Paulownia Research Center

The Schematic representation of a Stereolithography (SL) scalable manufacturing technology called additive manufacturing (AM) in oil and gas industry.

The Chinese chestnut (Castanea mollissima) seed provides a rich source of carbohydrates as food and feed. However, little is known about starch biosynthesis in the seeds. The objectives of this study were to determine seed composition profiles and identify genes involved in starch and sucrose metabolism. Metabolite analysis showed that starch was the major component and rapidly accumulated during seed endosperm development. Amylopectin was approximately 3-fold of amylose content in chestnut starch. Illumina platform-based transcriptome sequencing generated 56671 unigenes in two cDNA libraries from seed endosperms collected at 45 and 75 days after flowering (DAF). A total of 1537 unigenes showed expression differences ≥2-fold in the two stages of seeds including 570 up-regulated and 967 down-regulated unigenes. One hundred and fifty-two unigenes were identified as involved in starch and sucrose metabolism, including 1 for glycogenin glucosyltransferase, 4 for adenylate transporter (brittle1-type), 3 for ADP-glucose pyrophosphorylase (AGP, not brittle2- or shrunken2-type), 3 for starch synthase (SS), 2 for starch branching enzyme, 5 for starch debranching enzyme, 11 for sucrose synthase, and 3 for sucrose-phosphate synthase. Among them, 58 unigenes showed a ≥2-fold expression difference between the 45 and 75 DAF seeds including 11 up- and 47 down-regulated unigenes. The expression of 21 unigenes putatively coding for major enzymes in starch and sucrose metabolism was validated by qPCR using RNA from five seed stages. Expression profiles and correlation analysis indicated that the mRNA levels of AGP (large and small subunits), granule-bound SS2, and soluble SS1 and SS4 were well-correlated with starch accumulation in the seeds. This study suggests that the starch biosynthesis pathway in Chinese chestnut is similar to that of potato tuber/Arabidopsis leaf and differs from that of maize endosperm. The information provides valuable metabolite and genetic resources for future research in starch and sucrose metabolism in Chinese chestnut tree.

seedlings using quantitative mass spectrometry. In total, we identified 8963 proteins, 2893 acetylated proteins (5558 acetylation sites), and 1271 succinylated proteins (1970 succinylation sites), with 425 (533 sites) simultaneously acetylated and succinylated. Comparative analysis revealed that 276 proteins, 546 acetylated proteins (741 acetylation sites) and 5 succinylated proteins (5 succinylation sites) were regulated in response to phytoplasma infection, suggesting that acetylation may be more important than succinylation in PaWB. Enzymatic assays showed that acetylation of specific sites in protochlorophyllide reductase and RuBisCO, key enzymes in chlorophyll and starch biosynthesis, respectively, modifies their activity in phytoplasma-infected seedlings. On the basis of these results, we propose a model to elucidate the molecular mechanism of responses to PaWB and offer a resource for functional studies on the effects of acetylation on protein function.

This paper aims to establish evidence for available phosphorous (AP) binding with total nitrogen (N) in subtropical forest soils. Soil organic carbon (SOC), total N, total phosphorous (P) and AP concentration were measured for three contrasting forest types in southern China: Masson pine forest (MPF), coniferous and broadleaved mixed forest (CBMF) and monsoon evergreen broadleaved forest (MEBF). A pot experiment with N addition was conducted to confirm the dominant factor to affect on soil AP concentration. The results showed that mean soil total N concentration in 0-10 cm soil layer was 440 ± 50 for MPF, 840 ± 80 for CBMF and 1020 ± 50 mg kg(-1) for MEBF, respectively. The mean soil AP concentration in 0-10 cm soil layer was 2.67 ± 0.87 for MPF, 2.65 ± 0.58 for CBMF, 4.10 ± 0.29 mg kg(-1) for MEBF, respectively. The soil total N concentration could explain about 70% of the variations in soil AP concentration in the top 20 cm soil layers in the three forest types. A pot experiment with N addition also showed an increase of AP concentration from 2.56 to 5.63 mg kg(-1), when N addition increased from 5 g to 17 g NH4NO3. Our results therefore suggested that N addition significantly increased soil AP concentration, which might be beneficial for stabilizing the net primary production of subtropical forests that were limited by soil AP. This finding may provide a theory basis for tropical and subtropical forests management.

