水稻短根白化突变体sra1生理生化分析及基因定位
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摘要
叶色突变体是研究光合作用及叶绿素合成与降解途径的理想材料,有助于了解高等植物叶绿体发育和光合作用的调控机制。利用甲基磺酸乙酯(EMS)处理西农1B,获得一个短根白化突变体sra1 (short radicle and albino 1),从出芽至第三叶期叶片始终为白色,胚根较同时期野生型明显变短。对相同位置的叶片观察发现,西农1B叶肉细胞叶绿体含量丰富且膜系统发育完整,而sra1叶肉细胞液泡化严重,叶绿体数目明显减少或没有,基粒类囊体垛叠松散且稀少。生理生化分析发现sra1叶绿素a、叶绿素b和类胡萝卜素含量接近于零,净光合速率为负值。遗传分析表明该短根白化表型由单个隐性核基因控制,最终将SRA1定位于水稻第3染色体长臂InDel标记Z-20和Z-42间,物理距离约657 kb,是一个未报道的新基因。
The leaf color mutant is an ideal material for studying the process of photosynthesis and the pathways of chlorophyll synthesis and degradation.Studies on rice leaf color mutants are helpful to explain the gene network of chloroplast development and photosynthesis in higher plants.The mutant sra1,derived from the progeny of EMS-treated indica rice Xinong 1 B.Leaves of sra1 were white in color from budding to the third leaf,and the radicle of sra1 was significantly shorter than that of wild type in the same stage.The observation of leaves in the same position showed that in Xinong 1 B,the chloroplast of mesophyllous cells was abundant and the membrane system was fully developed,while in sra1,the vacuolarization of mesophyll cells was serious,the number of chloroplasts was significantly reduced or absent,and the granule thylakoids were loosely folded.The contents of chlorophyll a,chlorophyll b,and carotenoid in sra1 were close to zero,and the net photosynthetic rate was negative.Genetic analysis indicated that the short-root and albino phenotype was controlled by a single recessive nuclear gene,and SRA1 was localized between the long-arm InDel markers Z-20 and Z-42 of rice chromosome 3.The physical distance between the two InDel markers was about 657 kb.No genes related to chloroplast and root development have been reported in this interval,indicating that sra1 is a novel mutant.The sra1 is a novel mutant with albino and accompanying short roots,suggesting that SRA1 may be involved in regulating chloroplast and root development.
The leaf color mutant is an ideal material for studying the process of photosynthesis and the pathways of chlorophyll synthesis and degradation.Studies on rice leaf color mutants are helpful to explain the gene network of chloroplast development and photosynthesis in higher plants.The mutant sra1,derived from the progeny of EMS-treated indica rice Xinong 1 B.Leaves of sra1 were white in color from budding to the third leaf,and the radicle of sra1 was significantly shorter than that of wild type in the same stage.The observation of leaves in the same position showed that in Xinong 1 B,the chloroplast of mesophyllous cells was abundant and the membrane system was fully developed,while in sra1,the vacuolarization of mesophyll cells was serious,the number of chloroplasts was significantly reduced or absent,and the granule thylakoids were loosely folded.The contents of chlorophyll a,chlorophyll b,and carotenoid in sra1 were close to zero,and the net photosynthetic rate was negative.Genetic analysis indicated that the short-root and albino phenotype was controlled by a single recessive nuclear gene,and SRA1 was localized between the long-arm InDel markers Z-20 and Z-42 of rice chromosome 3.The physical distance between the two InDel markers was about 657 kb.No genes related to chloroplast and root development have been reported in this interval,indicating that sra1 is a novel mutant.The sra1 is a novel mutant with albino and accompanying short roots,suggesting that SRA1 may be involved in regulating chloroplast and root development.
引文
[1]Dario L.Chloroplast research in the genomic age.Trends Genet,2003,19:47-56.
[2]成浩,李素芳,陈明,虞富恋,晏静,刘益民,陈龙安.安吉白茶特异性状的生理生化本质.茶叶科学,1999,19:87-92.Cheng H,Li S F,Chen M,Yan F L,Qi J,Liu Y M,Chen L G.Physiological and biochemical properties of specificity of Anji white tea.Tea Sci,1999,19:87-92(in Chinese with English abstract).
