任佳佳,洪廷,张宁,黄翔鹄,李长玲

• 1:广东海洋大学水产学院
• 2:广东海洋大学深圳研究院
• 3:广东省藻类养殖及应用工程技术研究中心

摘要(Abstract)：

【目的】研究波吉卵囊藻(Oocystis borgei)在不同盐度下叶绿素荧光参数的变化和编码叶绿体D1蛋白的PsbA基因的表达特性。【方法】利用叶绿素荧光仪测定不同盐度条件下波吉卵囊藻叶绿素荧光参数;利用cDNA末端快速扩增技术(RACE)克隆获取波吉卵囊藻PsbA基因序列,并进行生物信息学分析;利用实时荧光定量PCR(qPCR)方法分析PsbA基因在不同盐度下的表达情况。【结果】高盐(盐度60)条件下PSⅡ最大光化学量子产量(F_v/F_m)及PSII实际光化学量子效率(ΦPSⅡ)显著下降(P <0.05)。克隆获得1 314 bp波吉卵囊藻PsbA基因cDNA序列,包含1 062 bp开放阅读框(ORF),预测编码353个氨基酸,该蛋白属于跨膜蛋白。qPCR结果表明,高盐(盐度60)条件下PsbA基因极显著下调(P <0.01)。【结论】高盐(盐度60)显著抑制波吉卵囊藻PsbA基因的表达和叶绿素荧光参数(F_v/F_m和ΦPSⅡ)的降低,表明波吉卵囊藻处于胁迫状态;而盐度10和30能促进PsbA基因的表达。

关键词(KeyWords)： 波吉卵囊藻;盐度;叶绿素荧光参数;PsbA基因

Abstract:

Keywords:

基金项目(Foundation): 活性微藻制品产业化关键技术的研究和示范(KY20180112)

