Zhi-Bin Luo

Nitrogen metabolism of two contrasting poplar species during acclimation to limiting nitrogen availability

To investigate N metabolism of two contrasting Populus species in acclimation to low N availability, saplings of slow-growing species (Populus popularis, Pp) and a fast-growing species (Populus alba × Populus glandulosa, Pg) were exposed to 10, 100, or 1000 μM NH4NO3. Despite greater root biomass and fine root surface area in Pp, lower net influxes of NH4 + and NO3 – at the root surface were detected in Pp compared to those in Pg, corresponding well to lower NH4 + and NO3 – content and total N concentration in Pp roots. Meanwhile, higher stable N isotope composition (δ15N) in roots and stronger responsiveness of transcriptional regulation of 18 genes involved in N metabolism were found in roots and leaves of Pp compared to those of Pg. These results indicate that the N metabolism of Pp is more sensitive to decreasing N availability than that of Pg. In both species, low N treatments decreased net influxes of NH4 + and NO3 –, root NH4 + and foliar NO3 – content, root NR activities, total N concentration in roots and leaves, and transcript levels of most ammonium (AMTs) and nitrate (NRTs) transporter genes in leaves and genes involved in N assimilation in roots and leaves. Low N availability increased fine root surface area, foliar starch concentration, δ15N in roots and leaves, and transcript abundance of several AMTs (e.g. AMT1;2) and NRTs (e.g. NRT1;2 and NRT2;4B) in roots of both species. These data indicate that poplar species slow down processes of N acquisition and assimilation in acclimation to limiting N supply.

Overexpression of bacterial γ-glutamylcysteine synthetase mediates changes in cadmium influx, allocation and detoxification in poplar.

Overexpression of bacterial γ-glutamylcysteine synthetase in the cytosol of Populus tremula × P. alba produces higher glutathione (GSH) concentrations in leaves, thereby indicating the potential for cadmium (Cd) phytoremediation. However, the net Cd(2+) influx in association with H(+) /Ca(2+) , Cd tolerance, and the underlying molecular and physiological mechanisms are uncharacterized in these poplars. We assessed net Cd(2+) influx, Cd tolerance and the transcriptional regulation of several genes involved in Cd(2+) transport and detoxification in wild-type and transgenic poplars. Poplars exhibited highest net Cd(2+) influxes into roots at pH 5.5 and 0.1 mM Ca(2+) . Transgenics had higher Cd(2+) uptake rates and elevated transcript levels of several genes involved in Cd(2+) transport and detoxification compared with wild-type poplars. Transgenics exhibited greater Cd accumulation in the aerial parts than wild-type plants in response to Cd(2+) exposure. Moreover, transgenic poplars had lower concentrations of O2 ˙(-) and H2 O2 ; higher concentrations of total thiols, GSH and oxidized GSH in roots and/or leaves; and stimulated foliar GSH reductase activity compared with wild-type plants. These results indicate that transgenics are more tolerant of 100 μM Cd(2+) than wild-type plants, probably due to the GSH-mediated induction of the transcription of genes involved in Cd(2+) transport and detoxification.

Ectomycorrhizas with Paxillus involutus enhance cadmium uptake and tolerance in Populus × canescens.

Ectomycorrhizas (EMs), which are symbiotic organs formed between tree roots and certain fungi, can mediate cadmium (Cd) tolerance of host plants, but the underlying physiological and molecular mechanisms are not fully understood. To investigate EMs mediated Cd tolerance in woody plants, Populusu2009×u2009canescens was inoculated with Paxillus involutus (strain MAJ) to establish mycorrhizal roots. Mycorrhizal poplars and non-mycorrhizal controls were exposed to 0 or 50u2009μM CdSO4 . EMs displayed higher net Cd(2+) influx than non-mycorrhizal roots. Net Cd(2+) influx was coupled with net H(+) efflux and inactivation of plasma membrane (PM) H(+) -ATPases reduced Cd(2+) uptake of EMs less than of non-mycorrhizal roots. Consistent with higher Cd(2+) uptake in EMs, in most cases, transcript levels of genes involved in Cd(2+) uptake, transport and detoxification processes were increased in EMs compared to non-mycorrhizal roots. Higher CO2 assimilation, improved nutrient and carbohydrate status, and alleviated oxidative stress were found in mycorrhizal compared to non-mycorrhizal poplars despite higher Cd(2+) accumulation. These results indicate that mycorrhizas increase Cd(2+) uptake, probably by an enlarged root volume and overexpression of genes involved in Cd(2+) uptake and transport, and concurrently enhance Po.u2009×u2009canescensu2005Cd tolerance by increased detoxification, improved nutrient and carbohydrate status and defence preparedness.

