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Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal R hodophyta. Here, the ef fects of salinity and light intensity on the quantum yield of photosystem II in P y. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under dif ferent stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The c DNA was synthesized and the expression levels of seven potential internal control genes were evaluated using q RT-PCR method. Then, we used ge Norm, a common statistical algorithm, to analyze the q RT-PCR data of seven reference genes. Potential genes that may constantly be expressed under dif ferent conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while t ubulin, glyceraldehyde-3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the ge Norm screening. Based on the results of the software analysis and absolute quantification, we proposed that e longation factor 3 and 18 S ribosomal RNA could be used as internal control genes when the P y. haitanensis blades were subjected to salinity stress, and that α-tubulin and 18 S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.  相似文献   

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Crossbreeding is an effective approach to manage the genetic decline in aquaculture.One-way hybrids of Crassostrea sikamea(♀)and Crassostrea gigas(♂)have advantages in growth traits and adaptation to high temperature.Here,we used high-throughput sequencing to analyze the molecular processes in the hybrids under and after thermal stress.The hybrids were cultured in the seawater with an increasing temperature from 25℃to 40℃during 10 hours,which is regarded as the thermal stress stage.Then the temperature decreased from 40℃to 25℃within 2 h,which is regarded as the recovery stage.In this study,1293 significant diffe-rentially expressed genes(DEGs)were obtained under thermal stress,of which 576 were upregulated and 717 were downregulated,and 740 significant differentially expressed genes(DEGs)were obtained in the recovery stage,of which the number of upregulated and downregulated genes was 409 and 331,respectively.The antigen processing and presentation,NOD-like,and NF-kappa B path-ways were significantly enriched during the thermal stress stage.The MAPK and PPAR signaling pathways were significantly enrich-ed during the recovery stage.The HSP70,HSP90,and CANX genes were strongly and rapidly upregulated in the control/thermal stress groups but were slightly less upregulated in the thermal stress/recovery group.These results indicate that the innate immune system or nonspecific immunity was deployed to protect interior tissues from thermal stress.In addition,85%of the mutual DEGs were involved in bidirectional regulation(up/down or down/up)when the oysters were removed from the thermal stress to recover.This study provides preliminary insight into the molecular response of C.sikamea(♀)and C.gigas(♂)hybrids to thermal stress and provides a basis for future studies on temperature-adaptation and the possible expansion of hybrid breeding.  相似文献   

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Anadara broughtonii is one of the main commercially important species of shellfish in northern China.A.broughtonii lives in relatively stable subtidal zone where the clam could re spond to environmental changes with minimum energy.Therefore,slight fluctuations in water environment may have a great impact on physiological processes such as feeding and metabolism.Large-scale mortality often occurs during the intermediate rearing processes,and high temperatures in summer are considered the leading cause of mortality.To understand the physiological and molecular response patterns of A.broughtonii at high temperatures and to estimate the appropriate metabolism temperature for A.broughtonii,the effects of temperature on the respiratory metabolism and food intake at different growth stages were studied.The response patterns of heat shock protein genes to heat stress and post-stress recovery were also explored.Results show that the temperature and growth stage of A.broughtonii were two important factors affecting metabolism and feeding.The optimum temperature for feeding and physiological activities in each shelllength group was 24℃.The temperature at which the peak metabolic rate occurred in each shell-length group increased with the increase in shell length.With the increase in heat stress,the expression of three heat shock protein genes(Abhsp60,Abhsp70,and Abhsp90) in the tissues of two size groups of A.broughtonii increased significantly when temperature was above 24℃ and reached a peak at 30℃.After heat shock at30℃ for 2 h,the expression of the three heat shock protein genes rapidly increased,peaked at 2 h after the heat shock,and then gradually decreased to the level of the control group at 48 h after the heat shock.The three heat shock protein genes respond rapidly to heat stress and can be used as indicators to the cellular stre ss response in A.broughtonii under an environmental stre ss.  相似文献   

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Qiu  Liping  Mao  Yunxiang  Tang  Lei  Tang  Xianghai  Mo  Zhaolan 《中国海洋湖沼学报》2019,37(3):1102-1112
Pyropia yezoensis(formerly Porphyra yezoensis) is an economically important red alga that is cultured extensively in China. The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses. An incidence of red rot disease was found in a P. yezoensis farm from mid-November to mid-December 2015 at Lianyungang, Jiangsu Province, China. Histopathological examination revealed that the naturally infected thalli were infected apparently by a pathogen, leading to red rot symptoms. The causative agent was isolated and identified as the oomycete Pythium chondricola by morphological analysis and sequence analysis of the internal transcribed spacer and cytochrome oxidase subunit 1(cox1). In artificial infection experiments on the P. yezoensis blades, the P. chondricola isolate was able to cause the same characteristic histopathology seen in natural infections. P. chondricola grew well at a wide range of temperatures in the range 8-31℃, salinities at 0-45 and pH 5-9. In an orthogonal test used to determine the effects of environmental factors(temperature, salinity, and zoospore concentration)on infection, the data revealed that temperature was the most important factor to affect red rot disease development, with the optimal conditions for disease expansion being 20℃, 35 salinity, and a zoospore concentration of 10~6 zoospores/mL. The results obtained from the present study prompted us to set up a comprehensive epidemiological study on Pyropia, which will provide support to maintain the healthy development of the Pyropia industry in China.  相似文献   

