Pinus Bungeana Zucc Research Articles

Calibration of Thermal Dissipation Probes for Xylem Sap Flow in the Wood of a Diffuse-Porous and a Conifer Species under Cyclic Heating

The most popular sap flow measurement technique uses thermal dissipation probes. Differences in wood characteristics and the natural temperature gradient between probes have affected the accuracy and applicability of the sap flow equation. In addition, the continued heat of the probe can also cause thermal damage to tree tissue. The objectives of this study were to use cyclic heating and calibrate the probes with two species: Pinus bungeana Zucc. And Salix matsudana Koidz., two typical diffuse-porous species. This experiment evaluated a thermal dissipation probe in three heating modes: continuous heating, 10 min heating and 50 min cooling (10/50), and 30 min heating and 30 min cooling (30/30). The heating modes were evaluated on two species. Temperature differences between the heating needle and the control needle under different heating modes and transpiration water consumption (whole-tree weighing method) were observed simultaneously. The sap flow estimation equation under cyclic heating mode was established by analyzing the relationship between the sap flow rate and the values obtained from whole-tree weighing. The results showed that the original equation underestimated sap flow rate of P. bungeana and S. matsudana by 67% and 60%. Under the cyclic heating modes, the modified equations were different from the original equation, and their accuracy was improved. After verification, the corrected equations [Fd = 0.0264K0.738 (P. bungeana, 30/30, R2 = 0.67), Fd = 0.0722K1.113(S. matsudana, 30/30, R2 = 0.60), Fd is the sap flow density, K is temperature coefficient] reduced the influence of the natural temperature gradient on the estimation of sap flow rate, thereby significantly improving the accuracy of sap flow rate estimation. The resulting equation may be more suitable for actual field observations of sap flow in the two tested species. The cyclic heating mode has the potential to measure plant transpiration for extended periods in the field.

Enhanced Root and Stem Growth and Physiological Changes in Pinus bungeana Zucc. Seedlings by Microbial Inoculant Application

Background and Objectives: As an extensively used tree species in landscaping and afforestation in China, lacebark pine (Pinus bungeana Zucc.) seedlings are in high demand. However, the small number of fine roots and the low growth rate of lacebark pine seedlings increase the risks encountered during transplant and extend the nursery time for outplanting. We aimed to find out whether a microbial inoculant would promote root growth and accordingly, shorten the nursery cultivation time. Materials and Methods: One-year-old lacebark pine seedlings were treated with the inoculant Bacillus subtilis 8–32 six times from June to September. At each application time, five treatments of undiluted microbial inoculants (UM), 30 times diluted microbial inoculants (30 DM), 40 times diluted microbial inoculants (40 DM), 50 times diluted microbial inoculants (50 DM), and distilled water as a control (CTRL) were administered to the seedlings. In the end, all the seedlings were harvested to measure the root growth, aboveground growth, and the physiological indices. Results: Root and stem growth was enhanced by the inoculants in terms of the increased number of root tips, the length and surface area of the roots, the biomass of the roots and stems, as well as the increase in height and basal stem diameter. The chlorophyll a/b of the needles was increased, in spite of the fact that the total chlorophyll content was decreased by the microbial inoculant treatments at the end of the growth phase. Meanwhile, the maximum photochemical efficiency (Fv/Fm) of the needles was increased by the inoculant treatments. The soluble sugar content was additionally translocated into the stems in the UM treatment, suggesting the change in carbon allocation. The content of available potassium, phosphorus, and ammonium nitrogen in the potting soil was increased in the 30 DM group, and the content of soil organic matter was increased in all the inoculant treatments. Conclusions: The microbial inoculant Bacillus subtilis 8–32, in appropriate concentrations, could be applied to promote root and shoot growth and improve the seedling quality of the lacebark pine during cultivation.

