Non-Coding RNAs Participate in the Regulation of CRY-DASH in the Growth and Early Development of Saccharina japonica (Laminariales, Phaeophyceae).

CRY-DASH, a new cryptochrome blue light receptor, can repair damaged DNA and regulate secondary metabolism and development of fungus. However, its role in regulation during the growth of Saccharina japonica is still unclear. After cloning the full-length of CRY-DASH from S. japonica (sjCRY-DASH), we deduced that its open reading frame was 1779 bp long and encoded 592 amino acids. sjCRY-DASH transcription was rapidly upregulated within 30 min in response to blue light and exhibited 24 h periodicity with different photoperiods. Moreover, sjCRY-DASH maintained the same periodicity in suitable growth temperature, suggesting a close relationship between this periodicity and circadian rhythm regulation. Novel-m3234-5p, which was targeted to sjCRY-DASH, decreased with increasing sjCRY-DASH transcription, acting as a negative modulator of sjCRY-DASH. Six long non-coding RNAs classified as long intergenic non-coding RNAs (lincRNAs) exhibited co-expression with sjCRY-DASH. A miRNA sjCRY DASH lincRNA network was consequently identified. By predicting the endogenous competing mRNAs of novel-m3234-5p, we found that sjCRY-DASH indirectly participated in the regulation of DNA damage repair, protein synthesis and processing, and actin transport. In conclusion, our results revealed that non-coding RNAs participate in the regulation of sjCRY-DASH, which played vital roles in the growth and early development of S. japonica.

The 40S Ribosomal Protein S6 Response to Blue Light by Interaction with SjAUREO in Saccharina japonica.

Blue light (BL) plays an important role in regulation of the growth and development of aquatic plants and land plants. Aureochrome (AUREO), the recent BL photoreceptor identified in photosynthetic stramenopile algae, is involved in the photomorphogenesis and early development of Saccharina japonica porophytes (kelp). However the factors that interact with the SjAUREO under BL conditions specifically are not clear. Here in our study, three high quality cDNA libraries with CFU over 5 × 106 and a recombination rate of 100% were constructed respectively through white light (WL), BL and darkness (DK) treatments to the juvenile sporophytes. Based on the constructed cDNA libraries, the interactors of SjAUREO were screened and analyzed. There are eighty-four genes encoding the sixteen predicted proteins from the BL cDNA library, sixty-eight genes encoding eighteen predicted proteins from the DK cDNA library, and seventy-four genes encoding nineteen proteins from the WL cDNA library. All the predicted proteins are presumed to interact with SjAUREO when co-expressed with SjAUREO seperately. The 40S ribosomal protein S6 (RPS6), which only exists in the BL treated cDNA library except for two other libraries, and which is essential for cell proliferation and is involved in cell cycle progression, was selected for detailed analysis. We showed that its transcription was up-regulated by BL, and was highly transcribed in the basal blade (meristem region) of juvenile sporophytes but less in the distal part. Taken together, our results indicated that RPS6 was highly involved in BL-mediated kelp cellular division and photomorphogenesis by interacting with SjAUREO.

The response of the early developmental stages of Laminaria japonica to enhanced ultraviolet-B radiation.

The responses of the early development of Laminaria japonica collected from Kiaochow Bay in China to enhanced ultraviolet-B radiation (UV-B, 280-320 nm) were studied in the laboratory. The low UV-B radiations (11.7-23.4 J x m(-2) x d(-1)) had no significant effects on zoospores attachment, but when the UV-B dose > 35.1 J x m(-2) x d(-1) the attachment decreased significantly compared with the control. Germination of embryospores was >93% under the low (11.7-35.1 J x m(-2) x d(-1)) doses, and in the range of 78.5%-88.5% under the high (46.8-70.2 J x m(-2) x d(-1)) UV-B doses, indicating a significant radiation effect. Under the higher UV-B exposure (35.1-70.2 J x m(-2) x d(-1)), all of the few gametophytes formed from embryospores died 120 h post-release. After exposure to the low UV-B radiation (11.7-23.4 J x m(-2) x d(-1)), the formation of sporophytes decreased and the female gametophyte clones increased compared with the control. However, the sex ratio and the relative growth of female gametophytes/sporophytes had not significantly changed. According to the results, enhanced UV-B radiation has a significant effect on the early development of L. japonica under laboratory conditions, suggesting that the UV-B radiation could not be overlooked as one of the important environmental factors influencing the ontogeny of macroalgae living in marine ecosystems.

Effects of ultraviolet radiation and temperature on the ultrastructure of zoospores of the brown macroalga Laminaria hyperborea.

The interactive effects of an 8 h exposure to UV radiation and altered temperatures on the ultrastructure and germination of zoospores of the sublittoral brown alga Laminaria hyperborea (Gunn.) Foslie were investigated for the first time. Spores were exposed to four temperatures (2, 7, 12 and 17 degrees C) and three light regimes (PAR, PAR + UV-A, PAR + UV-A+UV-B). Freshly-released spores of L. hyperborea lack a cell wall and contain a nucleus with fine granular nucleoplasm and a nucleolus, one chloroplast, several mitochondria, dictyosomes and an endoplasmatic reticulum. Further, several kinds of so-called adhesive vesicles, lipid globuli and physodes containing UV-absorbing phlorotannins are embedded in the cytoplasm. No eye-spot is present. Physodes were found but they were rare and small. After an 8 h exposure to UV-B, the nucleoplasm had a mottled structure, chloroplasts contained plastoglobuli, the structure of the mitochondria changed from crista- to sacculus-type and germination was strongly inhibited at all temperatures. UV-A only had an impact on the ultrastructure at the highest temperature tested. The strongest effects were found at 17 degrees C, where germination was reduced to 35%, 32% and 9% after exposure to PAR, PAR+UV-A and PAR + UV-A + UV-B, respectively. This study indicates that UV-B radiation has strong damaging effects on the physiology and ultrastructure of zoospores of L. hyperborea. The results are important for developing scenarios for the effect of enhanced UV radiation and increasing temperatures caused by global climate changes.

