Pyrus Species Research Articles

Epidemiological description of fire blight introduction patterns to Central Asia and the Caucasus region based on CRISPR spacer typing and genome analysis

In the last two decades, fire blight has progressively spread eastward from Europe and the Mediterranean area to several pome-fruit producing regions of Asia. Its causative agent, the bacterial pathogen Erwinia amylovora, was detected in several new countries, including Georgia, Kyrgyzstan, and Kazakhstan. In the latter two states, the disease creates a threat not only to the commercial apple and pear production, but also to the wild Malus and Pyrus species that constitute the basis of the local forest ecosystems. In this study, we investigated the genetic diversity of the pathogen in Central Asia and the Caucasus region utilizing CRISPR Repeat Regions (CRRs) genotyping and genome sequencing, with the aim to understand its dissemination patterns across the continent. Genome sequence analysis revealed that all strains from these two regions exclusively derived from the archetypal CRR1 genotype A. Our analysis revealed three main E. amylovora clades in Central Asia, with distinct yet partial overlapping geographical distributions. Genomic relationships among isolates indicate that Central Asian strains are genetically closest to those from the Persian region and the Middle East, while the Georgian population is genetically more distant and can align with strains from the Volga District in southern Russia and the Eastern Mediterranean area. Notably, this study also includes strains from the first confirmed occurrences of fire blight in Uzbekistan, Tajikistan, and China. Our findings highlight the importance of phylogenetic analysis and genome sequencing in understanding the phytopathogen epidemics and protecting key agricultural species and the genetic resources of their wild counterparts in the forest.

Chloroplast genome sequencing and divergence analysis of 18 Pyrus species: insights into intron length polymorphisms and evolutionary processes.

Pears constitute an essential temperate crop and are primarily produced through interspecific hybridization owing to self-incompatibility that complicates their breeding history. To address this, we sequenced the complete chloroplast (cp) genomes of 18 Pyrus and one Malus species using the Illumina HiSeq4000 platform. The cp genomes ranged from 159,885 bp to 160,153bp and exhibited a conserved circular DNA structure with an average GC content of 36.5%. Each cp genome contained 127 genes, including 83 protein-coding, 36 tRNA, and 8 rRNA genes. Divergence analysis with mVISTA showed high conservation in the coding regions and notable variations in the non-coding regions. All species shared 17 intron-containing genes, with ycf3 and clpP each having two introns. Five intron-containing genes (ndhB, rpl2, rps12, trnA-UGC, and trnE-UUC) were located in the inverted repeat regions, while trnL-UAA was located in the large single-copy region, with conserved intron lengths across Pomoideae. We identified polymorphic intron sequences in the rpl22, petB, clpP, ndhA, and rps16 genes and designed primers for these regions. Notably, the two Pyrus ussuriensis accessions Doonggeullebae and Cheongdangrori showed intron-length polymorphisms despite being classified as the same species. Phylogenetic analysis of the cp genome sequences revealed two major clusters, indicating distinct maternal lineages and evolutionary origins. This study underscores the importance of cp gene polymorphisms in P. fauriei, P. calleryana, P. ussuriensis, and P. pyrifolia, providing valuable insights into Pyrus evolution as well as aiding in the conservation and breeding of pear germplasm.

Pharmacognostical, Phytochemical Evaluation and Unlocking the Pharmacological Power of <i>Pyrus communis</i> L. Leaf Extract on DOCA Salt Induced Hypertension in Wistar Rats

