetramer, trigalloyl hexahydroxydiphenoyl (HHDP) glucose, and tetragalloyl hexoside. Fresh pecan kernels resulted in more pronounced changes in hydrolysable tannins (HTs), whereas dry kernels resulted in the most accentuated changes in condensed tannins (CTs). To the best of our knowledge, this is the first attempt to study individual phenolic changes during storage of pecan in such massive amounts. The results can offer a valuable theoretical basis for future control of pecan quality deterioration through phenolics during storage.

Do Nonstructural Carbohydrates Contribute to Pecan (Carya illinoinensis) Secondary Budburst?

Spring freeze is among the problems threatening pecan bloom and production. Pecan tree height and structure make them difficult to protect from spring freezes. Some cultivars can compensate because the secondary buds can produce healthy flowers if the primary buds freeze. The mechanism that precipitates secondary budbreak is unknown. Our results show a correlation between successful secondary budbreak and 1-year-old shoot carbohydrate levels. ‘Kanza’ and ‘Pawnee’, with the higher secondary budburst, also had higher carbohydrate levels than ‘Maramec’. This suggests higher carbohydrate levels in the bud-bearing 1-year-old shoots promote successful secondary bud burst after spring freeze destruction of the primary buds.

Random amplified microsatellites (RAMS) analysis ascertains genetic variation of Alternaria alternata causing black spot disease on Carya illinoinensis in South Africa.

Limited information regarding the occurrence of black spot disease of pecan (Carya illinoinensis), caused by A. alternata, in South Africa is known. The pecan industry is growing rapidly, so it is essential to understand the impact of the fungal pathogen to pecan health. In this study, the genetic variation of 364 A. alternata isolates was investigated by two RAMS primers (CCA5 and CGA5). In total, 6,525 alleles were produced, with a minimum of 3,182 alleles on the CGA5 primer and maximum of 3,343 alleles for CCA5 primer. Further analysis of the primers showed relatively low genetic diversity of A. alternata isolate populations, with mean values; (H = 0.12) and Shannon's information index (I = 0.20). The analysis of molecular variance (AMOVA) revealed significant differences between populations, with 88% of the genetic variation was found within populations (Nm = 3.59, PhiPT = 0.12), and were not significantly different (p > 0.001). While 12% variation was observed among populations (Nm = 2.89, PhiPT = 0.08) and the estimates were statistically significant (p < 0.001). STRUCTURE HARVESTER output showed that K value is K = 8, where ΔK cannot find the true number of populations because of less variation. The dendrogram cluster tree generated by Ward's analysis unveiled two main distinct clades and 10 sub-clades, revealing similar findings as those of PCoA analysis clusters. Therefore, it was evident that these analyses depicted no distinct relationship between the A. alternata isolates and their geographic locations or the prevalence of distribution among the populations.

Quantitative determination of specialised metabolites in different parts of Juglans regia Linn. and Carya illinoinensis (Wangenh.) K. Koch by using UHPLC-DAD-QTOF-MS/MS.

Juglans regia Linn. and Carya illinoinensis (Wangenh.) K. Koch are nut-producing plant species of the Juglandaceae family. Bioactive compounds like naphthoquinones, tetralones, and diarylheptanoids are dominant in these species. The study aimed to develop and validate a fast and sensitive analytical method by ultrahigh-performance liquid chromatography diode array detection mass spectrometry (UHPLC-DAD-MS) for quantification and identification of bioactive compounds in fruit pericarps and leaves of J.regia and C.illinoinensis collected from two different states of north India. The dried pericarps of J.regia and C.illinoinensis (500 mg) were extracted with ethyl acetate-methanol (50:50 v/v, 20 mL, 50°C, 30 min) by ultrasonication and analysed by ultrahigh-performance liquid chromatography diode array detection quadrupole time-of-flight tandem mass spectrometry (UHPLC-DAD-QTOF-MS/MS) for qualitative and quantitative examination of phytoconstituents. The method was validated according to International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human use (ICH) guidelines for linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ). Here, we report the quantification of dihydrophaseic acid (1), 4,5-dihydroxytetralone (2), 4,8-dihydroxytetralone (3), 5,8-dihydroxy-3-methoxytetralone (4), and juglanin A (5) in the pericarps and leaves of J.regia and C.illinoinensis. Furthermore, using the hyphenated analytical method, a total of 62 compounds were tentatively characterised in different samples. The principal component analysis (PCA) showed diversity between the analysed sample's composition. Also, the study evaluated the variation of bioactive compounds among different parts of J.regia and C.illinoinensis collected from different regions of northern India by UHPLC-DAD-QTOF-MS/MS. The developed method is simple, rapid, and selective for the identification and characterisation of bioactive molecules.