Pyrus ussriensis Maxim. is native to the northern part of China, but whose habitats are currently being destroyed by environmental changes and human deforestation. An investigation of population structure and genetic diversity of wild Ussurian pear is a priority in order to acquire fundamental knowledge for conservation. A total of 153 individuals of wild Ussurian pear from the main habitats, Heilongjiang, Jilin, and Inner Mongolia in China, possessed low genetic diversity as a result of habitat fragmentation. The genetic diversity of the populations in Inner Mongolia and north east of Heilongjiang was especially low and there was the possibility of inbreeding. Wild Ussurian pears were divided into 5 groups based on the Bayesian clustering method using 20 nuclear SSRs (nSSRs) and 5 groups by haplotype distributions using 16 chloroplast SSRs (cpSSRs), and the populations in Inner Mongolia and north east of Heilongjiang represented unique genotypes. AMOVA indicated there was a 20.05% variation in nSSRs and a 44.40% variation in cpSSRs among populations. These values are relatively high when compared to those of other tree species. Haplotype E, positioned in the center of the cpSSR analysis network and showed the largest number of connections with other haplotypes, represented the most important haplotype. Inner Mongolia and the north east of Heilongjiang are two areas that need urgent conservation because of their genetic vulnerability and peculiarity. We determined 4 conservation units based on the clustering by nSSRs and cpSSRs, and geographic factor. This information is helpful in deciding the conservation strategies for wild Ussurian pear in China.

Radopholus similis is a migratory plant-parasitic nematode that causes severe damage to many agricultural and horticultural crops. Calreticulin (CRT) is a Ca2+-binding multifunctional protein that plays key roles in the parasitism, immune evasion, reproduction and pathogenesis of many animal parasites and plant nematodes. Therefore, CRT is a promising target for controlling R. similis. In this study, we obtained the full-length sequence of the CRT gene from R. similis (Rs-crt), which is 1,527-bp long and includes a 1,206-bp ORF that encodes 401 amino acids. Rs-CRT and Mi-CRT from Meloidogyne incognita showed the highest similarity and were grouped on the same branch of the phylogenetic tree. Rs-crt is a multi-copy gene that is expressed in the oesophageal glands and gonads of females, the gonads of males, the intestines of juveniles and the eggs of R. similis. The highest Rs-crt expression was detected in females, followed by juveniles, eggs and males. The reproductive capability and pathogenicity of R. similis were significantly reduced after treatment with Rs-crt dsRNA for 36 h. Using plant-mediated RNAi, we confirmed that Rs-crt expression was significantly inhibited in the nematodes, and resistance to R. similis was significantly improved in transgenic tomato plants. Plant-mediated RNAi-induced silencing of Rs-crt could be effectively transmitted to the F2 generation of R. similis; however, the silencing effect of Rs-crt induced by in vitro RNAi was no longer detectable in F1 and F2 nematodes. Thus, Rs-crt is essential for the reproduction and pathogenicity of R. similis.