In-situ visualizing selective lignin dissolution of tracheids wall in reaction wood

As a renewable biological macromolecule with aromatic structure, lignin can serve as matrix substance to maintain cell wall integrity and is regarded as the natural biomass recalcitrance. Substantial differences in the cell wall lignin topochemistry between opposite (Ow) and compression wood (Cw) trachieds in Pinus bungeana Zucc. were visualized during [Emim][OAc] pretreatment at room temperature. The ionic liqiuds treatment induced a more obvious wall swelling for highly lignified Cw tracheids than that of Ow, while dynamic Raman spectra analysis indicated the higher lignin and carbohydrates removal for Ow tracheids. Raman imaging further revealed that both lignin and carbohydrates were dissolved simultaneously within the middle lamella and secondary wall of Ow and pretreatment has little effects on Cw tracheids wall. Moreover, it was demonstrated that lignin composition was the key factor to affect the composition dissolution. In particular, lignin G-units were selectively removed from cell corner middle lamella (52.3 %) and secondary wall (62.0 %) of Ow tracheids. When cotton fiber, as a reference was treated under the same conditions, lattice conversion moving from cellulose I to II occurred. The findings confirmed the important role of lignin compostion in the dissolution behavior of carbohydrate dominant tracheids wall.

Low genetic diversity and population connectivity fuel vulnerability to climate change for the Tertiary relict pine Pinus bungeana

AbstractEndemic species are important components of regional biodiversity and hold the key to understanding local adaptation and evolutionary processes that shape species distributions. This study investigated the biogeographic history of a relict conifer Pinus bungeana Zucc. ex Endl. confined to central China. We examined genetic diversity in P. bungeana using genotyping‐by‐sequencing and chloroplast and mitochondrial DNA markers. We performed spatial and temporal inference of recent genetic and demographic changes, and dissected the impacts of geography and environmental gradients on population differentiation. We then projected P. bungeana's risk of decline under future climates. We found extremely low nucleotide diversity (average π 0.0014), and strong population structure (global FST 0.234) even at regional scales, reflecting long‐term isolation in small populations. The species experienced severe bottlenecks in the early Pliocene and continued to decline in the Pleistocene in the western distribution, whereas the east expanded recently. Local adaptation played a small (8%) but significant role in population diversity. Low genetic diversity in fragmented populations makes the species highly vulnerable to climate change, particularly in marginal and relict populations. We suggest that conservation efforts should focus on enhancing gene pool and population growth through assisted migration within each genetic cluster to reduce the risk of further genetic drift and extinction.

Investigation of the Impact of Land-Use Distribution on PM2.5 in Weifang: Seasonal Variations.

As air pollution becomes highly focused in China, the accurate identification of its influencing factors is critical for achieving effective control and targeted environmental governance. Land-use distribution is one of the key factors affecting air quality, and research on the impact of land-use distribution on air pollution has drawn wide attention. However, considerable studies have mostly used linear regression models, which fail to capture the nonlinear effects of land-use distribution on PM2.5 (fine particulate matter with a diameter less than or equal to 2.5 microns) and to show how impacts on PM2.5 vary with land-use magnitudes. In addition, related studies have generally focused on annual analyses, ignoring the seasonal variability of the impact of land-use distribution on PM2.5, thus leading to possible estimation biases for PM2.5. This study was designed to address these issues and assess the impacts of land-use distribution on PM2.5 in Weifang, China. A machine learning statistical model, the boosted regression tree (BRT), was applied to measure nonlinear effects of land-use distribution on PM2.5, capture how land-use magnitude impacts PM2.5 across different seasons, and explore the policy implications for urban planning. The main conclusions are that the air quality will significantly improve with an increase in grassland and forest area, especially below 8% and 20%, respectively. When the distribution of construction land is greater than around 10%, the PM2.5 pollution can be seriously substantially increased with the increment of their areas. The impact of gardens and farmland presents seasonal characteristics. It is noted that as the weather becomes colder, the inhibitory effect of vegetation distribution on the PM2.5 concentration gradually decreases, while the positive impacts of artificial surface distributions, such as construction land and roads, are aggravated because leaves drop off in autumn (September–November) and winter (December–February). According to the findings of this study, it is recommended that Weifang should strengthen pollution control in winter, for instance, expand the coverage areas of evergreen vegetation like Pinus bungeana Zucc. and Euonymus japonicus Thunb, and increase the width and numbers of branches connecting different main roads. The findings also provide quantitative and optimal land-use planning and strategies to minimize PM2.5 pollution, referring to the status of regional urbanization and greening construction.