[3]Dong H,Fei G L,Wu C Y,Wu F Q,Sun Y Y,Chen M J,Ren Y L,Zhou K N,Cheng Z J,Wang J L,Jiang L,Zhang X,Guo X P,Lei C L,Su N,Wang H,Wan J M.A rice virescent-yellow leaf mutant reveals new insights into the role and assembly of plastid caseinolytic protease in higher plants.Plant Physiol,2013,162:1867-1880.
[4]李育红,王宝和,戴正元.水稻叶色突变体及其基因定位、克隆的研究进展.江苏农业科学,2011,39(2):34-39.Li Y H,Wang B H,Dai Z Y.Research progress of rice leaf color mutant and its gene localization and cloning.Jiangsu Agric Sci,2011,39(2):34-39(in Chinese with English abstract).
[5]Ishizaki Y,Tsunoyama Y,Hatano K,Ando K,Kato K,Shinmyo A,Kobori M,Takeba G,Nakahira Y,Shiina T A.Nuclear-encoded sigma factor,Arabidopsis SIG6,recognizes sigma-70 type chloroplast promoters and regulates early chloroplast development in cotyledons.Plant J,2005,42:133-144.
[6]Liere K,Maliga P.In vitro characterization of the tobacco rpoBpromoter reveals a core sequence motif conserved between phage-type plastid and plant mitochondrial promoters.EMBO J,1999,18:249-257.
[7]Wu H,Zhang L X.The PPR protein PDM1 is involved in the processing of rpoA pre-mRNA in Arabidopsis thaliana.Chin Sci Bull,2010,55:3485-3489.
[8]Asakura Y,Barkan A.A CRM domain protein functions dually in group I and group II intron splicing in land plant chloroplasts.Plant Cell,2008,19:3864-3875.
[9]Zhang Z,Tan J J,Shi Z Y,Xie Q J,Xing Y,Liu C Q,Chen Q H,Zhu H T,Wang J,Zhang J L,Zhang G Q.Albino leaf1,that encodes the sole octotricopeptide repeat protein is responsible for chloroplast development.Plant Physiol,2016,171:1182-1191.
[10]Liu C H,Zhu H T,Xing Y,Tan J J,Chen X H,Zhang J J,Peng HF,Xie Q J,Zhang Z M.Albino leaf 2 is involved in the splicing of chloroplast group I and II introns in rice.J Exp Bot,2016,67:5339-5347.
[11]Lin Q,Jiang K,Zheng S,Chen H,Zhou X,Gong J,Xu S,Teng Y.Mutation of the rice ASL2 gene encoding plastid ribosomal protein L21 causes chloroplast developmental defects and seedling death.Plant Biol,2015,17:599-607.
[12]Beale S I.Green genes gleaned.Trends Plant Sci,2005,10:309-312.
[13]Bollivar D W.Recent advances in chlorophyll biosynthesis.Photosynth Res,2006,90:173-194.
[14]Nakamura H M M,Hakata M,Ueno O,Nagamura Y,Hirochika H,Takano M,Ichikawa H.Ectopic overexpression of the transcription factor OsGLK1 induces chloroplast development in non-green rice cells.Plant Cell Physiol,2009,50:1933-1949.
[15]Stenbaek A,Jensen P E.Redox regulation of chlorophyll biosynthesis.Phytochemistry,2010,71:853-859.
[16]Eckhardt U,Grimm B,Hortensteiner S.Recent advances in chlorophyll biosynthesis and breakdown in higher plants.Plant Mol Biol,2004,56:1-14.
[17]Kumar A M,Soll D.Antisense HEMA1 RNA expression inhibits heme and chlorophyll biosynthesis in Arabidopsis.Plant Physiol,2000,122:49-56.
[18]Jung K H,Hur J,Ryu C H,Choi Y,Chung Y Y,Miyao A,Hirochika H,An G.Characterization of a rice chlorophyll-deficient mutant using the T-DNA gene-trap system.Plant Cell Physiol,2003,44:463-472.
[19]Zhang H,Li J,Yoo J H,Yoo S C,Cho S H,Koh H J,Seo H S,Paek N C.Rice Chlorina-1 and Chlorina-9 encode ChlD and ChlI subunits of Mg-chelatase,a key enzyme for chlorophyll synthesis and chloroplast development.Plant Mol Biol,2006,62:325-337.
[20]Yang Y L,Xu J,Huang L C,Leng Y J,Dai L P,Rao Y C,Chen L,Wang Y Q,Tu Z J,Hu J,Ren D Y,Zhang G H,Zhu L,Guo L B,Qian Q,Zeng D.PGL,encoding chlorophyllide a oxygenase 1,impacts leaf senescence and indirectly affects grain yield and quality in rice.J Exp Bot,2016,67:1297-1310.
