Abies Beshanzuensis Research Articles

Undescribed polyketides from endophytes associated with the critically endangered conifer Abies beshanzuensis

Four undescribed polyketides, beshanzones A (1) and B (2) as well as beshanhexanols A (3) and B (4), along with three known ones (5–7) were isolated from the rice fermentation of two endophytic fungi associated with the critically endangered Chinese endemic conifer Abies beshanzuensis. γ-Butyrolactone derivatives 1, 2, and 5 were isolated from Phomopsis sp. BSZ-AZ-2, an interesting strain that drawn our attention this time. The cyclohexanol derivatives 3, 4, 6, and 7 were obtained during a follow-up investigation on Penicillium commune BSZ-P-4-1. The chemical structures including absolute configurations of compounds 1–4 were determined by spectroscopic methods, Mo2(OAc)4 induced electronic circular dichroism (IECD), GIAO NMR calculations and DP4+ probability analyses. In particular, compound 2 contains a novel 5/5 bicyclic ring system, which might be biogenetically derived from the known compound 5 through hydrolysis followed by an Aldol reaction. All isolates were evaluated for their antimicrobial activities against a small panel of bacterial and fungal pathogens. Compounds 6 and 7 showed moderate inhibitory activities against Candida albicans, with MIC values of 16 and 32 μg/mL, respectively.

MYB6/bHLH13-AbSUS2 involved in sugar metabolism regulates root hair initiation of Abies beshanzuensis.

Root hair is regarded as a pivotal complementary survival tactic for mycorrhizal plant like Abies beshanzuensis when symbiosis is disrupted. Relatively little is known about the mechanism underlying root hair morphogenesis in plant species that are strongly dependent on mycorrhizal symbiosis. Many of these species are endangered, and this knowledge is critical for ensuring their survival. Here, a MYB6/bHLH13-sucrose synthase 2 (AbSUS2) module was newly identified and characterized in A. beshanzuensis using bioinformatics, histochemistry, molecular biology, and transgenesis. Functional, expression pattern, and localization analysis showed that AbSUS2 participated in sucrose synthesis and was involved in root hair initiation in A. beshanzuensis. Additionally, the major enzymatic product of AbSUS2 was found to suppress root hair initiation invitro. Our data further showed that a complex involving the transcription factors AbMYB6 and AbbHLH13 directly interacted with the promoter of AbSUS2 and strengthened its expression, thereby inhibiting root hair initiation in response to exogenous sucrose. Our findings offer novel insights into how root hair morphogenesis is regulated in mycorrhizal plants and also provide a new strategy for the preservation of endangered mycorrhizal plant species.

Testing a Method Based on an Improved UNet and Skeleton Thinning Algorithm to Obtain Branch Phenotypes of Tall and Valuable Trees Using Abies beshanzuensis as the Research Sample.

Sudden changes in the morphological characteristics of trees are closely related to plant health, and automated phenotypic measurements can help improve the efficiency of plant health monitoring, and thus aid in the conservation of old and valuable tress. The irregular distribution of branches and the influence of the natural environment make it very difficult to monitor the status of branches in the field. In order to solve the problem of branch phenotype monitoring of tall and valuable plants in the field environment, this paper proposes an improved UNet model to achieve accurate extraction of trunk and branches. This paper also proposes an algorithm that can measure the branch length and inclination angle by using the main trunk and branches separated in the previous stage, finding the skeleton line of a single branch via digital image morphological processing and the Zhang-Suen thinning algorithm, obtaining the number of pixel points as the branch length, and then using Euclidean distance to fit a straight line to calculate the inclination angle of each branch. These were carried out in order to monitor the change in branch length and inclination angle and to determine whether plant branch breakage or external stress events had occurred. We evaluated the method on video images of Abies beshanzuensis, and the experimental results showed that the present algorithm has more excellent performance at 94.30% MIoU as compared with other target segmentation algorithms. The coefficient of determination (R2) is higher than 0.89 for the calculation of the branch length and inclination angle. In summary, the algorithm proposed in this paper can effectively segment the branches of tall plants and measure their length and inclination angle in a field environment, thus providing an effective method to monitor the health of valuable plants.

