Different Patterns of mRNA Nuclear Retention during Meiotic Prophase in Larch Microsporocytes.

Recent studies show a crucial role of post-transcriptional processes in the regulation of gene expression. Our research has shown that mRNA retention in the nucleus plays a significant role in such regulation. We studied larch microsporocytes during meiotic prophase, characterized by pulsatile transcriptional activity. After each pulse, the transcriptional activity is silenced, but the transcripts synthesized at this time are not exported immediately to the cytoplasm but are retained in the cell nucleus and especially in Cajal bodies, where non-fully-spliced transcripts with retained introns are accumulated. Analysis of the transcriptome of these cells and detailed analysis of the nuclear retention and transport dynamics of several mRNAs revealed two main patterns of nuclear accumulation and transport. The majority of studied transcripts followed the first one, consisting of a more extended retention period and slow release to the cytoplasm. We have shown this in detail for the pre-mRNA and mRNA encoding RNA pol II subunit 10. In this pre-mRNA, a second (retained) intron is posttranscriptionally spliced at a precisely defined time. Fully mature mRNA is then released into the cytoplasm, where the RNA pol II complexes are produced. These proteins are necessary for transcription in the next pulse to occur.mRNAs encoding translation factors and SERRATE followed the second pattern, in which the retention period was shorter and transcripts were rapidly transferred to the cytoplasm. The presence of such a mechanism in various cell types from a diverse range of organisms suggests that it is an evolutionarily conserved mechanism of gene regulation.

Seasonal variations in carbon, nitrogen and phosphorus concentrations and C:N:P stoichiometry in different organs of a Larix principis-rupprechtii Mayr. plantation in the Qinling Mountains, China.

Understanding how concentrations of elements and their stoichiometry change with plant growth and age is critical for predicting plant community responses to environmental change. We used long-term field experiments to explore how the leaf, stem and root carbon (C), nitrogen (N) and phosphorous (P) concentrations and their stoichiometry changed with growth and stand age in a L. principis-rupprechtii Mayr. plantation from 2012-2015 in the Qinling Mountains, China. Our results showed that the C, N and P concentrations and stoichiometric ratios in different tissues of larch stands were affected by stand age, organ type and sampling month and displayed multiple correlations with increased stand age in different growing seasons. Generally, leaf C and N concentrations were greatest in the fast-growing season, but leaf P concentrations were greatest in the early growing season. However, no clear seasonal tendencies in the stem and root C, N and P concentrations were observed with growth. In contrast to N and P, few differences were found in organ-specific C concentrations. Leaf N:P was greatest in the fast-growing season, while C:N and C:P were greatest in the late-growing season. No clear variations were observed in stem and root C:N, C:P and N:P throughout the entire growing season, but leaf N:P was less than 14, suggesting that the growth of larch stands was limited by N in our study region. Compared to global plant element concentrations and stoichiometry, the leaves of larch stands had higher C, P, C:N and C:P but lower N and N:P, and the roots had greater P and C:N but lower N, C:P and N:P. Our study provides baseline information for describing the changes in nutritional elements with plant growth, which will facilitates plantation forest management and restoration, and makes a valuable contribution to the global data pool on leaf nutrition and stoichiometry.

Increased phosphate uptake but not resorption alleviates phosphorus deficiency induced by nitrogen deposition in temperate Larix principis-rupprechtii plantations.

The imbalance between nitrogen (N) and phosphorus (P) deposition may shift temperate ecosystems from N- to P-limitation. However, it is unclear how the imbalanced N : P input affects the strategies of plants to acquire P and, therefore, the growth of plants and the competition among species. We conducted a 4-yr N-addition experiment in young and mature larch (Larix principis-rupprechtii) stands. Plant growth and P acquisition strategies were assessed for larch and understorey vegetation. N addition stimulated the aboveground productivity of understorey vegetation in the young stand and larch in the mature stand, with other species unaffected. The competitive advantages of understorey vegetation in the young stand and larch in the mature stand were associated with their high stoichiometric homoeostasis. To maintain the N : P homoeostasis of these species, an increase in phosphatase activity but not P resorption efficiency increased the supply of P. Additionally, N addition accelerated P mineralization by decreasing the fungal-to-bacterial ratios and improved uptake of soil P by increasing the arbuscular mycorrhizas-to-ectomycorrhizas ratios. Our results suggest that plants with high stoichiometric homoeostasis could better cope with N deposition-induced P-deficiency. Although P resorption efficiency showed little plasticity in response, plants activated a variety of P-acquisition pathways to alleviate the P-deficiency caused by N deposition.

Application of ionic liquids in the microwave-assisted extraction of proanthocyanidins from Larix gmelini bark.

Ionic liquid based, microwave-assisted extraction (ILMAE) was successfully applied to the extraction of proanthocyanidins from Larix gmelini bark. In this work, in order to evaluate the performance of ionic liquids in the microwave-assisted extraction process, a series of 1-alkyl-3-methylimidazolium ionic liquids with different cations and anions were evaluated for extraction yield, and 1-butyl-3-methylimidazolium bromide was selected as the optimal solvent. In addition, the ILMAE procedure for the proanthocyanidins was optimized and compared with other conventional extraction techniques. Under the optimized conditions, satisfactory extraction yield of the proanthocyanidins was obtained. Relative to other methods, the proposed approach provided higher extraction yield and lower energy consumption. The Larix gmelini bark samples before and after extraction were analyzed by Thermal gravimetric analysis, Fourier-transform infrared spectroscopy and characterized by scanning electron microscopy. The results showed that the ILMAE method is a simple and efficient technique for sample preparation.