Abstract Background Persimmon ( Diospyros kaki Thunb.) has various labile sex types, and studying its sex differentiation can improve breeding efficiency. However, studies on sexual regulation patterns in persimmon have focused mainly on monoecy and dioecy, whereas little research has been published on andromonoecy. In order to reveal the sex differentiation regulation mechanism of andromonoecious persimmon, we performed histological and cytological observations, evaluated OGI and MeGI expression and conducted phytohormones assays and mRNA and small RNA transcriptome analyses of the male and hermaphroditic floral buds of the andromonoecious persimmon ‘Longyanyeshi 1’. Results Stages 2 and 4 were identified as the critical morphological periods for sex differentiation of ‘Longyanyeshi 1’ by histological and cytological observation. At both stages, OGI was differentially expressed in male and hermaphroditic buds, but MeGI was not. This was different from their expressions in dioecious and monoecious persimmons. Meantime, the results of phytohormones assays showed that high IAA, ABA, GA 3 , and JA levels at stage 2 may have promoted male floral bud differentiation. However, high JA levels at stage 4 and high ZT levels at stages 2 and 4 may have promoted hermaphroditic floral bud differentiation. In these phytohormone biosynthesis and signaling pathways, 52 and 54 differential expression genes (including Aux/IAA , ARFs , DELLA , AHP , A-ARR , B-ARR , CYP735A , CRE1 , PP2C , JAZ , MYC2 , COI1 , CTR1 , SIMKK , ACO, and MPK6 ) were identified, respectively. During the development of male floral buds, five metacaspases genes may have been involved in pistil abortion. In addition, MYB , FAR1 , bHLH, WRKY, and MADS transcription factors might play important roles in persimmon floral bud sex differentiation. Noteworthy, miR169v_1, miR169e_3, miR319_1, and miR319 were predicted to contribute to phytohormone biosynthesis and signaling pathways and floral organogenesis and may also regulate floral bud sex differentiation . Conclusion The present study revealed the differences in morphology and phytohormones content between male and hermaphroditic floral buds of ‘Longyanyeshi 1’ during the process of sex differentiation, and identified a subset of candidate genes and miRNAs putatively associated with its sex differentiation. These findings can provide a foundation for molecular regulatory mechanism researching on andromonoecious persimmon.

Eucommia ulmoides Oliv. is a traditional medical plant in Asia; however, it is still unknown whether Eucommia male flowers have an antihypertensive activity. In this study, we found that the aqueous extract of Eucommia ulmoides Oliv. male flowers can lower the blood pressure of SHR in a dose‐dependent manner. Mechanistic studies suggested that the aqueous extract of male flowers can promote the mRNA and protein expressions of ACE2 in the kidney of SHR. ELISA assay showed that the plasma levels of ANG II was decreased, while ANG‐(1–7) was increased in SHR treated with the aqueous extract of male flowers. ACE2 inhibitor DX600 can reverse the aqueous extract of Eucommia ulmoides Oliv. male flower‐induced downregulation of Ang II and upregulation of Ang‐(1–7), as well as the reduction of blood pressure in SHR. Moreover, Ang‐(1–7)‐Mas receptor antagonist A‐779 abolished the antihypertensive effects of the aqueous extract of Eucommia ulmoides Oliv. male flower in SHR. The aqueous extract of Eucommia ulmoides Oliv. male flowers exhibited an antihypertensive action through the activation of ACE2‐Ang‐(1–7)‐Mas signaling pathways in spontaneously hypertensive rats.

The mitogen-activated protein kinase (MAPK) cascades, which play crucial roles in plant development processes, are universal modules of signal transduction in eukaryotes and consist of a core module of three sequentially phosphorylated kinases: MAPK, MAPK kinase (MAPKK), and MAPKK kinase (MAPKKK). This is the first report on the identification and analysis of MAPK cascades in Eucommia ulmoides. We conducted a genome-wide screening and identified 13 EuMAPKs, five EuMAPKKs, and 57 EuMAPKKKs. The construction of phylogenetic trees revealed that EuMAPKs and EuMAPKKs were divided into four groups (A, B, C, and D), and EuMAPKKKs were divided into three subfamilies (MEKK, RAF, and ZIK). These subfamilies were further confirmed by conserved domain/motif analysis and gene structure analysis. Based on the expression profiles of all identified EuMAPK cascades in various organs at different developmental stages, three genes (EuRAF22-2, EuRAF34-1, and EuRAF33-2) with stable expression patterns at all stages of fruit or leaf development, three genes (EuRAF2-3, EuMPK11, and EuMEKK21) with differential expression patterns, and two highly expressed genes (EuZIK1 and EuMKK2) were screened and validated by qRT-PCR. Overall, our results could be used for further research on the precise role of MAPK cascades during organ development in E. ulmoides.