Comparative Transcriptome Analysis Reveals the Interaction of Sugar and Hormone Metabolism Involved in the Root Hair Morphogenesis of the Endangered Fir Abies beshanzuensis

Abies beshanzuensis, an extremely rare and critically endangered plant with only three wild adult trees globally, is strongly mycorrhizal-dependent, leading to difficulties in protection and artificial breeding without symbiosis. Root hair morphogenesis plays an important role in the survival of mycorrhizal symbionts. Due to the lack of an effective genome and transcriptome of A. beshanzuensis, the molecular signals involved in the root hair development remain unknown, which hinders its endangered mechanism analysis and protection. Herein, transcriptomes of radicles with root hair (RH1) and without root hair (RH0) from A. beshanzuensis in vitro plantlets were primarily established. Functional annotation and differentially expressed gene (DEG) analysis showed that the two phenotypes have highly differentially expressed gene clusters. Transcriptome divergence identified hormone and sugar signaling primarily involved in root hair morphogenesis of A. beshanzuensis. Weighted correlation network analysis (WGCNA) coupled with quantitative real-time PCR (qRT-PCR) found that two hormone-sucrose-root hair modules were linked by IAA17, and SUS was positioned in the center of the regulation network, co-expressed with SRK2E in hormone transduction and key genes related to root hair morphogenesis. Our results contribute to better understanding of the molecular mechanisms of root hair development and offer new insights into deciphering the survival mechanism of A. beshanzuensis and other endangered species, utilizing root hair as a compensatory strategy instead of poor mycorrhizal growth.

Subtropical forest vegetation development and climate change in Baishanzu area of Zhejiang Province, China, since the Holocene

Human activities and climate change pose major challenges for the sustainable development of terrestrial biological forests. The modern vegetation in the Baishanzu area is typical mixed subtropical evergreen broad-leaved, coniferous, and deciduous broad-leaved forest, with a vegetation combination sensitive to climate change. Higher resolution pollen records from the Shangyang Lake Wetland, located in the subtropical monsoon climate of the Baishanzu area in Zhejiang, China, were combined with environmental proxy data, such as data on loss on ignition and charcoal, to reconstruct the vegetation and climate changes since the Holocene. The results show that the climate was cool and dry during the early Holocene (11660–6580 cal. aBP), when the zonal vegetation changed from evergreen deciduous broad-leaved mixed forests dominated by evergreen Quercus (Quercus(E)), Castanopsis, deciduous Quercus (Quercus(D)), and Castanea to coniferous and broad-leaved mixed forests with rich Cryptomeria. Abies pollen spread to lower altitudes during the period 9620–8020 cal. aBP with Abies beshanzuensis covering large areas of land. The increased in the East Asian summer monsoon from 6580 to 2040 cal. aBP led to the Holocene Climate Optimum, when the climate was particularly warm and humid. The zonal vegetation was middle subtropical evergreen broad-leaved forest dominated by Quercus(E), Castanopsis, and Cyclobalanopsis with particularly high forest coverage and few fire events. For the last 2000 years, coniferous and broad-leaved mixed forests, mainly Pinus taiwanensis, have dominated the vegetation in the area. Sharp increase in such flora was observed after the year 980. Decreased temperature, cold climate, and increased anthropogenic disturbances have resulted in a substantial reduction in the arboreal pollen content, including Quercus, Castanopsis, Castanea, and Ilex, and an increase in Pinus and Poaceae. The results of this study will play a positive role in fully understanding the historical background and internal mechanisms of subtropical forest changes.

