Composition and storage of soil inorganic carbon as well as the controlling factors in coastal area of the northern Jiangsu, China.

The sequestration of soil inorganic carbon (SIC) especially pedogenic carbonate (PC) is one of the important pathways reducing the concentration of atmospheric carbon dioxide and thus mitigating climate change in coastal areas. Using the technology of 13C stable isotope, we analyzed the differences in the composition and storage of SIC, and explored the key physicochemical properties influencing soil PC storage in different horizons (0-10, 11-20, 21-40, 41-60, 61-80 and 81-100 cm) from Suaeda salsa wetland (SS), Spartina alterniflora wetland (SA), young poplar plantation (YP), and mature poplar plantation (MP) in coastal area of the northern Jiangsu Province. The results showed that except for the surface (0-10 cm) soil in MP, the SIC content was higher than SOC in all soil horizons. Overall, neither the soil PC to SIC ratio nor the SIC storage were significantly different in SA and SS soils. Compared to wetland soils (0-40 cm), the soil PC to SIC ratio was reduced by 32.7% and 54.1% and the PC storage was reduced by 40.5% and 59.2%, the lithogenic carbonate (LC) storage changed little, while the SIC storage was reduced by 21.0% and 17.9%, respectively in the YP and MP soils. Compared to the YP soils (0-100 cm), both the soil PC to SIC ratio and the PC storage were significantly reduced while the LC storage was significantly increased, especially at the 41-100 cm soil horizons, meanwhile, the SIC storage was not significantly changed in the MP soils. Results of the structural equation modeling (SEM) indicated that key factors influencing soil PC storage were the ratio of PC to SIC, followed by the SOC content and bulk density. SOC could inhibit the formation of soil PC. Generally, the coastal wetlands have greater SIC storage and sequestration potential than poplar plantations, and the PC sequestration can be regulated by modulating the ratio of PC to SIC and SOC content.

Comparative Genomics Analysis of the Populus Epidermal Pattern Factor (EPF) Family Revealed Their Regulatory Effects in Populus euphratica Stomatal Development.

Drought stress seriously threatens plant growth. The improvement of plant water use efficiency (WUE) and drought tolerance through stomatal regulation is an effective strategy for coping with water shortages. Epidermal patterning factor (EPF)/EPF-like (EPFL) family proteins regulate stomatal formation and development in plants and thus contribute to plant stress adaptation. Here, our analysis revealed the presence of 14 PeEPF members in the Populus euphratica genome, which exhibited a relatively conserved gene structure with 1-3 introns. Subcellular localisation prediction revealed that 9 PeEPF members were distributed in the chloroplasts of P. euphratica, and 5 were located extracellularly. Phylogenetic analysis indicated that PeEPFs can be divided into three clades, with genes within the same clade revealing a relatively conserved structure. Furthermore, we observed the evolutionary conservation of PeEPFs and AtEPF/EPFLs in certain domains, which suggests their conserved function. The analysis of cis-acting elements suggested the possible involvement of PeEPFs in plant response to multiple hormones. Transcriptomic analysis revealed considerable changes in the expression level of PeEPFs during treatment with polyethylene glycol and abscisic acid. The overexpression of PeEPF2 resulted in low stomatal density in transgenetic lines. These findings provide a basis for gaining insights into the function of PeEPFs in response to abiotic stress.

Populus trichocarpa EXPA6 Facilitates Radial and Longitudinal Transport of Na+ under Salt Stress.