Responses of Parameters for Electrical Impedance Spectroscopy and Pressure–Volume Curves to Drought Stress in Pinus bungeana Seedlings

Background and Objectives: Drought occurs more frequently in Northern China with the advent of climate change, which might increase the mortality of tree seedlings after afforestation due to hydraulic failure. Therefore, investigating water relations helps us understand the drought tolerance of tree seedlings. Electrical impedance spectroscopy (EIS) is widely used to assess the responses of plant tissues to stress factors and may potentially reveal the water relations of cells. The aim of this study is to reveal the relationships between EIS and water related parameters, produced by pressure–volume (PV) curves in lacebark pine (Pinus bungeana Zucc.) seedlings reacting to drought stress. Materials and Methods: Four-year-old pot seedlings were divided into three parts (0, 5, and 10 days of drought) before planting, the treated seedlings were then replanted, and finally exposed to post-planting drought treatments with the following soil relative water contents: (i) adequate irrigation (75%–80%), (ii) light drought (55%–60%), (iii) moderate drought (35%–40%), and (iv), severe drought (15%–20%). During the post-planting growth phase, the EIS parameters of needles and shoots, and the parameters of PV curves, were measured coincidently; thus, the correlations between them could be obtained. Results: The extracellular resistance (re) of needles and shoots were substantially reduced after four weeks of severe post-planting drought stress. Meanwhile, the osmotic potential at the turgor-loss point (ψtlp) and the saturation water osmotic potential (ψsat) of shoots decreased after drought stress, indicating an osmotic adjustment in acclimating to drought. The highest correlations were found between the intracellular resistance (ri) of the shoots and ψtlp and ψsat. Conclusions: EIS parameters can be used as a measure of drought tolerance. The change in intracellular resistance is related to the osmotic potential of the cell and cell wall elasticity. Extracellular resistance is a parameter that shows cell membrane damage in response to drought stress in lacebark pine seedlings.

Effect of Temperature, Light, and Storage Time on the Seed Germination of Pinus bungeana Zucc. ex Endl.: The Role of Seed-Covering Layers and Abscisic Acid Changes

Pinus bungeana Zucc. ex Endl. is an endemic conifer tree species in China with high ornamental value. In order to investigate favorable conditions for seed germination and explore the germination inhibition mechanism of this species at high temperatures, the effects of temperature, light, and storage on the mean germination time (MGT), speed of germination (SG), and total germination percentage (TGP) are evaluated here. Seeds that have either been kept still or entered into a state of dormancy at high temperature are assessed here by a recovery experiment. Furthermore, the contribution of covering layers on thermo-inhibition is analyzed here, including the way they work. This has been realized by the structural observation and via the determination of the abscisic acid (ABA) content. The results show that seeds germinate to a high percentage (approximately 90%) at temperatures of 15 or 20 °C, with or without light, whereas higher temperatures of 25 or 30 °C impeded radicle protrusion and resulted in the germination percentage decreasing sharply (within 5%). Inhibition at high temperatures was thoroughly reversed (bringing about approximately 80% germination) by placing the ungerminated seeds in favorable temperatures and incubating them for an additional 30 days. Dry cold storage did little to reduce the temperature request for germination. Embryo coverings, especially the nucellar membrane, and ABA levels both had a dominant role in seed germination regulation in response to temperature. Under favorable temperature conditions, the levels of ABA significantly decreased. Germination occurred when the levels dropped to a threshold of 15 ng/g (FW (Fresh Weight)). Incubation at a high temperature (25 °C) greatly increased ABA levels and caused the inhibition of radicle protrusion.

