水稻苗期低温白叶突变体cde2的鉴定和基因定位

doi:10.3969/j.issn.1001-7216.2014.02.001

摘要/Abstract

摘要： 从粳稻品种Asominori的组培后代中发现了一个温度敏感的叶绿素缺乏突变体。低温（23℃）条件下该突变体幼苗3叶期前表现为白化表型随后致死，但在正常温度条件下与野生型无明显差异（30℃）。与野生型相比，该突变体幼苗低温条件下叶绿素含量明显下降，叶绿体结构发育异常。遗传分析结果表明，该突变体受一对隐性核基因控制，定名为cde2（chlorophyll deficient 2）基因。从cde2与籼稻品种培矮64衍生的F2群体中挑选1064株表现为突变表型的单株进行基因定位，将该基因初步定位于水稻第1染色体的着丝粒附近，随后利用已有的SSR标记和自行开发的Indel标记，进一步将该基因定位在标记RM11041和Indel1之间，物理距离为365.6 kb。此外，对该突变体叶绿素合成、光合作用以及质体转录/翻译系统相关基因的表达量测定表明，CDE2突变后增加了与叶绿素合成和质体转录/翻译相关基因的表达，但降低了光合作用相关基因的表达。结果表明，CDE2在水稻叶绿素合成以及叶绿体的发育过程中起着重要的作用。

关键词: 水稻, 叶绿素缺乏, 温度敏感突变, 基因定位

Abstract: A thermosensitive chlorophyll deficient mutant was obtained from the progeny of tissue culture of a japonica rice cultivar Asominori. The mutant was characterized by albino leaves before 3leaf stage under low temperature （23℃）, while showing normal phenotype under high temperature（30℃）. The chlorophyll contents of the mutant were significantly lower than those of wildtype before 3leaf stage under low temperature, which is due to altered chloroplast development under low temperature. Genetic analysis showed that the mutation was controlled by a single recessive gene, which is tentatively designed as cde2（chlorophyll deficient 2）. Furthermore, using an F2 population derived from cde2/Peiai 64 and polymorphic markers between cde2 and Peiai 64, the cde2 was finally localized on the centromeric region of chromosome 1 and positioned between SSR markers RM11041 and Indel1, within the physical distance about 365.6 kb. In addition, the expression level of genes associated with chlorophyll biosynthesis, photosynthesis and plastidial transcription/translation apparatus was measured and the results shows that the cde2 mutation increased the transcript level of chlorophyll biosynthesis genes and the platidial transcription/translation apparatus related genes, but reduced the expression level of photosynthesis genes, indicating that CDE2 may involve in the chlorophyll biosynthesis and chloroplast development.

Key words: rice, chlorophyll deficient, thermosensitive mutant, gene mapping

中图分类号:

Q343.5
Q754

魏祥进1,#, 宋建1,2,#,刘胜1,邵高能1,圣忠华1,唐绍清1 ,胡培松1，*. 水稻苗期低温白叶突变体cde2的鉴定和基因定位[J]. 中国水稻科学, 2014, 28(2): 111-118.

WEI Xiangjin1, #, SONG Jian1, 2, #, LIU Sheng1, SHAO Gaoneng1, SHENG Zhonghua1, TANG Shaoqing1 , HU Peisong1,*. Identification and Genetic Mapping of a Thermosensitive White Leaf Mutant in Rice[J]. Chinese Journal of Rice Science, 2014, 28(2): 111-118.

参考文献

\[1\]Tanaka A, Tanaka R. Chlorophyll metabolism. Curr Opin Plant Biol, 2006, 9(3): 248255.

\[2\]Rudiger W. Chlorophyll metabolism: From outer space down to the molecular level. Phytochemistry, 1997, 46(7): 11511167.

\[3\]Eckhardt U, Grimm B, Hortensteiner S. Recent advances in chlorophyll biosynthesis and breakdown in higher plants. Plant Mol Biol, 2004, 56(1): 114.

\[4\]Liu W, Fu Y, Hu G, et al. Identification and fine mapping of a thermosensitive chlorophyll deficient mutant in rice (Oryza sativa L.). Planta, 2007, 226(3): 785795.