Expansins are cell wall (CW) proteins that mediate the CW loosening and regulate salt tolerance in a positive or negative way. However, the role of Populus trichocarpa expansin A6 (PtEXPA6) in salt tolerance and the relevance to cell wall loosening is still unclear in poplars. PtEXPA6 gene was transferred into the hybrid species, Populus alba × P. tremula var. glandulosa (84K) and Populus tremula × P. alba INRA '717-1B4' (717-1B4). Under salt stress, the stem growth, gas exchange, chlorophyll fluorescence, activity and transcription of antioxidant enzymes, Na+ content, and Na+ flux of root xylem and petiole vascular bundle were investigated in wild-type and transgenic poplars. The correlation analysis and principal component analysis (PCA) were used to analyze the correlations among the characteristics and principal components. Our results show that the transcription of PtEXPA6 was downregulated upon a prolonged duration of salt stress (48 h) after a transient increase induced by NaCl (100 mM). The PtEXPA6-transgenic poplars of 84K and 717-1B4 showed a greater reduction (42-65%) in stem height and diameter growth after 15 days of NaCl treatment compared with wild-type (WT) poplars (11-41%). The Na+ accumulation in roots, stems, and leaves was 14-83% higher in the transgenic lines than in the WT. The Na+ buildup in the transgenic poplars affects photosynthesis; the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT); and the transcription of PODa2, SOD [Cu-Zn], and CAT1. Transient flux kinetics showed that the Na+ efflux of root xylem and leaf petiole vascular bundle were 1.9-3.5-fold greater in the PtEXPA6-transgenic poplars than in the WT poplars. PtEXPA6 overexpression increased root contractility and extensibility by 33% and 32%, indicating that PtEXPA6 increased the CW loosening in the transgenic poplars of 84K and 717-1B4. Noteworthily, the PtEXPA6-promoted CW loosening was shown to facilitate Na+ efflux of root xylem and petiole vascular bundle in the transgenic poplars. We conclude that the overexpression of PtEXPA6 leads to CW loosening that facilitates the radial translocation of Na+ into the root xylem and the subsequent Na+ translocation from roots to leaves, resulting in an excessive Na+ accumulation and consequently, reducing salt tolerance in transgenic poplars. Therefore, the downregulation of PtEXPA6 in NaCl-treated Populus trichocarpa favors the maintenance of ionic and reactive oxygen species (ROS) homeostasis under long-term salt stress.

[Characteristics and influence factors of rainfall redistribution in eight typical plantations in the loess area in West Shanxi, China]. / 晋西黄土区8ç§å ¸åž‹æž—åˆ†é™é›¨å†åˆ†é ç‰¹å¾ä¸Žå½±å“å› ç´ .

Aiming for clarifying the potential distribution characteristics of canopy rainfall partitioning of the loess area, we explored the process of rainfall partitioning across eight typical forest stands (Pinus tabuliformis forest, Robinia pseudoacacia forest, Platycladus orientalis forest, mixed forest of Robinia pseudoacacia-Pinus tabuliformis, mixed forest of Platycladus orientalis-Robinia pseudoacacia, Quercus wutaishanica forest, Populus davidiana forest, mixed forest of Quercus wutaishanica-Populus davidiana), and used boosted regression trees (BRT) to quantify the relative influences of stand structures and meteorological environment factors. We established multiple regression relationships according to the most influential factors extracted by BRT, and applied to the dataset of mining to verify the performance of the BRT-derived predictive model. The results showed that the percentages of throughfall (TF), stemflow (SF), and canopy interception (Ic) in total precipitation were 24.5%-95.1%, 0-13.6%, and 0.7%-55.7% among eight typical forest stands, respectively. For the individual rainfall threshold of TF, coniferous forest (3.06±1.21 mm) was significantly higher than broad-leaved forest (1.97±0.52 mm), but there was no significant difference between coniferous forest and broad-leaved mixed forest (3.01±0.98 mm). There was no significant difference in the individual rainfall threshold of SF among different composition stands. BRT analysis showed that stand structure factors accounted for a relatively small proportion for TF and SF, respectively. By contrast, stand structure factors dominated the Ic. Rainfall was the most important factor in determining TF and SF. Tree height was the most important factor in determining Ic, followed by rainfall, canopy area, diameter at breast height, and stand density. Compared with the general linear function and the power function, the prediction effect of BRT prediction model constructed here on TF and SF had been further improved, and the prediction of canopy interception still needed to explore. In conclusion, the BRT model could better quantitatively evaluate the effects of stand structure and meteorological environmental factors on rainfall partitioning components, and the performance of the BRT predictive model could satisfy and lay the foundation for the optimization strategy for stand configuration.

Microbial colonisation rewires the composition and content of poplar root exudates, root and shoot metabolomes.

