Hybrid Poplar Research Articles

Understory vegetation diversity and composition in intensively managed plantations compared to extensively managed forests

The benefits of plantations for wood production are well known, but it is unclear whether they have a clear negative effect on biodiversity, compared to natural forests. It is also unclear how overstory species identity (i.e., exotic, compared to native) would interact with the simple plantation effects on understory vegetation biodiversity. Here, we assessed bryophyte, lichen, and vascular plant diversity and composition in mono-specific and mixed plantations of exotic and native hybrid poplars (Populus spp.) and exotic and native spruces (Picea spp.). We compared these plantations to reference forests with similar overstory types (coniferous, deciduous, mixed) to those of the plantations in the southern boreal forest of Quebec. We also assessed whether the overstory species identity in plantations influenced understory vegetation biodiversity compared to reference forests. Our results show that plantations were not biological deserts, since mixed plantations contained similar bryophyte and vascular plant diversity as reference mixed native forests. The presence of native spruces and exotic poplars in mixed plantations increased bryophyte and vascular plant diversity respectively. This suggests that the influence of plantations on understory species diversity, compared to reference forests, was linked to the overstory species identity. Plantations were associated with ruderal plant species, while reference forests were composed of forest species. Bryophyte composition was similar in plantations and reference forests, while lichen species were only present in plantations. Our findings highlight that plantations, especially mixtures of white spruce and exotic poplar, are a promising option for planting strategies to enhance biodiversity.

Breed Selection of Poplars Imposes Greater Selection Pressure on the Rhizosphere Bacterial Community

Breed selection alters the coevolution of plant–microbiome associations that have developed over long periods of natural evolution. We investigated the effects of breed selection on the rhizosphere microbiomes and metabolites of hybrid parents (I101 and 84K) and their offspring (Q1–Q5) using metagenomics and untargeted metabolomics. Rhizosphere archaeal, bacterial and fungal community β-diversity significantly differed among hybrid parents and offspring, but only the dominant bacterial phyla and bacterial community α-diversity revealed significant differences. Approximately 5.49%, 14.90% and 7.86% of the archaeal, bacterial and fungal species significantly differed among the poplar hybrid parents and offspring. Rhizosphere microbial functional genes and metabolites were both clustered into the following three groups: I101 and 84K; Q2 and Q4; and Q1, Q3 and Q5. Compared with the hybrid parents, 15 phytochemical compounds were enriched in the hybrid offspring and explained 7.15%, 18.24% and 6.68% of the total variation in the archaeal, bacterial and fungal community compositions, respectively. Rhizosphere metabolites significantly affected the bacterial community, rather than the archaeal and fungal communities. Our observations suggested that poplar breed selection imposed greater selection pressure on the rhizosphere bacterial community, which was mainly driven by metabolites.

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<2mm) samples were collected using soil cores at depths of 0-20, 20-40 and 40-60cm, 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 20-40cm 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 0-20cm 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.

Functional impacts of PtrMYB203 on phenylpropanoid pathway regulation and wood properties in hybrid poplar

The phenylpropanoid pathway is vital for plant growth and development, producing lignin and flavonoids. This study investigates PtrMYB203, a homolog of MYB repressors of proanthocyanidin (PA) biosynthesis in Populus trichocarpa, as a transcriptional repressor in the phenylpropanoid pathway of hybrid poplar (Populus alba x P. glandulosa). Overexpression of PtrMYB203 (35S::PtrMYB203) in hybrid poplar detrimentally impacted plant growth and development. Histological analysis revealed irregular xylem vessel formation and decreased lignin content, corroborated by Klason lignin assays. Moreover, 35S::PtrMYB203 transgenic poplars exhibited significant decreases in anthocyanin and PA accumulations in callus tissues, even under high light conditions. Quantitative RT-PCR analysis and protoplast-based transcriptional activation assay confirmed the downregulation of lignin and flavonoid biosynthesis genes. This genetic modification also alters the expression of several MYB transcription factors, essential for phenylpropanoid pathway regulation. Remarkably, saccharification efficiency in the 35S::PtrMYB203 poplar was improved by over 34% following hot water treatment alone. These findings suggest PtrMYB203 as a potential genetic target for enhancing wood properties for bioenergy production, providing valuable insights into the manipulation of metabolite pathways in woody perennials to advance wood biotechnology.