Physiological Perspective of Starch as a Carbon Source in Two Varieties of Carya illinoinensis Koch in Northern Mexico

Abstract The study took as a purpose to determine the Total Carbon (TC) content in the biomass, the Starch Carbon fraction (SC) and its annual dynamics in the biomass of perennial organs (stem and root) in adult trees of two of walnut tree (Carya illinoinensis). Four adult Western and Wichita tree stem and root samples were carried out monthly for a whole year. The TC was determined with an elemental analyser and the SC has gotten based on the molecular mass of the glucose (0.40 gC/gGlucose). t-Student test was performed between varieties per organ for the comparison of TC and SC through the program SPSS 15.0 with a significance of p ≤ 0.05. The results in Western variety were 160.02 kg TC and 4.90 kg SC to 7.54 kg SC in the stem; 64.58 kg TC and 1.74 kg SC to 3.09 kg SC in the root; in Wichita variety were presented 119.72 kg TC and 4.49 kg SC to 6.83 kg SC in the stem; 45.72 kg TC and 1.35 kg SC to 2.75 kg SC in the root. The root was the organ where the greatest amount of SC was stored in relation to the stem, due this latter constitutes a transport organ. Temperature has a marked inversely proportional influence on the accumulation of SC in both varieties. Global solar radiation and solar radiation proportionally influence the accumulation of SC.

Purification of pecan shell liquid smoke using the distillation method to separate polycyclic aromatic hydrocarbons (PAHs)

Pecan (Carya illinoinensis) is one of the macadamia nuts that is used as a source of oil and spices. Its shell is an organic waste that is not utilized optimally. Pecan shells can be the raw material to produce liquid smoke to minimize waste. Liquid smoke obtained from the pyrolysis pecan shell contains antibacterial and antioxidant compounds. Liquid smoke purification is performed to remove polycyclic aromatic hydrocarbons (PAHs) and tar to obtain clear yellow liquid smoke. The purification is carried out by a distillation process with temperature variations of 150 °C, 175 °C, and 200 °C. The distillation process time is varied by 30, 60, and 90 minutes. The phenol, acid, and carbonyl components determine the characteristics of pecan shell liquid smoke. At 175°C and 90 minutes, the best phenol content was 15.53%. Acid content of 7.65% was obtained at 175°C and 90 minutes. While the carbonyl content of 14.44% was obtained at 125°C for 60 minutes. Determination of liquid smoke quality by utilizing the distillation method produces grade 1 liquid smoke at 175°C for 90 minutes, grade 2 at 200°C for 30 minutes, and grade 3 at 200°C for 90 minutes.

Transfer of generic Escherichia coli and attenuated Salmonella enterica Typhimurium from the soil to the surface of in-shell pecans during harvest

During harvest pecan nuts are at risk of contamination with foodborne pathogens from extended contact with the ground. The objective of this study was to determine the potential transfer of Escherichia coli and Salmonella from the ground to in-shell pecans during the harvesting process. Plots (2 m2) were sprayed with 1 L of a rifampicin (rif) resistant strain of either E. coli TVS 353 or an attenuated Salmonella Typhimurium inoculum at a low (∼4 log CFU/ml), mid (∼6 log CFU/ml) or high (∼8 log CFU/ml) concentrations. The following day, nuts were mechanically harvested and samples from each plot were collected at 1 min, 4 h, and 24 h. Samples were enumerated for Salmonella and E. coli on tryptic soy agar supplemented with rif. The Salmonella levels in the soil from the inoculated plots were 2.0 ± 0.3, 4.1 ± 0.1, and 6.4 ± 0.2 log CFU/g for the low, mid, and high inocula, respectively. The E. coli levels in the soil from the inoculated plots were 1.5 ± 0.4, 3.7 ± 0.3, and 5.8 ± 0.1 log CFU/g for the low, mid, and high inocula, respectively. There was a significant difference in the average daily rainfall among the three trials. Trial 3 received 23.8 ± 9.2 cm, while trials 1 and 2 received much less (0.1 ± 0.1 0.0 ± 0.0 cm, respectively). Inoculation concentration and trial were significant (P<0.05) factors that influenced the transfer of E. coli and Salmonella to pecans. For the high inoculum treatment, bacterial transfer to pecans ranged from 0.7 ± 0.3 to 4.1 ± 0.2 for E. coli and 1.3 ± 0.7 to 4.3 ± 0.4 log CFU/g for Salmonella. For the medium inoculum treatment, transfer ranged from <0.3 to 1.5 ± 0.1 for E. coli and <0.3 to 1.9 ± 0.2 log CFU/g for Salmonella. For the low treatment, transfer ranged from <0.3 to 0.4 ± 0.2 and <0.3 to 0.5 ± 0.1 log CFU/g for E. coli and Salmonella, respectively. These results show the need for implementing agricultural practices that prevent potential transfer of foodborne pathogens onto the surface of in-shell pecans during harvest.