First Report of Diaporthe eres Causing Twig Dieback of White Bark Pine in China

White bark pine (Pinus bungeana Zucc.) is popularly cultivated as a garden ornamental tree in China because of the high ornamental value. During July 2017, twig dieback symptoms were found on white bark pine trees in the campus of Henan Agricultural University, Zhenzhou, Henan Province, China. The rate of diseased plants was 25% (up to 40% in 2018) in this campus. Early symptoms appeared as round or oval, red-brown lesions on the twigs, which enlarged progressively, leading to dieback. Two representative Diaporthe strains (zm170130 and zm180920) were isolated and purified from two specimens collected in 2017 and 2018 on potato dextrose agar (PDA). Pycnidia scattered in PDA, dark brown to black, globose to subglobose. Conidiogenous cells phialidic, cylindrical, tapering toward the apex, with periclinal thickening. Two kinds of conidia were produced: α-conidia, unicellular, hyaline elliptical or fusiform, two-guttulate, 5.2 to 9.0 × 2 to 3 µm (n = 50); β-conidia hyaline, aseptate, without guttules, filiform, curved, with obtuse ends, 15.5 to 31 × 0.8 to 1.5 µm (n = 50). These morphological characteristics of the isolates agreed with those of Diaporthe eres (Dissanayake et al. 2015; Liu and Tang 1995; Udayanga et al. 2014). The identity of the two fungal isolates was confirmed by DNA sequencing of the ITS rDNA region (GenBank accession nos. MK265702 and MK265701), TUB gene (GenBank accession nos. MK265704 and MK265703), and histone H3 gene (GenBank accession nos. MK265700 and MK265699), which were over 99% homologous to those consensus sequences of the ex-epitype strain D. eres (KJ210529, KJ420799, KJ210550) (Guarnaccia et al. 2018). The pathogenicity of the isolate zm180920 was tested on 2-year-old healthy twigs of white bark pine in July 2018. Five 2-year-old healthy twigs (30 cm long) cut from 5-year-old trees were used for each treatment. Fifteen mycelial plugs harvested from the periphery of a 7-day-old colony were placed on the surface of the wounded twigs, after the twigs had been surface sterilized with 75% ethanol for 2 min; an equal number of noncolonized plugs of PDA served as the control treatment. All test twigs were kept in a clear plastic box with wet sterilized cotton to maintain high relative humidity at 26 to 30°C in the greenhouse. Symptoms similar to those originally observed on the naturally infected twigs were observed 10 days after inoculation, and the same fungus was reisolated from each of the symptomatic twigs; control twigs remained asymptomatic, and no fungus was isolated. The isolates from the symptomatic twigs were identified as D. eres on the basis of the morphology and the same gene sequences. Koch’s postulates were repeated three times with the same results. The fungus has been reported on peach, grapevine, jujube, Red Robin, and so on in China (Dissanayake et al. 2015; Liu and Tang 1995; Song et al. 2019; Yang et al. 2018; Zhang et al. 2018). To our knowledge, this is the first report of D. eres causing twig dieback of white bark pine in China. Disease control strategies in the region are being further studied.