[Effects of short-term continuous shear stress on cells growth and recovery of Laminaria japonica gametophytic cells in photobioreactor].

Laminaria japonica gametophytic cells were cultivated in a photobioreactor under continuous shear stress (0-1000 r/min) in 60 hours and the following static cultivation within 23.5 days. The content of chlorophyll a reached the maximum value of 2.36 mg/L at the end of continuous shear stress when the agitation speed was 90 r/min, while the chlorophyll a (chl a) concentration decreased quickly and nitrogen and phosphorus were released under high shear force (270-1000 r/min). The cell injury ratio at 1000r/min was as 18 times as that of the control. During the recovery course, gametophytic cells showed themselves distinct recovery capability at all agitation speeds. Furthermore, the content of chl a is a more exact index as biomass than dry cells weight (DCW). Besides cell injury ratio, the liberation of phosphorus demonstrates the cells injury.

Exposure to ultraviolet radiation delays photosynthetic recovery in Arctic kelp zoospores.

Seasonal reproduction in some Arctic Laminariales coincides with increased UV-B radiation due to stratospheric ozone depletion and relatively high water temperatures during polar spring. To find out the capacity to cope with different spectral irradiance, the kinetics of photosynthetic recovery was investigated in zoospores of four Arctic species of the order Laminariales, the kelps Saccorhiza dermatodea, Alaria esculenta, Laminaria digitata, and Laminaria saccharina. The physiology of light harvesting, changes in photosynthetic efficiency and kinetics of photosynthetic recovery were measured by in vivo fluorescence changes of Photosystem II (PSII). Saturation irradiance of freshly released spores showed minimal I ( k ) values (photon fluence rate where initial slope intersects horizontal asymptote of the curve) values ranging from 13 to 18 micromol photons m(-2) s(-1) among species collected at different depths, confirming that spores are low-light adapted. Exposure to different radiation spectra consisting of photosynthetically active radiation (PAR; 400-700 nm), PAR+UV-A radiation (UV-A; 320-400 nm), and PAR+ UV-A+UV-B radiation (UV-B; 280-320 nm) showed that the cumulative effects of increasing PAR fluence and the additional effect of UV-A and UV-B radiations on photoinhibition of photosynthesis are species specific. After long exposures, Laminaria saccharina was more sensitive to the different light treatments than the other three species investigated. Kinetics of recovery in zoospores showed a fast phase in S. dermatodea, which indicates a reduction of the photoprotective process while a slow phase in L. saccharina indicates recovery from severe photodamage. This first attempt to study photoinhibition and kinetics of recovery in zoospores showed that zoospores are the stage in the life history of seaweeds most susceptible to light stress and that ultraviolet radiation (UVR) effectively delays photosynthetic recovery. The viability of spores is important on the recruitment of the gametophytic and sporophytic life stages. The impact of UVR on the zoospores is related to the vertical depth distribution of the large sporophytes in the field.

Thallus morphology and optical characteristics affect growth and DNA damage by UV radiation in juvenile Arctic Laminaria sporophytes.

Growth of young sporophytes of the brown algae Laminaria digitata, L. saccharina and L. solidungula from Spitsbergen were measured in the laboratory after being exposed for 21 days to either photosynthetically active radiation (PAR = P) or to full light spectrum (PAR + UV-A + UV-B = PAB) using of cutoff glass filters. The plants were grown at 8+/-2 degrees C and 16 h light : 8 h dark cycles with 6 h additional ultraviolet radiation (UVR) exposure in the middle of the light period. Growth was measured every 10 min using growth chambers with online video measuring technique. Tissue morphology and absorption spectra were measured in untreated young sporophytes while chlorophyll (Chl) a content and DNA damage were measured in treated thalli at the end of the experiment. In all species, growth rates were significantly higher in sporophytes exposed to P alone compared to sporophytes exposed to PAB. Tissue DNA damage is dependent on thallus thickness and absorption spectra characteristics of pigments and UV-absorbing compounds. In sporophytes exposed to UVR, energy demands for repair of DNA damage and synthesis of UV-absorbing compounds for protection effectively diverts photosynthate at the expense of growth. Photosynthetic pigment was not significantly different between treatments suggesting a capacity for acclimation to moderate UVR fluence. The general growth pattern in sporophytes exposed to P alone showed an increasing growth rate from the onset of light (0500-0900 hours) to a peak at the middle of the light phase (0900-1500 hours), a decline towards the end of the light phase (1500-2100 hours) and a minimum "low" growth in the dark (2100-0500 hours) relative to growth during the entire light phase. Under PAB, different growth patterns were observed such as growth compensation at night in L. digitata, delayed growth recovery in L. saccharina and minimal but continuous growth in L. solidungula. Growth as an integrative parameter of all physiological processes showed that the effect of UVR is correlated to the depth distribution of these species.

Optimal light regime for the cultivation of transgenic Laminaria japonica gametophytes in a bubble-column bioreactor.

Fluctuating light intensity had a more significant impact on growth of gametophytes of transgenic Laminaria japonica in a 2500 ml bubble-column bioreactor than constant light intensity. A fluctuating light intensity between 10 and 110 microE m(-2) s(-1), with a photoperiod of 14 h:10 h light:dark, was the best regime for growth giving 1430 mg biomass l(-1).