Objectives: Identification of the active ingredient in medication is greatly aided by pharmacognostical and phytochemical research, such as the macroscopic, GC-MS analysis and to study hypertensive treatment with ethanolic extract of Pyrus communis L. Methods: Transverse sections, powder microscopy, Histochemical analysis also performed preliminary phytochemical analysis and GC-MS of P. communis. The different groups of Wistar rats were administered 400mg/kg as a lower dose and 600mg/kg as a higher dose given DOCA salt-induced hypertension. Results: Histochemical analysis of this leaf shows the presence of cutin, mucilage cells, tannin, alkaloids, lignin, starch grains, calcium oxalates cluster crystals, and oils. Eleven components in the extract were reported via GC-MS. This report confirms the existence of various phytocomponents which are 4-pyridine methanol; N-Methylmaleimide; 4-acetyl-1h-pyrroline-2-carbaldehyde; Carbamic acid, 2-(Dimethyl amino) Ethyl Ester; 5-Acetyl-4-Amino-3-(2-Dimethylaminoethylthio)Thieno[3,2-C]Isothiazole; Cyclobutaneoctol; 1,3-Propane diamine, N, N’-Bis (3-Aminopropyl)-; Arginine; 9-Methyl-11-Oxo-1,6-Diaza tricyclo [7.2.0.0(6,8)] Undecane; Egtazic Acid; animal study significantly reduced the pressure compared to the control group. P. communis ethanolic extract possesses a significant (p≤0.05) reduction in hypertensive rats from measurements of control group as 157/137 mmHg systolic and diastolic blood pressure to 140/98 mmHg in 600 mg/kg Conclusions: These results will also be helpful to add to the advanced knowledge of P. communis standardisation and identification, which are essential components in separating real Pyrus species from adulterants and imitations in the herbal medicine formulations better for the treatment of hypertension. The empirical and phytochemical study of P. communis leaf holds significant potential for the development of novel herbal remedies for hypertensive patients.

Chloroplast Genome Profiling and Phylogenetic Insights of the “Qixiadaxiangshui” Pear (Pyrus bretschneideri Rehd.1)

The “Qixiadaxiangshui” pear (Pyrus bretschneideri Rehd.1) is a highly valued cultivar known for its crisp texture, abundant juice, and rich aroma. In this study, we reported the first complete chloroplast genome sequence of the “Qixiadaxiangshui” pear, which is 159,885 bp in length with a GC content of 36.58%. The genome exhibits a typical circular quadripartite structure, comprising a large single-copy region (LSC), a small single-copy region (SSC), and a pair of inverted repeat regions (IRs). A total of 131 genes were identified, including 84 protein-coding genes, 8 rRNA genes, and 37 tRNA genes. We also identified 209 simple sequence repeats (SSRs) and several mutation hotspots, such as ndhC-trnM-CAU and trnR-UCU-atpA, which can be applied in molecular identification and phylogenetic studies of Pyrus. Comparative genomic analysis showed high conservation among ten pear cultivars. Phylogenetic analysis indicated that the “Qixiadaxiangshui” pear is closely related to germplasm Dangshansuli, Wonwhang, and Yali, suggesting a recent common ancestor. These findings provided valuable insights into the genetic diversity and evolutionary dynamics of the Pyrus species and contribute to the conservation and breeding of pear germplasm resources.

Genetic and molecular regulation of chilling requirements in pear: breeding for climate change resilience.

Pear (Pyrus spp.) is a deciduous fruit tree that requires exposure to sufficient chilling hours during the winter to establish dormancy, followed by favorable heat conditions during the spring for normal vegetative and floral budbreak. In contrast to most temperate woody species, apples and pears of the Rosaceae family are insensitive to photoperiod, and low temperature is the major factor that induces growth cessation and dormancy. Most European pear (Pyrus Communis L.) cultivars need to be grown in regions with high chilling unit (CU) accumulation to ensure early vegetative budbreak. Adequate vegetative budbreak time will ensure suitable metabolite accumulation, such as sugars, to support fruit set and vegetative development, providing the necessary metabolites for optimal fruit set and development. Many regions that were suitable for pear production suffer from a reduction in CU accumulation. According to climate prediction models, many temperate regions currently suitable for pear cultivation will experience a similar accumulation of CUs as observed in Mediterranean regions. Consequently, the Mediterranean region can serve as a suitable location for conducting pear breeding trials aimed at developing cultivars that will thrive in temperate regions in the decades to come. Due to recent climatic changes, bud dormancy attracts more attention, and several studies have been carried out aiming to discover the genetic and physiological factors associated with dormancy in deciduous fruit trees, including pears, along with their related biosynthetic pathways. In this review, current knowledge of the genetic mechanisms associated with bud dormancy in European pear and other Pyrus species is summarized, along with metabolites and physiological factors affecting dormancy establishment and release and chilling requirement determination. The genetic and physiological insights gained into the factors regulating pear dormancy phase transition and determining chilling requirements can accelerate the development of new pear cultivars better suited to both current and predicted future climatic conditions.