Subcellular dissolution of xylan and lignin for enhancing enzymatic hydrolysis of microwave assisted deep eutectic solvent pretreated Pinus bungeana Zucc

The mechanism for enhancing enzymatic hydrolysis during microwave-assisted deep eutectic solvent (Mw-DES) pretreatment in deconstruction of plant cell wall was proposed by combining wet chemical analysis and microscopic measurements. Mw-DES pretreatment achieved significantly higher enzymatic conversion of 81.90% with lower lignin and comparable xylan removal (42.81% and 74.73%, respectively). While DES pretreated sample with higher lignin and xylan removal (66.59% and 74.93%, respectively) obtained limited sugar yield (45.67%). There were no significant differences with respect to chemical structures of lignin fraction between DES and Mw-DES pretreatment but primary discrepancies of topochemical and morphological changes were observed. Non- or low-substituted xylan was directly removed from secondary walls (SW) exposed more cellulose for enzyme attacking after Mw-DES pretreatment. Meanwhile, high-substituted xylan and lignin were synergistically dissolved from cell corner middle lamella (CCML). These topochemical changes of components resulted in cracked and porous cell wall structure, thus facilitating the accessibility of cellulose.

How Does Leaf Surface Micromorphology of Different Trees Impact Their Ability to Capture Particulate Matter?

Particulate matter (PM), including PM10 and PM2.5, has a major impact on air quality and public health. It has been shown that trees can capture PM and improve air quality. In this study, we used two-way ANOVA to investigate the significance of micro-morphological leaf surface characteristics of green trees in capturing PM at different parks in Beijing. The results show that leaf structure significantly impacts the ability of plants to capture PM. Pinus tabuliformis Carr. and Pinus bungeana Zucc. were mainly impacted by the density of stomata, waxy cuticle, and epidermis, while the major contributor to PM retention in other test trees, including Acer truncatum Bunge, Salix matsudana Koid., Populus tomentosa Carr. and Ginkgo biloba Linn. was leaf roughness. There were significant variations in leaf-droplet contact angle (representative of leaf wettability) and the ability of trees to capture PM (p < 0.05): the bigger the contact angle, the less able the plant was to capture particulate matter.

Investigating Seed Dormancy in Pinus bungeana Zucc. ex Endl.: Understanding the Contributions of Enclosing Tissues and Temperature on Germination

Pinus bungeana Zucc. ex Endl. is an endangered conifer tree species that is endemic to Western Central China. With the aim to confirm whether the Pinus bungeana seeds were dormant or not, an isolated embryo was cultured and seed coat interference with imbibition was examined by the water uptake test; the contribution of enclosing structures to germination inhibition and the effect of different temperatures (10 °C, 15 °C, 20 °C, 25 °C, and 30 °C) on seed germination were investigated by the germination test, then total germination percentage (TGP) and mean germination time (MGT) were calculated. Results showed that the Pinus bungeana seeds were non-dormant, seeds without any prechilling treatment germinated readily and achieved high germination (approximately 90%) at favorable temperatures (15 and 20 °C). At 25 °C, seed germination was inhibited and intact seeds exhibited 5% germination, but an interference with imbibition was not responsible for this result. In the seed tissue removal experiment, cracking the seed coat did not improve germination effectively, removing the seed coat and leaving the nucellar membrane either intact or having a quarter of it removed both elicited greater germination (34.7% and 40%, respectively). Meanwhile, removing the seed coat and removing either half or all of the nucellar membrane both promoted higher germination (approximately 80%), with germination rates that were nearly the same as that for the isolated embryos (86%). Germination inhibition was mainly induced by the enclosing structures and the nucellar membrane played an important role in inhibiting germination.

De Novo Sequencing and Assembly Analysis of Transcriptome in Pinus bungeana Zucc. ex Endl.

To enrich the molecular data of Pinus bungeana Zucc. ex Endl. and study the regulating factors of different morphology controled by apical dominance. In this study, de novo assembly of transcriptome annotation was performed for two varieties of Pinus bungeana Zucc. ex Endl. that are obviously different in morphology. More than 147 million reads were produced, which were assembled into 88,092 unigenes. Based on a similarity search, 11,692 unigenes showed significant similarity to proteins from Picea sitchensis (Bong.) Carr. From this collection of unigenes, a large number of molecular markers were identified, including 2829 simple sequence repeats (SSRs). A total of 158 unigenes expressed differently between two varieties, including 98 up-regulated and 60 down-regulated unigenes. Furthermore, among the differently expressed genes (DEGs), five genes which may impact the plant morphology were further validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The five genes related to cytokinin oxidase/dehydrogenase (CKX), two-component response regulator ARR-A family (ARR-A), plant hormone signal transduction (AHP), and MADS-box transcription factors have a close relationship with apical dominance. This new dataset will be a useful resource for future genetic and genomic studies in Pinus bungeana Zucc. ex Endl.