Auxiliary Seed Treatment Is Necessary to Increase Recruitment of a Critically Endangered Species, Abies beshanzuensis (Pinaceae)

Disordered germination is widely observed in plant species with extremely small populations (PSESPs) in China. Abies beshanzuensis M. H. Wu, a PSESP (with only three extant adults in Zhejiang Province, China) that also has poor seed germination in the field, belongs to the Pinaceae family. Pinaceae generally tend toward symbiosis with ectomycorrhizal (ECM) fungi to alleviate climate change-induced stresses. Therefore, exploring how to improve seed germination of A. beshanzuensis and whether it is an ECM species will contribute to increasing recruitment for conducting further conservation research. In this study, four temperature regimes (10/15 °C, 15/20 °C, 20/25 °C, and 25/30 °C) were selected to explore the response of seed germination to rising temperature. Based on the microscopic features of fine roots, together with molecular techniques, the mycorrhizal type of this species was identified. The seed germination of A. beshanzuensis was increased from 1–2% to around 4.5% by 14-day cold stratification and under 20/25 °C fluctuating temperature conditions. Our findings indicated that A. beshanzuensis may be endangered as a result of insufficient seedling recruitment due to poor germination under the current climate. A. beshanzuensis was confirmed as an ECM fungi-associated tree species. This study highlights the necessity of incorporating auxiliary seed treatment into population recovery programs of A. beshanzuensis, thereby better conserving the species under ongoing environmental changes.

Beshanzuamide A, an unprecedented prenylated indole alkaloid produced by Aspergillus sp. Y-2 from the critically endangered conifer Abies beshanzuensis.

A structurally unprecedented prenylated indole alkaloid, beshanzuamide A (1), together with five known analogues (2–6) were isolated and identified from the endophytic fungus derived from the needles of the critically endangered conifer Abies beshanzuensis. The new structure was determined by extensive spectroscopic methods and quantum chemical calculations of NMR and electronic circular dichroism (ECD) data. Compound 1 features a unique N,O-spiroketal/δ-lactone motif connected to a pyranoindole-derived bicyclo[2.2.2]diazaoctane ring. A plausible biogenetic pathway for the assembly of 1 was proposed.

Genomic landscape of a relict fir-associated fungus reveals rapid convergent adaptation towards endophytism.

Comparative and pan-genomic analyses of the endophytic fungus Pezicula neosporulosa (Helotiales, Ascomycota) from needles of the relict fir, Abies beshanzuensis, showed expansions of carbohydrate metabolism and secondary metabolite biosynthetic genes characteristic for unrelated plant-beneficial helotialean, such as dark septate endophytes and ericoid mycorrhizal fungi. The current species within the relatively young Pliocene genus Pezicula are predominantly saprotrophic, while P. neosporulosa lacks such features. To understand the genomic background of this putatively convergent evolution, we performed population analyses of 77 P. neosporulosa isolates. This revealed a mosaic structure of a dozen non-recombining and highly genetically polymorphic subpopulations with a unique mating system structure. We found that one idiomorph of a probably duplicated mat1-2 gene was found in putatively heterothallic isolates, while the other co-occurred with mat1-1 locus suggesting homothallic reproduction for these strains. Moreover, 24 and 81 genes implicated in plant cell-wall degradation and secondary metabolite biosynthesis, respectively, showed signatures of the balancing selection. These findings highlight the evolutionary pattern of the two gene families for allowing the fungus a rapid adaptation towards endophytism and facilitating diverse symbiotic interactions.

Abieshanesides A and B, two unique ent-18,19-dinoricetexane diterpenoid glycosides from Abies beshanzuensis M.H. Wu

Two unprecedented ent-18,19-dinoricetexane diterpenoid glycosides, named abieshanesides A (1) and B (2), together with seven known compounds, have been isolated from the dead trunks and branches of Abies beshanzuensis M.H. Wu. To our knowledge, abieshanesides A and B represent the first ent-18,19-dinoricetexane diterpenoid glycosides found in natural sources. Their structures and absolute configurations were elucidated by using a combination of spectroscopic techniques and comparison of experimental and calculated electronic circular dichroism (ECD) data. The MTT experiments showed that (E)-resveratrol (7) could inhibit viability of MH7A cells with the IC50 value of 12.5 μM. Compound 7 was able to block MH7A cell proliferation and was associated with G0/G1-phase cell cycle arrest. Flow cytometric analysis showed that the treatment by 7 significantly induced the proliferation of MH7A cells in a dose-dependent manner.

