Comprehensive Proteome and Acetylome Analysis of Needle Senescence in Larix gmelinii.

Leaf senescence is essential for the growth and development of deciduous trees in the next season. Larix gmelinii, a deciduous coniferous tree, exhibits its most distinctive feature by turning yellow in the autumn and eventually shedding its leaves, resulting in significant changes in its appearance during the fall. Lysine acetylation plays an important role in diverse cellular processes; however, limited knowledge is available regarding acetylations in the needle senescence of L. gmelinii. In this study, the proteomics and acetylated modification omics of two phenotypic leaves, yellow and green (senescent and non-senescent) needles, were analyzed before autumn defoliation. In total, 5022 proteins and 4469 unique acetylation sites in 2414 lysine acylated proteins were identified, and this resulted in the discovery of 1335 differentially expressed proteins (DEPs) and 605 differentially expressed acetylated proteins (DAPs) in yellow versus green needles. There are significant differences between the proteome and acetylome; only 269 proteins were found to be DEP and DAP, of which 136 proteins were consistently expressed in both the DEP and DAP, 91 proteins were upregulated, and 45 proteins were down-regulated. The DEPs participate in the metabolism of starch and sucrose, while the DAPs are involved in glycolysis and the tricarboxylic acid cycle. Among them, DEPs underwent significant changes in glycolysis and citric acid cycling. Most of the enzymes involved in glycolysis and the citrate cycle were acetylated. DAPs were down-regulated in glycolysis and up-regulated in the citrate cycle. In all, the results of this study reveal the important role of lysine acetylation in the senescence of L. gmelinii needles and provide a new perspective for understanding the molecular mechanism of leaf senescence and tree seasonal growth.

[Effects of litter thickness on natural regeneration of Larix principis-rupprechtii plantations]. / 枯落物厚度对华北落叶松人工林天然更新的影响.

In this study, we explored the thickness influence of undecomposed litter layer and semi-decomposed litter layer on the natural regeneration in an artificial pure forest of Larix principis-rupprechtii in the forest area of Guandi Mountain. We divided the litter into an undecomposed layer and a semi-decomposed layer, which was further divided into eight groups based on the thickness. The results showed that when the thickness of undecomposed layer was 0.32-0.83 cm, and that of semi-decomposed layer was 0.18-0.89 cm, the regeneration index was larger (≥0.15), and the regeneration was better. When the thickness of undecomposed layer was more than 1.1 cm and that of semi-decomposed layer was more than 0.5 cm, the regeneration index was smaller (≤0.07), and the rege-neration of understory was worse. Results of redundancy analysis showed that the undecomposed layer thickness of litter had a high and stable explanatory ability for natural regeneration, with a contribution rate of 38.7%, while the semi-decomposed layer thickness had no significant effect on natural regeneration. Structural equation modeling revealed that the thickness of undecomposed layer of litter increased the mechanical resistance to seed germination which had a negative direct effect on natural regeneration (-0.617), and a positive indirect effect on natural rege-neration by influencing the content of alkali-hydrolyzed nitrogen and available phosphorus (+0.178). The combined effects (-0.439) showed an inhibitory effect on the natural regeneration. In conclusion, the thickness of undecomposed layer of litter under L. principis-rupprechtii was most closely related to natural regeneration, and the thickness of semi-decomposed layer had a minimal effect on natural regeneration.

[Age estimation model for individual tree in natural Larix gmelinii forest based on random forest model]. / åŸºäºŽéšæœºæ£®æž—çš„å ´å®‰è½å¶æ¾å¤©ç„¶æž—å•æœ¨å¹´é¾„é¢„ä¼°æ¨¡åž‹.

To accurately estimate the age of individual tree and to achieve full-cycle sustainable management of natural Larix gmelinii forest in Great Xing'an Mountains of northeastern China, we constructed individual tree age prediction model using stepwise regression and random forest algorithms based on 44 fixed plots data and 280 stan-dard tree cores obtained from the Pangu Forest Farm. We analyzed the influence of stand structure, site conditions, and competition index on the accuracy of model prediction. The model was evaluated by the coefficient of determination (R2), root mean square error (RMSE), and mean absolute error (MAE). The results showed that the random forest model had the highest prediction accuracy when number of decision trees was 1500 and number of node con-tention variables was 8. The random forest model had better accuracy and prediction ability than the stepwise regression model, with R2, RMSE and MAE of 0.5882, 9.9259 a, 8.1155 a. Diameter at breast height was the most important factor affecting age prediction (83.8%), followed by tree height (34.4%), elevation (17.9%), and basal area per hectare (17.5%). The random forest algorithm exhibited better adaptability and modeling effect on constructing a predictive model for individual tree age. This research contributed to improving the accuracy of growth and harvest estimation for L. gmelinii, and could provide a reference for other scientific studies related to tree age estimation in forests.