BACKGROUND: Trees are associated with a broad range of microorganisms colonising the diverse tissues of their host. However, the early dynamics of the microbiota assembly microbiota from the root to shoot axis and how it is linked to root exudates and metabolite contents of tissues remain unclear. Here, we characterised how fungal and bacterial communities are altering root exudates as well as root and shoot metabolomes in parallel with their establishment in poplar cuttings (Populus tremula x tremuloides clone T89) over 30 days of growth. Sterile poplar cuttings were planted in natural or gamma irradiated soils. Bulk and rhizospheric soils, root and shoot tissues were collected from day 1 to day 30 to track the dynamic changes of fungal and bacterial communities in the different habitats by DNA metabarcoding. Root exudates and root and shoot metabolites were analysed in parallel by gas chromatography-mass spectrometry. RESULTS: Our study reveals that microbial colonisation triggered rapid and substantial alterations in both the composition and quantity of root exudates, with over 70 metabolites exclusively identified in remarkably high abundances in the absence of microorganisms. Noteworthy among these were lipid-related metabolites and defence compounds. The microbial colonisation of both roots and shoots exhibited a similar dynamic response, initially involving saprophytic microorganisms and later transitioning to endophytes and symbionts. Key constituents of the shoot microbiota were also discernible at earlier time points in the rhizosphere and roots, indicating that the soil constituted a primary source for shoot microbiota. Furthermore, the microbial colonisation of belowground and aerial compartments induced a reconfiguration of plant metabolism. Specifically, microbial colonisation predominantly instigated alterations in primary metabolism in roots, while in shoots, it primarily influenced defence metabolism. CONCLUSIONS: This study highlighted the profound impact of microbial interactions on metabolic pathways of plants, shedding light on the intricate interplay between plants and their associated microbial communities. Video Abstract.

Influence of surface water and groundwater gradient on spatial distribution of typical vegetation in the hinterland of Taklamakan desert.

Water resources are essential for desert oases and are key drivers of local ecological processes critical to the growth of desert vegetation. In this study, the oasis in the hinterland of the Taklamakan Desert, China, was selected as the research subject. Using high-precision classification of oasis vegetation through machine learning, surface water within the oasis was identified and extracted from multi-year Landsat remote sensing data. The spatial distribution patterns of the main community-building species, Populus euphratica and Tamarix ramosissima, were studied under different moisture gradients using environmental covariates and measured groundwater depth to invert its spatial distribution and K-mean clustering to construct surface water and groundwater moisture gradients. The results indicated that the classification accuracy for the two species reached 0.917. Gradients 1-5 were used to categorize the water resources, dividing surface water and groundwater into five gradients. Gradient 3 exhibited the optimal moisture conditions, with a high surface water distribution frequency (0.017) and shallow groundwater depth (3.158 m), while Gradient 4 showed the least optimal moisture conditions, characterized by a low surface water distribution frequency (0.008) and deep groundwater depth (4.820 m). The water gradient decreased in the following order: Gradient 3 > Gradient 5 > Gradient 1 > Gradient 2 > Gradient 4. The optimum gradients for growth of P. euphratica and T. ramosissima were gradients 5, 1, and 2. The normalized vegetation index spatial distribution patterns of the two species were consistent with that of the moisture gradient. Tamarix ramosissima was found to be more tolerant to salinity and drought than P. euphratica. Overall, this study provides valuable information on the effect of the spatial distribution of water resource gradients on oasis vegetation and can guide future water delivery policies in oases.

How is tree growth rate linked to root functional traits in phylogenetically related poplar hybrids?

Fine roots play a crucial role in soil nutrient and water acquisition, significantly contributing to tree growth. Fine roots with a high specific root length (SRL) and small diameter are often considered to help trees grow fast. However, inconsistencies in the literature do not provide a clear basis on the effect of root functional traits, such as SRL or root mass density (RMD), on tree growth rate in phylogenetically related trees. Our aim was to examine relationships between tree growth rate and root functional traits, using clones displaying different growth rates in a hybrid poplar plantation located in New Liskeard, ON, Canada. Fine roots (diameter < 2 mm) samples were collected using soil cores at depths of 0-20, 20-40 and 40-60 cm, and analyzed for morphological, chemical and architectural traits. High SRL and thin fine roots were associated with the least productive clones, which is not consistent with the root economics spectrum (RES) theory. However, the most productive clone had larger fine root diameter and higher root lignin concentrations, probably reducing root construction and maintenance costs and carbon losses. Therefore, at the 0-20 and 20-40 cm depths, tree growth rates showed positive correlations with root diameter and root lignin concentrations, but negative correlations with SRL and root soluble compounds concentration. Increasing RMD at the 0-20 cm depth promoted tree growth rates, showing the importance of soil exploration in the topsoil for tree growth. We conclude that fine root variation does not always follow the RES hypothesis and argue that the rapid growth rate of trees may also be driven by fine root growth in diameter and mass in phylogenetically related trees.