A chromosome-scale assembly for 'd'Anjou' pear.

Cultivated pear consists of several Pyrus species with Pyrus communis (European pear) representing a large fraction of worldwide production. As a relatively recently domesticated crop and perennial tree, pear can benefit from genome-assisted breeding. Additionally, comparative genomics within Rosaceae promises greater understanding of evolution within this economically important family. Here, we generate a fully phased chromosome-scale genome assembly of P. communis 'd'Anjou.' Using PacBio HiFi and Dovetail Omni-C reads, the genome is resolved into the expected 17 chromosomes, with each haplotype totaling nearly 540 Megabases and a contig N50 of nearly 14 Mb. Both haplotypes are highly syntenic to each other and to the Malus domestica 'Honeycrisp' apple genome. Nearly 45,000 genes were annotated in each haplotype, over 90% of which have direct RNA-seq expression evidence. We detect signatures of the known whole-genome duplication shared between apple and pear, and we estimate 57% of d'Anjou genes are retained in duplicate derived from this event. This genome highlights the value of generating phased diploid assemblies for recovering the full allelic complement in highly heterozygous crop species.

DNA base analysis: An effective tool to resolve taxonomic uncertainty and establish phylogenetic relationship in cryptic genotypes of Pyrus (pear), Rosaceae

In the current study, a total of 16 Pyrus (pear) were collected from the Biayari Valley, Allai, KP, Pakistan. A good quality gDNA was extracted, target region of DNA of each genotype was amplified through PCR, amplified region of DNA was extracted from agarose gel and was sequenced. Among 16 sequences, good quality 09 query sequences were analyzed through different bioinformatics tools and techniques for establishment of phylogenetic tree. Molecular analysis showed that the query genotype-B1"Tora tangai" possess 86% identity and 100% query cover with P. sinkiangensis, P. communis, and P. pyrifolia, while the query genotype-B2 “Kado batang” showed 83 % identity and 98 % query cover with P. communis, P. ussuriensis, and P. pyrifolia. Similarly, the query genotype-B6 “Shakar batang” possess 86% identity and 69% query cover with P. Malus rockii, Malus rocki, P. pyrifolia, and P. communis. The query genotype-B7 “kado batang” showed 92% identity and 13% query cover with; P. sinkiangensis, P. pyrifolia, P. communis, P. domestica The query genotypeB9 "Mamosai" revealed 98% identity and 3% query cover with genotype P. pyrifolia, Malus rokii, P. ussuriensis, and P. sinkiangensis. Similarly, BLAST search result showed that query genotype-C1 “Kado batang” possess 95% identity and 91% query cover with P. calleryana, P. pyrifolia, P. pyrifolia, P. pyrifolia, and P. pyraster. Initial search result revealed that the query genotype-C4 “Nashpati” has 42% query cover and 95% identity with; P. dimorphophylla, P. dimorphophylla, P. pashia, P. pashia P. ussuriensis. and similarly, genotype-D1 “Nashpati” showed 93% identity and 9% query cover with, Pyrus pyrifolia and Pyrus x domestica. Over all, BLAST search result showed that all the query genotypes possess maximum identity and query cover with different species of the genus Pyrus (pear), family Rosaceae. Further, phylogenetic tree showed that the query genotypes B1, B2, B6, B7, B8, C1, C2 and C4 fall independently due to its slight genetic variation, an indication for development of sub-species and new varieties. This genetic variation is due cross pollination and their availability in the wild form decades. These finding promote horticulture especially in development of new varieties in pear.

Comparative Evaluation of Pyrus Species to Identify Possible Resources of Interest in Pear Breeding

Pear is one of the most important fruit species grown in the temperate zones of the globe. Besides fruit production, pear species are highly valued in forestry and agroforestry systems; in landscaping, as ornamental features; as fruits of ecological value, and in other areas. The Pyrus species, obtained from a gene bank, were evaluated for the different morphological traits of the trees, leaves, flowers, and fruits, as well as their responses to attacks from principal diseases and pests. Phenotypic data were examined using correlation and multivariate analyses, and a dendrogram of morphological traits was completed via molecular investigations at the DNA level using the RAPD markers. The findings revealed the complexities of the phenotypic and genetic connections among Pyrus species, as well as the difficulty in establishing phylogenetic relationships among pear species. The findings also demonstrated that the wide variability between species with different geographical origins, and their multiple peculiarities of interest, represents a cornerstone as the source of genes of great utility for pear breeding or for utilizing trees for different edible crops and for silvocultural, landscape, or ecological purposes.