[21]Cao Y Y,Hua D,Yang L N,Wang Z Q,Zhou S C,Yang J C.Effect of heat-stress during meiosis on grain yield of rice cultivars differing in heat-tolerance and its physiological mechanism.Acta Agron Sin,2008,34:2134-2142.
[22]Fang J,Chai C,Qian Q,Li C,Tang J,Sun L,Huang Z,Guo X,Sun C,Liu M.Mutations of genes in synthesis of the carotenoid precursors of ABA lead to pre-harvest sprouting and photo-oxidation in rice.Plant J,2008,54:177-189.
[23]Cheng H,Chen M,Yu F L,Li S F.The variation of pigment-protein complexes in the albescent stage of tea.Plant Physiol,2000,36:300-304.
[24]洪建,徐颖,徐正.植物病毒侵染寄主叶绿体的形态结构变化.电子显微学报,2000,19:335-336Hong J,Xu Y,Xu Z.Morphological changes of chloroplasts by plant vituses.Microscopy,2000,19:335-336(in Chinese with English abstract)
[25]Wellburn A R.The spectral determination of chlorophylls a and b,as well as total carotenoids,using various solvents with spectrophotometers of different resolution.Plant Physiol,1994,144:307-313.
[26]简在友,王文全,孟丽,许桂芳,王秋玲,李卫东,俞敬波.芍药组内不同类群间光合特性及叶绿素荧光特性比较.植物生态学报,2010,34:1463-1471.Jian Z Y,Wang W Q,Meng L,Xu G F,Wang Q L,Li W D,Yu J B.Comparison of photosynthetic characteristics and chlorophyll fluorescence among different taxa in Paeonia lactiflora group.J Plant Ecol-UK,2010,34:1463-1471(in Chinese with English abstract).
[27]唐永凯,贾永义.荧光定量PCR数据处理方法的探讨.生物技术,2008,18(3):89-91.Tang Y K,Jia Y Y.Discussion on data processing method of fluorescence quantitative PCR.Biotechnology,2008,18(3):89-91(in Chinese with English abstract).
[28]Chen T,Zhang Y,Zhao L,Zhu Z,Lin J,Zhang S,Wang C.Fine mapping and candidate gene analysis of a green-revertible albino gene in rice.J Genet Genomics,2009,36:117-123.
[29]孙萌萌,余庆波,张慧琦.控制水稻叶绿体发育基因Os ALB23的定位.植物生理与分子生物学报,2006,32:433-437.Sun M M,Yu Q B,Zhang H Q.Controlling the location of chloroplast developmental gene OsALB23 in rice.J Plant Physiol Mol Biol,2006,32:433-437(in Chinese with English abstract).
[30]程世超,刘合芹,冯世座,赵辉,汪得凯,陶跃之,翟国伟.水稻白化致死突变体abl4的鉴定和基因定位.中国水稻科学,2013,27:240-246.Cheng S C,Liu H Q,Feng S Z,Zhao H,Wang D K,Tao Y Z,Qi G W.Identification and gene mapping of lethal mutant abl4 in rice albino.Chin J Rice Sci,2013,27:240-246(in Chinese with English abstract).
[31]Gong X D,Jiang Q,Xu J L,Zhang J H,Teng S,Lin D Z,Dong YJ.Disruption of the rice plastid ribosomal protein S20 leads to chloroplast developmental defects and seedling lethality.Genes Genome Genet,2013,3:1769-1777.
[32]Jia L Q,Zhang B T,Mao C Z,Li J H,Wu Y R,Wu P,Wu Z C.OsCYT-INV1 for alkaline/neutral invertase is involved in root cell development and reproductivity in rice(Oryza sativa L.).Planta,2008,228:51-59.
[33]Yao S G,Kodama R,Wang H,Ichii M,Taketa S,Yoshida H.Analysis of the rice SHORT-ROOT5 gene revealed functional diversification of plant neutral/alkaline invertase family.Plant Sci,2009,176:627-634.
[2]成浩,李素芳,陈明,虞富恋,晏静,刘益民,陈龙安.安吉白茶特异性状的生理生化本质.茶叶科学,1999,19:87-92.Cheng H,Li S F,Chen M,Yan F L,Qi J,Liu Y M,Chen L G.Physiological and biochemical properties of specificity of Anji white tea.Tea Sci,1999,19:87-92(in Chinese with English abstract).