Transcriptome Analysis Revealed the Paternal Importance to Vegetative Growth Heterosis in Populus.

Parental selection is important for heterosis formation during crossbreeding of Populus. However, in poplar hybrids, the effect of parents on vegetative growth heterosis is not well understood. In this study, one female parent (P. simonii XY4) and two male parents (P. nigra OH and P. deltoides × P. nigra BJLY3#) were used to produce two progenies (Hyb1 and Hyb2). Vegetative growth investigation showed that both Hyb1 and Hyb2 performed heterosis in plant growth and ground diameter. The vegetative growth of hybrids was strongly correlated with the male parents but not with the female parents. The gene expression levels in the hybrids were more biased toward the male parents. In Hyb1 and Hyb2, 51.93% and 45.03% of the expressed genes showed the non-additive effect, respectively, and over 65% of the non-additively expressed genes showed the dominant effect. It is noteworthy that genes of paternal expression dominant effect (ELD_♂) account for the majority of dominantly expressed genes, suggesting the paternal contribution to heterosis. KEGG enrichment analysis indicated that a large number of non-additively expressed genes were enriched in the plant hormone signal transduction pathway. WGCNA analysis showed that MEcyan was significantly correlated with the traits of hybrids, and 12 plant hormone signal transduction pathway genes were enriched in this module. Transcription factors (TFs) MYB88, LHY, and TCP4 may be involved in the regulation of these pathway genes. This finding supported that the male parents play an important role in the formation of vegetative growth heterosis of Populus. In addition, the non-additively expressed genes of the signal transduction pathway and the regulation of TFs related to these pathway genes may be one of the reasons for the generation of heterosis.

Planting trees in buffer strips for attenuating phosphorus runoff in Jiangsu Province, China

AbstractIn the last four decades, the Jiangsu province, like the rest of China, has undergone rapid economic development coupled with a rapid increase in environmental pollution. Freshwater ecosystems have been particularly affected. In Lake Tai, China's third‐largest freshwater lake by volume, water quality has been severely reduced. There is a renewed interest in establishing riparian buffer strips in China as a conservation practice for mitigating agricultural non‐point source pollution. To study the effect of riparian buffers on phosphorus attenuation, eight 50 m × 20 m plots were established between a rice farm and the shore of Lake Tai in China, with the 20 m width facing the lake shore. They were planted with hybrid poplar (Populus deltoides × euramericana), hybrid cypress (Taxodium ascendens × mucronatum), or a mixture of both at densities of 2 m × 3 m, 2 m × 5 m, or 5 m × 5 m, while one plot was left as a control. Samples collected from soil, tree leaves, and groundwater during 2014–2018 were analyzed for multiple forms of phosphorus. Results indicated that riparian buffer width and time (year) were the most significant factors of phosphorus attenuation on all plots. Tree density had a minor effect, with medium density being the most effective, and we found negligible differences among hybrid tree species on attenuating phosphorus. Farmers should use densely vegetated buffer strips at least 15 m wide to ensure a significant reduction in nutrient runoff. Species selection should be based on market value and ecological benefits.

INFRARED SPECTRUM CHANGE OF UV IRRADIATED AND NATURAL WOOD SAMPLES DURING 12 YEARS OF STORAGE IN TOTAL DARKNESS

The stability of the surface of UV-irradiated wood samples was investigated after 12 years of storage in total darkness at room temperature. The investigated specimens were earlywood and latewood of Scots pine sapwood, earlywood and latewood of spruce, earlywood of ash, beech and hybrid poplar. The thin (1 mm tick) samples contained only earlywood or latewood, and the tangential surfaces were used for infrared spectrum measurement. For comparison, the non-irradiated natural surface (back side) of the specimens was used for infrared spectrum measurement. Natural wood surfaces were stable during the storage. Only ether linkages in hemicelluloses showed minor degradation at 1175 cm-1 wavenumber. Lignin molecules remained stable during the 12-year storage period on both UV irradiated and non-irradiated side of the specimens. In contrast, UV irradiated samples suffered alterations during the 12 years of thermal treatment at low temperature (20-25°C). Hemicelluloses in photodegraded surface layers underwent thermal degradation and oxidation processes, generating new carbonyl groups. Extractives also presented absorption increase in the conjugated carbonyl region.