[Responses of radial growth of Larix principis-rupprechtii at different densities to drought events]. / 不同密度华北落叶松径向生长对干旱事件的响应.

In recent years, a surge in drought occurrences has dramatically impacted tree growth worldwide. We examined the ecological resilience of Larix principis-rupprechtii plantations with varying densities (1950, 2355, and 2595 trees·hm-2) at the Saihanba Mechanical Forest Farm, by extracting the increment cores using the standard dendrochronological method to measure individual-tree basal area increments (BAI) as part of our assessment of ecological resilience, including resistance (Rt), recovery (Rc), and resilience (Rs). The results showed that drought events occurred in 2006-2010, 2015, and 2018. The Rt for L. principis-rupprechtii plantations varied from 0.76 to 2.01 across three drought events, indicating generally high resistance, except for the plantation with 2355 trees·hm-2 during the second dry year (Rt=0.69). The Rt for the plantation with 2595 trees·hm-2 significantly decreased across all drought events, while no significant change was observed in the plantations with 1950 and 2355 trees·hm-2. The Rc showed no differences in response to a single drought event across plantation densities, with a significant upward trend for all the densities with each occurrence of drought event. There was no significant difference in the resilience of different densities of L. principis-rupprechtii to the first drought event, whereas the plantation with 2595 trees·hm-2 exhibited significantly lower Rs during the second and third drought events compared with 1950 and 2355 trees·hm-2, respectively. During the 2015 drought event, plantation with 2595 trees·hm-2 experienced a significant growth decline (radial growth change rate was -26.5%), while no such decline was observed in the plantations with 1950 and 2355 trees·hm-2. Overall, the plantation with 2595 trees·hm-2 demonstrated the lowest resilience to drought events.

[Response of radial growth of different timberline species to climate change in Yading Nature Reserve, Sichuan, China]. / å››å·äºšä¸è‡ªç„¶ä¿æŠ¤åŒºä¸åŒæž—çº¿æ ‘ç§å¾„å‘ç”Ÿé•¿å¯¹æ°”å€™å˜åŒ–çš„å“åº”.

The radial growth of trees in alpine timberline is particularly sensitive to climate change. We sampled and disposed tree-ring cores of three coniferous tree species including Juniperus saltuaria, Abies forrestii, and Larix potaninii at alpine timberline in Yading Nature Reserve. The standard tree-ring chronology was used to explore the response of radial growth of different timberline species to climate change. The results showed that radial growth of L. potaninii increased after 2000, while that of A. forrestii declined after 2002, and J. saltuaria showed a significant decreasing growth trend in the past 10 years. Such results indicated divergent growth responses to climate factors among the three tree species at alpine timberline. The radial growth of J. saltuaria was sensitive to temperature, and was positively correlated with the minimum temperature from previous October to current August, the mean tempera-ture from previous November to current April and from current July to October, but was negatively associated with the relative humidity from current July to October. The radial growth of A. forrestii showed negative correlation with mean temperature and the maximum temperature from May to June in the current year, while it exhibited positive association with the relative humidity and the Palmer drought severity index from May to June in the current year. L. potaninii radial growth was positively associated with mean temperature and the maximum temperature of November-December in the previous year, the maximum temperature of current March and mean temperature of current August. The temporal stability of climate-growth relationship varied among different timberline species. The positive correlation between radial growth of A. forrestii and J. saltuaria and temperature gradually decreased, while the posi-tive relationship of L. potaninii radial growth and temperature gradually increased. Under the background of climate warming, rapid rise in surface air temperatures may promote the radial growth of L. potaninii, while inhibit that of J. saltuaria and A. forrestii, which may change the position of regional timberline.

Contrasting water-use strategies to climate warming in white birch and larch in a boreal permafrost region.