Comparative Transcriptome Analysis of Male and Female Conelets and Development of Microsatellite Markers in Pinus bungeana, an Endemic Conifer in China.

The sex determination in gymnosperms is still poorly characterized due to the lack of genomic/transcriptome resources and useful molecular genetic markers. To enhance our understanding of the molecular mechanisms of the determination of sexual recognition of reproductive structures in conifers, the transcriptome of male and female conelets were characterized in a Chinese endemic conifer species, Pinus bungeana Zucc. ex Endl. The 39.62 Gb high-throughput sequencing reads were obtained from two kinds of sexual conelets. After de novo assembly of the obtained reads, 85,305 unigenes were identified, 53,944 (63.23%) of which were annotated with public databases. A total of 12,073 differentially expressed genes were detected between the two types of sexes in P. bungeana, and 5766 (47.76%) of them were up-regulated in females. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enriched analysis suggested that some of the genes were significantly associated with the sex determination process of P. bungeana, such as those involved in tryptophan metabolism, zeatin biosynthesis, and cysteine and methionine metabolism, and the phenylpropanoid biosynthesis pathways. Meanwhile, some important plant hormone pathways (e.g., the gibberellin (GA) pathway, carotenoid biosynthesis, and brassinosteroid biosynthesis (BR) pathway) that affected sexual determination were also induced in P. bungeana. In addition, 8791 expressed sequence tag-simple sequence repeats (EST-SSRs) from 7859 unigenes were detected in P. bungeana. The most abundant repeat types were dinucleotides (1926), followed by trinucleotides (1711). The dominant classes of the sequence repeat were A/T (4942) in mononucleotides and AT/AT (1283) in dinucleotides. Among these EST-SSRs, 84 pairs of primers were randomly selected for the characterization of potential molecular genetic markers. Finally, 19 polymorphic EST-SSR primers were characterized. We found low to moderate levels of genetic diversity (NA = 1.754; HO = 0.206; HE = 0.205) across natural populations of P. bungeana. The cluster analysis revealed two distinct genetic groups for the six populations that were sampled in this endemic species, which might be caused by the fragmentation of habitats and long-term geographic isolation among different populations. Taken together, this work provides important insights into the molecular mechanisms of sexual identity in the reproductive organs of P. bungeana. The molecular genetic resources that were identified in this study will also facilitate further studies in functional genomics and population genetics in the Pinus species.

500-yr. precipitation variability in Southern Taihang Mountains, China, and its linkages to ENSO and PDO

An annually resolved and absolutely dated ring-width chronology spanning 657 yrs. is constructed with Whitebark pine (Pinus bungeana Zucc.) samples from the southern Taihang Mountains, Eastern China. On the basis of a significant correlation between the tree-ring width index and observed instrumental data, precipitation in current May is reconstructed for the region since AD 1510, with predictor variables accounting for 37.9 % of the variance in precipitation data. In agreement with other drought reconstructions, notable dry spells occur in the 1630s–1650s, 1680s–1700s, and 1770s–1800s, whereas wet periods prevail in the 1530s–1570s, 1840s–1870s, and 1950s-present. Wavelet analysis reveals clear 2–8, 20–40, and 80–130 yrs cycles at the 95 % confidence level for the reconstructed series over the past 500 yrs, suggesting possible linkages with the El Nino-Southern Oscillation (ENSO) and the Pacific Decadal Oscillation (PDO). Correlation analysis between the tree-ring series, ENSO, and PDO index further demonstrates that precipitation is negatively correlated with PDO and ENSO in the long term.