Exogenous abscisic acid alleviates zinc uptake and accumulation in Populus × canescens exposed to excess zinc

A greenhouse experiment was conducted to study whether exogenous abscisic acid (ABA) mediates the responses of poplars to excess zinc (Zn). Populusu2009×u2009canescens seedlings were treated with either basal or excess Zn levels and either 0 or 10 μm ABA. Excess Zn led to reduced photosynthetic rates, increased Zn accumulation, induced foliar ABA and salicylic acid (SA), decreased foliar gibberellin (GA3 ) and auxin (IAA), elevated root H2 O2 levels, and increased root ratios of glutathione (GSH) to GSSG and foliar ratios of ascorbate (ASC) to dehydroascorbate (DHA) in poplars. While exogenous ABA decreased foliar Zn concentrations with 7u2009d treatments, it increased levels of endogenous ABA, GA3 and SA in roots, and resulted in highly increased foliar ASC accumulation and ratios of ASC to DHA. The transcript levels of several genes involved in Zn uptake and detoxification, such as yellow stripe-like family protein 2 (YSL2) and plant cadmium resistance protein 2 (PCR2), were enhanced in poplar roots by excess Zn but repressed by exogenous ABA application. These results suggest that exogenous ABA can decrease Zn concentrations in P.u2009×u2009canescens under excess Zn for 7u2009d, likely by modulating the transcript levels of key genes involved in Zn uptake and detoxification.

Phosphorus and nitrogen physiology of two contrasting poplar genotypes when exposed to phosphorus and/or nitrogen starvation.

Phosphorus (P) and nitrogen (N) are the two essential macronutrients for tree growth and development. To elucidate the P and N physiology of woody plants during acclimation to P and/or N starvation, we exposed saplings of the slow-growing Populus simonii Carr (Ps) and the fast-growing Populusu2005×u2005euramericana Dode (Pe) to complete nutrients or starvation of P, N or both elements (NP). P.u2005×u2005euramericana had lower P and N concentrations and greater P and N amounts due to higher biomass production, thereby resulting in greater phosphorus use efficiency/N use efficiency (PUE/NUE) compared with Ps. Compared with the roots of Ps, the roots of Pe exhibited higher enzymatic activities in terms of acid phosphatases (APs) and malate dehydrogenase (MDH), which are involved in P mobilization, and nitrate reductase (NR), glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH), which participate in N assimilation. The responsiveness of the transcriptional regulation of key genes encoding transporters for phosphate, ammonium and nitrate was stronger in Pe than in Ps. These results suggest that Pe possesses a higher capacity for P/N uptake and assimilation, which promote faster growth compared with Ps. In both poplars, P or NP starvation caused significant decreases in the P concentrations and increases in PUE. Phosphorus deprivation induced the activity levels of APs, phosphoenolpyruvate carboxylase and MDH in both genotypes. Nitrogen or NP deficiency resulted in lower N concentrations, amino acid levels, NR and GOGAT activities, and higher NUE in both poplars. Thus, in Ps and Pe, the mRNA levels of PHT1;5, PHT1;9, PHT2;1, AMT2;1 and NR increased in the roots, while PHT1;9, PHO1;H1, PHO2, AMT1;1 and NRT2;1 increased in the leaves during acclimation to P, N or NP deprivation. These results suggest that both poplars suppress P/N uptake, mobilization and assimilation during acclimation to P, N or NP starvation.

Global poplar root and leaf transcriptomes reveal links between growth and stress responses under nitrogen starvation and excess

Nitrogen (N) starvation and excess have distinct effects on N uptake and metabolism in poplars, but the global transcriptomic changes underlying morphological and physiological acclimation to altered N availability are unknown. We found that N starvation stimulated the fine root length and surface area by 54 and 49%, respectively, decreased the net photosynthetic rate by 15% and reduced the concentrations of NH4+, NO3(-) and total free amino acids in the roots and leaves of Populus simonii Carr. in comparison with normal N supply, whereas N excess had the opposite effect in most cases. Global transcriptome analysis of roots and leaves elucidated the specific molecular responses to N starvation and excess. Under N starvation and excess, gene ontology (GO) terms related to ion transport and response to auxin stimulus were enriched in roots, whereas the GO term for response to abscisic acid stimulus was overrepresented in leaves. Common GO terms for all N treatments in roots and leaves were related to development, N metabolism, response to stress and hormone stimulus. Approximately 30-40% of the differentially expressed genes formed a transcriptomic regulatory network under each condition. These results suggest that global transcriptomic reprogramming plays a key role in the morphological and physiological acclimation of poplar roots and leaves to N starvation and excess.