Eucommia ulmoides Oliver, the only member of the Eucommiaceae family, is a rare and valuable tree used to produce a highly valued traditional Chinese medicine and contains α-linolenic acid (ALA) up to 60% of the total fatty acids in the kernels (embryos). Glycolysis provides both cellular energy and the intermediates for other biosynthetic processes. However, nothing was known about the molecular basis of the glycolytic pathway in E. ulmoides kernels. The purposes of this study were to identify novel genes of E. ulmoides related to glycolytic metabolism and to analyze the expression patterns of selected genes in the kernels. Transcriptome sequencing based on the Illumina platform generated 96,469 unigenes in four cDNA libraries constructed using RNAs from 70 and 160 days after flowering kernels of both low- and high-ALA varieties. We identified and characterized the digital expression of 120 unigenes coding for 24 protein families involved in kernel glycolytic pathway. The expression levels of glycolytic genes were generally higher in younger kernels than in more mature kernels. Importantly, several unigenes from kernels of the high-ALA variety were expressed more than those from the low-ALA variety. The expression of 10 unigenes encoding key enzymes in the glycolytic pathway was validated by qPCR using RNAs from six kernel stages of each variety. The qPCR data were well consistent with their digital expression in transcriptomic analyses. This study identified a comprehensive set of genes for glycolytic metabolism and suggests that several glycolytic genes may play key roles in ALA accumulation in the kernels of E. ulmoides.

Camellia oleifera is an important edible oil woody plant in China. Lack of useful molecular markers hinders current genetic research on this tree species. Transcriptome sequencing of developing C. oleifera seeds generated 69,798 unigenes. A total of 6949 putative microsatellites were discovered among 6042 SSR-containing unigenes. Then, 150 simple sequence repeats (SSRs) were evaluated in 20 varieties of C. oleifera. Of these, 52 SSRs revealed polymorphism, with the number of alleles per locus ranging from 2 to 15 and expected heterozygosity values from 0.269 to 0.888. The polymorphic information content varied from 0.32 to 0.897. Cross-species transferability rates in Camellia chekangoleosa and Camellia japonica were 90.4 and 78.8%, respectively. The 52 polymorphic unigene-derived SSR markers serve to enrich existing microsatellite marker resources for C. oleifera and offer potential for applications in genetic diversity evaluation, molecular fingerprinting, and genetic mapping in C. oleifera, C. chekangoleosa, and C. japonica.

Paper mulberry, a vigorous pioneer species used for ecological reclamation and a high-protein forage plant for economic development, has been widely planted in China. To further develop its potential value, it is necessary to explore the regulatory mechanism of nitrogen metabolism for rational nitrogen utilization. In this study, we investigated the morphology, physiology and transcriptome of a paper mulberry hybrid (Broussonetia kazinoki × B. papyrifera) in response to different nitrogen concentrations. Moderate nitrogen promoted plant growth and biomass accumulation. Photosynthetic characteristics, concentration of nitrogenous compounds and activities of enzymes were stimulated under nitrogen treatment. However, these enhancements were slightly or severely inhibited under excessive nitrogen supply. Nitrite reductase and glutamate synthase were more sensitive than nitrate reductase and glutamine synthetase and more likely to be inhibited under high nitrogen concentrations. Transcriptome analysis of the leaf transcriptome identified 161,961 unigenes. The differentially expressed genes associated with metabolism of nitrogen, alanine, aspartate, glutamate and glycerophospholipid showed high transcript abundances after nitrogen application, whereas those associated with glycerophospholipid, glycerolipid, amino sugar and nucleotide sugar metabolism were down-regulated. Combined with weighted gene coexpression network analysis, we uncovered 16 modules according to similarity in expression patterns. Asparagine synthetase and inorganic pyrophosphatase were considered two hub genes in two modules, which were associated with nitrogen metabolism and phosphorus metabolism, respectively. The expression characteristics of these genes may explain the regulation of morphological, physiological and other related metabolic strategies harmoniously. This multifaceted study provides valuable insights to further understand the mechanism of nitrogen metabolism and to guide utilization of paper mulberry.