Enhancing monolignol ferulate conjugate levels in poplar lignin via OsFMT1

BackgroundThe phenolic polymer lignin is one of the primary chemical constituents of the plant secondary cell wall. Due to the inherent plasticity of lignin biosynthesis, several phenolic monomers have been shown to be incorporated into the polymer, as long as the monomer can undergo radicalization so it can participate in coupling reactions. In this study, we significantly enhance the level of incorporation of monolignol ferulate conjugates into the lignin polymer to improve the digestibility of lignocellulosic biomass.ResultsOverexpression of a rice Feruloyl-CoA Monolignol Transferase (FMT), OsFMT1, in hybrid poplar (Populus alba x grandidentata) produced transgenic trees clearly displaying increased cell wall-bound ester-linked ferulate, p-hydroxybenzoate, and p-coumarate, all of which are in the lignin cell wall fraction, as shown by NMR and DFRC. We also demonstrate the use of a novel UV–Vis spectroscopic technique to rapidly screen plants for the presence of both ferulate and p-hydroxybenzoate esters. Lastly we show, via saccharification assays, that the OsFMT1 transgenic p oplars have significantly improved processing efficiency compared to wild-type and Angelica sinensis-FMT-expressing poplars.ConclusionsThe findings demonstrate that OsFMT1 has a broad substrate specificity and a higher catalytic efficiency compared to the previously published FMT from Angelica sinensis (AsFMT). Importantly, enhanced wood processability makes OsFMT1 a promising gene to optimize the composition of lignocellulosic biomass.

Drought stress impacts soil microbial nutrient limitation more strongly than O3 pollution

Summer drought conditions generally coincide with exposure to high ground-level concentrations of ozone (O3). These two stressors compromise the availability of water, energy, and nutrients to vegetation and microbial communities present within the soil. However, relatively little is known regarding the responses of rhizosphere and non-rhizosphere soil microbial nutrient acquisition to conditions of drought and O3 exposure. Here, we used ecoenzymatic stoichiometry modeling and vector analyses to decipher the resource constraints on rhizosphere and non-rhizosphere soil microbial metabolic limitation within the soil of hybrid poplars exposed to two watering regimens (well-watered or reduced water of 40 %) and two chronic O3 treatments (charcoal-filtered air or ambient air +40 ppb of O3). Results show that, simulated drought stress resulted in stronger effects on the structure of soil microbial communities and associated enzymatic activity relative to exposure to elevated O3 levels. Exposure to these two stressors induced a microbial phosphorus (P) limitation response, with these limitation effects being more pronounced in the rhizosphere soil relative to non-rhizosphere soil. Structural equation modeling revealed that drought-associated declines in soil water availability and pH values were the primary factors influencing this microbial P limitation. Reduced soil water levels resulted in the exacerbation of microbial P limitation as a consequence of the inhibition of P mineralization and availability of C and N, whereas declining soil pH values had the opposite effect, alleviating microbial P limitations by reducing soil alkalinity and thereby aiding in the dissolution of the insoluble P present in alkaline soil. These results emphasize the complex responses of rhizosphere and non-rhizosphere soil microbial metabolic activity to elevated O3 levels and drought stress, providing invaluable insight into the importance of elemental stoichiometry-driven microbial metabolic variability of nutrient cycling activity at the root-soil interface in O3-polluted and drought-exposed ecosystems.

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.

