Corylus mandshurica, also known as pilose hazelnut, is an economically and ecologically important species in China. In this study, ten polymorphic simple sequence repeat (SSR) markers were applied to evaluate the genetic diversity and population structure of 348 C. mandshurica individuals among 12 populations in China. The SSR markers expressed a relatively high level of genetic diversity (Na = 15.3, Ne = 5.6604, I = 1.8853, Ho = 0.6668, and He = 0.7777). According to the coefficient of genetic differentiation (F_st = 0.1215), genetic variation within the populations (87.85%) were remarkably higher than among populations (12.15%). The average gene flow (Nm = 1.8080) significantly impacts the genetic structure of C. mandshurica populations. The relatively high gene flow (Nm = 1.8080) among wild C. mandshurica may be caused by wind-pollinated flowers, highly nutritious seeds and self-incompatible mating system. The UPGMA (unweighted pair group method of arithmetic averages) dendrogram was divided into two main clusters. Moreover, the results of STRUCTURE analysis suggested that C. mandshurica populations fell into two main clusters. Comparison of the UPGMA dendrogram and the Bayesian STRUCTURE analysis showed general agreement between the population subdivisions and the genetic relationships among populations of C. mandshurica. Group I accessions were located in Northeast China, while Group II accessions were in North China. It is worth noting that a number of genetically similar populations were located in the same geographic region. The results further showed that there was obvious genetic differentiation among populations from Northeast China to North China. Results from the Mantel test showed a weak but still significant positive correlation between Nei’s genetic distance and geographic distance (km) among populations (r = 0.419, P = 0.005), suggesting that genetic differentiation in the 12 C. mandshurica populations might be related to geographic distance. These data provide comprehensive information for the development of conservation strategies of these valuable hazelnut resources.

Corylus L. is an economically and phylogenetically important genus in the family Betulaceae. Taxonomic and phylogenetic relationships of Corylus species have long been controversial for lack of effective molecular markers. In this study, the complete chloroplast (cp) genomes of six Corylus species were assembled and characterized using next-generation sequencing. We compared the genome features, repeat sequences, sequence divergence, and constructed the phylogenetic relationships of the six Corylus species. The results indicated that Corylus cp genomes were typical of the standard double-stranded DNA molecule, ranging from 160,445 base pairs (bp) (C. ferox var. thibetca) to 161,621 bp (C. yunnanensis) in length. Each genome contained a pair of inverted repeats (IRs), a large single-copy (LSC) region and a small single-copy (SSC) region. Each of the six cp genomes possessed 113 unique genes arranged in the same order, including 80 protein-coding, 29 tRNA, and 4 rRNA genes. C. yunnanensis contained the highest number of repeat sequences, and the richest SSRs in six cp genomes were A/T mononucleotides. Comparative analyses of six Corylus cp genomes revealed four hotspot regions (trnH-psbA, rpoB-trnC, trnF-ndhJ, and rpl32-trnL) that could be used as potential molecular markers. Phylogenetic analyses of the complete chloroplast genomes and 80 protein-coding genes exhibited nearly identical topologies that strongly supported the monophyly of Corylus and simultaneously revealed the generic relationships among Betulaceae. The availability of these genomes can offer valuable genetic information for further taxonomy, phylogeny, and species delimitation in Corylus or even Betulaceae plants.

Background: Previous phylogenetic conclusions on the family Betulaceae were based on either morphological characters or traditional single loci, which may indicate some limitations. The chloroplast genome contains rich polymorphism information, which is very suitable for phylogenetic studies. Thus, we sequenced the chloroplast genome sequences of three Betulaceae species and performed multiple analyses to investigate the genome variation, resolve the phylogenetic relationships, and clarify the divergence history. Methods: Chloroplast genomes were sequenced using the high-throughput sequencing. A comparative genomic analysis was conducted to examine the global genome variation and screen the hotspots. Three chloroplast partitions were used to reconstruct the phylogenetic relationships using Maximum Likelihood and Bayesian Inference approaches. Then, molecular dating and biogeographic inferences were conducted based on the whole chloroplast genome data. Results: Betulaceae chloroplast genomes consisted of a small single-copy region and a large single copy region, and two copies of inverted repeat regions. Nine hotspots can be used as potential DNA barcodes for species delimitation. Phylogenies strongly supported the division of Betulaceae into two subfamilies: Coryloideae and Betuloideae. The phylogenetic position of Ostryopsis davidiana was controversial among different datasets. The divergence time between subfamily Coryloideae and Betuloideae was about 70.49 Mya, and all six extant genera were inferred to have diverged fully by the middle Oligocene. Betulaceae ancestors were probably originated from the ancient Laurasia. Discussions: This research elucidates the potential of chloroplast genome sequences in the application of developing molecular markers, studying evolutionary relationships and historical dynamic of Betulaceae.It also reveals the advantages of using chloroplast genome data to illuminate those phylogenies that have not been well solved yet by traditional approaches in other plants.