Spatial Distribution Characteristics of Suitable Planting Areas for Pyrus Species under Climate Change in China.

Planting suitability determines the distribution and yield of crops in a given region which can be greatly affected by climate change. In recent years, many studies have shown that carbon dioxide fertilization effects increase the productivity of temperate deciduous fruit trees under a changing climate, but the potential risks to fruit tree planting caused by a reduction in suitable planting areas are rarely reported. In this study, Maxent was first used to investigate the spatial distribution of five Pyrus species in China, and the consistency between the actual production area and the modeled climatically suitable area under the current climatic conditions were determined. In addition, based on Coupled Model Intercomparison Project Phase 6, three climate models were used to simulate the change in suitable area and the migration trend for different species under different emission scenarios (SSP1-2.6, SSP2-4.5, SSP3-7.0 and SSP5-8.5). The results showed that the suitable area for pear was highly consistent with the actual main production area under current climate conditions. The potential planting areas of P. ussuriensis showed a downward trend under all emission paths from 2020 to 2100; other species showed a trend of increasing first and then decreasing or slowing down and this growth effect was the most obvious in 2020-2040. Except for P. pashia, other species showed a migration trend toward a high latitude, and the trend was more prominent under the high emission path. Our results emphasize the response difference between species to climate change, and the method of consistency analysis between suitable planting area and actual production regions cannot only evaluate the potential planting risk but also provide a reasonable idea for the accuracy test of the modeled results. This work has certain guiding and reference significance for the protection of pear germplasm resources and the prediction of yield.

Comparative Analysis of Phenotypic and Molecular Data on Response to Main Pear Diseases and Pest Attack in a Germplasm Collection

The pear is an important fruit tree in temperate areas, but due to its sensitivity, fruit yield and quality are often affected by disease and pest attacks. Pear genotypes from a germplasm collection comprising 13 Pyrus species, 17 Romanian varieties, and 50 non-Romanian varieties from a worldwide assortment were investigated in this study. Throughout four years, response to attack of the principal pathogens and pests was investigated phenotypically under natural conditions of infection and infestation. SSR markers were used to analyze the genetic diversity of the genotypes. A standardized method for the evaluation of responses to biotic stressors was proposed, which highlighted significant differences between genotypes. The species and varieties with the lowest degrees of attack (DA%), calculated based on the frequency and intensity of attack, were identified for pear scab (Venturia pyrina), septoria (Septoria pyricola), fire blight (Erwinia amylovora), and psyllids (Psylla sp.). These accessions could provide valuable sources of genes of interest to develop resistant varieties in new pear breeding programs. By combining phenotypic and molecular analyses, significant information was obtained that can be exploited to generate high variability for selection through artificial hybridization by harnessing accessions with complementary molecular fingerprints and high genetic distances.

Comprehensive Assessment of Intergeneric Cross-compatibility of Six Fruit Tree Species in the Tribe Maleae (Rosaceae) Based on <i>In Vivo</i> Pollen Tube Growth and Field Pollination

The development of intergeneric hybrids for horticultural crops has been attempted to introduce new quality and resistance traits and to enlarge the gene pool. Interspecific and intergeneric hybridization are often hindered by incompatibility reactions occurring at various stages of hybridization, from early pollination to initial growth, and the reproductive stages of the progeny. In this study, we investigated intergeneric and interspecific cross-compatibility among six species in the tribe Maleae (Rosaceae), namely, Pyrus communis (European pear), P. pyrifolia (Japanese pear), Malus × domestica (apple), Eriobotrya japonica (loquat), Cydonia oblonga (quince), and Pseudocydonia sinensis (Chinese quince). In vivo pollen tube growth tests showed the presence of a postmating, prezygotic barrier in many cross-combinations, in which cross-compatibility was regulated by both genetic distance and crossing direction. Strong hybridization barriers were observed in pollen tube growth and fruit setting when intergeneric hybridization was performed with E. japonica, a species phylogenetically distant from the others studied. Different compatibility reactions in reciprocal crosses were observed in some intergeneric hybridizations; C. oblonga as a pollen donor was incompatible with P. sinensis, whereas the reciprocal cross was compatible, resulting in the development of hybrid seedlings. Furthermore, the pollen tube growth rate differed among Pyrus species when pollinated on the apple pistils, suggesting divergence of cross-compatibility response in a specific linage. Factors affecting intergeneric hybridization are discussed with reference to the genetic distance between species and morphological characteristics such as pistil length. Our comprehensive assessment of intergeneric cross-compatibility will help provide a way to overcome crossing barriers and develop new hybrid crops in the tribe Maleae.