Quantifying the characteristics of particulate matters captured by urban plants using an automatic approach

It is widely accepted that urban plant leaves can capture airborne particles. Previous studies on the particle capture capacity of plant leaves have mostly focused on particle mass and/or size distribution. Fewer studies, however, have examined the particle density, and the size and shape characteristics of particles, which may have important implications for evaluating the particle capture efficiency of plants, and identifying the particle sources. In addition, the role of different vegetation types is as yet unclear. Here, we chose three species of different vegetation types, and firstly applied an object-based classification approach to automatically identify the particles from scanning electron microscope (SEM) micrographs. We then quantified the particle capture efficiency, and the major sources of particles were identified. We found (1) Rosa xanthina Lindl (shrub species) had greater retention efficiency than Broussonetia papyrifera (broadleaf species) and Pinus bungeana Zucc. (coniferous species), in terms of particle number and particle area cover. (2) 97.9% of the identified particles had diameter ≤10μm, and 67.1% of them had diameter ≤2.5μm. 89.8% of the particles had smooth boundaries, with 23.4% of them being nearly spherical. (3) 32.4%–74.1% of the particles were generated from bare soil and construction activities, and 15.5%–23.0% were mainly from vehicle exhaust and cooking fumes.

Nucleotide polymorphism and phylogeographic history of an endangered conifer species Pinus bungeana

Pinus bungeana Zucc. ex Endl. (Pinaceae), is an endangered conifer tree species and is endemic to western central China. In this study, we detected the nucleotide polymorphism and phylogeographic history of P. bungeana based on three nuclear genes. We collected 16 natural populations and covered the entire distributional ranges of P. bungeana. We found that this endangered species have extremely low level of nucleotide diversity (πt = 0.00098, πsil = 0.00229) compared with that of other conifers. Meanwhile, hierarchical AMOVA revealed that genetic variations of two nuclear loci (010054_01 and 0_12329_02) mainly occurred among populations (54% and 55%), and another locus (0_14221_01) existed within populations (64%). This finding may be caused by random fixation of nuclear genes. Neutrality tests and mismatch analysis suggested that P. bungeana may have experienced the recent range expansion. In addition, high genetic diversity and unique haplotypes were identified in the west areas of Qinling-Daba and Lvliang Mountains, and these areas may be the glacial refuges of P. bungeana. After the glaciation, the species expanded or migrated outward from these refuges, and the founder effect or bottleneck effect in this process may have resulted in the low level of nucleotide polymorphism of P. bungeana.

The complete plastid genome of Bunge’s pine Pinus bungeana (Pinaceae)

The complete nucleotide sequence of Bunge’s pine Pinus bungeana Zucc. ex Endl. chloroplast genome (cp DNA) was determined in this study. The cpDNA was 117 861 bp in length, containing a pair of 475 bp inverted repeat regions (IRa and IRb), which were separated by large and small single copy regions (LSC and SSC) of 65 373 and 51 538 bp, respectively. The cpDNA contained 111 genes, including 71 protein-coding genes (71 PCG species), 4 ribosomal RNA genes (4 rRNA species) and 36 tRNA genes (32 tRNA species). In these genes, 13 harbored a single intron and 1 (ycf3) contained a couple of introns. The overall AT content of Bunge’s pine cpDNA is 61.2%, while the corresponding values of the LSC, SSC and IR regions are 61.9%, 60.2% and 62.5%, respectively. A phylogenetic reconstruction based on the maximum parsimony analysis suggested that all the sampled Pinus species clustered a monophyletic clade and have a high bootstrap support, and the cpDNA of P. bungeana is closely related to that of congeneric P. gerardiana.