Beshanzoides A-D, unprecedented cycloheptanone-containing polyketides from Penicillium commune P-4-1, an endophytic fungus of the endangered conifer Abies beshanzuensis.

A number of previously undescribed (1–7) and structurally related known (8–17) isobenzofuran-type polyketides were obtained from the fermentation of Penicillium commune P-4-1, an endophytic fungus isolated from the fresh trunk bark of the critically endangered conifer Abies beshanzuensis. Beshanzoides A–D (1–4, resp.) feature a cycloheptanone-containing isobenzofuran ring system hitherto unknown, which might be biosynthesized via two steps of aldol reactions starting from a common co-occurring isobenzofuran-type polyketide as the precursor. The new structures were elucidated by spectroscopic methods, electronic circular dichroism data, and single crystal X-ray diffraction analyses. Beshanzoide E (5) showed antimicrobial activity (MIC: 16 μg mL−1) against Staphylococcus aureus, whereas (±)-strobide A (10) inhibited (MIC: 16 μg mL−1) Candida albicans. Cyclopaldic acid (12) and 3-O-methyl-cyclopaldic acid (13) exhibited inhibitory effects against acetyl-CoA carboxylase 1 (ACC1) with IC50 values of 0.96 and 11.77 μM, respectively. Compound 12 also inhibited (IC50: 7.56 μM) ATP-citrate lyase (ACL).

Chromenopyridin A, a new N-methoxy-1-pyridone alkaloid from the endophytic fungus Penicillium nothofagi P-6 isolated from the critically endangered conifer Abies beshanzuensis

A new N-methoxy-1-pyridone alkaloid [chromenopyridin A (1)] and four known compounds (2–5) were isolated and identified from the endophytic fungus Penicillium nothofagi P-6, which was derived from the bark of the critically endangered conifer Abies beshanzuensis. Their structures were elucidated by extensive spectroscopic analyses and single-crystal X-ray diffraction. Among the isolates, compound 1 showed considerable cytotoxicities against the A549 and Hela human cancer cell lines, with IC50 values of 14.7 and 11.3 μM. In addition, compounds 1 and 4 exhibited potent antibacterial activity against Staphylococcus aureus with MIC values of 62.5 and 15.6 μg/mL, respectively.

Aflatoxins from the endophytic fungus Aspergillus sp. Y-2 isolated from the critically endangered conifer Abies beshanzuensis

Two new [asperxins A (1) and B (2)] and four known (3–6) aflatoxins were isolated and identified from the endophytic fungus Aspergillus sp. Y-2, which was derived from the needles of the critically endangered conifer Abies beshanzuensis. Their structures were elucidated by extensive spectroscopic methods. Among the isolates, compounds 1 and 5 showed considerable cytotoxicities against the A549 and Hela human cancer cell lines, with IC50 values in the range of 7.5–16.8 μM.

The complete chloroplast genome sequence of Abies beshanzuensis, a highly endangered fir species from south China

Abies beshanzuensis is critically endangered and endemic to Zhengjiang province of China, with only three surviving individuals. In present study, we reported the complete chloroplast (cp) genome of Abies beshanzuensis. The complete chloroplast genome size is 121,399 bp. In total, 114genes were identified, including 68 peptide-encoding genes, 35 tRNA genes, four rRNA genes, six open reading frames and one pseudogene. Loss of ndh genes was also identified in the genome of A. beshanzuensis like other genomes in the family Pinaceae. Thirteen genes contain one (11 genes) or two (rps12 and ycf3 genes) introns. Inverted repeat sequences located in 42-kb inversion points (1186 bp) include trnS-psaM-ycf12-trnG genes. In phylogenetic analysis, the tree confirms that the four Abies species are strongly supported as monophyletic. The complete plastome of A. beshanzuensis will provide potential genetic resources for further conservation and evolutionary studies of this highly endangered species.