PagMYB128 regulates anthocyanin biosynthesis by directly activating PagMYB116 in Populus alba × Populus glandulosa

MYB transcription factors regulate the biosynthesis of anthocyanin, a crucial flavonoid in plants. In this research, we identified PagMYB128 as a new activator of anthocyanin production in the hybrid poplar (Populus alba × Populus glandulosa). The PagMYB128 protein was located in the nucleus and has higher expression level in stems. Dominant repression of PagMYB128 led to a reduction of anthocyanin content in the petiole and stems. Overexpression of PagMYB128 resulted in the opposite phenotype. We found that PagMYB128 promoted anthocyanin biosynthesis by activating PagCHS1, PagF3’5’H1, PagDFR2, and PagANR1, which encoded the essential enzymes of anthocyanin biosynthesis. Further, PagMYB116 was identified as a potential direct target of PagMYB128 by ChIP-seq. Through ChIP-qPCR, Y1H, and Split-LUC assays, it was demonstrated that PagMYB128 could directly interact with the MYB-site located in the promoter region of PagMYB116, thereby exerting control over the transcription of PagMYB116. Moreover, through the poplar transient transformation method, we found that PagMYB116 could activate anthocyanin biosynthetic enzyme genes PagF3’5’H1 and PagANR1, through direct binding to their promoters. These findings indicate that the PagMYB128-MYB116 module for transcriptional regulation plays a crucial part in the synthesis of anthocyanin, enriching our comprehension of the networks responsible for regulating anthocyanin transcription.

ENERGY CROPS – OUTLOOK FOR ENVIRONMENTAL FUTURE FOR HUMANITY AND THE PLANET EARTH

The growing risk of irreversible depletion of fossil energy sources like petrol, natural gas and coal, and the emissions of gasses consequence from their combustion, is causing the necessity of the introduction of energy crops for the manufacture of energy products. These include annual or perennial plants, fast-growing tree types – hybrid willows, poplars and others, and water crops – red algae, giant kelp, seaweed etc. Most preferred in Europe and the US are Paulownia sp. (P. elongata и P. tomentosa), giant reed (Arundo donax), elephant grass (Miscanthus giganteus), weeping willow (Salix babylonica) and wicker (Salix viminalis). So far in Bulgaria are preferred herbaceous energy crops such as corn, canola, sorghum, etc., but also were introduced some kind of timber. The global development of the sector would help to address the environmental problems associated with the release of huge quantities of greenhouse gasses. In the processing and utilization of energy crops, the amount of waste products is reduced to near zero, thereby avoiding environmental pollution.

Elevated CO2 concentration alleviates the negative effect of vapour pressure deficit and soil drought on juvenile poplar growth

Abstract The growth performance of short-rotation woody coppice (SRWC) is strongly influenced by successful establishment in the initial months after planting. Future climates, expected to be warmer due to elevated atmospheric CO2 (eCO2), may bring about more frequent soil droughts alongside increased vapour pressure deficit (eVPD). Hence, this growth chamber experiment aimed to explore the interactive effects of eVPD, eCO2, and soil drought on growth and physiology traits of juvenile hybrid poplars under warmer climates. Our findings with juvenile hybrid poplar J-105 revealed that eVPD resulted in reductions in leaf area (–21%), root (–20%) and stem biomass (–9%), as well as in net assimilation (–15%), stomatal conductance (–26%), and transpiration (–13%). However, these decreases were relatively minor compared to the compensating effect of eCO2, which generally exerted a stronger influence than eVPD. While soil drought emerged as the primary growth-limiting factor in our study, elevated VPD is not expected to pose a significant additional threat to central European SRWC plantations of juvenile hybrid poplars under future conditions of ongoing climate change.

PagMYB180 regulates adventitious rooting via a ROS/PCD-dependent pathway in poplar

The formation of adventitious roots (AR) is an essential step in the vegetative propagation of economically woody species. Reactive oxygen species (ROS) function as signaling molecules in regulating root growth and development. Here, we identified an R2R3-MYB transcription factor PagMYB180 as a regulator of AR formation in hybrid poplar (Populus alba × Populus glandulosa). PagMYB180 was specifically expressed in the vascular tissues of poplar roots, stems and leaves, and its protein was localized in the nucleus and acted as a transcriptional repressor. Both dominant repression and overexpression of PagMYB180 resulted in a significant reduction of AR quantity, a substantial increase of AR length, and an elevation of both the quantity and length of lateral roots (LR) compared to the wild type (WT) plants. Furthermore, PagMYB180 regulates programmed cell death (PCD) in root cortex cells, which is associated with elevated levels of ROS. Transcriptome and reverse transcription-quantitative PCR (RT-qPCR) analyses revealed that a series of differentially expressed genes are related to ROS, PCD and ethylene synthesis. Taken together, these results suggest that PagMYB180 may regulate AR development via a ROS/PCD-dependent pathway in poplar.