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Pyropia yezoensis, belongs to the genus of Porphyra before 2011, inhabit on intertidal zone rocks where irradiation changes dramatically, implying that the seaweed has gained certain mechanisms to survive a harsh environment. Based on the photosynthetic parameters with or without the inhibitors determined by a Dual-PAM-100 apparatus, we investigated the photosynthetic performance and the changes in electron flow that occurred during the algae were stressed with different light intensities previously. When the irradiation saturation was approaching, the CEF around PS I became crucial since the addition of inhibitors usually led to an increase in non-photochemical quenching. The inhibitor experiments showed that there were at least three different CEF pathways in Py. yezoensis and these pathways compensated each other. In addition to maintaining a proper ratio of ATP/NAD(P)H to support efficient photosynthesis, the potential roles of CEF might also include the regulation of different photoprotective mechanisms in Py. yezoensis. Under the regulation of CEF, chlororespiration is thought to transport electrons from the reduced plastoquinone (PQ) pool to oxygen in order to mitigate the reduction in the electron transfer chain. When irradiation was up to the high-grade stress conditions, the relative value of CEF began to decrease, which implied that the NADP+ pool or PQ+ pool was very small and that the electrons were transferred from reduced PS I to oxygen. The scavenging enzymes might be activated and the water-water cycle probably became an effective means of removing the active oxygen produced by the irradiation stressed Py. yezoensis. We believe that the different mechanisms could make up the photoprotective network to allow Py. yezoensis for survival in a highly variable light stress habitat, which may enlighten scientists in future studies on irradiance stress in other algae species.  相似文献   

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Members of the DnaJ family are proteins that play a pivotal role in various cellular processes, such as protein folding, protein transport and cellular responses to stress. In the present study, we identified and characterized the full-length DnaJ cDNA sequence from expressed sequence tags of Pyropia yezoensis (PyDnaJ) via rapid identification of cDNA ends. This cDNA encoded a protein of 429 amino acids, which shared high sequence similarity with other identified DnaJ proteins, such as a heat shock protein 40/DnaJ from Pyropia haitanensis. The relative mRNA expression level of PyDnaJ was investigated using real-time PCR to determine its specific expression during the algal life cycle and during desiccation. The relative mRNA expression level in sporophytes was higher than that in gametophytes and significantly increased during the whole desiccation process. These results indicate that PyDnaJ is an authentic member of the DnaJ family in plants and red algae and might play a pivotal role in mitigating damage to P. yezoensis during desiccation.  相似文献   

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The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure line was cloned and sequenced. Sequence analysis was executed for this sequence and other 22 sequences retrieved from GenBank. A phylogenetic tree was constructed using the neighbor-joining method. The results revealed a high diversity of 18S rDNA sequences in genus Porphyra and the considerable variation of 18S rDNA sequences in different strains of the same species P. yezoensis and P. tenera. Significant difference of 18S rDNA sequence was observed between P. yezoensis from Qingdao, China, and the two strains of P. yezoensis from Japan, but the three strains of P. yezoensis formed a stable clade in the phylogenetic tree. These results indicate the possibility of interspecies and intraspecies discrimination of Porphyra using the 18S rDNA sequences.  相似文献   

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A two-dimensional spectral-difference mode (with vorticity and density equations) of internal tides isdeveloped for studying the genration and propagration of internal tides generated at the continentalshelf/slope. In general, internal tides propagate seaward in deep sea regions and shoreward on the shelf,and are dissipated rapidly. When the Vaisala frequency decreases vertically, waves may be mostly limited to thecontinental slope region. in deep sea region, motions may have either boam-like structure or modal structure,depending on the stratification strerigth and structure, whereas a modal structure may always exist onthe shelf. Waves show strong bottom intensification on the slope when strong stratification exists on thebottom. The barotropic tidal advection may affed the temporal character of internal tides at thecontinental slope, shelf break and shelf regions. but may have little influence on the energy density and energy flux of internal tides. ln the case of strong stratification, waverforms of  相似文献   

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In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.  相似文献   

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Fluorescence-based maximal photochemical efficiency, Fv/Fm, is widely used as an indicator to photosynthetic competence in marine systems. It has been considered a useful parameter diagnosing the nutrient stress on phytoplankton photosynthesis, but many studies argue its usefulness. In the present study, we try to find a temporal relationship between Fv/Fm and nitrogen concentration, and provide a possible explanation on the controversy. We continuously measured Fv/Fm and nitrogen concentration once every 10 days from September 2003 to March 2004 at two stations in Jiaozhou Bay, northern China. It was found that Fv/ffm did not significantly correlate to synchronous nitrogen concentration, but the variation (i.e. the change between two adjacent cruises) of nitrogen concentration of the previous cruise and the variation ofFv/Fm of the current cruise were strongly correlated. This result indicates that a time lag exists between the variation of nutrient status and the subsequent Fv/Fm response. Length of the time lag seems just matched the interval of our measurements (10 days). In the field, direct dependence of Fv/Fm on nitrogen concentration may not be found because of the lagged response of Fv/Fm to nitrogen concentration variations or physiological acclimation. Our results provide a possible way to explain the previously reported conflicting results on the relationship between Fv/Fm and nutrient status. To give a more-accurate estimate about the length of the time lag, an investigation that includes more frequent measurements is needed.  相似文献   

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