RNA-seq reveals the gene expression in patterns in Populus × euramericana 'Neva' plantation under different precision water and fertilizer-intensive management.

BACKGROUND: Populus spp. is a crucial fast-growing and productive tree species extensively cultivated in the mid-latitude plains of the world. However, the impact of intensive cultivation management on gene expression in plantation remains largely unexplored. RESULTS: Precision water and fertilizer-intensive management substantially increased key enzyme activities of nitrogen transport, assimilation, and photosynthesis (1.12-2.63 times than CK) in Populus × euramericana 'Neva' plantation. Meanwhile, this management approach had a significant regulatory effect on the gene expression of poplar plantations. 1554 differential expression genes (DEGs)were identified in drip irrigation (ND) compared with conventional irrigation. Relative to ND, 2761-4116 DEGs, predominantly up-regulated, were identified under three drip fertilization combinations, among which 202 DEGs were mainly regulated by fertilization. Moreover, drip irrigation reduced the expression of cell wall synthesis-related genes to reduce unnecessary water transport. Precision drip and fertilizer-intensive management promotes the synergistic regulation of carbon and nitrogen metabolism and up-regulates the expression of major genes in nitrogen transport and assimilation processes (5 DEGs), photosynthesis (15 DEGs), and plant hormone signal transduction (11 DEGs). The incorporation of trace elements further enhanced the up-regulation of secondary metabolic process genes. In addition, the co-expression network identified nine hub genes regulated by precision water and fertilizer-intensive management, suggesting a pivotal role in regulating the growth of poplar. CONCLUSION: Precision water and fertilizer-intensive management demonstrated the ability to regulate the expression of key genes and transcription factor genes involved in carbon and nitrogen metabolism pathways, plant hormone signal transduction, and enhance the activity of key enzymes involved in related processes. This regulation facilitated nitrogen absorption and utilization, and photosynthetic abilities such as light capture, light transport, and electron transport, which faintly synergistically regulate the growth of poplar plantations. These results provide a reference for proposing highly efficient precision intensive management to optimize the expression of target genes.

A molecular module connects abscisic acid with auxin signals to facilitate seasonal wood formation in Populus.

Perennial trees have a recurring annual cycle of wood formation in response to environmental fluctuations. However, the precise molecular mechanisms that regulate the seasonal formation of wood remain poorly understood. Our prior study indicates that VCM1 and VCM2 play a vital role in regulating the activity of the vascular cambium by controlling the auxin homoeostasis of the cambium zone in Populus. This study indicates that abscisic acid (ABA) affects the expression of VCM1 and VCM2, which display seasonal fluctuations in relation to photoperiod changes. ABA-responsive transcription factors AREB4 and AREB13, which are predominantly expressed in stem secondary vascular tissue, bind to VCM1 and VCM2 promoters to induce their expression. Seasonal changes in the photoperiod affect the ABA amount, which is linked to auxin-regulated cambium activity via the functions of VCM1 and VCM2. Thus, the study reveals that AREB4/AREB13-VCM1/VCM2-PIN5b acts as a molecular module connecting ABA and auxin signals to control vascular cambium activity in seasonal wood formation.

Identification and transcriptome analysis of a photosynthesis deficient mutant of Populus davidiana Dode.

Photosynthesis is the main source of energy for plants to sustain growth and development. Abnormalities in photosynthesis may cause defects in plant development. The elaborate regulatory mechanism underlying photosynthesis remains unclear. In this study, we identified a natural mutant from the Greater Khingan Mountains and named it as "1-T". This mutant had variegated leaf with irregular distribution of yellow and green. Chlorophyll contents and photosynthetic capacity of 1-T were significantly reduced compared to other poplar genotypes. Furthermore, a transcriptome analysis revealed 3269 differentially expressed genes (DEGs) in 1-T. The products of the DEGs were enriched in photosystem I and photosystem II. Three motifs were significantly enriched in the promoters of these DEGs. Yeast one-hybrid, Electrophoretic mobility shift assays and tobacco transient transformation experiments indicated that PdGLKs may bind to the three motifs. Further analysis indicated that these photosystem related genes were also significantly down-regulated in PdGLK-RNAi poplars. Therefore, we preliminarily concluded that the down-regulation of PdGLKs in 1-T may affect the expression of photosystem-related genes, resulting in abnormal photosystem development and thus affecting the growth and development. Our results provide new insights into the molecular mechanism of photosynthesis regulating plant growth.