The effects of rising atmospheric CO2 concentrations (Ca) with climate warming on intrinsic water-use efficiency and radial growth in boreal forests are still poorly understood. We measured tree-ring cellulose δ13C, δ18O, and tree-ring width in Larix dahurica (larch) and Betula platyphylla (white birch), and analyzed their relationships with climate variables in a boreal permafrost region of northeast China over past 68 years covering a pre-warming period (1951-1984; base period) and a warm period (1985-2018; warm period). We found that white birch but not larch significantly increased their radial growth over the warm period. The increased intrinsic water-use efficiency in both species was mainly driven by elevated Ca but not climate warming. White birch but not larch showed significantly positive correlations between tree-ring δ13C, δ18O and summer maximum temperature as well as vapor pressure deficit in the warm period, suggesting a strong stomatal response in the broad-leaved birch to temperature changes. The climate warming-induced radial growth enhancement in white birch is primarily associated with a conservative water-use strategy. In contrast, larch exhibits a profligate water-use strategy. It implies an advantage for white birch over larch in the warming permafrost regions.

Stem moisture content prediction model for Larix olgensis based on beta regression.

To investigate the longitudinal variation patterns of sapwood, heartwood, bark and stem moisture content along the trunk of artificial Larix olgensis, we constructed mixed effect models of moisture content based on beta regression by combining the effects of sampling plot and sample trees. We used two sampling schemes to calibrate the model, without limiting the relative height (Scheme Ⅰ) and with a limiting height of less than 2 m (Scheme II). The results showed that sapwood and stem moisture content increased gradually along the trunk, heartwood moisture content decreased slightly and then increased along the trunk, and bark moisture content increased along the trunk and then levelled off before increasing. Relative height, height to crown base, stand area at breast height per hectare, age, and stand dominant height were main factors driving moisture content of L. olgensis. Scheme Ⅰ showed the stable prediction accuracy when randomly sampling moisture content measurements from 2-3 discs to calibrate the model, with the mean absolute percentage error (MAPE) of up to 7.2% for stem moisture content (randomly selected 2 discs), and the MAPE of up to 7.4%, 10.5% and 10.5% for sapwood, heartwood and bark moisture content (randomly selected 3 discs), respectively. Scheme Ⅱ was appropriate when sampling moisture content measurements from discs of 1.3 and 2 m height and the MAPE of sapwood, heartwood, bark and stem moisture content reached 7.8%, 11.0%, 10.4% and 7.1%, respectively. The prediction accuracies of all mixed effect beta regression models were better than the base model. The two-level mixed effect beta regression models, considering both plot effect and tree effect, would be suitable for predicting moisture content of each part of L. olgensis well.

Carbon stock projection for four major forest plantation species in Japan.

Carbon sequestration via afforestation and forest growth is effective for mitigating global warming. Accurate and robust information on forest growth characteristics by tree species, region, and large-scale land-use change is vital and future prediction of forest carbon stocks based on this information is of great significance. These predictions allow exploring forestry practices that maximize carbon sequestration by forests, including wood production. Forest inventories based on field measurements are considered the most accurate method for estimating forest carbon stocks. Japan's national forest inventories (NFIs) provide stand volumes for all Japanese forests, and estimates from direct field observations (m-NFIs) are the most reliable. Therefore, using the m-NFI from 2009 to 2013, we selected four major forest plantation species in Japan: Cryptomeria japonica, Chamaecyparis obtusa, Pinus spp., and Larix kaempferi and presented their forest age-carbon density function. We then estimated changes in forest carbon stocks from the past to the present using the functions. Next, we investigated the differences in the carbon sequestration potential of forests, including wood production, between five forestry practice scenarios with varying harvesting and afforestation rates, until 2061. Our results indicate that, for all four forest types, the estimates of growth rates and past forest carbon stocks in this study were higher than those considered until now. The predicted carbon sequestration from 2011 to 2061, assuming that 100 % of harvested carbon is retained for a long time, twice the rate of harvesting compared to the current rate, and a 100 % afforestation rate in harvested area, was three to four times higher than that in a scenario with no harvesting or replanting. Our results suggest that planted Japanese forests can exhibit a high carbon sequestration potential under the premise of active management, harvesting, afforestation, and prolonging the residence time of stored carbon in wood products with technology development.

Boreal conifers maintain carbon uptake with warming despite failure to track optimal temperatures.