PagWOX11/12a from hybrid poplar enhances drought tolerance by modulating reactive oxygen species

WOX11/12 is a homeobox gene of WOX11 and WOX12 in Arabidopsis that plays important roles in crown root development and growth. It has been reported that WOX11/12 participates in adventitious root (AR) formation and different abiotic stress responses, but the downstream regulatory network of WOX11/12 in poplar remains to be further investigated. In this study, we found that PagWOX11/12a is strongly induced by PEG-simulated drought stress. PagWOX11/12a-overexpressing poplar plantlets showed lower oxidative damage levels, greater antioxidant enzyme activities and reactive oxygen species (ROS) scavenging capacity than non-transgenic poplar plants, whereas PagWOX11/12a dominant repression weakened root biomass accumulation and drought tolerance in poplar. RNA-seq analysis revealed that several differentially expressed genes (DEGs) regulated by PagWOX11/12a are involved in redox metabolism and drought stress response. We used RT-qPCR and yeast one-hybrid (Y1H) assays to validate the downstream target genes of PagWOX11/12a. These results provide new insights into the biological function and molecular regulatory mechanism of WOX11/12 in the abiotic resistance processes of poplar.

Tissue and cellular localization of condensed tannins in poplar roots and potential association with nitrogen uptake.

Condensed tannins are common in vegetative tissues of woody plants, including in roots. In hybrid poplar (Populus tremula x alba; also known as P. x canescens) CT assays indicated they were most concentrated in younger white roots and at the root tip. Furthermore, CT-specific staining of embedded tissue sections demonstrated accumulation in root cap cells and adjacent epidermal cells, as well as a more sporadic presence in cortex cells. In older, brown roots as well as roots with secondary growth (cork zone), CT concentration was significantly lower. The insoluble fraction of CTs was greatest in the cork zone. To determine if CT accumulation correlates with nutrient uptake in poplar roots, a microelectrode ion flux measurement (MIFE™) system was used to measure flux along the root axis. Greatest NH4 + uptake was measured near the root tip, but NO3- and Ca2+ did not vary along the root length. In agreement with earlier work, providing poplars with ample nitrogen led to higher accumulation of CTs across root zones. To test the functional importance of CTs in roots directly, CT-modified transgenic plants could be important tools.

PagPXYs improve drought tolerance by regulating reactive oxygen species homeostasis in the cambium of Populus alba × P. glandulosa

PXY (Phloem intercalated with xylem) is a receptor kinase required for directional cell division during the development of plant vascular tissue. Drought stress usually affects plant stem cell division and differentiation thereby limiting plant growth. However, the role of PXY in cambial activities of woody plants under drought stress is unclear. In this study, we analyzed the biological functions of two PXY genes (PagPXYa and PagPXYb) in poplar growth and development and in response to drought stress in a hybrid poplar (Populus alba × P. glandulosa, ‘84K’). Expression analysis indicated that PagPXYs, similar to their orthologs PtrPXYs in Populus trichocarpa, are mainly expressed in the stem vascular system, and related to drought. Interestingly, overexpression of PagPXYa and PagPXYb in poplar did not have a significant impact on the growth status of transgenic plants under normal condition. However, when treated with 8 % PEG6000 or 100 mM H2O2, PagPXYa and PagPXYb overexpressing lines consistently exhibited more cambium cell layers, fewer xylem cell layers, and enhanced drought tolerance compared to the non-transgenic control ‘84K’. In addition, PagPXYs can alleviate the damage caused by H2O2 to the cambium under drought stress, thereby maintaining the cambial division activity of poplar under drought stress, indicating that PagPXYs play an important role in plant resistance to drought stress. This study provides a new insight for further research on the balance of growth and drought tolerance in forest trees.

