Morphological growth performance and genetic parameters on Korean pine in Northeastern China

Korean pine (Pinus koraiensis) is an economically valuable species owing to its excellent timber quality and nuts useful for various purposes. But few studies have been made on growth performance, and aspects combining the genetic gain and classification method on phenotypic similarity in the selection process of superior families. Thus, the present study aimed at analyzing the genetic variation and highlight suitable morphological traits for family selection; establishing trait correlations and families' ordination based on similarities in phenotypic characters, and selecting elite families and suitable parent trees. Full-sib families from 28 crosses established in randomized complete block design from Naozhi orchard in Northeast China were used, and 11 morphological traits were investigated. Significant differences were observed among families for all traits. The traits coefficients of variation ranged from 6.07 to 56.25 % and from 0.029 to 15.213 % in phenotype and genotypic variation, respectively. A moderate level of inherited genetic control was observed (broad sense heritability H-2, varied from 0.155 to 0.438). Traits related to stem growth were highly positively correlated to each other whereas crown traits showed a weak correlation with stem traits (Pearson correlation r, ranged from -0.161 to 0.956). Based on multi-trait comprehensive analysis, we selected six elite families and six parents, which resulted in a genetic gain of 5.6 %, 16.9 %, and 36.4 % in tree height, diameter at breast height, and volume, respectively. These results make a theoretical basis for selecting excellent families and establish orchards of Korean pine from improved seeds

Isolation and Characterization of Saponin-Producing Fungal Endophytes from Aralia elata in Northeast China

The purpose of this study was to investigate the diversity of endophytic fungi of Aralia elata distributed in Northeast China as well as their capacity to produce saponins. Ninety-six strains of endophytic fungi were isolated, and polymerase chain reaction (PCR) and sequencing were employed to identify the isolates. The saponin concentrations of the culture filtrates of representative strains were measured. The agar diffusion method was used to test antimicrobial activity, while high-performance liquid chromatography (HPLC) was employed to identify the saponins produced by representative strains. Alternaria, Botryosphaeria, Camarosporium, Cryptosporiopsis, Diaporthe, Dictyochaeta, Penicillium, Fusarium, Nectria, Peniophora, Schizophyllum, Cladosporium and Trichoderma species were isolated in this study. Overall, 25% of the isolates belonged to Diaporthe (Diaporthe eres), and 12.5% belonged to Alternaria. The highest concentration of saponins was produced by G22 (2.049 mg/mL). According to the results of the phylogenetic analysis, G22 belonged to the genus Penicillium. The culture filtrate of G22 exhibited antibacterial activity against Staphylococcus aureus, and ginsenosides Re and Rb2 were detected in G22 culture filtrates by HPLC

Callose Deposition Is Required for Somatic Embryogenesis in Plasmolyzed Eleutherococcus senticosus Zygotic Embryos

Abstract: Dynamic changes in callose content, which is deposited as a plant defense response to physiological changes, were analyzed during somatic embryogenesis in Eleutherococcus senticosus zygotic embryos plasmolyzed in 1.0 M mannitol. During plasmolysis, callose deposition was clearly observed inside the plasma membrane of zygotic embryo epidermal cells using confocal laser scanning microscopy. The callose content of zygotic embryos gradually increased between 0 and 12 h plasmolysis and remained stable after 24 h plasmolysis. During eight weeks induction of somatic embryogenesis, the callose content of explants plasmolyzed for 12 h was slightly higher than explants plasmolyzed for 6 or 24 h, with the largest differences observed after 6 weeks culture, which coincided with the maximum callose content and highest number of globular somatic embryos. The highest frequency of somatic embryo formation was observed in explants plasmolyzed for 12 h. The somatic embryo induction rate and number of somatic embryos per explant were markedly different in zygotic embryos pretreated with plasmolysis alone (78.0%, 43 embryos per explant) and those pretreated with plasmolysis and the callose synthase inhibitor 2-deoxy-D-glucose (11.5%, 8 embryos per explant). This study indicates tha