Warming shifts the thermal optimum of net photosynthesis (ToptA) to higher temperatures. However, our knowledge of this shift is mainly derived from seedlings grown in greenhouses under ambient atmospheric carbon dioxide (CO2) conditions. It is unclear whether shifts in ToptA of field-grown trees will keep pace with the temperatures predicted for the 21st century under elevated atmospheric CO2 concentrations. Here, using a whole-ecosystem warming controlled experiment under either ambient or elevated CO2 levels, we show that ToptA of mature boreal conifers increased with warming. However, shifts in ToptA did not keep pace with warming as ToptA only increased by 0.26-0.35 °C per 1 °C of warming. Net photosynthetic rates estimated at the mean growth temperature increased with warming in elevated CO2 spruce, while remaining constant in ambient CO2 spruce and in both ambient CO2 and elevated CO2 tamarack with warming. Although shifts in ToptA of these two species are insufficient to keep pace with warming, these boreal conifers can thermally acclimate photosynthesis to maintain carbon uptake in future air temperatures.

The shifts in the structure of the prokaryotic community of mountain-grassland soil under the influence of artificial larch plantations.

Creation of artificial forest plantations on a global scale is one of the ways to mitigate the negative effects of climate change on ecosystems, at the same time providing soil protection from erosion, regulation of the hydrological regime and carbon sequestration in soils of different natural and climatic zones. However, the change of the dominant plant community cause significant ecosystem changes, reflecting at the structure and functioning of the soil microbial complex as well. The shifts in prokaryotic community of the meadow soil resulting from the conversion of the native meadow (further grassland) phytocenosis to the artificial forest plantations was investigated with the use of NGS sequencing technology and metabarcoding approach-amplicon sequencing of V4 region of 16 S rRNA (performed on Illumina Miseq platform). The identified shifts in taxonomic structure and diversity may be the result of changes in the physic-chemical conditions of soils and, on the other hand, may serve as indicators of such changes. Cultivation of larch led to an increase in the diversity of the prokaryotic community and its stratification by depth. The acidifying effect of larch manifested itself in an increase in the proportion and diversity of acidobacteria, in the abundance of oligotrophic microorganisms of phyla Chloroflexi, Firmicutes, and a simultaneous comparative decrease in the bacteria of Verrucomicrobia phylum, alphaproteobacteria of or. Rhizobiales and Burkholderiales. The absence of clearly expressed dominants in the prokaryotic community, as well as a significant increase in alpha-diversity indices, compared with the control plot of native mountain-meadow soil under grassland vegetation, suggests a transitional nature of the soil ecosystem of artificial forest plantations.

Embryogenic callus induction from immature zygotic embryos and genetic transformation of Larix kaempferi 3x Larix gmelinii 9.

To date, there are few reports of the successful genetic transformation of larch and other conifers, mainly because it is difficult to transform and integrate exogenous genes. In this study, hybrid larch Larix kaempferi 3x Larix gmelinii 9 cones were collected on June 27, July 1, July 4, July 7 and July 16, 2017. Embryogenic callus induction was studied using a combination of different plant growth regulators and concentrations. The results showed that July 1 was the best stage; the highest induction rate was 10.83%, which cultured in BM medium (Button medium, which formula was listed in S1 Table) with 1.0 mg/L 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.2 mg/L KT(kinetin). When cultured on a proliferation medium for 12 days, proliferation was the fastest, reaching 323.08%, which could also maintain the freshness and vitality. The suitable pre-culture medium for somatic embryogenesis was 1/4 BM medium containing 10 g/L inositol and 60 g/L sucrose. The combination of 45 mg/L ABA (abscisic acid) and 75 g/L PEG4000 (Polyethyene glycol 4000) could promote the number of somatic embryos, and reached the maximum, 210 140 per 1 g FW. The genetic transformation was carried out by the Agrobacterium-mediated transformation method with embryogenic callus cultured for 12 days. The results showed the optimal OD600 of the infection solution(suspension of A. tumefaciens) was 0.5, co-culture time was 2 days, and screening concentration of Hyg (hygromycin B) was 4 mg/L. In this study, the transformation rate of resistance callus was 32.1%. It provides a reference for low genetic transformation efficiency of larch at present. This study could be beneficial for the innovation and breeding of larch by genetic engineering and provides a certain basis for rapid propagation of excellent larch germplasm resources and genetic engineering breeding of larch and other conifers.

Root foraging of birch and larch in heterogeneous soil nutrient patches under water deficit.