Properties of Hybrid Plywood Produced by Utilisation of Peeler Cores

The aim of this research work was to investigate the feasibility of upcycling poplar (Populus spp.) peeler cores as a by-product from the production of plywood in manufacturing lightweight hybrid poplar and beech plywood panels, containing different ratios (60%, 80%, and 100%) of peeler core sections in the core layer. This corresponds to effective percentages of panel surface glued with peeler core slides of 80%, 64%, and 48%, respectively. The physical properties (density, water absorption, and thickness swelling) and mechanical properties (bending strength (MOR) and modulus of elasticity (MOE)) of the laboratory-fabricated hybrid panels were determined in accordance with the applicable European standards (EN 310, EN 317, and EN 323). The highest MOE and MOR values of 3575 N·mm−2 and 28.1 N·mm−2, respectively, were obtained for the hybrid poplar plywood panels with a thickness of 20 mm and 100% peeler core sections. The use of beech veneer in the face layers did not lead to a significant increase in the mechanical properties. In this case, the MOE and MOR values of the hybrid plywood panels with a thickness of 20 mm and 80% peeler core sections were 5954 N·mm−2 and 35.2 N·mm−2, respectively.

Overexpressing GLUTAMINE SYNTHETASE 1;2 maintains carbon and nitrogen balance under high-ammonium conditions and results in increased tolerance to ammonium toxicity in hybrid poplar.

The glutamine synthetase/glutamic acid synthetase (GS/GOGAT) cycle plays important roles in N metabolism, growth, development, and stress resistance in plants. Excess ammonium (NH4+) restricts growth, but GS can help to alleviate its toxicity. In this study, the 84K model clone of hybrid poplar (Populus alba × P. tremula var. glandulosa), which has reduced biomass accumulation and leaf chlorosis under high-NH4+ stress, showed less severe symptoms in transgenic lines overexpressing GLUTAMINE SYNTHETASE 1;2 (GS1;2-OE), and more severe symptoms in RNAi lines (GS1;2-RNAi). Compared with the wild type, the GS1;2-OE lines had increased GS and GOGAT activities and higher contents of free amino acids, soluble proteins, total N, and chlorophyll under high-NH4+ stress, whilst the antioxidant and NH4+ assimilation capacities of the GS1;2-RNAi lines were decreased. The total C content and C/N ratio in roots and leaves of the overexpression lines were higher under stress, and there were increased contents of various amino acids and sugar alcohols, and reduced contents of carbohydrates in the roots. Under high-NH4+ stress, genes related to amino acid biosynthesis, sucrose and starch degradation, galactose metabolism, and the antioxidant system were significantly up-regulated in the roots of the overexpression lines. Thus, overexpression of GS1;2 affected the carbon and amino acid metabolism pathways under high-NH4+ stress to help maintain the balance between C and N metabolism and alleviate the symptoms of toxicity. Modification of the GS/GOGAT cycle by genetic engineering is therefore a potential strategy for improving the NH4+ tolerance of cultivated trees.

PagARGOS promotes low-lignin wood formation in poplar.

Wood formation, which occurs mainly through secondary xylem development, is important not only for supplying raw material for the 'ligno-chemical' industry but also for driving the storage of carbon. However, the complex mechanisms underlying the promotion of xylem formation remain to be elucidated. Here, we found that overexpression of Auxin-Regulated Gene involved in Organ Size (ARGOS) in hybrid poplar 84 K (Populus alba × Populus tremula var. glandulosa) enlarged organ size. In particular, PagARGOS promoted secondary growth of stems with increased xylem formation. To gain further insight into how PagARGOS regulates xylem development, we further carried out yeast two-hybrid screening and identified that the auxin transporter WALLS ARE THIN1 (WAT1) interacts with PagARGOS. Overexpression of PagARGOS up-regulated WAT1, activating a downstream auxin response promoting cambial cell division and xylem differentiation for wood formation. Moreover, overexpressing PagARGOS caused not only higher wood yield but also lower lignin content compared with wild-type controls. PagARGOS is therefore a potential candidate gene for engineering fast-growing and low-lignin trees with improved biomass production.