Hazelnut has gained economic value in China in recent years, but its large-scale planting and research started later than other countries. Conducting basic research on hazelnut trees requires studying their related microorganisms. Here, we used high-throughput DNA sequencing to quantify the fungal communities in the root endospheres and rhizosphere soil of four hazelnut species. Fungal diversity in the rhizosphere soil was significantly higher than that in the root endospheres. Rhizosphere soil had more Mortierellomycota, and the fungal community compositions differed among the four hazelnut species. The root endospheres, especially those of the Ping’ou (Corylus heterophylla×Corylus avellana) trees, contained more ectomycorrhizal fungi. The co-occurrence networks in the rhizosphere soil were more sophisticated and stable than those in the root endospheres, even when the root endospheres had higher modularity, because the structural differentiation of the root endospheres differed from that of the rhizosphere soil. Two-factor correlation network analysis and linear regression analysis showed that the total organic carbon was the main environmental factor affecting the fungal communities. Our study revealed the community compositions, functional predictions, and co-occurrence network structural characteristics of fungi in hazelnut root endospheres and rhizosphere soil. We also examined the potential keystone taxa, and analyzed the environmental factors of the dominant fungal community compositions. This study provides guidance for the growth of hazelnut and the management of hazelnut garden, and provides an insight for future development of fungal inoculants to be used in hazelnut root.

Hazelnut is one of the four major nuts in the world and has high nutritional and economic value. This study employed Illumina sequencing of ITS rDNA and 16S rRNA genes to identify the seasonal changes in soil microbial community, the predominant environmental factors driving microbial community composition, and the differences in soil microbial composition among different species of the genus Corylus. We found that the soil microbial community composition of species of Corylus changed significantly with the change in seasons. Corylus heterophylla and Corylus kweichowensis had more ectomycorrhiza in their soil compared to Corylus avellane. The main factor influencing fungal community composition in soil was the available potassium, while that of bacteria was the total phosphorus content. Co-occurrence network analysis revealed that the ratio of positive interaction to negative interaction in soil of C. heterophylla and Ping’ou (C. heterophylla × C. avellane) was higher, while the negative interaction of soil community structure in C. avellane was greater. The bacterial community was more stable than the fungal community according to microbial diversity and co-occurrence network analyses. The findings of this research may facilitate improvements to the production and soil system management in hazel planting processes.

Living cover is an important management measure for orchards in China, and has certain influences on soil properties, microorganisms, and the micro-ecological environment. However, there are few studies on the effects of living cover on the soil changes in hazelnut orchards. In this study, we compared the soils of living cover treatments with Vulpia myuros and the soils of no cover treatments, and analyzed the observed changes in soil properties, microorganisms, and microbial functions by using high-throughput ITS rDNA and 16S rRNA gene Illumina sequencing. The results demonstrated that the total organic carbon content in the 20–40 cm deep soils under the living cover treatments increased by 32.87 and 14.82% in May and July, respectively, compared with those under the no cover treatments. The living cover treatment with V. myuros also significantly increased the contents of total phosphorus (TP), total nitrogen (TN), available phosphorus (AP), and available potassium (AK) in the soil samples. Moreover, the influence of seasons was not as significant as that of soil depth. The living cover treatment also significantly improved the soil enzyme activity levels. The results demonstrated that Ascomycota, Mortierellomycota and Basidiomycota were the dominant fungal phyla in all samples, while Proteobacteria, Actinobacteria, Acidobacteria, Firmicutes, and Chloroflexi were the dominant bacterial phyla, but the different treatments impacted the compositions of fungal and bacterial communities. Principal component analysis (PCA) showed that living cover with V. myuros significantly changed the soil fungal community structures whereas the bacterial community structures may be more sensitive to seasonal changes. At the microbial functional level, the living cover treatment increased the fungal operational taxonomic units (OTUs) of symbiotrophs and decreased that of pathotrophs. According to this study, we believe that the application of a living cover with V. myuros has a favorable regulating influence on soil properties, microbial communities and microbial function. This treatment can also reduce the use of herbicides, reduce the cost of orchard management, and store more carbon underground to achieve sustainable intensification of production in hazelnut orchards, so it can be considered as a management measure for hazelnut orchards.