Annual and intra-annual growth dynamics of <Emphasis Type="Italic">Myricaria elegans</Emphasis> shrubs in arid Himalaya

Key message

To disentangle complex drivers of Myricaria elegans growth in arid Himalaya, we combined tree-ring analysis with detailed dendrometer records. We found that the combination of winter frost, summer floods, and strong summer diurnal temperature fluctuations control annual and intra-annual growth dynamics. The relative importance of these drivers is, however, changing with ongoing climate change.

Abstract

High-mountain areas are among the most sensitive environments to climate change. Understanding how different organisms cope with ongoing climate change is now a major topic in the ecology of cold environments. Here, we investigate climate drivers of the annual and intra-annual growth dynamics of Myricaria elegans, a 3–6 m tall tree/shrub, in a high-elevation cold desert in Ladakh, a rapidly warming region in the NW Himalayas. As Myricaria forms narrow stands around glacier streams surrounded by the desert, we hypothesized that its growth between 3800 and 4100 m will be primarily limited by low temperatures and summer floods. We found that warmer and less snowy conditions in April and May enhance earlywood production. Latewood formation is mostly driven by the June–July temperatures (T). The positive effect of warmer summers on both annual and intra-annual growth is related to fluctuating daily T (from +30 to 0 °C). In particular, dendrometer measurements over a 2-year period showed that net daily growth increments increased when the summer night T remained above 6 °C. While high night T during generally cold desert nights promoted growth, high daytime T caused water stress and growth inhibition. The growth–temperature dependency has gradually weakened due to accelerated warming since the 1990s. In addition, positive latewood responses to high March precipitation during the colder 1960s–1980s have become negative during the warmer 1990s–2000s, reflecting an intensification of summer floods. Latewood width increased while earlywood width decreased from the 1990s, indicating a prolonged growing season and a higher risk of drought-induced embolism in earlywood vessels. Due to a multiplicity of environmental drivers including winter frost, intensified floods and strong summer diurnal T fluctuations, Myricaria growth is not controlled by a single climate parameter. Similar results are increasingly reported from other Himalayan treelines, showing that ongoing climate change will trigger complex and probably spatially variable responses in tree growth. Our study showed that these complex climatic signals can be disentangled by a combination of long-term data from tree-rings with detailed, but short-term, records from dendrometers.

     

The Root-Associated Microbial Community of the World’s Highest Growing Vascular Plants

Upward migration of plants to barren subnival areas is occurring worldwide due to raising ambient temperatures and glacial recession. In summer 2012, the presence of six vascular plants, growing in a single patch, was recorded at an unprecedented elevation of 6150 m.a.s.l. close to the summit of Mount Shukule II in the Western Himalayas (Ladakh, India). Whilst showing multiple signs of stress, all plants have managed to establish stable growth and persist for several years. To learn about the role of microbes in the process of plant upward migration, we analysed the root-associated microbial community of the plants (three individuals from each) using microscopy and tagged amplicon sequencing. No mycorrhizae were found on the roots, implying they are of little importance to the establishment and early growth of the plants. However, all roots were associated with a complex bacterial community, with richness and diversity estimates similar or even higher than the surrounding bare soil. Both soil and root-associated communities were dominated by members of the orders Sphingomonadales and Sphingobacteriales, which are typical for hot desert soils, but were different from communities of temperate subnival soils and typical rhizosphere communities. Despite taxonomic similarity on the order level, the plants harboured a unique set of highly dominant operational taxonomic units which were not found in the bare soil. These bacteria have been likely transported with the dispersing seeds and became part of the root-associated community following germination. The results indicate that developing soils act not only as a source of inoculation to plant roots but also possibly as a sink for plant-associated bacteria.     

Sensitivity to radiation of human normal, hyperthyroid, and neoplastic thyroid epithelial cells in primary culture

Samples of thyroid tissue removed surgically from 63 patients were cultured in vitro and exposed to X irradiation to investigate the radiosensitivities of various types of thyroid epithelial cells. A total of 76 samples were obtained, including neoplastic cells from patients with papillary carcinoma (PC) or follicular adenoma (FA), cells from hyperthyroidism (HY) patients, and normal cells from the surgical margins of PC and FA patients. Culturing of the cells was performed in a manner which has been shown to yield a predominance of epithelial cells. Results of colony formation assays indicated that cells from HY and FA patients were the least radiosensitive: when adjusted to the overall geometric mean plating efficiency of 5.5%, the average mean lethal dose Do was 97.6 cGy for HY cells and 96.7 and 94.3 cGy, respectively for neoplastic and normal cells from FA patients. Cells from PC patients were more radiosensitive, normal cells having an adjusted average Do of 85.0 cGy and PC cells a significantly (P = 0.05) lower average Do of 74.4 cGy. After allowing for this variation by cell type, in vitro radiosensitivity was not significantly related to age at surgery (P = 0.82) or sex (P = 0.10). These results suggest that malignant thyroid cells may be especially radiosensitive.     