Flavonoids are natural antioxidants in plants that affect the color of plant tissues. Flavonoids can be divided into eight subgroups, including flavonols, anthocyanins, and proanthocyanidins. The mechanisms of flavonoid synthesis in model plants have been widely studied. However, there are a limited number of reports on the synthesis of flavonoids in the red leaf varieties of woody plants. In this study, we combined morphological observation, pigment content determination, metabolomics, and transcriptomics to investigate the metabolites and gene regulation present in the red and green leaves of Eucommia ulmoides Oliv. The results showed that the red leaves have a lower chlorophyll content and a higher anthocyanin content. Metabonomic analysis identified that the relative content of most flavonoids is up-regulated in red leaves based on UPLC-ESI-MS/MS, which included the apigenin class, quercetin class, kaempferol class, and procyanidins. Transcriptome data suggested that the differentially up-regulated genes are enriched in flavonoid and anthocyanin synthesis pathways, ABC transport, and GST pathways. The integrative analysis of the transcriptome and metabolome showed that the up-regulation of flavonoid metabolism and a high expression of chlorophyll degradation with the down-regulation of chlorophyll biosynthesis genes are detected in E. ulmoides red leaves compared with green leaves. In addition, the co-expression networks implied that cyanidin 3-5-O-diglucoside, EuDR5, EuPAL2, EuDFR11, Eu3MaT1, and EuF3′H are likely associated with the red leaf coloration of E. ulmoides. In summary, this research provided a reference for studying the mechanism of red leaf coloration in woody plants and the use of E. ulmoides red leaves as feedstock for bioactive products.

• The co-evolutionary arms race between plants and pathogens. • Pathogens manipulate host epigenetic machinery to suppress defenses. • Plants deploy countermeasures to maintain immune resilience. • Advances in high-throughput sequencing and CRISPR-based tools are shedding light on these complex interactions. • There is need to explore the interplay between these epigenetic mechanisms under multifaceted environmental stresses. Epigenetic modifications are emerging as pivotal regulatory mechanisms in plant-pathogen interactions, especially under changing environmental conditions. This review systematically examines the application of epigenetic processes (DNA methylation, histone modifications, non-coding RNAs, and RNA methylation) in shaping the intricate dynamics between plants and pathogens. DNA methylation influences plant immunity by modulating gene expression and chromatin accessibility, while histone modifications, such as acetylation and methylation, enable plants to establish robust defenses by fine-tuning stress-responsive genes. Small RNAs (miRNAs) and long non-coding RNAs (lncRNAs) act as post-transcriptional regulators, orchestrating immune signaling and pathogen counter-defense strategies. RNA methylation, particularly N6-methyladenosine (m6A), adds another layer of regulation, controlling the stability and translation of transcripts involved in host immunity and pathogen virulence. The review highlights the co-evolutionary arms race between plants and pathogens, where pathogens manipulate host epigenetic machinery to suppress defenses, while plants deploy countermeasures to maintain immune resilience. Advances in high-throughput sequencing and CRISPR-based tools are shedding light on these complex interactions, offering opportunities to engineer crops with enhanced disease resistance. Furthermore, the review emphasizes the need to explore the interplay between these epigenetic mechanisms under multifaceted environmental stresses. By integrating recent findings, this review underscores the potential of epigenetic research to revolutionize crop protection strategies, ensuring agricultural sustainability in a rapidly changing world.