Comparative growth, hydraulic conductivity, and sap flow responses of seedlings of two pyrus species to drought stress

Drought is one of the major abiotic stresses limiting the advancement of the pear industry in the arid and semiarid regions of China. Currently, the production of pear seedlings mainly depends on the grafting propagation of rootstocks from Pyrus betulaefolia Bunge and Pyrus calleryana Decne. However, limited information is available on the drought stress potential of these rootstocks. Here, we conducted pot experiments with controlled watering conditions on both rootstocks' seedlings and compared their growth, plant hydraulic conductivity, and stem sap flow velocity. Both rootstocks' seedlings were challenged with three drought regimes i.e. light (LD), moderate (MD), and heavy (HD) drought stress. The results indicated that P. betulaefolia performed better in growth than P. calleryana under the same drought regimes. Under LD, MD, and HD the relative growth of the new shoots, leaf-specific canopy hydraulic conductivity, leaf, stem, and root hydraulic conductivity, and stem sap flow velocity of P. betulaefolia exhibited reduction as compared to control (CK). Similar reduction trends were noted for P. calleryana, with slight variations in the case of stem-specific canopy hydraulic conductivity.

Pest categorisation of Xylella taiwanensis.

The EFSA Plant Health Panelperformed a pest categorisation of Xylella taiwanensis, a Gram-negative bacterium belonging to the Xanthomonadaceae. The pathogen is a well-defined taxonomic entity, and it is the causal agent of the pear leaf scorch. X. taiwanensis is present in subtropical and temperate areas of the island of Taiwan, where it affects low chilling pear cultivars of the species Pyrus pyrifolia (Asian pear). No other plant species are reported to be affected by the pathogen. The pathogen is not known to be present in the EU territory and it is not included in the Commission Implementing Regulation (EU) 2019/2072. The main pathway for the entry of the pathogen into the EU territory is host plants for planting (except seeds); another possible pathway might be represented by putative insect vectors, though their identity remains unknown. The cultivated area of P. pyrifolia in the EU territory is very limited. Conversely, the genetically related P. communis is widely cultivated in most EU Member States and there is no information so far on the susceptibility of its several cultivars. Should the pest establish in the EU, economic impact is expected, provided that suitable insect vectors are present and P. communis is as susceptible to infection as P. pyrifolia. Phytosanitary measures are available to prevent the introduction and spread of the pathogen into the EU, since plants for planting from Taiwan is a closed pathway; nonetheless, putative vectors, if confirmed and identified, may represent an additional risk of the pathogen's introduction and spread. The lack of knowledge on whether X. taiwanensis can infect P. communis, the identity and presence of suitable vectors in the EU lead to key uncertainties on entry, establishment, spread and impact. X. taiwanensis satisfies the criteria that are within the remit of EFSA to assess for this species to be regarded as a potential Union quarantine pest.

Micromorphological study on the seeds of wild species of Pyrus (Rosaceae) in Iran and their taxonomic implication

Micromorphological study on the seeds of wild species of Pyrus (Rosaceae) in Iran and their taxonomic implication

Functional characterization of a terpene synthase responsible for (E)-β-ocimene biosynthesis identified in Pyrus betuleafolia transcriptome after herbivory.