First Report of Diaporthe pseudophoenicicola causing Leaf Blight on Pecan in China.

Pecan (Carya illinoinensis) is one of the important economic forest crops widely cultivated in Jiangsu Provinces, China. From August to September in both 2021 and 2022, a foliar blight was observed in 7-ha and 6-ha pecan orchards in Changzhou (31°58'9.6″ N, 119°48'33.84″ E), and Jurong (31°52'15.46″ N, 119°9'24.62″ E), Jiangsu Province. The disease severity was about 32% with 8% incidence on 120 surveyed trees of the two orchards. Typical symptoms were lesions with a dark-brown color, which later became brown. We collected eighteen pecan leaves with typical symptoms in the surveyed pecan orchards and took them back to the laboratory for identification. Small fragments (approximately 9 mm2) from the necrotic borders of infected leaves were surfaced sterilized, plated on potato dextrose agar (PDA) and then incubated in darkness at 25°C. Pure cultures were obtained by single-spore culture. Thirty-three isolates with similar characteristics were obtained from the infected leaves (isolation frequency 85%), and the colonies surface on PDA was ochreous with patchs of olivaceous-yellow and sparse aerial mycelium. Observing from the back of the plate, the colonies were cream-yellow. Two types of single-cell conidia were produced on PDA. Alpha-conidia were 7.4 (range, 5.9 to 8.8) × 2.1 (range, 1.6 to 2.8) μm (n = 100), aseptate, smooth, fusiform, straight and tapering towards both ends. Beta-conidia were 25.1 (range, 19.1 to 36.2) × 1.3 (range, 1.0 to 2) µm (n = 100), filiform, hyaline, aseptate and curved at one end. The morphological features of these isolates agreed with those of Diaporthe sp. (Gomes et al. 2013; Gao et al. 2017). To further identify the isolates, the regions of internal transcribed spacer (ITS, OR214967 to OR214969), calmodulin (CAL, OR228558 to OR228560), translation elongation factor 1-α (EF1a, OR228561 to OR228563), histone H3 (HIS, OR228564 to OR228566), and beta-tubulin 2 (TUB2, OR228567 to OR228569) were amplified and sequenced from genomic DNA for the three representative isolates (LSM1, LSM2 and LSM3), respectively (Gomes et al. 2013). Multilocus phylogenetic analysis revealed that the three isolates and D. pseudophoenicicola were clustered in the same clade. Based on the results of morphological and molecular analysis, these isolates were identified as D. pseudophoenicicola. The pathogenicity of three isolates were tested on leaves of pecan seedlings. The conidial suspension (1 × 105 conidia/ml) of each isolate was sprayed evenly on the surface of leaves of three healthy seedlings. Sterilized distilled water was used for negative controls. Finally, all inoculated plants were kept in a greenhouse at 28°C under 100% relative humidity. Two weeks after inoculation, the inoculated plants developed symptoms similar to those of the original diseased plants, while controls remained asymptomatic. D. pseudophoenicicola were re-isolated from from inoculated plants. The pathogenicity experiment was repeated three times. Previously, D. pseudophoenicicola has been reported to cause stem-end browning disease in ripe mango (Takushi et al. 2016; Xu et al 2022). To our knowledge, this is the first report of D. pseudophoenicicola causing leaf blight on pecan . This study provides important information for developing effective pecan disease management practices.

Prospects for developing allergen-depleted food crops.