\[5\]Nagata N, Tanaka R, Satoh S, et al. Identification of a vinyl reductase gene for chlorophyll synthesis in Arabidopsis thaliana and implications for the evolution of Prochlorococcus species. Plant Cell, 2005, 17(1): 233240.

\[6\]Jung K H, Hur J, Ryu C H, et al. Characterization of a rice chlorophylldeficient mutant using the TDNA genetrap system. Plant Cell Physiol, 2003, 44(5): 463472.

\[7\]Zhang H, Li J, Yoo J H, et al. 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(3): 325337.

\[8\]Wang P, Gao J, Wan C, et al. Divinyl chlorophyll (ide) a can be converted to monovinyl chlorophyll (ide) a by a divinyl reductase in rice. Plant Physiol, 2010, 153(3): 994.

\[9\]Wu Z, Zhang X, He B, et al. A chlorophylldeficient rice mutant with impaired chlorophyllide esterification in chlorophyll biosynthesis. Plant Physiol, 2007, 145(1): 29.

\[10\]Lee S, Kim J H, Yoo E S, et al. Differential regulation of chlorophyll a oxygenase genes in rice. Plant Mol Biol, 2005, 57(6): 805818.

\[11\]Su N, Hu M L, Wu D X, et al. Disruption of a rice pentatricopeptide repeat protein causes a seedlingspecific albino phenotype and its utilization to enhance seed purity in hybrid rice production. Plant Physiol, 2012, 159(1): 227238.

\[12\]Miyoshi K, Ito Y, Serizawa A, et al. OsHAP3 genes regulate chloroplast biogenesis in rice. Plant J, 2003, 36(4): 532540.

\[13\]Kusumi K, Yara A, Mitsui N, et al. Characterization of a rice nuclearencoded plastid RNA polymerase gene OsRpoTp. Plant Cell Physiol, 2004, 45(9): 11941201.

\[14\]Gothandam K M, Kim E S, Cho H J, et al. OsPPR1, a pentatricopeptide repeat protein of rice is essential for the chloroplast biogenesis. Plant Mol Biol, 2005, 58(3): 421433.

\[15\]Chi Y H, Moon J C, Park J H, et al. Abnormal chloroplast development and growth inhibition in rice thioredoxin m knockdown plants. Plant Physiol, 2008, 148(2): 808817.

\[16\]Alberte R S, Hesketh J D, Hofstra G, et al. Composition and activity of the photosynthetic apparatus in temperaturesensitive mutants of higher plants. Proc Natl Acad Sci USA, 1974, 71(6): 24142418.

\[17\]Markwell J P, Danko S J, Bauwe H, et al. A temperaturetensitive thlorophyll bdeficient mutant of sweetclover (Melilotus alba). Plant Physiol, 1986, 81(2): 329334.

\[18\]Markwell J, Osterman J C. Occurrence of temperaturesensitive phenotypic plasticity in chlorophylldeficient mutants of Arabidopsis thaliana. Plant Physiol, 1992, 98(1): 392394.

\[19\]Galova E, Bohmova B, Sevcovicova A. Analysis of some barley chlorophyll mutants and their response to temperature stress. Photosynthetica, 2000, 38(1): 2935.

\[20\]Pasini L, Bruschini S, Bertoli A, et al. Photosynthetic performance of coldsensitive mutants of maize at low temperature. Physiol Plant, 2005, 124(3): 362370.

\[21\]Kusumi K, Sakata C, Nakamura T, et al. A plastid protein NUS1 is essential for buildup of the genetic system for early chloroplast development under cold stress conditions. Plant J, 2011, 68(6): 10391050.

\[22\]Sugimoto H, Kusumi K, Noguchi K, et al. The rice nuclear gene, VIRESCENT 2, is essential for chloroplast development and encodes a novel type of guanylate kinase targeted to plastids and mitochondria. Plant J, 2007, 52(3): 512527.

\[23\]Yoo S C, Cho S H, Sugimoto H, et al. Rice virescent3 and stripe1 encoding the large and small subunits of ribonucleotide reductase are required for chloroplast biogenesis during early leaf development. Plant Physiol, 2009, 150(1): 388401.