PagMYB128 regulates secondary cell wall formation by direct activation of cell wall biosynthetic genes during wood formation in poplar.

The biosynthesis of cellulose, lignin, and hemicelluloses in plant secondary cell walls (SCWs) is regulated by a hierarchical transcriptional regulatory network. This network features orthologous transcription factors shared between poplar and Arabidopsis, highlighting a foundational similarity in their genetic regulation. However, knowledge on the discrepant behavior of the transcriptional-level molecular regulatory mechanisms between poplar and Arabidopsis remains limited. In this study, we investigated the function of PagMYB128 during wood formation and found it had broader impacts on SCW formation compared to its Arabidopsis ortholog, AtMYB103. Transgenic poplar trees overexpressing PagMYB128 exhibited significantly enhanced xylem development, with fiber cells and vessels displaying thicker walls, and an increase in the levels of cellulose, lignin, and hemicelluloses in the wood. In contrast, plants with dominant repression of PagMYB128 demonstrated the opposite phenotypes. RNA sequencing and reverse transcription - quantitative polymerase chain reaction showed that PagMYB128 could activate SCW biosynthetic gene expression, and chromatin immunoprecipitation along with yeast one-hybrid, and effector-reporter assays showed this regulation was direct. Further analysis revealed that PagSND1 (SECONDARY WALL-ASSOCIATED NAC-DOMAIN PROTEIN1) directly regulates PagMYB128 but not cell wall metabolic genes, highlighting the pivotal role of PagMYB128 in the SND1-driven regulatory network for wood development, thereby creating a feedforward loop in SCW biosynthesis.

PagKNAT2/6b regulates tension wood formation and gravitropism by targeting cytokinin metabolism.

Tension wood is a specialized xylem tissue associated with gravitropism in angiosperm trees. However, few regulators of tension wood formation have been identified. The molecular mechanisms underpinning tension wood formation remain elusive. Here, we report that a Populus KNOTTED-like homeobox gene, PagKNAT2/6b, is involved in tension wood formation and gravity response. Transgenic poplar plants overexpressing PagKNAT2/6b displayed more sensitive gravitropism than controls, as indicated by increased stem curvature. Microscopic examination revealed greater abundance of fibre cells with a gelatinous cell wall layer (G-layer) and asymmetric growth of secondary xylem in PagKNAT2/6b overexpression lines. Conversely, PagKNAT2/6b dominant repression plants exhibited decreased tension wood formation and reduced response to gravity stimulation. Moreover, sensitivity to gravity stimulation showed a negative relationship with development stage. Expression of genes related to growth and senescence was affected in PagKNAT2/6b transgenic plants. More importantly, transcription activation and electrophoretic mobility shift assays suggested that PagKNAT2/6b promotes the expression of cytokinin metabolism genes. Consistently, cytokinin content was increased in PagKNAT2/6b overexpression plants. Therefore, PagKNAT2/6b is involved in gravitropism and tension wood formation, likely via modulation of cytokinin metabolism.

MicroRNA257 promotes secondary growth in hybrid poplar.

Populus, a significant fast-growing tree species with global afforestation and energy potential, holds considerable economic value. The abundant production of secondary xylem by trees, which serves as a vital resource for industrial purposes and human sustenance, necessitates the orchestration of various regulatory mechanisms, encompassing transcriptional regulators and microRNAs (miRNAs). Nevertheless, the investigation of microRNA-mediated regulation of poplar secondary growth remains limited. In this study, we successfully isolated a novel microRNA (Pag-miR257) from 84 K poplar and subsequently integrated it into the 35 S overexpression vector. The overexpression of Pag-miR257 resulted in notable increases in plant height, stem diameter, and fresh weight. Additionally, the overexpression of Pag-miR257 demonstrated a significant enhancement in net photosynthetic rate. The findings from the examination of cell wall autofluorescence indicated a substantial increase in both xylem area and the number of vessels in poplar plants overexpressing Pag-miR257. Furthermore, the cell wall of the Pag-miR257 overexpressing plants exhibited thickening as observed through transmission electron microscopy. Moreover, the Fourier Transforms Infrared (FTIR) analysis and phloroglucinol-HCl staining revealed an elevation in lignin content in Pag-miR257 overexpressing poplar plants. The findings of this study suggest that microRNA257 may play a role in the control of secondary growth in poplar stems, thereby potentially enhancing the development of wood engineering techniques for improved material and energy production.