(E)-β-ocimene, a ubiquitous monoterpene volatile in plants, is emitted from flowers to attract pollinators and/or from vegetative tissues as part of inducible defenses mediated by complex signaling networks when plants are attacked by insect herbivores. Wild pear species Pyrus betuleafolia used worldwide as rootstock generally displays valuable pest-resistant traits and is a promising genetic resource for pear breeding. In the current study, transcriptional changes in this wild pear species infested with a polyphagous herbivore Spodoptera litura and the underlying molecular mechanisms were fully investigated. A total of 3,118 differentially expressed genes (DEGs) were identified in damaged pear leaf samples. Spodoptera litura larvae infestation activated complex phytohormonal signaling networks in which jasmonic acid, ethylene, brassinosteroids, cytokinin, gibberellic acid and auxin pathways were induced, whereas salicylic acid and abscisic acid pathways were suppressed. All DEGs associated with growth-related photosynthesis were significantly downregulated, whereas most DEGs involved in defense-related early signaling events, transcription factors, green leaf volatiles and volatile terpenes were significantly upregulated. The PbeOCS (GWHGAAYT028729), a putative (E)-β-ocimene synthase gene, was newly identified in P. betuleafolia transcriptome. The upregulation of PbeOCS in S. litura-infested pear leaves supports a potential role for PbeOCS in herbivore-induced plant defenses. In enzyme-catalyzed reaction, recombinant PbeOCS utilized only geranyl pyrophosphate but not neryl diphosphate, farnesyl pyrophosphate or geranylgeranyl diphosphate as a substrate, producing (E)-β-ocimene as the major product and a trace amount of (Z)-β-ocimene. Moreover, as a catalytic product of PbeOCS, (E)-β-ocimene showed repellent effects on larvae of S. litura in dual-choice bioassays. What is more, (E)-β-ocimene increased mortalities of larvae in no-choice bioassays. These findings provide an overview of transcriptomic changes in wild pears in response to chewing herbivores and insights into (E)-β-ocimene biosynthesis in pear plants, which will help elucidate the molecular mechanisms underlying pear-insect interactions.

Rhizosphere Acidification as the Main Trait Characterizing the Differential In Vitro Tolerance to Iron Chlorosis in Interspecific Pyrus Hybrids

Physiological responses of different interspecific Pyrus hybrids and an open pollinated Pyrus communis ‘Williams’ (Pcw) grown under in vitro culture conditions simulating lime induced chlorosis were studied. The hybrids were derived from crosses between the ‘Pyriam’ pear rootstock and four Pyrus species of the Mediterranean region, namely P. amygdaliformis Vill. (Pa), P. amygdaliformis persica Bornme. (Pap), P. communis cordata (Desv.) Hook. (Pcc), and P. elaeagrifolia Pall (Pe), all known for their higher field tolerance to iron-chlorosis than P. communis. Twenty hybrids and one open pollinated Pcw were micropropagated, and plantlets were in vitro characterized for their physiological responses to iron-deficiency conditions. Rooted plantlets were transferred to a culture medium with 2 µM Fe3+ DTPA and 10 or 20 mM NaHCO3. These physiological responses were scored at 1, 3, 7, and 28 days from the start of the in vitro assay. Leaf total chlorophyll content, the capacity of roots to acidify the medium, reduced iron, and exudates of phenolic acids and organic acids were analyzed in each media and time sample. Leaf chlorophyll levels for the clones derived from Pcc were the highest, especially under the highest bicarbonate concentration, followed by those derived from Pap and Pa. The higher chlorophyll content of Pcc clones were related with their higher capacity to acidify the media but not with their iron reduction capacity at the root level. On the other hand, hybrid clones derived from Pe showed a higher Fe3+ reduction ability than clones from all the other species during the whole assay but only when the bicarbonate concentration was lower. The exudation of phenolic acids by the roots was higher in Pcw than in the other species, and this response might explain why the total chlorophyll levels in Pcw clones are similar to those of Pe and Pa ones. These results with Pyrus spp. bring more evidence in support of the idea that iron reduction capacity at the root level is not directly related with a higher tolerance to iron deficiency caused by the high pH of calcareous soils. Instead, the ability to acidify the rhizosphere is the trait of choice for the selection of the pear hybrid clones better adapted to lime induced chlorosis. In addition, the in vitro assay to select the Pyrus clones for tolerance to iron chlorosis could be shortened to one week of culture in 10 mM NaHCO3, measuring the leaf chlorophyll level, acidification of the culture medium, and exudation of phenolic acids as the physiological responses to predict tolerance to lime-induced chlorosis.

Seed Transmission of Three Viruses in Two Pear Rootstock Species Pyrus betulifolia and P. calleryana.