Background In the Chinese health care industry, male Eucommia ulmoides Oliv. flowers are newly approved as a raw material of functional food. Core collections have been constructed from conserved germplasm resources based on phenotypic traits and molecular markers. However, little is known about these collections’ phytochemical properties. This study explored the chemical composition of male E. ulmoides flowers, in order to provide guidance in the quality control, sustainable cultivation, and directional breeding of this tree species. Methods We assessed the male flowers from 22 core collections using ultra-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) non-targeted metabolomics, and analyzed them using multivariate statistical methods including principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). Results We annotated a total of 451 and 325 metabolites in ESI+ and ESI− modes, respectively, by aligning the mass fragments of the secondary mass spectra with those in the database. Four chemotypes were well established using the ESI+ metabolomics data. Of the 29 screened biomarkers, 21, 6, 19, and 5 markers corresponded to chemotypes I, II, III, and IV, respectively. More than half of the markers belonged to flavonoid and amino acid derivative classes. Conclusion Non-targeted metabolomics is a suitable approach to the chemotype classification and biomarker screening of male E. ulmoides flower core collections. We first evaluated the metabolite profiles and compositional variations of male E. ulmoides flowers in representative core collections before establishing possible chemotypes and significant biomarkers denoting the variations. We used genetic variations to infer the metabolite compositional variations of male E. ulmoides flower core collections instead of using the geographical origins of the germplasm resources. The newly proposed biomarkers sufficiently classified the chemotypes to be applied for germplasm resource evaluation.

Quercus variabilis Blume is one of the most ecologically valuable tree species in China and is known to have adaptive mechanisms to climate change. Our objective was to quantify the variation pattern in the fruit morphology of Q. variabilis. Fruit samples were collected from 43 natural populations in autumn of 2019. Our results indicated that the coefficient of variation (CV) of the fruit length (FL) and fruit width (FW) were 10.08% and 11.21%, respectively. There were significant differences in the FL, FW, and fruit length-to-width ratios (FL/FW) among the studied populations. Also, there was a significant positive correlation between the FW and FL. The FL decreased with increasing precipitation in the wettest quarter (PWQ). A concave trend was observed in the variations in FL with the equivalent latitude (ELAT), longitude (LON), annual mean air temperature (MAT), and annual precipitation (AP). A similar concave trend was observed for the FL/FW with LON, MAT, and AP. A positive correlation was observed between the FW, FL and FL/FW, and the ELAT. The cluster analysis revealed five groups of the 43 natural populations. Our study findings suggests that Q. variabilis has high levels of phenotypic plasticity for geographical and climatic factors.

Paulownia catalpifolia is an important, fast-growing timber species known for its high density, color and texture. However, few transcriptomic and genetic studies have been conducted in P. catalpifolia. In this study, single-molecule real-time sequencing technology was applied to obtain the full-length transcriptome of P. catalpifolia leaves treated with varying degrees of drought stress. The sequencing data were then used to search for microsatellites, or simple sequence repeats (SSRs). A total of 28.83 Gb data were generated, 25,969 high-quality (HQ) transcripts with an average length of 1624 bp were acquired after removing the redundant reads, and 25,602 HQ transcripts (98.59%) were annotated using public databases. Among the HQ transcripts, 16,722 intact coding sequences, 149 long non-coding RNAs and 179 alternative splicing events were predicted, respectively. A total of 7367 SSR loci were distributed throughout 6293 HQ transcripts, of which 763 complex SSRs and 6604 complete SSRs. The SSR appearance frequency was 28.37%, and the average distribution distance was 5.59 kb. Among the 6604 complete SSR loci, 1-3 nucleotide repeats were dominant, occupying 97.85% of the total SSR loci, of which mono-, di- and tri-nucleotide repeats were 44.68%, 33.86% and 19.31%, respectively. We detected 112 repeat motifs, of which A/T (42.64%), AG/CT (12.22%), GA/TC (9.63%), GAA/TTC (1.57%) and CCA/TGG (1.54%) were most common in mono-, di- and tri-nucleotide repeats, respectively. The length of the repeat SSR motifs was 10-88 bp, and 4997 (75.67%) were ≤ 20 bp. This study provides a novel full-length transcriptome reference for P. catalpifolia and will facilitate the identification of germplasm resources and breeding of new drought-resistant P. catalpifolia varieties.