Growth and photosynthetic traits of hybrid larch F1 (Larix gmelinii var. japonica x L. kaempferi) under elevated CO2 concentration with low nutrient availability.

The hybrid larch F(1) (Larix gmelinii var. japonica × Larix kaempferi) is considered one of the most important tree species not only for timber production but also as an afforestation material for severe conditions such as infertile soil. To predict the ability of hybrid larch F(1) as an afforestation material under potential climates in the future, it is important to understand the response of hybrid larch F(1) to elevated CO(2) concentration ([CO(2)]) under low nutrient availability. Three-year-old seedlings of hybrid larch F(1) were grown under two different levels of [CO(2)], 360 (ambient) and 720 µmol mol(-1) (elevated), in combination with two different levels of nitrogen (N) supply (0 and 30 kg ha(-1)) for one growing season. Elevated [CO(2)] reduced the maximum rates of carboxylation and electron transport in the needles. Net photosynthetic rates at growth [CO(2)] (i.e., 360 and 720 µmol mol(-1) for ambient and elevated treatment, respectively) did not differ between the two CO(2) treatments. Reductions in N content and N use efficiency to perform photosynthetic functions owing to the deficiency of nutrients other than N, such as P and K, and/or increase in cell wall mass were considered factors of photosynthetic down-regulation under elevated [CO(2)], whereas stomatal closure little affected the photosynthetic down-regulation. Although we observed strong down-regulation of photosynthesis, the dry matter increase of hybrid larch F(1) seedlings was enhanced under elevated [CO(2)]. This is mainly attributable to the increase in the amount of needles on increasing the number of sylleptic branches. These results suggest that elevated CO(2) may increase the growth of hybrid larch F(1) even under low nutrient availability, and that this increase may be regulated by changes in both crown architecture and needle photosynthesis, which is mainly affected not by stomatal limitation but by biochemical limitation.

Localisation pattern of homogalacturonan and arabinogalactan proteins in developing ovules of the gymnosperm plant Larix decidua Mill.

We have identified and characterised the temporal and spatial distribution of the homogalacturonan (HG) and arabinogalactan proteins (AGP) epitopes that are recognised by the antibodies JIM5, JIM7, LM2, JIM4, JIM8 and JIM13 during ovule differentiation in Larix decidua Mill. The results obtained clearly show differences in the pattern of localisation of specific HG epitopes between generative and somatic cells of the ovule. Immunocytochemical studies revealed that the presence of low-esterified HG is characteristic only of the wall of megasporocyte and megaspores. In maturing female gametophytes, highly esterified HG was the main form present, and the central vacuole of free nuclear gametophytes was particularly rich in this category of HG. This pool will probably be used in cell wall building during cellularisation. The selective labelling obtained with AGP antibodies indicates that some AGPs can be used as markers for gametophytic and sporophytic cells differentiation. Our results demonstrated that the AGPs recognised by JIM4 may constitute molecules determining changes in ovule cell development programs. Just after the end of meiosis, the signal detected with JIM4 labelling appeared only in functional and degenerating megaspores. This suggests that the antigens bound by JIM4 are involved in the initiation of female gametogenesis in L. decidua. Moreover, the analysis of AGPs distribution showed that differentiation of the nucellus cells occurs in the very young ovule stage before megasporogenesis. Throughout the period of ovule development, the pattern of localisation of the studied AGPs was different both in tapetum cells surrounding the gametophyte and in nucellus cells. Changes in the distribution of AGPs were also observed in the nucellus of the mature ovule, and they could represent an indicator of tissue arrangement to interact with the growing pollen tube. The possible role of AGPs in fertilisation is also discussed.

[Root decomposition and nutrient release of Fraxinus manshurica and Larix gmelinli plantations].

A two-year buried bag experiment with different sizes of Fraxinus mandshurica and Larix gmelinii roots showed that the coarse (5 to approximately 10 mm), medium (2 to approximately 5 mm) and fine ( <2 mm) roots of F. mandshurica had an annual decomposition coefficient of 0.3649, 0.4381 and 0.2720, while those of L. gmelinii had the coefficient of 0.1967, 0.1955 and 0.2464, respectively. During root decomposition, large amounts of carbon and nutrients released. After buried into soil for 150 days, more than 90% of soluble sugar in all sizes of the two species roots was released, and about 40%, 71% and 95% of potassium was released from the coarse and medium, and fine roots of F. mandshurica, and all sizes of L. gmelinii roots, respectively. In the second year of the experiment, about 50% of nitrogen and 40% of phosphorus were released from the coarse and medium roots of the two species, and 60% of nitrogen and phosphorus were released from their fine roots. In the study of forest ecosystem's carbon and nutrient cycles, root decomposition shouldn't be ignored.

[Utilization of insoluble phosphate by Larix gmelini seedlings under phosphorus starvation].

The utilization of insoluble phosphate by 2 years old larch(Larix gmelini) seedlings was studied in greenhouse. The seedlings were grown in sand and AlPO4 was supplied as a source of P. The results showed that larch seedlings could utilize some amount of AlPO4. Supplied with AlPO4 but not incubated with mycorrhizal-fungi, the seedlings could absorb 35.1% and 64.9% of the phosphorus uptake when supplied with soluble phosphate. The effects of various mycorrhizal-fungi on utilization of phosphate were different, i.e., the utilization rate of AlPO4 was higher when incubated with Suillus granulatus, but was similar to control when incubated with Suillus grevillei. The proportion of root/crown became higher under P-starvation, and larch seedlings could gain more P by enlarging their root growth.