Viral seed transmission causes the spread of many plant viral diseases. Pyrus betulifolia and P. calleryana are important rootstock germplasms for pear production in China. This study revealed the widespread infection of apple stem grooving virus (ASGV), apple chlorotic leaf spot virus (ACLSV), and apple stem pitting virus (ASPV) in maternal trees of P. betulifolia and P. calleryana by nested multiplex reverse transcription-polymerase chain reaction (nmRT-PCR) assays. Seeds from eight P. betulifolia and two P. calleryana trees had positive rates of 15.9–73.9%, 0–21.2%, and 40.4% for ASGV, ASPV, and ACLSV, respectively. At the cotyledon and 6–8 true leaf stages, seedlings grown from seeds of infected trees gave positive rates of 5.4% and 9.3% for ASGV, 6.7% and 15.6% for ACLSV, and 0% and 2.7% for ASPV, respectively. Incidence in nursery P. betulifolia seedlings of 10.1%, 5.3%, and 3.5% were determined for ASGV, ACLSV, and ASPV, respectively. The nucleotide sequences of coat protein (CP) and movement protein coding genes of both ASGV and ASPV, and CP gene of ACLSV from maternal trees, seeds, and seedlings were analyzed. Sequence identities and phylogenetic comparison with corresponding sequences from GenBank demonstrated that molecular variation occurred within ASGV, ACLSV, and ASPV isolates, with most sequences determined here had close relationships with reported isolates infecting pear or formed independent clades. This is the first report on the seed transmission and the molecular characteristics of these viruses infecting two rootstock species. These findings provided important evidence in management effort for pear viral diseases.

Determination of some chemical properties of wild pear (Pyrus spinosa Forsk.)

Wild pear plant species Pyrus spinosa Forsk., naturally grown in woodlands in Turkey, are one of the most important wild fruits consumed by local people as food, as well as used for medical purposes. In this study, some chemical properties were determined such as fruit seed and fruit pulp antioxidant and antimicrobial activities, fruit seed fatty acid components of Pyrus spinosa determined, which grow naturally in Ödemiş, İzmir, in the south of Turkey. The qualitative and quantitative analyses of fatty acid components of fruit seed of P. spinosa were determined by GC/MS analysis. A total of 16 fatty acids in the fruit seed were found in the analysis results and 44.8% linoleic acid (C18:2n6c), 40.5% oleic acid (C18:1n9c), and 7.6% palmitic acid (C16:0) components were found in higher proportions. Free radical removal effects of fruit seed and fruit pulp extracts were determined using the DPPH method. The antimicrobial activity of fruit pulp extract of P. spinosa was determined by the disk diffusion method against E. coli, S. aureus, and C. albicans. This study revealed that the fruit pulp and fruit seed of P. spinosa, which had a significant level of antioxidant activity, antimicrobial activity, and fruit seed had high levels fatty acids components.

First Report of Postharvest Blue Mold decay caused by Penicillium expansum on Lemon (Citrus limon) fruit in China.