In addition to the challenge of meeting global demand for food production, there are increasing concerns about food safety and the need to protect consumer health from the negative effects of foodborne allergies.Certain bio-molecules (usually proteins) present in food can act as allergens that trigger unusual immunological reactions, with potentially life-threatening consequences. The relentless working lifestyles of the modern era often incorporate poor eating habits that include readymade prepackaged and processed foods, which contain additives such as peanuts, tree nuts, wheat, and soy-based products, rather than traditional home cooking. Of the predominant allergenic foods (soybean, wheat, fish, peanut, shellfish, tree nuts, eggs, and milk), peanuts (Arachis hypogaea) are the best characterized source of allergens, followed by tree nuts (Juglans regia, Prunus amygdalus, Corylus avellana, Carya illinoinensis, Anacardium occidentale, Pistacia vera, Bertholletia excels), wheat (Triticum aestivum), soybeans (Glycine max), and kidney beans (Phaseolus vulgaris). The prevalence of food allergies has risen significantly in recent years including chance of accidental exposure to such foods. In contrast, the standards of detection, diagnosis, and cure have not kept pace and unfortunately are often suboptimal. In this review, we mainly focus on the prevalence of allergies associated with peanut, tree nuts, wheat, soybean, and kidney bean, highlighting their physiological properties and functions as well as considering research directions for tailoring allergen gene expression. In particular, we discuss how recent advances in molecular breeding, genetic engineering, and genome editing can be used to develop potential low allergen food crops that protect consumer health.

Optimisation Using Response Surface Methodology of Quality, Nutritional and Antioxidant Attributes of ‘Wichita’ Pecan Nuts Roasted by Microwaves

Pecan (Carya illinoinensis) nuts are rich in functional compounds (unsaturated fatty acids, phytosterols, polyphenols, and tocopherols) associated with various health benefits. Commercially, pecan nuts are roasted to enhance their physical, chemical, and sensory properties. In the present study, response surface methodology (RSM) was used to optimise the quality and nutritional and antioxidant attributes of ‘Wichita’ pecan nuts roasted by using a microwave process with a range of microwave power (96.45–803.55 W) and roasting time (1.37–5.62 min). The microwave-roasted pecan nuts were analysed for hardness, total colour difference (TCD), and radical scavenging activity and modelled using the central composite design. The results showed that microwave power and roasting time significantly (p &lt; 0.05) influenced the quality attributes of the pecan. The quadratic model adequately described the changes in TCD and hardness, respectively, while the 2FI model adequately described the changes in DPPH radical scavenging activity. To obtain the desired pecan nuts quality attributes (TCD = 1863.391; hardness = 28.755 N and DPPH radical scavenging activity = 33.877 mmol Trolox/g), the determined conditions were 700 W and 2.24 min, with a desirability of 0.557. The primary unsaturated fatty acids, including cis-oleic, cis-linoleic, α-linolenic, and stearic acids, were not affected (p &lt; 0.05) by microwave roasting the pecan nuts at determined conditions. Volatile compounds, such as alcohols, aldehydes, ketones, lactones, hydrocarbons, and carboxylic acids, were identified in both raw and microwave-roasted pecan nuts, with limonene, which possesses various health properties, being the major volatile compound. It can be concluded that microwave roasting may be optimised using response surface methodology to produce quality pecan nuts that can be used as snacks or as an ingredient in other snack products.

Bacterial community diversity for three selected land use systems as affected by soil moisture regime

Despite their important role in soil nutrient cycling, soil bacterial communities' response to land management practices and soil hydrologic fluctuations are not sufficiently studied. The effects of land use and soil moisture regime on soil bacterial community structure and diversity in the Missouri River Floodplain have been investigated in this study. Three land use systems represented riparian forest (RF), pecan (Carya illinoinensis) orchard/hay agroforestry system (AF), and row crop corn [Zea mays L.]/soybean [Glycine max (L.) Merr.] agriculture system (AG). Microbial diversity indices and the abundance of selected bacterial phyla were investigated in bulk soil samples by performing 16 s rRNA amplicon sequencing. Soil samples were collected from three selected land use systems in September 2020 and May 2021. To simulate the hydrological dynamics of the riparian systems, intact soil cores were subjected to two different soil moisture regimes: saturated (FD) and alternately saturated and field capacity (FL) for 8 weeks of incubation. A control sample was maintained at field capacity for the entire duration of the experiment. Soil physicochemical properties substantially differed among land use. Soil organic matter (OM%) was significantly greater in AF and RF than in AG. Proteobacteria, Actinobacteriota, and Acidobacteriota were found to be the most dominant bacterial phyla across all samples. The bacterial richness and Shannon indices were not considerably different among land use systems. Soil FD conditions considerably increased the abundance of Actinobacteriota in AF soils. The abundance of these phyla was strikingly higher in AF with the greatest organic matter relative to AG and RF under all three water regimes. Further work may be needed to assess the effects of hydrological events on floodplain soils' microbial community indices.