Water and nutrient are two critical factors that limit plant growth to spatial-temporal extents. Tree root foraging behavior has not received adequate attention in heterogeneous soil environments in temperate forest under drought pressure. In this study, birch (Betula platyphylla) and larch (Larix olgensis) seedlings were raised in pots in a split-root system with artificially heterogeneous soil environments to study the root foraging response to drought. Potted space was split into two halves where substrates were mixed with fertilizers in 67.5 mg nitrogen (N) plant-1 (N-P2O5-K2O, 14-13-13) to both halves as to create a homogeneous condition. Otherwise, a rate of 135 mg N plant-1 of fertilizers was delivered to a random half to create a heterogeneous condition. Half of seedlings were fully sub-irrigated every three days with the other half received the drought treatment by being watered every six days. Both birch and larch seedlings showed greater net shoot growth and biomass increment in well-watered condition, while root morphology was promoted by drought. Both species placed more fine roots with higher root N concentration in nutrient-enriched patches. In the heterogeneous pattern, birch showed a higher foraging precision assessed by biomass and greater foraging plasticity assessed in morphology and physiology. In contrast, larch seedlings had higher root N concentration in the well-watered condition. Neither species showed a significant response of N utilization to the heterogeneous pattern, but both used more N when water supply was improved. Overall, birch is better at acclimating to heterogeneous soil conditions, but its ability to seize N was lower than larch when drought was alleviated.

Spatial and Temporal Distribution of Arabinogalactan Proteins during Larix decidua Mill. Male Gametophyte and Ovule Interaction.

The role of ArabinoGalactan Proteins (AGPs) in the sexual reproduction of gymnosperms is not as well documented as that of angiosperms. In earlier studies, we demonstrated that AGPs play important roles during ovule differentiation in Larix decidua Mill. The presented results encouraged us to carry out further studies focused on the functions of these unique glycoproteins during pollen/pollen tube and ovule interactions in Larix. We identified and analyzed the localization of AGPs epitopes by JIM4, JIM8, JIM13 and LM2 antibodies (Abs) in male gametophytes and ovule tissue during pollination, the progamic phase, and after fertilization and in vitro growing pollen tubes. Our results indicated that (1) AGPs recognized by JIM4 Abs play an essential role in the interaction of male gametophytes and ovules because their appearance in ovule cells is induced by physical contact between reproductive partners; (2) after pollination, AGPs are secreted from the pollen cytoplasm into the pollen wall and contact the extracellular matrix of stigmatic tip cells followed by micropylar canal cells; (3) AGPs synthesized in nucellus cells before pollen grain germination are secreted during pollen tube growth into the extracellular matrix, where they can directly interact with male gametophytes; (4) in vitro cultured pollen tube AGPs labeled with LM2 Abs participate in the germination of pollen grain, while AGPs recognized by JIM8 Abs are essential for pollen tube tip growth.

Cavitation fatigue in conifers: a study on eight European species.

After drought-induced embolism and repair, tree xylem may be weakened against future drought events (cavitation fatigue). As there are few data on cavitation fatigue in conifers available, we quantified vulnerability curves (VCs) after embolism/repair cycles on eight European conifer species. We induced 50% and 100% loss of conductivity (LC) with a cavitron, and analyzed VCs. Embolism repair was obtained by vacuum infiltration. All species demonstrated complete embolism repair and a lack of any cavitation fatigue after 50% LC . After 100% LC, European larch (Larix decidua), stone pine (Pinus cembra), Norway spruce (Picea abies), and silver fir (Abies alba) remained unaffected, while mountain pine (Pinus mugo), yew (Taxus baccata), and common juniper (Juniperus communis) exhibited 0.4-0.9 MPa higher vulnerability to embolism. A small cavitation fatigue observed in Scots pine (Pinus sylvestris) was probably biased by incomplete embolism repair, as indicated by a correlation of vulnerability shifts and conductivity restoration. Our data demonstrate that cavitation fatigue in conifers is species-specific and depends on the intensity of preceding LC. The lack of fatigue effects after moderate LC, and relevant effects in only three species after high LC, indicate that conifers are relatively resistant against cavitation fatigue. This is remarkable considering the complex and delicate conifer pit architecture and may be important considering climate change projections.

Long-term effects of mixed planting on arbuscular mycorrhizal fungal communities in the roots and soils of Juglans mandshurica plantations.

