Heat stress response in the closest algal relatives of land plants reveals conserved stress signaling circuits.

All land plants (embryophytes) share a common ancestor that likely evolved from a filamentous freshwater alga. Elucidating the transition from algae to embryophytes - and the eventual conquering of Earth's surface - is one of the most fundamental questions in plant evolutionary biology. Here, we investigated one of the organismal properties that might have enabled this transition: resistance to drastic temperature shifts. We explored the effect of heat stress in Mougeotia and Spirogyra, two representatives of Zygnematophyceae - the closest known algal sister lineage to land plants. Heat stress induced pronounced phenotypic alterations in their plastids, and high-performance liquid chromatography-tandem mass spectroscopy-based profiling of 565 transitions for the analysis of main central metabolites revealed significant shifts in 43 compounds. We also analyzed the global differential gene expression responses triggered by heat, generating 92.8 Gbp of sequence data and assembling a combined set of 8905 well-expressed genes. Each organism had its own distinct gene expression profile; less than one-half of their shared genes showed concordant gene expression trends. We nevertheless detected common signature responses to heat such as elevated transcript levels for molecular chaperones, thylakoid components, and - corroborating our metabolomic data - amino acid metabolism. We also uncovered the heat-stress responsiveness of genes for phosphorelay-based signal transduction that links environmental cues, calcium signatures and plastid biology. Our data allow us to infer the molecular heat stress response that the earliest land plants might have used when facing the rapidly shifting temperature conditions of the terrestrial habitat.

Nutraceutical profile and evidence of alleviation of oxidative stress by Spirogyra porticalis (Muell.) Cleve inhabiting the high altitude Trans-Himalayan Region.

The high altitude trans-Himalayan region indeed is hostile domain for survival. Algae inhabiting  this  hostile terrain have evolutionarily developed mechanisms to produce unique adaptogenic molecules against climatic stressors. The present study has focused on the high altitude alga Spirogyra porticalis (Muell.) Cleve- a filamentous Charophyte, and reports the estimation of amino acids (AAs), fatty acids (FAs), vitamins and their efficacy against oxidative stress. Reverse phase-HPLC, GC-FID and rapid resolution-LC/tandem mass spectrometry were used for analysis of AAs, FAs and vitamins. Analysis of the alga  revealed the presence of 19 AAs (239.51 ± 8.57 to 13102.40 ± 11.08 µg/g), dominated by alanine, proline and lysine. Enriched phenylalanine, cysteine-HCl and high lysine:arginine ratio could also have beneficial impact against hypoxia -induced cognitive impairment. A total of 9 FAs were detected (0.43 ± 0.00% to 34.76 ± 0.52%). Polyunsaturated and monounsaturated FAs were found to be dominant. The alga showed the presence of 8 vitamins within the range of 39.654 ± 3.198 to 5468.184 ± 106.859 µg/Kg, wherein Vitamin B5, B3 and B2 were dominant. 600 µg/ml of methanolic extract showed recovery of GSH and trolox equivalent antioxidants in rat blood/hemolysate, while 400 µg/ml of extract showed revival in superoxide dismutase (SOD) activity. The present study concludes that the alga S. porticalis has immense potential to counter oxidative stress as a nutraceutical supplement.

Two-Track Control of Cellular Machinery for Photomovement in Spirogyra varians (Streptophyta, Zygnematales).

Plants and freshwater algae devoid of flagella evolved various photomovements to optimize their photosynthetic efficiency. The filaments of Spirogyra varians exhibit complex swaying and undulating movement and form a compact mat which enables them to adjust their light exposure. Photomovement of filament fragments (1-10 cells) was analyzed using various photoreceptor and cytoskeleton inhibitors under monochromatic light. Different patterns of movement were observed under red and blue light. The filaments showed positive phototropism under blue light. Under red light, the filaments bent to undulating shape, but rapidly became unbent by a short exposure to far-red light suggesting the involvement of phytochrome in this movement. The mechanical effector for the red-light response was microtubules; the movement was inhibited effectively by the microtubule inhibitor, oryzalin. The blue-light movement was partially inhibited by the single treatment of either cytochalasin D or oryzalin, but was completely blocked when both chemicals were applied together. Phototropin-signaling inhibitors, wortmannin and LY294002, reversibly inhibited the blue-light movement. Caffeine treatment reversibly stopped the blue-light movement, while the red-light movement was not affected by calcium inhibitors. Our results suggest that the complex photomovement of S. varians is the result of a two-track control of microtubules and microfilaments signaled by the combination of phytochrome and phototropin-like receptors.

A promising new wavelength region for three-photon fluorescence microscopy of live cells.

We report three-photon laser scanning microscopy (3PLSM) using a bi-directional pumped optical parametric oscillator (OPO) with signal wavelength output at λ= 1500 nm. This novel laser was used to overcome the high optical loss in the infrared spectral region observed in laser scanning microscopes and objective lenses that renders them otherwise difficult to use for imaging. To test our system, we performed 3PLSM auto-fluorescence imaging of live plant cells at λ= 1500 nm, specifically Spirogyra, and compared performance with two-photon excitation (2PLSM) imaging using a femtosecond pulsed Ti:Sapphire laser at λ= 780 nm. Analysis of cell viability based on cytoplasmic organelle streaming and structural changes of cells revealed that at similar peak powers, 2PLSM caused gross cell damage after 5 min but 3PLSM showed little or no interference with cell function after 15 min. The λ= 1500 nm OPO is thus shown to be a practical laser source for live cell imaging.