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1 脱水对条斑紫菜叶状体 psba 和 psbd 基因表达量的影响 1, 2, 2, 2, 1 (1., ; 2., ) 摘要 : 从处于不同脱水复水阶段的条斑紫菜 (Porphyra yezoensis Ueda) 叶状体中提取总 RNA, 用荧光定量 PCR 技术检测条斑紫菜不同脱水和复水过程中 psba 和 psbd 基因的转录水平上的相对表达量的变化实验结果表明, psba 和 psbd 基因转录水平上的相对表达量的变化趋势相似但变化幅度不一致在相对含水量约为 71% 时两种基因转录水平上的相对表达量分别增加了 6.18 倍和 2.13 倍说明在条斑紫菜叶状体脱水过程中两种基因的转录并非同比例完成 ; 在轻度脱水状态下 psba 和 psbd 基因转录水平上的相对表达量的快速增大可能是藻体暴露在空气中时适应陆地环境的初始响应关键词 : 条斑紫菜 (Porphyra yezoensis Ueda); 脱水复水 ; psba; psbd; 转录水平上的相对表达量中图分类号 : S917.3 文献标识码 : A 文章编号 : (2011) ,, ;,,,,,,, [1] ; (Porphyra yezoensis Ueda),, [2] [3], ( ) [2],,,, [2], 20, 20%, 12 [2],,, [4], [3, 5-8] ; [9], [10] [4],,,,, : ; : : ( , B ); (2006BAD09A04); 863 (2006AA10A402, 2007AA09Z406, 2006AA10A413) : (1985-),,,, : , yangfang051228@163.com;,,,, : , gcwang@qdio.ac.cn Marine Sciences / Vol. 35, No. 4 /

2 ,, [11] I ( PS ) ( PS ) PS P680, PS PS, [12] PS D1 D2 psba psbd, psba psbd,, psba psbd, psba psbd, [13] psba psbd, psba psbd, PS (D1, D2 ) 1 材料与方法 1.1 样品的采集及制备 4,,, -80, 100%, 15 min 60 min 105 min, min 样品相对含水量的测定及定量 PCR 分析 :, 1.1.2, 15 min 1 1, 60, ( 48 h),, (3 ) (R WC ) : Wt Wd RWC (%) = 100 W0 Wd, W 0 (g), W t (g), W d (60, 48 h) RNA 100 mg, 1.5 ml 1 ml Trizol(Invitrogen), 5 min g, 4, 1 0 min 200 μl, 15 s, 2~3 min g, 4, 15 min 500 μl, ~30 min g, 4, 10 min, 1 ml 75% g, 4, 5 min, 5 min DEPC RNA, -80 RNA RNA DNA RNA RQ1 Rnase-Free Dnase (Promega), 37, 30 min DNA 4 μl Stop Solution, min, cdna M-MLV (Promega), (100 mol/l)2.7 μl, RNA μl 70, 5 min 2 min : M-MLV 5 Reaction Buffer 5μL, dntp mix(10 mmol/l)5 μl, RNase Inhibitor 0.65 μl, M-MLV RT 1μL 36, 60 min PCR, PCR PCR [13], 3, CT [14] psba psbd, t 2 结果 2.1 条斑紫菜叶状体相对含水量测定结果 15 min 62 / 2011 / 35 / 4

3 ,,, 15 min 60 min 105 min 100% 71% 34% 21% 2.2 样品总 RNA 的检测 RNA ( 1) 28S, 18S rrna, 1, 2, 3, 4, 5 5S, 18S, 28S rrna RNA,, psba 0.68 (P < 0.01); psbd 0.30 (P < 0.01);, psba psbd 2.25 (P < 0.01) 2 Real-time PCR psba, psbd Fig. 2 Variations in psba and psbd transcript levels using real-time PCR. Total RNA was isolated from samples with various relative water content (RWC) of rehydration 3 讨论 1 RNA Fig. 1 Agarose gel electrophoresis of RNA extracted from Porphyra yezoensis with various relative water content (RWC) of rehydration M, DNA Marker 3; % 71% 34% 21% 30 min M. DNA Marker 3; 1. sample with relative water content (RWC) 100%; 2. sample with RWC 71%; 3.sample with RWC 34%; 4. sample with RWC 21%; 5. samples after 30 min of rehydration 2.3 psba psbd 相对表达量的变化 2, psba psbd, psba psbd 71%,, psba ( 100%) 7.18 (P < 0.01); psbd 3.13 (P < 0.01); psba psbd 2.29 (P < 0.05),, 21%, (P < 0.01), RNA Trizol RNA 18S 28S rrna,, RNA PCR DNA SYBR Green, (psba psbd 18S rrna ) [13],, PS, [15], PS [16-17] psba psbd 4 5, PS (psba psbd) [18] ; Yuan Shu [19], PS (psba psbd) Marine Sciences / Vol. 35, No. 4 /