[3]Dong H,Fei G L,Wu C Y,Wu F Q,Sun Y Y,Chen M J,Ren Y L,Zhou K N,Cheng Z J,Wang J L,Jiang L,Zhang X,Guo X P,Lei C L,Su N,Wang H,Wan J M.A rice virescent-yellow leaf mutant reveals new insights into the role and assembly of plastid caseinolytic protease in higher plants.Plant Physiol,2013,162:1867-1880.
[4]李育红,王宝和,戴正元.水稻叶色突变体及其基因定位、克隆的研究进展.江苏农业科学,2011,39(2):34-39.Li Y H,Wang B H,Dai Z Y.Research progress of rice leaf color mutant and its gene localization and cloning.Jiangsu Agric Sci,2011,39(2):34-39(in Chinese with English abstract).
[5]Ishizaki Y,Tsunoyama Y,Hatano K,Ando K,Kato K,Shinmyo A,Kobori M,Takeba G,Nakahira Y,Shiina T A.Nuclear-encoded sigma factor,Arabidopsis SIG6,recognizes sigma-70 type chloroplast promoters and regulates early chloroplast development in cotyledons.Plant J,2005,42:133-144.
[6]Liere K,Maliga P.In vitro characterization of the tobacco rpoBpromoter reveals a core sequence motif conserved between phage-type plastid and plant mitochondrial promoters.EMBO J,1999,18:249-257.
[7]Wu H,Zhang L X.The PPR protein PDM1 is involved in the processing of rpoA pre-mRNA in Arabidopsis thaliana.Chin Sci Bull,2010,55:3485-3489.
[8]Asakura Y,Barkan A.A CRM domain protein functions dually in group I and group II intron splicing in land plant chloroplasts.Plant Cell,2008,19:3864-3875.
[9]Zhang Z,Tan J J,Shi Z Y,Xie Q J,Xing Y,Liu C Q,Chen Q H,Zhu H T,Wang J,Zhang J L,Zhang G Q.Albino leaf1,that encodes the sole octotricopeptide repeat protein is responsible for chloroplast development.Plant Physiol,2016,171:1182-1191.
[10]Liu C H,Zhu H T,Xing Y,Tan J J,Chen X H,Zhang J J,Peng HF,Xie Q J,Zhang Z M.Albino leaf 2 is involved in the splicing of chloroplast group I and II introns in rice.J Exp Bot,2016,67:5339-5347.
[11]Lin Q,Jiang K,Zheng S,Chen H,Zhou X,Gong J,Xu S,Teng Y.Mutation of the rice ASL2 gene encoding plastid ribosomal protein L21 causes chloroplast developmental defects and seedling death.Plant Biol,2015,17:599-607.
[12]Beale S I.Green genes gleaned.Trends Plant Sci,2005,10:309-312.
[13]Bollivar D W.Recent advances in chlorophyll biosynthesis.Photosynth Res,2006,90:173-194.
[14]Nakamura H M M,Hakata M,Ueno O,Nagamura Y,Hirochika H,Takano M,Ichikawa H.Ectopic overexpression of the transcription factor OsGLK1 induces chloroplast development in non-green rice cells.Plant Cell Physiol,2009,50:1933-1949.
[15]Stenbaek A,Jensen P E.Redox regulation of chlorophyll biosynthesis.Phytochemistry,2010,71:853-859.
[16]Eckhardt U,Grimm B,Hortensteiner S.Recent advances in chlorophyll biosynthesis and breakdown in higher plants.Plant Mol Biol,2004,56:1-14.
[17]Kumar A M,Soll D.Antisense HEMA1 RNA expression inhibits heme and chlorophyll biosynthesis in Arabidopsis.Plant Physiol,2000,122:49-56.
[18]Jung K H,Hur J,Ryu C H,Choi Y,Chung Y Y,Miyao A,Hirochika H,An G.Characterization of a rice chlorophyll-deficient mutant using the T-DNA gene-trap system.Plant Cell Physiol,2003,44:463-472.
[19]Zhang H,Li J,Yoo J H,Yoo S C,Cho S H,Koh H J,Seo H S,Paek N C.Rice Chlorina-1 and Chlorina-9 encode ChlD and ChlI subunits of Mg-chelatase,a key enzyme for chlorophyll synthesis and chloroplast development.Plant Mol Biol,2006,62:325-337.