Biliary excretion and organ distribution of 14C radioactivity after 14C-2-ethylhexyl acrylate administration in rats

Rats were intravenously administered (14C)-2-ethylhexyl acrylate at the dose 10 mg/kg or 50 mg/kg b. w. Biliary excretion of 14C-radioactivity was followed in 1-3 hour intervals within the first 24 hours after administration. The rats were then sacrificed and distribution of 14C-radioactivity was followed in some organs. Highest radioactivity was found in liver, less in the kidneys and the least in the brain. A significant increase of bile flow was observed. In the 24-hour intervals 2.2% of the dose was eliminated via bile at both dosages, most of it (83%) during the first 3 hours.     

Differentiation of dicyclohexylcarbodiimide reactive sites of the ATPase complex bovine heart mitochondria

1. In isolated bovine heart mitochondria, the 14C-labelled dicyclohexylcarbodiimide (DCCD) induced inhibition of the ATPase activity is accompanied by labelling of three polypeptides of Mx 9000, 16 000 and 33 000. Of these, only the 9000 polypeptide reacts with [14C]DCCD proportionally to the inhibitory effect, being saturated when the enzyme is maximally inhibited. 2. The 9000 and 16 000 polypeptides are extracted by neutral chloroform/methanol (2 : 1 v/v) while the 33 000 polypeptide remains in the non-extractable residue. No disaggregation of the polypeptides takes place during the extraction. 3. In the ATPase complex immunoprecipitated with antibody against F1, the 9000 and 16 000 polypeptides are present, but the 33 000 polypeptide is absent. 4. The results obtained indicate that the 33 000 polypeptide is not a component of the ATPase complex. As far as F0 is concerned, two types of the binding sites for DCCD were demonstrated, corresponding to the 9000 and 16 000 polypeptides. Their existence is explained by a non-random arrangement among individual monomers of the DCCD-binding protein.     

Topological and functional characterization of the F0I subunit of the membrane moiety of the mitochondrial H+-ATP synthase

Using isolated polypeptides of the F0 sector of bovine heart mitochondrial H+-ATPase, antisera were developed detecting specifically two components of F0. These two components were identified as F0I and oligomycin-sensitivity-conferring protein (OSCP) respectively. Both F0I and OSCP were digested by mild trypsin treatment of submitochondrial particles depleted of the catalytic part of H+-ATPase (USMP). Proteolysis was largely prevented by binding of F1 to F0. Proteolysis of F0I resulted in the formation of three immunoreactive, membrane-bound fragments of apparently 26 kDa, 25.5 kDa and 18 kDa, respectively, indicating that F0I contains trypsin-accessible Arg or Lys residues located close to the end and the middle part of the protein, respectively, which are in intimate contact with F1. Digestion of USMP with trypsin resulted in depression of passive H+ conduction through F0 which could be ascribed to proteolysis of F0I.     

A Hybrid Non-Ribosomal Peptide/Polyketide Synthetase Containing Fatty-Acyl Ligase (FAAL) Synthesizes the β-Amino Fatty Acid Lipopeptides Puwainaphycins in the Cyanobacterium Cylindrospermum alatosporum

A putative operon encoding the biosynthetic pathway for the cytotoxic cyanobacterial lipopeptides puwainphycins was identified in Cylindrospermum alatosporum. Bioinformatics analysis enabled sequential prediction of puwainaphycin biosynthesis; this process is initiated by the activation of a fatty acid residue via fatty acyl-AMP ligase and continued by a multidomain non-ribosomal peptide synthetase/polyketide synthetase. High-resolution mass spectrometry and nuclear magnetic resonance spectroscopy measurements proved the production of puwainaphycin F/G congeners differing in FA chain length formed by either 3-amino-2-hydroxy-4-methyl dodecanoic acid (4-methyl-Ahdoa) or 3-amino-2-hydroxy-4-methyl tetradecanoic acid (4-methyl-Ahtea). Because only one puwainaphycin operon was recovered in the genome, we suggest that the fatty acyl-AMP ligase and one of the amino acid adenylation domains (Asn/Gln) show extended substrate specificity. Our results provide the first insight into the biosynthesis of frequently occurring β-amino fatty acid lipopeptides in cyanobacteria, which may facilitate analytical assessment and development of monitoring tools for cytotoxic cyanobacterial lipopeptides.