Anatomical, physiological and transcriptional responses of two contrasting poplar genotypes to drought and re-watering

Populus × euramericana (Pe) displays higher stable carbon isotope composition (δ(13)C) and intrinsic water use efficiency (WUEi) than Populus cathayana (Pc) under unlimited water conditions, rendering us to hypothesize that Pe is better acclimated to water deficiency than Pc. To examine this hypothesis, saplings of Pc and Pe were exposed to drought and subsequently re-watered. Pc and Pe exhibited distinct anatomical, physiological and transcriptional responses in acclimation to drought and re-watering, mainly due to stronger responsiveness of transcriptional regulation of genes encoding plasma membrane intrinsic proteins (PIPs), higher starch accumulation, δ(13)C, stable nitrogen isotope composition (δ(15)N) and WUEi , and lower reactive oxygen species (ROS) accumulation and scavenging in Pe. In acclimation to drought, both poplar genotypes demonstrated altered anatomical properties, declined height growth, differential expression of PIPs, activation of ABA signaling pathway, decreased total soluble sugars and starch, increased δ(13)C, δ(15)N and WUEi , and shifted homeostasis of ROS production and scavenging, and these changes can be recovered upon re-watering. These data indicate that Pe is more tolerant to drought than Pc, and that anatomical, physiological and transcriptional acclimation to drought and re-watering is essential for poplars to survive and grow under projected dry climate scenarios in the future.

Changes in carbon, nutrients and stoichiometric relations under different soil depths, plant tissues and ages in black locust plantations

To investigate influences of forest plantations on soil nutrient properties, biomass accumulation, major nutrient elements (NPK) and their stoichiometric couplings in different tissues and aged plants, and correlations between major nutrient contents in soils and in foliage of plants, 5-, 10-, 15- and 20-year-old plantations of black locust (Robinia pseudoacacia L.) and farmland were selected. Black locust plantations increased soil organic carbon (SOC) and N stocks by 23–327 and 23–119xa0%, respectively, in the 0–10xa0cm top soil layer compared to those in farmland. Soil C:N, C:P, C:K, N:P, N:K and P:K ratios were 10.1, 22.9, 0.7, 2.2, 0.7 and 0.03, respectively. These ratios were higher in the 0–10xa0cm soil layer than those in the 10–20xa0cm soil layer and increased under older plantations. Higher C contents in stem, N contents in leaf, the largest C pools in stem and N pools in root in 20-year-old plantation were observed. Correspondingly, the highest C:N, C:P and C:K and the lowest N:P and N:K ratios in stem, decreased C:N and C:P ratios in older trees were found. No strong correlations were observed between element contents in soils and in leaves of black locust trees. These results suggest that black locust plantations can increase soil nutrient concentrations, SOC and N stocks resulting in changes in element stoichiometric relations. CNPK contents and their stoichiometries vary with tissues and tree ages of black locust. No strong coupling relations exist between major nutrient element contents in the top soil and in foliage of black locust.

Physiological and transcriptional regulation in poplar roots and leaves during acclimation to high temperature and drought

To elucidate the physiological and transcriptional regulatory mechanisms that underlie the responses of poplars to high temperature (HT) and/or drought in woody plants, we exposed Populus albau2009×u2009Populus tremula var. glandulosa saplings to ambient temperature (AT) or HT under 80 or 40% field capacities (FC), or no watering. HT increased the foliar total carbon (C) concentrations, and foliar δ(13) C and δ(18) O. HT triggered heat stress signaling via increasing levels of abscisic acid (ABA) and indole-3-acetic acid (IAA) in poplar roots and leaves. After perception of HT, poplars initiated osmotic adjustment by increasing foliar sucrose and root galactose levels. In agreement with the HT-induced heat stress and the changes in the levels of ABA and carbohydrates, we detected increased transcript levels of HSP18 and HSP21, as well as NCED3 in the roots and leaves, and the sugar transporter gene STP14 in the roots. Compared with AT, drought induced greater enhancement of foliar δ(13) C and δ(18) O in poplars at HT. Similarly, drought caused greater stimulation of the ABA and foliar glucose levels in poplars at HT than at AT. Correspondingly, desiccation led to greater increases in the mRNA levels of HSP18, HSP21, NCED3, STP14 and INT1 in poplar roots at HT than at AT. These results suggest that HT has detrimental effects on physiological processes and it induces the transcriptional regulation of key genes involved in heat stress responses, ABA biosynthesis and sugar transport and HT can cause greater changes in drought-induced physiological and transcriptional responses in poplar roots and leaves.