作者(Author): 任佳佳,洪廷,张宁,黄翔鹄,李长玲

参考文献(References)：

• [1]彭益全.碱蓬和三角叶滨藜幼苗生长、光合特性对不同盐度的响应[J].草业学报, 2012, 21(6):64-74.
• [2]丁兰平,孙国栋,黄冰心,等.温度和盐度对刺枝鱼栖苔(Acanthophora spicifera)(红藻门,松节藻科)生长及其几种光合色素的影响[J].海洋与湖沼, 2013, 44(4):913-918.
• [3] PERCEY W J, MCMINN A, BOSE J, et al. Salinity effects on chloroplast PSII performance in glycophytes and halophytes1[J]. Functional Plant Biology, 2016,43(11):1003-1015.
• [4]刘永慧. NaCl胁迫下二色补血草光保护机制的研究[D].济南：山东师范大学, 2009.
• [5] XIA J R, LI Y J, ZOU D H. Effects of salinity stress on PSII in Ulva lactuca as probed by chlorophyll fluorescence measurements[J]. Aquatic Botany, 2004,80(2):129-137.
• [6]王斌,郑洲,缪锦来,等.南极冰藻类囊体膜光系统研究进展[J].海洋科学进展, 2005, 23(z1):90-94.
• [7] MASLENKOVA L T, ZANEV Y, POPOVA L P.Adaptation to salinity as monitored by PSII oxygen evolving reactions in barley thylakoids[J]. Journal of Plant Physiology, 1993, 142(5):629-634.
• [8]袁霞,张宜辉,黄冠闽,等.不同盐度对鱼藤幼苗生长及光合作用的影响[J].生态科学, 2010, 29(2):102-108.
• [9]李惠芳.莫莫格湿地扁秆藨草对水盐交互作用的生理生态响应特征[D].长春：中国科学院研究生院(东北地理与农业生态研究所), 2013.
• [10]徐严,王文磊,许凯,等.不同盐度胁迫对坛紫菜叶状体生理指标的影响[J].应用海洋学学报, 2018,37(3):380-386.
• [11] APOSTOLOVA E L, RASHKOV G, DANKOV K, et al.Influence of the sanosil-induced oxidative stress on the photosynthetic apparatus of different strains of green algae and cyanobacteria[J]. Indian Journal of Plant Physiology, 2015, 20(4):333-338.
• [12] MARIN K, YU K, DMITRY A L, et al. Gene expression profiling reflects physiological processes in salt acclimation of Synechocystis sp. strain PCC 6803[J].Plant Physiology, 2004, 136(2):3290-3300.
• [13] ALLAKHVERDIEV S I, KINOSHITA M, INABA M, et al. Unsaturated fatty acids in membrane lipids protect the photosynthetic machinery against salt-induced damage in Synechococcus[J]. Plant Physiology, 2001,125(2):1842-1853.
• [14]韩志国.环境胁迫（盐胁，热胁，渗透胁迫）对两种海洋浮游植物的影响[D].广州：暨南大学, 2002.
• [15]邹晓娟.盐胁迫对淡水螺旋藻的生理效应[D].南京:南京农业大学, 2015.
• [16] MULO P, SAKURAI I, ARO E M. Strategies for psb A gene expression in cyanobacteria, green algae and higher plants:From transcription to PSII repair[J].Biochimica et Biophysica Acta(BBA)-Bioenergetics,2012, 1817(1):247-257.
• [17] ARO E M, VIRGIN I, ANDERSSON B. Photoinhibition of Photosystem II. inactivation, protein damage and turnover[J]. Biochimica et Biophysica Acta(BBA)-Bioenergetics, 1993, 1143(2):113-134.
• [18] BAENA-GONZALEZ E, ARO E M. Biogenesis, assembly and turnover of photosystem II units[J]. Philosophical Transactions of the Royal Society of London. Series B:Biological Sciences, 2002, 357(1426):1451-1460.
• [19] NIXON P J, MICHOUX F, YU J, et al. Recent advances in understanding the assembly and repair of photosystem II[J]. Annals of Botany, 2010, 106(1):1-16.
• [20] MAUL J E, LILLY J W, CUI L, et al. The Chlamydomonas reinhardtii plastid chromosome:islands of genes in a sea of repeats[J]. The Plant Cell, 2002, 14(11):2659-2679.
• [21] SMITH D R, LEE R W, CUSHMAN J C, et al. The Dunaliella salina organelle genomes:large sequences,inflated with intronic and intergenic DNA[J]. BMC Plant Biology, 2010, 10(1):83-96.
• [22] BENNOUN P, SPIERER-HERZ M, ERICKSON J, et al.Characterization of photosystem II mutants of Chlamydomonas reinhardii lacking the psbA gene[J].Plant Molecular Biology, 1986, 6(3):151-160.
• [23]吕文兵,刘红涛,薛乐勋.不同盐浓度和光照强度对杜氏盐藻psb A基因表达的影响[J].华中师范大学学报（自然科学版）, 2010, 44(2):116-120.
• [24] ALLAKHVERDIEV S I, NISHIYAMA Y, MIYAIRI S,et al. Salt stress inhibits the repair of photodamaged photosystem II by suppressing the transcription and translation of psbA genes in Synechocystis[J]. Plant Physiology, 2002, 130(3):1443-1453.