Eucommia ulmoides Oliver is a woody perennial dioecious species native to China and has great economic value. However, little is known about flower bud development in this species. In this study, the transcriptomes of female and male flower buds were sequenced using the Illumina platform, a next-generation sequencing technology that provides cost-effective, highly efficient transcriptome profiling. In total, 11,558,188,080 clean reads were assembled into 75,065 unigenes with an average length of 1011 bp by de novo assembly using Trinity software. Through similarity comparisons with known protein databases, 47,071 unigenes were annotated, 146 of which were putatively related to the floral development of E. ulmoides. Fifteen of the 146 unigenes had significantly different expression levels between the two samples. Additionally, 24,346 simple sequence repeats were identified in 18,565 unigenes with 12,793 sequences suitable for the designed primers. In total, 67,447 and 58,236 single nucleotide polymorphisms were identified in male and female buds, respectively. This study provides a valuable resource for further conservation genetics and functional genomics research on E. ulmoides.

The Ussurian pear is the most important cultivated pear in the northern part of China. Cultivated Ussurian pears are considered to have derived from Pyrus ussuriensis Maxim. which is native to the northeast of China. In Japan, two varieties of P. ussuriensis, P. ussuriensis var. aromatica and var. hondoensis are native to the northern area and the central area of the main island respectively. In order to reveal the origin of Pyrus ussuriensis var. aromatica distributed in the northern area of main island of Japan, more than 40 explorations have been performed in Japan and in China, and more than 30 natural habitats were recognized. These natural habitats are at risk of extinction because of human development and forest degradation caused by climate change. Population structure and genetic diversity of P. ussuriensis in China and P. ussuriensis var. aromatica in Japan have been investigated using both morphological and molecular markers in order to define appropriate conservation units, and to provide a good focus for conservation management. Distant evolutionary relationships between P. ussuriensis Maxim. in China and P. ussuriensis var. aromatica in Japan inferred from population genetic structure and phylogenetic analysis are also discussed.

Eucommia ulmoides (E. ulmoides) is a significant national strategic resource in China. It is a natural high quality rubber resource, with great development potential. We found large differences in rooting ability during adventitious root (AR) formation in two E. ulmoides improved varieties. Therefore, we used two improved varieties of E. ulmoides, ‘Huazhong 6’ (H6, with rooting rate 85.3%) and ‘Huazhong 8’ (H8, with rooting rate 22.5%) to explore the cutting rooting mechanism. In this study, we mainly determined the morphological development process of E. ulmoides cutting rooting, and compared the rooting-related indexes of the two improved varieties, and the changes in physiological indexes closely related to rooting, which include endogenous hormones, oxidases and nutrients in the phloem of the basal stem. The results showed that indole–3–acetic acid (IAA), zeatin riboside (ZR), IAA/ZR and indoleacetic acid oxidase (IAAO) were the key factors that caused big differences in rooting ability between the two E. ulmoides improved varieties. The increase in endogenous hormone IAA content and IAA/ZR value were necessary for the formation of AR. The increase in IAA content was beneficial to AR formation. The activity of IAAO was significantly negatively correlated with the rooting ability of the E. ulmoides cuttings. The high IAAO activity of the H8 cuttings led to the consumption of IAA. Although the content of IAA increased, the rooting conditions were not reached. The accumulation of nutrients before rooting also has an important effect on rooting; it is easy for cuttings to root when the carbon–nitrogen ratio (C/N) value is high. This research provides an improved understanding of the cellular and physiological underpinnings of the AR process in woody plants. In addition, it provides a theoretical basis and foundation for subsequent research on E. ulmoides cuttage technology.