Lignans from the shed trunk barks of the critically endangered plant Abies beshanzuensis and their anti-neuroinflammatory activities

During a further and comprehensive phytochemical investigation on the shed trunk barks of the critically endangered plant Abies beshanzuensis, one new (1) and ten known (2–11) lignans with diverse structures were isolated. On the basis of spectroscopic methods, the new structure was established to be (7S,8R,8′R)-4′-methoxyl-α-conidendrin (1). Among the isolated lignans, (−)-matairesinol (5) and (−)-arctigenin (6) showed significant anti-neuroinflammatory activities by inhibiting the overproduction of nitric oxide in lipopolysaccharide-stimulated murine BV-2 microglial cells, with IC50 values of 11.5 and 19.0 μM, respectively.

ChemInform Abstract: Rare Sesquiterpenoids from the Shed Trunk Barks of the Critically Endangered Plant Abies beshanzuensis and Their Bioactivities.

AbstractBoth beshanzuenones A (Ia) and B (Ib) show considerable activity against the H3N2 virus, whereas beshanzuenones C (IIa) and D (IIb) exhibit significant inhibition on the human PTP1B enzyme.

Rare Sesquiterpenoids from the Shed Trunk Barks of the Critically Endangered Plant Abies beshanzuensis and Their Bioactivities

AbstractTwo rare rearranged cuparane‐type sesquiterpenoid C‐10 epimers [beshanzuenones A (1) and B (2)] and two bisabolane‐type sesquiterpenoid spirolactones [beshanzuenones C (3) and D (4)] featuring an unusual [6/6/5]‐fused tricyclic ring system were isolated from the shed trunk barks of Abies beshanzuensis, one of the critically endangered plant species in the world. The structures of these four new enone‐containing sesquiterpenoids were determined by spectroscopic analyses and single‐crystal X‐ray diffraction or electronic circular dichroism (ECD) calculations. Compounds 1 and 2 showed considerable activity against the H3N2 virus, whereas 3 and 4 exhibited significant inhibition on PTP1B.

China: home for the most endangered plants of the world?

If you ask to a western botanist: “Which is the world’s most endangered plant species?” he will probably answer the Florida torreya (Torreya taxifolia Arn.), the Wollemi pine (Wollemia nobilis W. G. Jones, K. D. Hill & J. M. Allen), or perhaps the Nebrodi fir (Abies nebrodensis Mattei), if the interviewed is of european origin. In contrast, if you make the same question to a Chinese (or at least to somebody familiar with the Chinese flora) you may get a quite different reply. Much less known than these emblematic trees, a large handful of Chinese plant taxa is currently in a more critical situation, with just a very few individuals surviving (He, 2009; Lopez-pujol & zhang, 2009). A rare cypress (Cupressus chengiana S. Y. Hu var. jiangeensis (N. zhao) Silba1) is arguably the most threatened gymnosperm on earth because only one individual tree occurs in Longmen (“Dragon’s Gate”) Mountains, in northern Sichuan. The baishanzu fir (Abies beshanzuensis M. H. Wu var. beshanzuensis1) from zhejiang, is not in a much wealthier situation, with only three representatives alive. Moreover, China has also the dubious honour of harbouring some of the surely world’s most endangered angiosperms, such as Carpinus putoensis W. C. Cheng (one fenced, old individual is remaining in the wild in putuo Island, in the east China Sea), Gleditsia japonica Miq. var. velutina L. Chu Li1 and Sinopora hongkongensis (N. H. Xia, Y. F. Deng & K. L.Yip) J. Li, N. H. Xia & H. W. Li (with two individuals each one), or Acer yangbiense Y. S. Chen & Q. e. Yang (with four representatives). These few conspicuous examples of rare plants, however, may represent just the iceberg’s tip of a flora facing a situation of extreme risk: of the approximately 4200 angiosperm taxa assessed in the first issue of the China Species Red List (which represents just 14% of the angiosperm flora of China), as many as 651 were listed as CR (“critically endangered”) following the 2001 IuCN criteria (Xie & Wang, 2007), that is, in the verge of extinction. The disproportionate number of extremely threatened plants in China may have resulted from the combination of natural and human-induced factors (Lopez-pujol & zhang, 2009). The existence of numerous refuge areas during the Quaternary glacial periods in China (Qian & Ricklefs, 2000; Lopezpujol, 2008; Lopez-pujol & Ren, 2010) yielded a large number of narrow endemics through survival of relict (pre-Quaternary) lineages as well as by differentiation and speciation in these favourable pockets, most of them located in the southern mountainous regions (Fig. 1). The huge destruction and deterioration of ecosystems, largely reported in China during recent decades (e.g. Lopez-pujol et al., 2006; He, 2009), must have decimated many of these “naturally rare” species, in some cases driving them to the brink of extinction (such as the abovementioned examples) or