Combined effects of cadmium and simulated acid rain on soil microbial communities in the early cultivation of Populus beijingensis seedlings.

The combined cadmium (Cd) and acid rain pollution poses a significant threat to the global ecological environment. Previous studies on the combined adverse effects have predominantly focused on the aboveground plant physiological responses, with limited reports on the microbial response in the rhizosphere soil. This study employed Populus beijingensis seedlings and potting experiments to simulate the impacts of combined mild acid rain (pH=4.5, MA) or highly strong acid rain (pH=3.0, HA), and soil Cd pollution on the composition and diversity of microbial communities, as well as the physiochemical properties in the rhizosphere soil. The results showed that Cd decreased the content of inorganic nitrogen, resulting in an overall decrease of 49.10 % and 46.67 % in ammonium nitrogen and nitrate nitrogen, respectively. Conversely, acid rain was found to elevate the content of total potassium and soil organic carbon by 4.68 %-6.18 % and 8.64-19.16 %, respectively. Additionally, simulated acid rain was observed to decrease the pH level by 0.29-0.35, while Cd increased the pH level by 0.11. Moreover, Cd alone reduced the rhizosphere bacterial diversity, however, when combined with acid rain, regardless of its intensity, Cd was observed to increase the diversity. Fungal diversity was not influenced by the acid rain, but Cd increased fungal diversity to some extend under HA as observed in bacterial diversity. In addition, composition of the rhizosphere bacterial community was primarily influenced by the inorganic nitrogen components, while the fungal community was driven mainly by soil pH. Furthermore, "Metabolism" was emerged as the most significant bacterial function, which was markedly affected by the combined pollution, while Cd pollution led to a shift from symbiotroph to other trophic types for fungi. These findings suggest that simulated acid rain has a mitigating effect on the diversity of rhizosphere bacteria affected by Cd pollution, and also alters the trophic type of these microorganisms. This can be attributed to the acid rain-induced direct acidic environment, as well as the indirect changes in the availability or sources of carbon, nitrogen, or potassium.

[Radial growth response of Populus euphratica to climate change in the Cele desert oasis ecotone, China]. / ç­–å‹’æ²™æ¼ ç»¿æ´²è¿‡æ¸¡å¸¦èƒ¡æ¨å¾„å‘ç”Ÿé•¿å¯¹æ°”å€™å˜åŒ–çš„å“åº”.

Populus euphratica is an important tree species in the arid regions of Northwest China, which is sensitive to climate changes. Climate of the Northwest China is changing to be "warm and humid", but how it would affect the regional forest growth is not clear. In this study, the radial growth response of P. euphratica to major climatic factors and their temporal changes during 1984-2021 were analyzed by using dendrochronology method in the desert oasis ecotone of Cele in the southern Tarim basin. The results showed that tree-ring width index of P. euphratica had a significant negative correlation with temperature in September of the previous year, and in February and May of current year, had significant positive correlation with precipitation in September of previous year and March and May of current year, and had significant positive correlations with SPEI in February and May of current year. The relationships between tree-ring width index and combined month climatic factors were more obvious. The results of moving correlation analysis showed that the correlation between tree-ring width index and temperature in the growing season tended to be strengthened in recent years, while the correlation between tree-ring width index and precipitation, SPEI tended to be declined or remain stable. The variations of the relationships between tree-ring width index and combined month climatic factors were more obvious compared that with single month. Current regional climate is conducive to the growth and development, as well as the improvement of ecological shelter function of P. euphratica forest in the desert oasis ecotone of Cele.

PagJAZ5 regulates cambium activity through coordinately modulating cytokinin concentration and signaling in poplar.