• [25] LIU M, HUANG X, ZHANG R, et al. Uptake of urea nitrogen by Oocystis borgei in prawn(Litopenaeus vannamei)aquaculture ponds[J]. Bulletin of Environmental Contamination and Toxicology, 2018,101(5):586-591.
• [26]江东,李长玲,黄翔鹄,等.波吉卵囊藻(Oocystis borgei)对Cu2+、Zn2+吸附研究[J].广东海洋大学学报,2011, 31(6):50-54.
• [27]黄翔鹄,刘梅,周美华,等.波吉卵囊藻对养殖水体溶解态氮吸收规律的研究[J].现代渔业化, 2012,39(3):38-43.
• [28] HUANG X H, LI X J, WANG Y Q, et al. Effects of environmental factors on the uptake rates of dissolved nitrogen by a salt-water green alga(Oocystis borgei Snow)[J]. Bulletin of Environmental Contamination and Toxicology, 2012, 89(4):905-909.
• [29]张晓琴,张宁,黄翔鹄,等.波吉卵囊藻遗传转化体系选择标记的筛选[J].广东海洋大学学报, 2018,38(6):38-44.
• [30]王帅,梁英.盐胁迫对盐生杜氏藻生长及叶绿素荧光特性的影响[J].中国海洋大学学报, 2015, 45(3):33-38.
• [31]欧阳叶新,施定基,黄开耀,等.鱼腥藻7120响应NaCl胁迫的光合特性[J].水生生物学报, 2003(1):74-77.
• [32]葛江丽,姜闯道.盐胁迫下钾离子对甜高粱碳同化和光系统Ⅱ的影响[J].东北农业大学学报, 2012, 43(7):70-74.
• [33]刘家尧,衣艳君,张承德,等.活体叶绿素荧光诱导动力学及其在植物抗盐生理研究中的应用[J].曲阜师范大学学报, 1997, 23(4):80-83.
• [34]赵会杰,邹琦,于振文.叶绿素荧光分析技术及其在植物光合机理研究中的应用[J].河南农业大学学报,2000, 34(3):248-251.
• [35] KUMAR K S, HAN T. Toxicity of single and combined herbicides on PSII maximum efficiency of an aquatic higher plant, Lemna sp.[J]. Toxicology Environmental Health Sciences, 2011, 3(2):97-105.
• [36] BAKER N R. Chlorophyll fluorescence:A probe of photosynthesis in vivo[J]. Annu Review of Plant Biology, 2008, 59(1):89-113.
• [37] LI G, WAN S W, ZHOU J, et al. Leaf chlorophyll fluorescence, hyperspectral reflectance, pigments content, malondialdehyde and proline accumulation responses of castor bean(Ricinus communis L.)seedlings to salt stress levels[J]. Industrial Crops Products, 2010, 31(1):13-19.
• [38]毕永红,邓中洋,胡征宇,等.发状念珠藻对盐胁迫的响应[J].水生生物学报, 2005, 29(2):125-129.
• [39]欧阳叶新,施定基,黄开耀,等.鱼腥藻7120响应NaCl胁迫的光合特性[J].水生生物学报, 2003, 27(1):74-77.
• [40]李晓,冯伟,曾晓春.叶绿素荧光分析技术及应用进展[J].西北植物学报, 2006, 26(10):2186-2196.
• [41] JUNEAU P, DEWEZ D, MATSUI S, et al. Evaluation of different algal species sensitivity to mercury and metolachlor by PAM-fluorometry[J]. Chemosphere,2001, 45(4-5):589-598.
• [42]江灵芝,孙雪,王玮蔚,等.盐度对蛋白核小球藻生长、叶绿素荧光参数及代谢酶的影响[J].宁波大学学报, 2013, 26(3):6-10.
• [43]冯力霞.环境胁迫对4株微藻叶绿素荧光特性的影响[D].青岛：中国海洋大学, 2006.
• [44]梁英,冯力霞,田传远,等.盐胁迫对塔胞藻生长及叶绿素荧光动力学的影响[J].中国海洋大学学报,2006, 36(5):726-732.
• [45]黄翔鹄.对虾高位池水环境养殖污染和浮游微藻生态调控机制研究[D].上海：东华大学, 2013.
• [46]于姗姗,王青林,孟宪亮,等.盐度骤变对仿刺参hsp70及hsp90基因表达的影响[J].中国海洋大学学报, 2012, 42(9):22-27.
• [47]钟珍萍,吴乃虎.植物光合基因psb A的结构及表达调控综述[J].福建农业大学学报, 1997, 26(3):257-261.
• [48] EVANS T M, TUTHILL D E, BROWN G K, et al.Monophyly and phylogenetic relationships in Lymania(Bromeliaceae:Bromelioideae)based on morphology and chloroplast DNA sequences[J]. Systematic Botany,2007, 32(2):264-270.
• [49] SUMMERFIELD T C, TOEPEL J, SHERMAN L A.Low-oxygen induction of normally cryptic psbA genes in Cyanobacteria[J]. Biochemistry, 2008, 47(49):12939-12941.
• [50] KóS P B, DEáK Z, CHEREGI O, et al. Differential regulation of psb A and psb D gene expression, and the role of the different D1 protein copies in the cyanobacterium Thermosynechococcus elongatus BP-1[J]. Biochimica et Biophysica Acta, 2008, 1777(1):74-83.
• [51]孙雪,吴晓微,徐年军,等.蛋白核小球藻psb A基因的克隆及其在自养和异养培养的表达变化[J].水产学报, 2011, 35(10):1469-1474.
• [52] LJUNGDAHL S M. Investigating light-induced psbA translation in Chloroplasts[D]. University of Oregon,2019.
• [53] CHOTEWUTMONTRI P, WILLIAMS-CARRIER R,BARKAN A. Exploring the Link between Photosystem II Assembly and Translation of the Chloroplast psb A m RNA[J]. Plants, 2020, 9(2):152.
• [54]魏慧敏.光抑制诱导的D1蛋白聚合与磷酸化对弱光下光抑制菠菜叶D1蛋白降解的影响[D].成都：四川大学, 2003.