Lemon (Citrus limon) is one of the most important commercial (both dried and fresh) citrus fruits in China. In the spring of 2019, postharvest blue mold decay was observed at an incidence of 3-5% on lemon fruit at the local markets in Beijing, China. Fruit lesions were circular, brown, soft, and watery, and rapidly expanded at 25°C. To isolate the causal organism, small pieces (2 mm3) were cut from the lesions, surface-sterilized for 1 min in 1.5% NaOCl, rinsed three times with sterilized water, dried with sterile filter paper, placed onto potato dextrose agar (PDA) medium, and incubated at 25°C for 6 days. Eight morphologically similar single-colony fungal isolates were recovered from six lemon fruit. Colony surfaces were bluish-green on the upper surface and cream to yellow-brown one the reverse. Hyphae on colony margins were entirely subsurface and cream in color. Mycelium was highly branched, septate, and colorless, and conidiophores were 250 to 450 × 3.0 to 4.0 µm in size. Stipe of conidiophores were smooth-walled, bearing terminal penicilli, typically terverticillate or less commonly birverticillate, rami occurring singly, 16 to 23 × 3.0 to 4.0 µm, metulae in 3 to 6, measuring 12 to 15 × 3.0 to 4.0 µm. Phialides were ampulliform to almost cylindrical, in verticils of 5 to 8, measuring 8 to 11 × 2.5 to 3.2 µm with collula. Conidia were smooth-walled, ellipsoidal, measuring 3.0 to 3.5 × 2.5 to 3.0 µm. According to morphological characteristics, the fungus was identified as Penicillium expansum (Visagie et al. 2014). For molecular identification, genomic DNA of eight fungal isolates was extracted, regions of the beta-tubulin (TUB), and calmodulin (CAL) genes and ITS region, were amplified using Bt2a/Bt2b, CAL-228F/ CAL-737, and ITS1/ITS4 primers respectively. Obtained sequences of all isolates were identical to sequences of the representative isolate YC-IK12, which was submitted in the GenBank. BLAST results of YC-IK12 sequences (ITS; MT856700: TUB; MT856958: CAL; MT856959) showed 98 to 100% similarity with P. expansum accessions (NR-077154, LN896428, JX141581). For pathogenicity tests, 10 μl of conidial suspension (10 × 105 conidia/ml) from seven-day-old YC-IK12 culture was inoculated using a sterilized needle into the surface of each five asymptomatic disinfected lemons. As a control, three lemons were inoculated using sterile distilled water. All inoculated lemons were placed in plastic containers and incubated at 25°C for 7 days. Decay lesions, identical to the original observations, developed on all inoculated lemons, while control lemons remained asymptomatic. Fungus re-isolated from the inoculated lemon was identified as P. expansum on the basis morphology and Bt2a/Bt2b, CAL-228F/ CAL-737, and ITS1/ITS4 sequences. Previously, Penicillium spp. including P. expansum have been reported as post-harvest pathogens on various Citrus spp. (Louw & Korsten 2015). However, P. digitatum has been reported on lemons and P. expansum has been reported on stored Kiwifruit (Actinidia arguta), Malus, and Pyrus species in China (Tai, 1979; Wang et al. 2015). To our knowledge, this is the first report of blue mold caused by P. expansum on lemons in China. References Louw, J. P., Korsten, L. 2015. Plant Dis. 99:21-30. Tai, F.L. 1979. Sylloge Fungorum Sinicorum. Sci. Press, Acad. Sin., Peking, 1527 pages. 8097 Visagie, C.M. et al. 2014. Studies. Mycol.78: 343. Wang, C. W. et al. 2015. Plant Dis. 99:1037.

The Distribution and Origins of Pyrus hopeiensis-"Wild Plant With Tiny Population" Using Whole Genome Resequencing.

Pyrus hopeiensis is a valuable but endangered wild resource in the genus Pyrus. It has been listed as one of the 120 wild species with tiny population in China. The specie has been little studied. A preliminary study of propagation modes in P. hopeiensis was performed through seed propagation, hybridization, self-crossing trials, bud grafting, branch grafting, and investigations of natural growth. The results showed that the population size of P. hopeiensis was very small, the distribution range was limited, and the habitat was extremely degraded. In the wild population, natural hybridization and root tiller production were the major modes of propagation. Whole genome re-sequencing of the 23 wild and cultivated accessions from Pyrus species collected was performed using an Illumina HiSeq sequencing platform. The sequencing depth range was 26.56x−44.85x and the average sequencing depth was 32x. Phylogenetic tree and principal component analyses (PCA) based on SNPs showed that the wild Pyrus species, such as PWH06, PWH07, PWH09, PWH10, PWH13, and PWH17, were closely related to both P. hopeiensis HB-1 and P. hopeiensis HB-2. Using these results in combination with morphological characteristics, it speculated that P. hopeiensis populations may form a natural hybrid group with frequent gene exchanges between and within groups. A selective elimination analysis on the P. hopeiensis population were performed using Fst and π radio and a total of 381 overlapping genes including SAUR72, IAA20, HSFA2, and RKP genes were obtained. These genes were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) function enrichment. And four KEGG pathways, including lysine degradation, sphingolipid metabolism, other glycan degradation, and betaine biosynthesis were significantly enriched in the P. hopeiensis population. Our study provided information on genetic variation, evolutionary relationships, and gene enrichment in P. hopeiensis population. These data will help reveal the evolutionary history and origin of P. hopeiensis and provide guidelines for subsequent research on the locations of functional genes.