From pattern to process: species and functional diversity in fungal endophytes of Abies beshanzuensis

The biodiversity–functional relationship in fungal ecology was recently developed and debated, but has rarely been addressed in endophytes. In this study, an integrative culture system was designed to capture a rich fungal consortium from the conifer Abies beshanzuensis. Results indicate an impressive diversity of fungal lineages (a total of 84 taxa classified in Dikarya) and a relatively high proportion of hitherto unknown species (27.4 %). The laccase gene was used as a functional marker due to its involvement in lignocellulose degradation. Remarkable diversity of laccase genes was found across a wide range of taxa, with at least 35 and 19 distinct sequences in ascomycetes and basidiomycetes respectively, were revealed. Many groups displayed variable ability to decompose needles. Furthermore, many ascomycetes, including three volatile-producing Muscodor species ( Xylariaceae), showed the ability to inhibit pathogens. Notably, most laccase-producing species showed little or no antibiosis and vice versa. Clavicipitalean and ustilaginomycetous fungi, specifically toxic to insects, were inferred from taxonomic information. Intra-specific physiological variation in Pezicula sporulosa, a second dominant species, was clearly high. We conclude that a suite of defensive characteristics in endophytes contributes to improving host fitness under various stresses and that a diversity of laccase genes confers an ecological advantage in competition for nutrients. Intra-specific diversity may be of great ecological significance for ecotypic adaptation. These findings suggest a fair degree of functional complementarity rather than redundancy among endemic symbionts of natural plant populations.

Lignin characteristics of Abies beshanzuensis, a critically endangered tree species

As one of the major components of plant cell walls, lignin structural features are closely related with taxonomical and genetic classification of plants. In this study, the structural features of lignin of Abies beshanzuensis were investigated. Abies firma, which is genetically the closest species to A. beshanzuensis, Cryptomeria japonica, a typical gymnosperm tree species, and Phyllostachys pubescens (bamboo), which includes p-hydroxyphenyl nuclei in arylglycerol-β-aryl intermonomer linkages, were also analyzed to compare lignin features with those of A. beshanzuensis. The lignin content of A. beshanzuensis (39.2%) was significantly higher than that of A. firma (33.7%). The high value may be due to the adaptation of A. beshanzuensis to environmental stresses in surviving the Riss glacial period. Alkaline nitrobenzene oxidation, ozonation, acidolysis, and 1H nuclear magnetic resonance spectra showed that the structural features of lignin of A. beshanzuensis were similar to those of A. firma, which is genetically the closest species of A. beshanzuensis. The results of this study suggest that A. firma would be a suitable mother tree species for grafting A. beshanzuensis on the basis of their lignin characteristics.