Determination of the Water use Efficiency of a Pecan Nut Orchard based on the Irrigation Scheduling and the Evapotranspiration Rate in Northern Mexico

Background: Adult pecan trees require a significant water supply for high yields and nut quality. Pecan nut orchards are established in semi-arid regions where water is the primary limiting resource for agriculture. Therefore, in these regions, improving water use efficiency is essential. Objective: Evaluate the water use efficiency of a pecan tree orchard based on the comparison of irrigation scheduling with the evapotranspiration rate data. Methods: The study was conducted in a pecan nut (Carya illinoinensis) orchard of 7.2 ha with 7-year-old Western and Wichita cv tress. The volume of water applied to each tree (drip irrigation) was converted to a daily irrigation depth and compared against the daily rate of actual (measured with an Eddy covariance system) and the FAO Penman-Monteith evapotranspiration. Results: Comparing the monthly water depth applied to each tree against the monthly FAO-Penman Monteith ET, surplus or deficit irrigation episodes were detected. Because the canopy trees only covered 18.7% of the orchard area, the daily rate of actual evapotranspiration during the months of the trees growing was very small (1 to 2 mm) compared with the orchards of mature pecan trees. The corresponding monthly crop coefficients (Kc) were also small (0.315 on average). Conclusion: This study showed that by comparing the rate of ETref against the irrigation depth applied to the trees in the irrigation scheduling, it is possible to reveal cases of surplus or deficit of water supplied to a pecan nut orchard.

First report of leaf spot on pecan (Carya illinoinensis) caused by Stemphylium eturmiunum in China.

Pecan [Carya illinoinensis (Wangenh.) K. Koch] is an important nut tree species, which has been widely planted in Jiangsu and Anhui Provinces of China in recent years (Mo et al. 2018). In May 2022, a new leaf spot disease was observed on both young and old leaves of pecan trees in the Luhe area, Nanjing, Jiangsu Province. Approximately 30% of pecan trees suffered from the disease, which affected the growth of young trees and nut production to cause economic loss. Initially, the leaf spots were grayish black and round. Then, disease spots enlarged and joined together, forming irregular lesions with uneven edges. In the last stage, the leaflets were withered. To isolate the pathogen, three symptomatic leaves were collected from each of three different pecan trees. Leaf sections (3 to 4 mm) were excised from the margin of spots, surface sterilized in 75% alcohol for 30 s, then sterilized in 1.5% NaClO for 90 s. After rinsing three times with sterile distilled water, leaf sections were placed on potato dextrose agar (PDA) medium and incubated at 25 °C in a dark environment for 5 days. Pure cultures were obtained by monosporic isolation. A total of 20 isolates were obtained, and 12 isolates were identified as Stemphylium sp. with the same morphological features and ITS sequences. A representative isolate, named LH3-3, was selected for further study. Colonies on PDA were light yellow with dense mycelium and were brownish yellow on the reverse side. Conidia were 16.3 to 34.4 × 8.1 to 16.3 µm) (n=35), muriform, brown, with transverse and longitudinal septa, lightly deformed at the transverse septa. Ascomata were not observed. The morphological characteristics were consistent with the description of Stemphylium eturmiunum (Simmons 2001). The internal transcribed spacer region (ITS) and portions of genes for calmodulin (cmdA) and glyceraldehyde-3-phosphate dehydrogenase (gpd) were amplified and sequenced with the primers ITS1/ITS4 (White et al. 1990), CALDF1/CALDR2 (Xu et al. 2022) and GPD-F/R (Xie et al. 2019), respectively. Sequences were deposited in GenBank under accessions OP482492 (ITS), OP495734 (cmdA), and OP495735 (gpd). BLAST analysis showed that the sequences had 99.67-100% homology to ITS (525/525 bp) of S. eturmiunum strain ST14 (MH843733), cmdA (694/694 bp) of strain CBS122124 (KU850832), and gpd (299/300 bp) of isolate UMSe0030 (MK336876). MEGA 7.0 was used to construct a phylogenetic analysis based on concatenated sequences of ITS, cmdA, and gpd using the neighbor-joining method. The results showed that LH3-3 clustered on the branch of S. eturmiunum, and the support rate was 100%. A spore suspension in sterile water was made from strain LH3-3 grown on PDA, and adjusted to 1×106 spores/mL with a hemocytometer. To test pathogenicity, 20 µl drops of the spore suspension were placed on the left sides of four healthy detached leaflets of mature pecan trees and leaves of three 3-month-old seedlings. The right side of each leaflet was inoculated with 20 µl drops of sterile distilled water as the control. All inoculated seedlings and detached leaflets were covered with a transparent plastic bag and cultured in a greenhouse at 25 °C, 80% relative humidity, and a 12 h light cycle until symptom appeared. The experiment was repeated three times. After 7 days of inoculation, grayish black lesions appeared on all inoculation sites with the spore suspension but not in the controls. The leaf spot symptoms were similar to those observed in orchards. The same fungus, identified by morphological characteristics and sequencing of ITS, cmdA, and gpd, was re-isolated from the diseased spots of the inoculated leaflets to complete Koch's postulates. S. eturmiunum has been reported to infect garlic (Dumin et al. 2022) and tomato (Prencipee et al. 2021), but this is the first report of S. eturmiunum causing leaf spot of C. illinoinensis. This study provides a basis for further study on the biology, epidemiology, and management of the disease.