BACKGROUND: Establishing mixed plantations is an effective way to improve soil fertility and increase forest productivity. Arbuscular mycorrhizal (AM) fungi are obligate symbiotic fungi that can promote mineral nutrient absorption and regulate intraspecific and interspecific competition in plants. However, the effects of mixed plantations on the community structure and abundance of AM fungi are still unclear. Illumina MiSeq sequencing was used to investigate the AM fungal community in the roots and soils of pure and mixed plantations (Juglans mandshurica × Larix gmelinii). The objective of this study is to compare the differential responses of the root and rhizosphere soil AM fungal communities of Juglans mandshurica to long-term mixed plantation management. RESULTS: Glomus and Paraglomus were the dominant genera in the root samples, accounting for more than 80% of the sequences. Compared with that in the pure plantation, the relative abundance of Glomus was higher in the mixed plantation. Glomus, Diversispora and Paraglomus accounted for more than 85% of the sequences in the soil samples. The relative abundances of Diversispora and an unidentified genus of Glomeromycetes were higher and lower in the pure plantation, respectively. The Root_P samples (the roots in the pure plantation) had the highest number of unique OTUs (operational taxonomic units), which belonged mainly to an unidentified genus of Glomeromycetes, Paraglomus, Glomus and Acaulospora. The number of unique OTUs detected in the soil was lower than that in the roots. In both the root and soil samples, the forest type did not have a significant effect on AM fungal diversity, but the Sobs value and the Shannon, Chao1 and Ace indices of AM fungi in the roots were significantly higher than those in the soil. CONCLUSIONS: Mixed forest management had little effect on the AM fungal community of Juglans mandshurica roots and significantly changed the community composition of the soil AM fungi, but not the diversity.

Transcription factor LkWOX4 is involved in adventitious root development in Larix kaempferi.

WUSCHEL-related homeobox4 (WOX4) plays important roles in vascular formation and adventitious root (AR) development. Here, we cloned the WOX4 from the AR of Larix kaempferi, whose cDNA is 1452 bp in length and encodes 483 amino acids. LkWOX4 is mainly expressed in the layer formation area of the stem at 10 days after cutting and its expression levels in the middles and ends of the ARs were higher than that in the AR tips. The fused protein LkWOX4-GFP localized in the nucleus. The heterologous overexpression of LkWOX4 in 84 K poplar significantly increased AR numbers and decreased AR lengths. In LkWOX4 plants, the endogenous jasmonic acid and abscisic acid contents significantly decreased in stems, while the auxin, jasmonic acid and abscisic acid contents significantly increased in ARs. RNA-Seq of those LkWOX4 overexpression poplar plants showed that the expression of plant hormone signaling genes (ARF2, ARF3, ARF7 and ARF18), rooting-related transcription factors (WOX5, LBD29 and SCR) and root development-related genes (CYCD3, GRF1 and TAA1) were affected. Moreover, we found that LkWOX4 interacts with LkPAT18, LkACBP6, and LkCIP7 using yeast two hybrid screening. Thus, we found LkWOX4 involves in the AR initiation and development, which might be regulated through the IAA, JA and ABA signaling pathways.

Nitrogen loading increases the ozone sensitivity of larch seedlings with higher sensitivity to nitrogen loading.

Larch (Larix sp.) tree is a critical species for the future afforestation in Northeast Asia. The impacts of elevated concentrations of ground-level ozone (O3) and nitrogen (N) deposition are raising concerns. However, knowledge of the combined effects of elevated O3 and N loading are still limited. We investigated whether nitrogen loading mitigates the negative impacts of ozone on two larch species: the Japanese larch (L. kaempferi) and its hybrid larch F1 (L. gmelinii var. japonica × L. kaempferi) or not. We used open-top cambers and compared responses of the larch seedlings. Results showed the N loading mitigated the negative effects of O3 on Japanese larch. However, in hybrid larch F1, N loading did not mitigate O3-induced inhibition of growth and photosynthetic capacity. Mitigation effect of N loading on negative O3 impacts may vary between the two Larix spp., Hybrid larch F1 could be more affected by the combined effects of O3 and N loading due to its higher growth response to N loading. Elevated O3 also reduced leaf nitrogen/phosphorus (N/P) ratio by elevated O3, with significant effects in hybrid larch F1, particularly under N loading. In the present study, leaf N/P ratio was utilized to validate the hypothesis that a positive effect of N loading may be observed if O3 does not induce P limitation in Larix spp. We demonstrated a potential leaf N/P ratio function, which could reflect responses to O3 and N loading in hybrid larch F1.