4 ; He [20], (Triticum aestivum L. cv. Longchun No. 10) psba psbd mrna ; Collett [21] Xerophyta humilis psbr, ChlP, psba psbp ; Zhang [22] Ammopiptanthus mongolicus cab, psba, PCR psba ( D1) psbd ( D2) 15 min, 60 min, 105 min( 71%, 34%, 21%) 3, psba psbd 18S rrna ( ) ( 2), PS : PS,, ( ) :, CO 2,,, PS psba psbd, PS, psba psbd,,, PS, PS, psba psbd, PS D1 D2 mrna, psba psbd ( 2) Kapoor [23], psba psbd,, D1 D2 PS, psba psbd mrna, D1, D1, [24-25], D1, PS D1 Synechocystis sp. PCC6803, PS [26], psba mrna D1 [27-28], psba mrna psba psbd, D1, psba mrna D1, psba psbd mrna,, Dl PS psbd Chlamydomonas reinhardtii, D1 mrna, D2, Dl [29] ;, psba Syneehocystis 6803, Dl, D2 [30] PS, (RWC 71%) (RWC 34%) psba psbd 2.29 (P < 0.05) 5 (P < 0.01), psbd, PS psba mrna, psbd mrna psba psbd : [1] Noda H. Health benefits and nutritional properties of 64 / 2011 / 35 / 4

5 nori [J]. J Appl Phycol, 1993, 5: [2],. [M]. :, 1996: [3] Lipkin Y, Beer S, Eshel A. The ability of Porphyra linearis (Rhodophyta) to tolerate prolonged periods of desiccation [J]. Bot mar, 1993, 36: [4] Lin A P, Wang G C, Yang F, et al. Photosynthetic parameters of sexually different parts of Porphyra katadai var. hemiphylla (Bangiales, Rhodophyta) during dehydration and re-hydration [J]. Planta, 2009, 229: [5] Quadir A, Harrison P J, Dewreede R E. The effects of emergence and submergence on the photosynthesis and respiration of marine macrophytes [J]. Phycologia, 1979, 18: [6] Johnston A M, Raven J A. The analysis of photosynthesis in air and water of Ascophyllum nodosum (L.) Le Jol. [J]. Oecologia, 1986, 69: [7] Madsen T V, Maberly S C. A comparison of air and water as environments for photosynthesis by the intertidal alga Fucus spiralis (Phaeophyta) [J]. J Phycol, 1990, 26(1): [8] Sampath-Wiley P, Neefus C D, Jahnke L S. Seasonal effects of sun exposure and emersion on intertidal macroalgae physiology: Fluctuations in antioxidant contents, photosynthetic pigments and photosynthetic efficiency in the red alga Porphyra umbilicalis Kützing (Rhodophyta, Bangiales) [J]. J Exp Mar Biol Ecol, 2008, 361(2): [9] Hodgson L M. Photosynthesis of the red alga Gastroclonium coulteri (Rhodophyta) in response to changes in temperature, light intensity, and desiccation[j]. J Phycol, 1981, 17(1): [10] Zou D H, Gao K S. Effects of desiccation and CO 2 concentrations on emersed photosynthesis in Porphyra haitanensis (Bangiales, Rhodophyta), a species farmed in China[J]. Eur J Phycol, 2002, 37: [11] [D]. :, 2007: [12] Buchanan B B. Biochemistry and Molecular Biology of Plants [M]. Beijing: Science Press, 2000: [13],. psba psbd [J]., 2008, 32(7): [14] Livak K J, Schmittgen T D. Analysis of relative gene expression data using Real-time quantitative PCR and the 2 -ΔΔCT method [J]. Methods, 2001, 25: [15] Havaux M, Canani O, Malkin S. Photosynthetic responses to water stress in leaves expressed by photoacoustics and related methods. II. The effect of rapid drought on the electron transport and relative activities of the two photosystems [J]. Plant Physiol, 1986, 82: [16] Cornic G. Drought stress inhibits photosynthesis by decreasing stomatal aperture not by affecting ATP synthesis [J]. Trends Plant Sci, 2000, 5: [17] Havaux M. Stress tolerance of photosystem II in vivo: antagonistic effects of water, heat, and photoinhibition stresses [J]. Plant Physiol, 1992, 100: [18],,,. psba, psbd cab [J]., 2005, 25(4): [19] Yuan S, Liu W J, Zhang N H, et al. Effects of water stress on major photosystem II gene expression and protein metabolism in barley leaves [J]. Physiol Plantarum, 2005, 125(4): [20] He J X, Wen J Q, Chong K, et al. Changes in transcript levels of chloroplast psba and psbd genes during water stress in wheat leaves [J]. Physiol Plantarum, 2002, 102(1): [21] Collett H, Butowt R, Smith J, et al. Photosynthetic genes are differentially transcribed during the dehydration-rehydration cycle in the resurrection plant, Xerophyta humilis [J]. J Exp Bot, 2003, 54(392): [22] Zhang L X, Li W R, Bi Y R, et al. Water availability affects photosynthetic gene expression in the desert plant Ammopiptanthus mongolicus [J]. Israel J Plant Sci, 2002, 50: [23] Kapoor S, Maheshwari S C, Tyagi A K. Developmental and light-dependent cues interact to establish steady-state levels of transcripts for photosynthesis-related genes(psba, psbd, psaa and rbcl) in rice(oryza sativa L.) [J]. Curr Genet, 1994, 25: [24] Anderson J M, Park Y I, Chow W S. Photoinhibition and photoprotection of photosystem in nature [J] Physiol Plant, 1997, 100: 2l [25] Niyogi K. Photoprotection revisited: genetic and molecular approaches [J]. Annu Rev Plant Physiol Plant Mol Biol, 1999, 50: [26] Yokota A, Takahara K, Akashi K. Water stress. Madhava Rao K V, Raghavendra A S, Janardhan Reddy K (eds.). Physiology and Molecular Biology of Stress Tolerance in Plants[M], Springer Netherlands, 2006: Marine Sciences / Vol. 35, No. 4 /