[20]Yang Y L,Xu J,Huang L C,Leng Y J,Dai L P,Rao Y C,Chen L,Wang Y Q,Tu Z J,Hu J,Ren D Y,Zhang G H,Zhu L,Guo L B,Qian Q,Zeng D.PGL,encoding chlorophyllide a oxygenase 1,impacts leaf senescence and indirectly affects grain yield and quality in rice.J Exp Bot,2016,67:1297-1310.
[21]Cao Y Y,Hua D,Yang L N,Wang Z Q,Zhou S C,Yang J C.Effect of heat-stress during meiosis on grain yield of rice cultivars differing in heat-tolerance and its physiological mechanism.Acta Agron Sin,2008,34:2134-2142.
[22]Fang J,Chai C,Qian Q,Li C,Tang J,Sun L,Huang Z,Guo X,Sun C,Liu M.Mutations of genes in synthesis of the carotenoid precursors of ABA lead to pre-harvest sprouting and photo-oxidation in rice.Plant J,2008,54:177-189.
[23]Cheng H,Chen M,Yu F L,Li S F.The variation of pigment-protein complexes in the albescent stage of tea.Plant Physiol,2000,36:300-304.
[24]洪建,徐颖,徐正.植物病毒侵染寄主叶绿体的形态结构变化.电子显微学报,2000,19:335-336Hong J,Xu Y,Xu Z.Morphological changes of chloroplasts by plant vituses.Microscopy,2000,19:335-336(in Chinese with English abstract)
[25]Wellburn A R.The spectral determination of chlorophylls a and b,as well as total carotenoids,using various solvents with spectrophotometers of different resolution.Plant Physiol,1994,144:307-313.
[26]简在友,王文全,孟丽,许桂芳,王秋玲,李卫东,俞敬波.芍药组内不同类群间光合特性及叶绿素荧光特性比较.植物生态学报,2010,34:1463-1471.Jian Z Y,Wang W Q,Meng L,Xu G F,Wang Q L,Li W D,Yu J B.Comparison of photosynthetic characteristics and chlorophyll fluorescence among different taxa in Paeonia lactiflora group.J Plant Ecol-UK,2010,34:1463-1471(in Chinese with English abstract).
[27]唐永凯,贾永义.荧光定量PCR数据处理方法的探讨.生物技术,2008,18(3):89-91.Tang Y K,Jia Y Y.Discussion on data processing method of fluorescence quantitative PCR.Biotechnology,2008,18(3):89-91(in Chinese with English abstract).
[28]Chen T,Zhang Y,Zhao L,Zhu Z,Lin J,Zhang S,Wang C.Fine mapping and candidate gene analysis of a green-revertible albino gene in rice.J Genet Genomics,2009,36:117-123.
[29]孙萌萌,余庆波,张慧琦.控制水稻叶绿体发育基因Os ALB23的定位.植物生理与分子生物学报,2006,32:433-437.Sun M M,Yu Q B,Zhang H Q.Controlling the location of chloroplast developmental gene OsALB23 in rice.J Plant Physiol Mol Biol,2006,32:433-437(in Chinese with English abstract).
[30]程世超,刘合芹,冯世座,赵辉,汪得凯,陶跃之,翟国伟.水稻白化致死突变体abl4的鉴定和基因定位.中国水稻科学,2013,27:240-246.Cheng S C,Liu H Q,Feng S Z,Zhao H,Wang D K,Tao Y Z,Qi G W.Identification and gene mapping of lethal mutant abl4 in rice albino.Chin J Rice Sci,2013,27:240-246(in Chinese with English abstract).
[31]Gong X D,Jiang Q,Xu J L,Zhang J H,Teng S,Lin D Z,Dong YJ.Disruption of the rice plastid ribosomal protein S20 leads to chloroplast developmental defects and seedling lethality.Genes Genome Genet,2013,3:1769-1777.
[32]Jia L Q,Zhang B T,Mao C Z,Li J H,Wu Y R,Wu P,Wu Z C.OsCYT-INV1 for alkaline/neutral invertase is involved in root cell development and reproductivity in rice(Oryza sativa L.).Planta,2008,228:51-59.
[33]Yao S G,Kodama R,Wang H,Ichii M,Taketa S,Yoshida H.Analysis of the rice SHORT-ROOT5 gene revealed functional diversification of plant neutral/alkaline invertase family.Plant Sci,2009,176:627-634.