Isolation and Characterization of Saponin-Producing Fungal Endophytes from Aralia elata in Northeast China

The purpose of this study was to investigate the diversity of endophytic fungi of Aralia elata distributed in Northeast China as well as their capacity to produce saponins. Ninety-six strains of endophytic fungi were isolated, and polymerase chain reaction (PCR) and sequencing were employed to identify the isolates. The saponin concentrations of the culture filtrates of representative strains were measured. The agar diffusion method was used to test antimicrobial activity, while high-performance liquid chromatography (HPLC) was employed to identify the saponins produced by representative strains. Alternaria, Botryosphaeria, Camarosporium, Cryptosporiopsis, Diaporthe, Dictyochaeta, Penicillium, Fusarium, Nectria, Peniophora, Schizophyllum, Cladosporium and Trichoderma species were isolated in this study. Overall, 25% of the isolates belonged to Diaporthe (Diaporthe eres), and 12.5% belonged to Alternaria. The highest concentration of saponins was produced by G22 (2.049 mg/mL). According to the results of the phylogenetic analysis, G22 belonged to the genus Penicillium. The culture filtrate of G22 exhibited antibacterial activity against Staphylococcus aureus, and ginsenosides Re and Rb2 were detected in G22 culture filtrates by HPLC

Genome-Wide Identification of <i>AP2/ERF</i> Superfamily Genes in <i>Juglans mandshurica</i> and Expression Analysis under Cold Stress

Juglans mandshurica has strong freezing resistance, surviving temperatures as low as −40 °C, making it an important freeze tolerant germplasm resource of the genus Juglans. APETALA2/ethylene responsive factor (AP2/ERF) is a plant-specific superfamily of transcription factors that regulates plant development, growth, and the response to biotic and abiotic stress. In this study, phylogenetic analysis was used to identify 184 AP2/ERF genes in the J. mandshurica genome, which were classified into five subfamilies (JmAP2, JmRAV, JmSoloist, JmDREB, and JmERF). A significant amount of discordance was observed in the 184 AP2/ERF genes distribution of J. mandshurica throughout its 16 chromosomes. Duplication was found in 14 tandem and 122 segmental gene pairs, which indicated that duplications may be the main reason for JmAP2/ERF family expansion. Gene structural analysis revealed that 64 JmAP2/ERF genes contained introns. Gene evolution analysis among Juglandaceae revealed that J. mandshurica is separated by 14.23 and 15 Mya from Juglans regia and Carya cathayensis, respectively. Based on promoter analysis in J. mandshurica, many cis-acting elements were discovered that are related to light, hormones, tissues, and stress response processes. Proteins that may contribute to cold resistance were selected for further analysis and were used to construct a cold regulatory network based on GO annotation and JmAP2/ERF protein interaction network analysis. Expression profiling using qRT-PCR showed that 14 JmAP2/ERF genes were involved in cold resistance, and that seven and five genes were significantly upregulated under cold stress in female flower buds and phloem tissues, respectively. This study provides new light on the role of the JmAP2/ERF gene in cold stress response, paving the way for further functional validation of JmAP2/ERF TFs and their application in the genetic improvement of Juglans and other tree species

Expression of <i>PnSS</i> Promotes Squalene and Oleanolic Acid (OA) Accumulation in <i>Aralia elata</i> via Methyl Jasmonate (MeJA) Induction