First report of Fusarium concentricum as a causal agent of Fusarium leaf blotch on pecan (Carya illinoinensis) in Southeast China.

The economically important nut crop pecan (Carya illinoinensis (Wangenh.) K. Koch) is seriously affected by increasing incidence of fungal disease worldwide (Xiao et al 2021). The top leaves of the pecan variety 'Pawnee' in the orchard of Zhejiang A&F University, Zhejiang, China were damaged by massive dark brown plaques in summer to autumn 2021. The causal agent was isolated from leaves with target plaques following the steps: sterilized with 70% alcohol (30 s × 2), rinsed with sterilized water (3 ×) before and after 5% sodium hypochlorite (30 s), excised the plaques, and placed on PDA medium at 28℃ in a dark incubator for 3-d. The mycelium on the edge of each colony was transferred to fresh SNA medium in dark for 2 weeks to induce conidia formation. A few conidia-germinated mycelia were transferredand inoculated on new plates containing fresh PDA medium to obtain the purified cultures. Koch's postulates were applied to validate the pathogenicity of the purified isolates. Non-woundedly healthy leaves (disinfected with 5% sodium hypochlorite) of 'Pawnee' seedlings were inoculated with 5 mm 7-d old purified cultures. Dark-brown spots appeared on the leaves 2 days post inoculation at 25℃. The spots became larger accompanied by partially cracking and slight deformation of inoculated leaves from day 2 to day 4, while the control leaves remained asymptomatic. A re-isolated strain ZJ-6 from these infected leaves was identified as the pathogenic isolate with the same symptom as the previous one. Morphologically, aerial mycelia of the pathogenic isolate ZJ-6 cashmere and white. The reverse of colony orange. The edge of the colony appeared gradually thinner, the aerial mycelia loose and flocky, and the matrix mycelium whitened. Hyphae were septate, translucent with smooth wall and 1.47-7.14 µm in width. Microconidia (n = 20) obovoid to fusoid, mainly with 0-septate, 4.45-7.78×4.79-16.25 µm. Macroconidia (n = 20) sickle, mainly with 3-5 septa, 5.56-10.28×56.67-114.54 µm. Simultaneous of monophialidic and polyphialidic conidiophores. Conidiophore width 1.47-3.68 µm, slightly smaller than vegetative hyphae. The morphological characteristics matched with previous descriptions of Fusarium species (Nirenberg and O'Donnell 1998; Wang et al 2013). The identity of ZJ-6 was confirmed by phylogenetic reconstruction using the concatenated sequences of the ATP citrate lyase (ACL1), Calmodulin (CaM), the internal transcribed spacer (ITS) rDNA region, ribosomal RNA gene (LSU), the largest subunit of DNA-dependent RNA polymerase II (RPB1), partial translation elongation factor-1 alpha (TEF) and β-Tubulin (TUB). To this end, the genomic DNA of ZJ-6 was extracted by the M5 hipermix-MF859 (Mei5 Biotechnology) and submitted to GenBank under the accession numbers OP933646, OP933647, OP925890, OP925889, OP933396, OP933648, and OP933397, respectively. The obtained sequences of ZJ-6 were used for nucleotide BLAST against thetandard databases, respectively, and the strains with sequence identification values above 98% were selected to construct multiple alignment for building a phylogenetic tree. This analyses allowed the identification of ZJ-6 as Fusarium concentricum Nirenberg & O'Donnell, a species with few reports that can cause serious damage to the fruits and branches of other hosts (Hasan et al 2020; Huda-Shakirah et al 2020; Wang et al 2013). This is the first report of pathogenic F. concentricum on pecan in Southeast China that caused no harvest of infected plants.