Wood is resulted from the radial growth paced by the division and differentiation of vascular cambium cells in woody plants, and phytohormones play important roles in cambium activity. Here, we identified that PagJAZ5, a key negative regulator of jasmonate (JA) signaling, plays important roles in enhancing cambium cell division and differentiation by mediating cytokinin signaling in poplar 84K (Populus alba × Populus glandulosa). PagJAZ5 is preferentially expressed in developing phloem and cambium, weakly in developing xylem cells. Overexpression (OE) of PagJAZ5m (insensitive to JA) increased cambium activity and xylem differentiation, while jaz mutants showed opposite results. Transcriptome analyses revealed that cytokinin oxidase/dehydrogenase (CKXs) and type-A response regulators (RRs) were downregulated in PagJAZ5m OE plants. The bioactive cytokinins were significantly increased in PagJAZ5m overexpressing plants and decreased in jaz5 mutants, compared with that in 84K plants. The PagJAZ5 directly interact with PagMYC2a/b and PagWOX4b. Further, we found that the PagRR5 is regulated by PagMYC2a and PagWOX4b and involved in the regulation of xylem development. Our results showed that PagJAZ5 can increase cambium activity and promote xylem differentiation through modulating cytokinin level and type-A RR during wood formation in poplar.

Different ectomycorrhizal fungal species impact poplar growth but not phosphorus utilization under low P supply.

Tree growth is often limited by phosphorus (P) availability. The trade-off between P homeostasis and growth is unknown. Ectomycorrhizal fungi (EMF) facilitate P availability but this trait varies among different fungal species and isolates. Here, we tested the hypotheses that (i) colonization with EMF boosts plant growth under P-limited conditions and that (ii) the poplars show P homeostasis because increased P uptake is used for growth and not for P accumulation in the tissues. We used two P treatments (high phosphate [HP]: 64 µM Pi, low phosphate [LP]: 0.64 µM Pi in the nutrient solution) and four fungal treatments (Paxillus involutus MAJ, Paxillus involutus NAU, Laccaria bicolor dikaryon LBD, Laccaria bicolor monokaryon LBM) in addition to non-inoculated poplar plants (NI) to measure growth, biomass, gas exchange and P contents. High phosphate (HP) stimulated growth compared with LP conditions. Poplars colonized with MAJ, NAU and NI showed higher growth and biomass production than those with LBD or LBM. Photosynthesis rates of poplars with lower biomass production were similar to or higher than those of plants with higher growth rates. The tissue concentrations of P were higher under HP than LP conditions and rarely affected by ectomycorrhizal colonization. Under LP, the plants produced 44% greater biomass per unit of P than under HP. At a given P supply, the tissue concentration was stable irrespective of the growth rate indicating P homeostasis. Laccaria bicolor caused growth inhibition, irrespective of P availability. These results suggest that in young poplars distinct species-specific ectomycorrhizal traits overshadowed potential growth benefits.

WUSCHEL-RELATED HOMEOBOX genes are crucial for normal vascular organization and wood formation in poplar.

Vascular cambium in tree species is a cylindrical domain of meristematic cells that are responsible for producing secondary xylem (also called wood) inward and secondary phloem outward. The poplar (Populus trichocarpa) WUSCHEL (WUS)-RELATED HOMEOBOX (WOX) family members, PtrWUSa and PtrWOX13b, were previously shown to be expressed in vascular cambium and differentiating xylem cells in poplar stems, but their functions remain unknown. Here, we investigated roles of PtrWUSa, PtrWOX13b and their close homologs in vascular organization and wood formation. Expression analysis showed that like PtrWUSa and PtrWOX13b, their close homologs, PtrWUSb, PtrWUS4a/b and PtrWOX13a/c, were also expressed in vascular cambium and differentiating xylem cells in poplar stems. PtrWUSa also exhibited a high level of expression in developing phloem fibers. Expression of PtrWUSa fused with the dominant EAR repression domain (PtrWUSa-DR) in transgenic poplar caused retarded growth of plants with twisted stems and curled leaves and a severe disruption of vascular organization. In PtrWUSa-DR stems, a drastic proliferation of cells occurred in the phloem region between vascular cambium and phloem fibers and they formed islands of ectopic vascular tissues or phloem fiber-like sclerenchyma cells. A similar proliferation of cells was also observed in PtrWUSa-DR leaf petioles and midveins. On the other hand, overexpression of PtrWOX4a-DR caused ectopic formation of vascular bundles in the cortical region, and overexpression of PtrWOX13a-DR and PtrWOX13b-DR led to a reduction in wood formation without affecting vascular organization in transgenic poplar plants. Together, these findings indicate crucial roles of PtrWUSa and PtrWOX13a/b in regulating vascular organization and wood formation, which furthers our understanding of the functions of WOX genes in regulating vascular cambium activity in tree species.