Isolation and characterization of larch BABY BOOM2 and its regulation of adventitious root development.

The BABY BOOM2 gene, designated LkBBM2, and its promoter were isolated from hybrid larch (Larix kaempferi × L. olgensis). The open reading frame of LkBBM2 was 2574 bp, encoding 857 amino acids. The LkBBM2 protein contains two AP2 DNA binding domains and a BBM specific motif, but lacks the euANT5 motif common to AP2 family members. The LkBBM2 promoter contains several hormone response and root-specific expression elements. LkBBM2 expression was significantly higher in larch adventitious roots (ARs) than in stems, leaves or stem tips, and increased after auxin treatment. The fused protein LkBBM2-GFP was localized in both the nucleus and cytoplasm whereas LkBBM1-GFP was only localized in the nucleus. Over-expression of LkBBM2 and LkBBM1 in Arabidopsis significantly elongated the roots. Furthermore, over-expression those two genes in the hybrid poplar (Populus alba × P. glandulosa) significantly increased ARs number. We speculated that these two genes regulate AR development.

Microbial biomass and enzymatic responses to temperate oak and larch forest thinning: Influential factors for the site-specific changes.

Microbial biomass and enzyme activity are essential for ecosystem function in managed forests; however, uncertainty remains because microbial biomass and enzymatic responses to thinning highly differ with case studies. This study addressed the drivers for the site-specific responses of microbial biomass and enzyme activity to thinning. Study sites included two oak and three larch forests; each had un-thinned control, intermediate thinning (15-23% basal area reduction), and heavy thinning treatments (30-44% basal area reduction). Soil properties (temperature, water content, pH, total and inorganic nitrogen, and total carbon/nitrogen ratio), microbial biomass, enzyme (ß-glucosidase, N-acetylglucosaminidase, leucyl aminopeptidase, acid phosphatase, and phenol oxidase) activity, and soil carbon storage were determined 6 years after thinning. Compared to the control, microbial biomass carbon and nitrogen were higher under the intermediate and the heavy thinning by 13.9 and 24.4% and 11.5 and 29.9% at one oak forests, respectively, and higher under the intermediate thinning by 53.7 and 70.7% at one larch forests. There were the post-thinning changes in leucyl aminopeptidase activity by -46.9% and by 150.0-210.0% at an oak and larch forest, respectively, acid phosphatase activity by 60.0% at one oak forest, and phenol oxidase activity by 355.0% at one oak forest. The effect sizes of thinning for soil properties explained 94% and 77% of variance of the effect sizes for microbial biomass and enzyme activity. Especially, the effect sizes for soil water content, NH4+, total carbon/nitrogen ratio, and temperature were the most influential. Furthermore, the effect size for soil carbon storage was parabolically related to the effect size for microbial biomass carbon (R2 = 0.66). These findings highlight that inconsistent thinning effects on soil properties varied microbial biomass and enzymatic responses to thinning, which differentiated the change in soil carbon storage across sites. Future studies should consider such inconsistencies when examining the effects of forest management.

Deciphering Hybrid Larch Reaction Norms Using Random Regression.

The link between phenotypic plasticity and heterosis is a broad fundamental question, with stakes in breeding. We report a case-study evaluating temporal series of wood ring traits of hybrid larch (Larix decidua × L. kaempferi and reciprocal) in relation to soil water availability. Growth rings record the tree plastic responses to past environmental conditions, and we used random regressions to estimate the reaction norms of ring width and wood density with respect to water availability. We investigated the role of phenotypic plasticity on the construction of hybrid larch heterosis and on the expression of its quantitative genetic parameters. The data came from an intra-/interspecific diallel mating design between both parental species. Progenies were grown in two environmentally contrasted sites, in France. Ring width plasticity with respect to water availability was confirmed, as all three taxa produced narrower rings under the lowest water availability. Hybrid larch appeared to be the most plastic taxon as its superiority over its parental species increased with increasing water availability. Despite the low heritabilities of the investigated traits, we found that the expression of a reliable negative correlation between them was conditional to the water availability environment. Finally, by means of a complementary simulation, we demonstrated that random regression can be applied to model the reaction norms of non-repeated records of phenotypic plasticity bound by a family structure. Random regression is a powerful tool for the modeling of reaction norms in various contexts, especially perennial species.