First report of Colletotrichum plurivorum causing anthracnose on pecan (Carya illinoinensis) in China.

Pecan (Carya illinoinensis) is an economically important nut crop worldwide (Xiao et al 2021). Anthracnose symptoms were found on pecan fruits and leaves in plantations in Anhui and Jiangsu provinces, China in August 2019. Irregular, dark brown or black spotted lesions firstly appeared on the surface and inside of fruits, and spread to all leaves. The symptoms resulted in 30% to 50% leaf drop and nearly a half of fruit decay in almost all trees of the susceptible cv. Wichita. The causal agent were isolated from fruits with target symptoms following the steps: surface disinfected with 75% ethanol (2×, 30 s), rinsed with sterile deionized water (3×), ~ 0.5 cm small fragments of the fruits excised and plated on potato dextrose agar (PDA) medium and incubated at 28 °C in dark for 3-d. Mycelium of each colony was picked and incubated on fresh PDA at 25 °C with a 12-hour light/dark cycle for 6-d to induce conidia formation. One 5-mm hyphal plug produced from each single spore isolate was transferred onto fresh PDA to obtain the pure cultures. Koch's postulates was employed for pathogenicity determination of the isolates. Non-wounded healthy leaves from seedlings of the disease susceptible cv. Pawnee were disinfected with 1% NaClO and inoculated with 5-mm 5-d hyphal of each isolate at 25 ℃. Tiny lesion spots were visible on the leaves after 2 days post inoculation (DPI) with isolate W-6 (the only pathogenic one among all isolates), and expanded over time until to the leaves withered, while the control leaves and leaves inoculated with other isolates remained asymptomatic. The pathogenicity of W-6 were confirmed using leaves and fruits of living Pawnee trees growing in Linglong Mountain Plantation, Lin'an, Hangzhou, China (119⁰38'51″E, 30⁰12'39″N, elevation: 119m). Three experimental replicates were conducted separately with three bio-replicates for all pathogenetic testing. The same symptoms were observed on both detached leaves and leaves and fruits of living trees.. The colony of W-6 have round cottony mycelium with complete edges and showed the fastest growth rate 3 - 4 DPI. After 7 DPI, white aerial mycelium turned yellowish brown and formed Acervulus in the mycelium. Conidia (n=50) one-celled, 12.0 - 20.0 μm × 3.5 to 6.0 μm width. Hyaline cylindrical with slightly rounded ends and two or three large guttulate at the centre. Most Acervulus dark brown and slightly irregular in shape, 12.70 × 18.79 μm (n=10). Setae were dark brown in color with average length around 34.10 μm (n=10). These characteristics matched previous descriptions of Colletotrichum orchidearum species complex, including C. plurivorum (Damm et al 2019). The identity of W-6 was confirmed by multi-locus phylogenetic analysis using the internal transcribed spacer (ITS) rDNA region and partial sequences of the conserved genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), beta-tubulin 2 (TUB2), and chitin synthase (CHS). The sequences of W-6 were used for Basic Local Alignment Search Tool (BLAST) against NCBI GenBank and the sequences with 100% identity to that of W-6 were achieved, respectively. The concatenated sequences of the ACT-CHS-GAPDH-ITS-TUB2 was used for building a phylogenetic tree. The molecular analyses allowed the identification of the pathogen as C. plurivorum. It was known that 9 of the 11 Colletotrichum species causing pecan anthrax worldwide were reported in southern China (Brenneman 1989; Oh et al 2021). This is the first report of C. plurivorum as causal agent of pecan in China.

Lipidomic analyses of five Carya illinoinensis cultivars.