C2H2 zinc finger protein PagIDD15A regulates secondary wall thickening and lignin biosynthesis in poplar.

Wood production is largely determined by the activity of cambial cell proliferation, and the secondary cell wall (SCW) thickening of xylem cells determines the wood property. In this study, we identified an INDETERMINATE DOMAIN (IDD) type C2H2 zinc finger transcription factor PagIDD15A as a regulator of wood formation in Populus alba × Populus glandulosa. Downregulation of PagIDD15A expression by RNA interference (RNAi) inhibited xylem development and xylem cell secondary wall thickening. RNA-seq analysis showed that PagPAL1, PagCCR2 and PagCCoAOMT1 were downregulated in the differentiating xylem of the PagIDD15A-RNAi transgenic plants, showing that PagIDD15A may regulate SCW biosynthesis through inhibiting lignin biosynthesis. The downregulation of PagVND6-B2, PagMYB10 and PagMYC4 and upregulation of PagWRKY12 in the differentiating xylem of RNAi transgenic plants suggest that PagIDD15A may also regulate these transcription factor (TF) genes to affect SCW thickening. RT-qPCR analysis in the phloem-cambium of RNAi transgenic demonstrates that PagIDD15A may regulate the expression of the genes associated with cell proliferation, including, PagSHR (SHORTROOT), PagSCR (SCARECROW), PagCYCD3;1 (CYCLIN D3;1) and PagSMR4 (SIAMESE-RELATED4), to affect the cambial activity. This study provides the knowledge of the IDD-type C2H2 zinc finger protein in regulating wood formation.

Impacts of forest age on soil characteristics and fertility quality of Populus simonii shelter forest at the southern edge of the Horqin Sandy Land, China.

The sand fixing shelter forests in the Horqin Sandy Land are a key area in the "3-North" Shelter Forest Program in China, which has a history of over 50 years of artificial afforestation. Populus simonii Carr is one of the most dominant silvicultural species in the region. The aim of this study is to understand the soil characteristics and soil fertility of Populus simonii shelter forests at different growth stages and to establish a scientific basis for soil nutrient regulation and sustainable management of Populus simonii shelter forests at the southern edge of the Horqin Sandy Land. Sample plots were selected for young (≤15 a), middle-aged (16-25 a), near-mature (26-30 a), mature (31-40 a), and over-mature (≥41 a) forests. Each forest studied was in a state of natural restoration with uniform stand conditions and no artificial fertilizer was applied. These sites were selected to study changes in the soil characteristics in soil depths of 0-20, 20-40, and 40-60 cm. In order to avoid the problem of multicollinearity between soil variables and to reduce redundancy, principal component analysis (PCA), Pearson's correlation analysis, and Norm value calculation were used to select the least correlated indicators with the highest factor loadings. This was used to establish the minimum data set. The soil fertility quality of these shelterbelts in different forest ages was quantified using the soil quality index (SQI). In the growth stage from young to nearly mature forests, the soil bulk weight and pH decreased with increasing forest age. Soil capillary porosity, noncapillary porosity, total porosity, water content, field water holding capacity, and organic carbon content increased with increasing forest age and soil nutrient content gradually improved. At the stage of near-mature to over-mature forests, the effect of forest age on soil bulk density was not significant and all other soil characteristics decreased to varying degrees as the forest age increased. The soil also developed from alkaline to neutral. The SQI of the total data set and the SQI of the minimum data set consistently showed that near-mature forests (NMF) > middle-aged forests (MAF) > mature forests (MF) > over-mature forests (OMF) > young forests (YF). The results of the two evaluation systems showed a significant positive correlation (P < 0.05, R 2 = 0.8263) indicating that it is feasible to use the minimum data set to evaluate the soil fertility of shelter forests of different forest ages. The age of the forest has an obvious effect on the soil characteristics and overall soil fertility of shelter forests. The Populus simonii shelter forests on the southern edge of the Horqin Sandy Land have great soil development at the early stage of afforestation and the soil nutrient content gradually increases. The soil fertility reaches a peak when the forest is nearly mature and the soil fertility declines after the age of the forest reaches 30 years.