Dilute acid pretreatment differentially affects the compositional and architectural features of Pinus bungeana Zucc. compression and opposite wood tracheid walls

Compression wood represents a unique challenge for biochemical processing of sustainable biomass to produce biofuels and bioproducts on account of its highly lignified tracheids relative to opposite wood. In the current work, differentiating response of Pinus bungeana Zucc. compression wood and opposite wood tracheid walls to dilute acid pretreatment was elucidated by combining chemical and microscopic approaches. Dilute acid pretreatment released greater amount of hemicelluloses from opposite wood (91.0%) than from compression wood (31.2%). The dissolution of hemicelluloses was not uniform and varied across the tracheid wall. In addition, for both compression wood and opposite wood the heterogeneity in lignin distribution was enhanced, probably attributed to the lignin migration and relocalization. The ultrastructural arrangement of the cell wall was also altered with the pretreated opposite wood tracheid wall exposing more apparent cellulose microfibril bundles and less abundant globular structures compared to compression wood. The dilute acid pretreatment demonstrated that the greater recalcitrance of compression wood probably originated from the higher lignin content and the resulting more compact structure of the cell wall.

2, 6-Dichlorobenzonitrile causes multiple effects on pollen tube growth beyond altering cellulose synthesis in Pinus bungeana Zucc.

Cellulose is an important component of cell wall, yet its location and function in pollen tubes remain speculative. In this paper, we studied the role of cellulose synthesis in pollen tube elongation in Pinus bungeana Zucc. by using the specific inhibitor, 2, 6-dichlorobenzonitrile (DCB). In the presence of DCB, the growth rate and morphology of pollen tubes were distinctly changed. The organization of cytoskeleton and vesicle trafficking were also disturbed. Ultrastructure of pollen tubes treated with DCB was characterized by the loose tube wall and damaged organelles. DCB treatment induced distinct changes in tube wall components. Fluorescence labeling results showed that callose, and acidic pectin accumulated in the tip regions, whereas there was less cellulose when treated with DCB. These results were confirmed by FTIR microspectroscopic analysis. In summary, our findings showed that inhibition of cellulose synthesis by DCB affected the organization of cytoskeleton and vesicle trafficking in pollen tubes, and induced changes in the tube wall chemical composition in a dose-dependent manner. These results confirm that cellulose is involved in the establishment of growth direction of pollen tubes, and plays important role in the cell wall construction during pollen tube development despite its lower quantity.

Physiological and Protein Responses to Drought in Four Pine Seedlings

Abstract Physiological and protein responses to drought stress in four pine seedlings, Pinus armandi Franch. (Pa), Pinus tabulaeformis Carr. (Pt), Pinus bungeana Zucc. ex Endl. (Pb) and Pinus sylvestris L. var. mongolica Litv. (Ps), were investigated using differential proteomics and water physiological indices. Firstly, the water physiological data showed that the decline rate of net photosynthesis rate, stomatal conductance, leaf water potential, turgor pressure except for under moderate drought stress was as follows: Pa>Ps>Pt>Pb. Pb and Pa always maintained the highest and lowest swelling pressure, respectively. Secondly, cluster analysis of 343 proteins indicated that the four pine species were classified into three groups with a genetic distance coefficient of 0.065. That is, five-needle-pine group (Pa), three-needle-pine group (Pb), two-needle-pine group (Pt and Ps), and the genetic distance between Pb and Pa was the farthest. The result was consistent with the declined rate in above physiological indices. Finally, for the differential proteomics analyzed, a total of 13 different proteins (P values < 0.01) changed significantly, the number of differentially expressed proteins was more in Pa (accounting for 46.2%) than the other three species, and 8 proteins were identified using matrix-assisted laser desorption/ionization time -of-flight mass spectrometry (MALDI-TOF-MS). These proteins are quite diverse in their functions and involved in photosynthesis, osmotic regulation and functioning as signal transduction. These results suggested that the sensitivities of the four pine species to drought were possibly related to genetic distance.