Carya illinoinensis (Wangenh.) K. Koch, nuts are a renowned health food. However, there are many cultivars of this nut tree, and their mature kernel lipid composition has not been thoroughly studied. Therefore, we used liquid chromatography-mass spectrometry (LC-MS) to analyze the lipid composition of mature nuts of five C. illinoinensis cultivars. In the mature kernels of all cultivars, there were 58 lipid types which were mainly composed of glycerolipids (c. 65%) and phospholipids (>30%). Triacylglycerol (TG) accounted for the largest proportion of mature nuts of all cultivars, exceeding 50%; and diacylglycerol (DG), ceramide (Cer), phosphatidylcholine (PC), and phosphatidylethanolamine (PE) were also relatively high. Additionally, nuts contain fatty acids, mainly oleic, linoleic, and linolenic acids. Our research provides a new perspective for the processing and utilization of plant and edible oils, and for the use of C. illinoinensis kernels in the development of medicine and food science.

Changes in the diversity of pecan (Carya illinoinensis) rhizosphere microbial community with different nitrogen fertilization, a case study in Oklahoma pecan orchard

Pecan (Carya illinoinensis) is primarily native to North America and is known as the leading nut produced in Oklahoma. Since the prices of fertilizers have increased and the activity of soil microbes may greatly affect fertilizer utilization, it is important to investigate the effect of fertilization on microbes. Samples were collected from pecan trees receiving full nitrogen (70 lbs/acre), half nitrogen (35 lbs/acre), and no nitrogen (0 lbs/acre) fertilization in an orchard. Microbes were characterized by using the next generation sequencing and annotated into functional groups (trophic mode and nitrogen related) by FUNGuild and FAPROTAX. Our results showed the core fungal genera in this orchard were Tuber and Hymenoscyphus, while the core bacterial families were Chitinophagaceae, Xanthobacteraceae, and Chthoniobacteraceae. Among them, the pecan truffle Tuber lyonii was the dominant fungus. In association with an increase of nitrogen fertilization, the Simpson Index (SI) of fungi in roots was decreased, and the presence of pathotroph (such as Monographella cucumerina and Lectera longa) was increased, implying that the risk of plant infection by pathogens was increased. The pecan with half rate of nitrogen fertilizer had the highest colonization of symbiotrophic fungi, the lowest presence of pathotrophic fungi, and the highest relative abundance of Tuber. The relative abundances of bacteria related to nitrate reduction, nitrogen respiration, and nitrate respiration were found to be the highest in pecan with half rate of nitrogen fertilizer, which might imply the highest efficiency of nitrogen fertilizer utilization in this treatment. It revealed that applying half the recommended nitrogen rate of fertilizer could improve beneficial microbe presence, perhaps increasing nitrogen uptake and decreasing nitrogen needed for optimum production.

Genome-wide identification of KCS gene family in Carya illinoinensis and their roles under abiotic stress conditions

Pecan (Carya illinoinensis) is an important economic tree species of genus Carya in the Juglandaceae. Its nuts are rich in nutrients, becoming one of the consumer's favorite ones. The pecan production is facing serious threats from drought, soil salinization and acidification. β-Keto-acyl-CoA synthase (KCS) is an important enzyme responsible for the formation of very long chain fatty acid (VLCFA) and its derivatives. However, the characteristic of CiKCS genes family in pecan and their function under abiotic stress remain greatly unknown. Here, 30 CiKCSs were identified and distributed in the 14 chromosomes, subjected to gene duplication and strong purification selection. The cis-regulatory elements in the 2 kb promoters of these CiKCSs mainly correlated with plant growth, development and stress response. Notably, CiKCS3/4/5/6/24/28 genes expressions were significantly up-regulated under drought, salt and acid stresses. Interestingly, the expression of CiKCS12 under acid stress were down-regulated. CiKCS3/5/6/12 were further shown to be mainly located in the cytoplasm, had no self-activation activity and could not interact with each other. Heterologous expression of these four genes improved VLCFA levels in tobacco and mediated the tolerance of yeast to osmotic, salt, and acid treatments. These results preliminarily suggested that CiKCS3/5/6/12 might function in the cytoplasm through monomeric forms and improve the tolerance of pecan to abiotic stress mainly by mediating VLCFA synthesis. These findings will deepen the understanding of the characteristic and function of CiKCS genes, and provide new direction and reference for the stress-resistant breeding of pecan.