6 [27] Staub J M, Maliga P. Translation of psba mrna is regulated by light via the 5 -untranslated region in tobacco plastids [J]. Plant J, 1994, 6(4): [28] Hirose T, Sugiura M. Cis-acting elements and trans-acting factors for accurate translation of chloroplast psba mrnas: development of an in vitro translation system from tobacco chloroplasts [J]. EMBO J, 1996, 15(7): [29] Erickson J M, Rahire M, Malnoë P, et al. Lack of the D2 protein in a Chlamydomonas reinhardtii psbd mutant affects photosystem II stability and D1 expression [J]. EMBO J, 1986, 5(8): [30] Nilsson F, Andersson B, Jansson C. Photosystem II characteristics of a constructed Synechocysfis 6803 mutant lacking synthesis of the D1 polypeptide [J]. Plant Mol Biol, 1990, 14: Effect of dehydration on the expressions of psba and psbd genes of Porphyra yezoensis YANG Fang 1, WANG Guang-ce 2, ZHANG Bao-yu 2, CHEN Zhang-fan 2, PAN Guang-hua 1 (1. College of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin ,China; 2. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao , China) Received: Oct., 26, 2010 Key words: Porphyra yezoensis, dehydration and rehydration, psba, psbd, the relative quantification of transcript level Abstract: The transcription changes of psba (encoding D1 protein) and psbd (encoding D2 protein) genes of Porphyra yezoensis during dehydration were determined with real-time PCR. The results showed that the relative quantification of transcript levels of psba and psbd genes generally appeared to vary in a similar manner but their variation ranges differed to some extent. The moderate dehydration of about 71% R WC (relative water content) increases the relative transcript levels for psba and psbd by about 6.18 and 2.13-fold, respectively, suggesting that D1 protein and D2 protein might be differentially synthesized. The significant increases of the relative quantification of transcript levels may be the initial response of P. yezoensis to the air to adjust to the terrestrial environment. ( 本文编辑 : 梁德海 ) 66 / 2011 / 35 / 4

PS II PS II GZX-250BS - III 2 d μmol / m 2 s 12 L /12 D FV MV 1 1 cm 2 50 ml

PS II PS II GZX-250BS - III 2 d μmol / m 2 s 12 L /12 D FV MV 1 1 cm 2 50 ml 34 3 2011 9 JOURNAL OF NANJING NORMAL UNIVERSITY Natural Science Edition Vol 34 No 3 Sept 2011 1 1 2 2 2 2 1 210046 2 215500 Porphyra katadai var hemiphylla FV MV 5 ~ 20 10 80 μmol / m 2 s 20 d Yield F

Microsoft Word - 杨 芳_news_.doc

Microsoft Word - 杨芳_news_.doc