Aralia elata is an important herb due to the abundance of pentacyclic triterpenoid saponins whose important precursors are squalene and OA. Here, we found that MeJA treatment promoted both precursors accumulation, especially the latter, in transgenic A. elata, overexpressing a squalene synthase gene from Panax notoginseng(PnSS). In this study, Rhizobium-mediated transformation was used to express the PnSS gene. Gene expression analysis and high-performance liquid chromatography (HPLC) were used to identify the effect of MeJA on squalene and OA accumulation. The PnSS gene was isolated and expressed in A. elata. Transgenic lines showed a very high expression of the PnSS gene and farnesyl diphosphate synthase gene (AeFPS) and a slightly higher squalene content than the wild-type, but endogenous squalene synthase (AeSS), squalene epoxidase (AeSE), and β-amyrin synthase (Aeβ-AS) gene were decreased as well as OA content. Following one day of MeJA treatment, the expression levels of PeSS, AeSS, and AeSE genes increased significantly. On day 3, the maximum content of both products reached 17.34 and 0.70 mg·g−1, which increased 1.39- and 4.90-fold than in the same lines without treatment. Transgenic lines expressing PnSS gene had a limited capability to promote squalene and OA accumulation. MeJA strongly activated their biosynthesis pathways, leading to enhance yield

Genome-Wide Identification and Analysis of the EIN3/EIL Transcription Factor Gene Family in Doubled Haploid (DH) Poplar

Ethylene (ET) is an important phytohormone that regulates plant growth, development and stress responses. The ethylene-insensitive3/ethylene-insensitive3-like (EIN3/EIL) transcription factor family, as a key regulator of the ET signal transduction pathway, plays an important role in regulating the expression of ET-responsive genes. Although studies of EIN3/EIL family members have been completed in many species, their role in doubled haploid (DH) poplar derived from another culture of diploid Populus simonii × P. nigra (donor tree, DT) remains ambiguous. In this study, a total of seven EIN3/EIL gene family members in the DH poplar genome were identified. Basic physical and chemical property analyses of these genes were performed, and these proteins were predicted to be localized to the nucleus. According to the phylogenetic relationship, EIN3/EIL genes were divided into two groups, and the genes in the same group had a similar gene structure and conserved motifs. The expression patterns of EIN3/EIL genes in the apical buds of different DH poplar plants were analyzed based on transcriptome data. At the same time, the expression patterns of PsnEIL1, PsnEIN3, PsnEIL4 and PsnEIL5 genes in different tissues of different DH plants were detected via RT-qPCR, including the apical buds, young leaves, functional leaves, xylem, cambium and roots. The findings presented above indicate notable variations in the expression levels of PsnEIL genes across various tissues of distinct DH plants. Finally, the PsnEIL1 gene was overexpressed in DT, and the transgenic plants showed a dwarf phenotype, indicating that the PsnEIL1 gene was involved in regulating the growth and development of poplar. In this study, the EIN3/EIL gene family of DH poplar was analyzed and functionally characterized, which provides a theoretical basis for the future exploration of the EIN3/EIL gene function

Phenotypic Variability and Genetic Diversity in a Pinus koraiensis Clonal Trial in Northeastern China

Combining phenotypic and genetic characteristics in a genetic variation study is of paramount importance to effectively orient the selection of producers&rsquo; elite trees in a seed orchard. In total, 28 phenotypic characteristics and 16 microsatellite loci were used to analyze the clonal genetic variation, to characterize the genetic diversity, and to refine the genetic classifications of 110 Pinus koraiensis clones grown in the Naozhi orchard in northeastern China. All clones were significantly different in most traits. Most of the phenotypic characteristics showed great genetic variation among clones, while the genotypic differentiation was weak between the selection sites of clones. The SSR markers showed a relatively high level of genetic diversity (Na = 4.67 &plusmn; 0.43, Ne = 2.916 &plusmn; 0.18, I = 1.15 &plusmn; 0.07, Ho = 0.69 &plusmn; 0.04, He = 0.62 &plusmn; 0.02, and mean polymorphic information content (PIC) of 0.574), with higher heterozygosity as an indication of a lower probability of inbreeding events in the orchard. Despite weak correlation coefficients between dissimilarity matrices (r(A/B), range equal to 0.022, p-value &lt; 0.001), the genetic and phenotypic classifications congruently subdivided all the clones into three major groups. The patterns of phenotypic trait variations and genetic diversity are valuable to effectively select materials in breeding programs of P. koraiensis