\[24\]Lichtenthaler H K. Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes. Method Enzymol, 1987, 148：350382.

\[25\]李超, 林冬枝, 董彦君, 等. 一个水稻苗期温敏感白色条斑叶突变体的遗传分析及基因定位. 中国水稻科学, 2010, 24(3): 223227.

\[26\]Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res, 1980, 8(19): 43214325.

\[27\]Hirochika H, Guiderdoni E, An G, et al. Rice mutant resources for gene discovery. Plant Mol Biol, 2004, 54(3): 325334.

\[28\]Kurata N, Miyoshi K, Nonomura K, et al. Rice mutants and genes related to organ development, morphogenesis and physiological traits. Plant Cell Physiol, 2005, 46(1): 4862.

\[29\]Iba K, Takamiya K I, Toh Y, et al. Formation of functionally active chloroplasts is determined at a limited stage of leaf development in virescent mutants of rice. Dev Genet, 1991, 12(5): 342348.

\[30\]Yatou O, Cheng X Y. Temperature sensitive chlorophyll mutations. Rice Genet Newsl, 1989, 6：131.

\[31\]董彦君, 董文其, 张小明, 等. 突变体 Fan5 苗色低温敏感性状的遗传分析. 中国水稻科学, 1995, 9(4): 249250.

\[32\]舒庆尧, 刘贵付, 夏英武. 温敏水稻叶色突变体的研究. 核农学报, 1996, 10(1): 610.

\[33\]Kusumi K, Mizutani A, Nishimura M, et al. A virescent gene V1 determines the expression timing of plastid genes for transcription/translation apparatus during early leaf development in rice. Plant J, 1997, 12(6): 12411250.

\[34\]Sugimoto H, Kusumi K, Tozawa Y, et al. The virescent2 mutation inhibits translation of plastid transcripts for the plastid genetic system at an early stage of chloroplast differentiation. Plant Cell Physiol, 2004, 45(8): 985996.

\[35\]Hajdukiewicz P T, Allison L A, Maliga P. The two RNA polymerases encoded by the nuclear and the plastid compartments transcribe distinct groups of genes in tobacco plastids. Embo J, 1997, 16(13): 40414048.

\[36\]Maliga P. Two plastid RNA polymerases of higher plants: An evolving story. Trends Plant Sci, 1998, 3:46

\[37\]Krause K, Maier R, Kofer W, et al. Disruption of plastidencoded RNA polymerase genes in tobacco: Expression of only a distinct set of genes is not based on selective transcription of the plastid chromosome. Mol Gene Genet, 2000, 263(6): 10221030.

\[38\]Legen J, Kemp S, Krause K, et al. Comparative analysis of plastid transcription profiles of entire plastid chromosomes from tobacco attributed to wildtype and PEPdeficient transcription machineries. Plant J, 2002, 31(2): 171188.

\[39\]Serino G, Maliga P. RNA Polymerase subunits encoded by the plastid rpogenes are not shared with the nucleusencoded plastid enzyme. Plant Physiol, 1998, 117(4): 11651170.

\[40\]Pfalz J, Liere K, Kandlbinder A, et al. pTAC2, 6, and12 are components of the transcriptionally active plastid chromosome that are required for plastid gene expression. Plant Cell Online, 2006, 18(1): 176197.

\[41\]Hajdukiewicz P T, Allison L A, Maliga P. The two RNA polymerases encoded by the nuclear and the plastid compartments transcribe distinct groups of genes in tobacco plastids. Embo J, 1997, 16(13): 40414048.

\[42\]Silhavy D, Maliga P. Mapping of promoters for the nucleusencoded plastid RNA polymerase (NEP) in the iojap maize mutant. Curr Genet, 1998, 33(5): 340344.

\[43\]Liere K, Maliga P. In vitro characterization of the tobacco rpoB promoter reveals a core sequence motif conserved between phagetype plastid and plant mitochondrial promoters. Embo J, 1999, 18(1): 249257.

\[44\]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(30): 34853489.

\[45\]Zhou W B, Cheng Y X, Yap A, et al. The Arabidopsis gene YS1 encoding a DYW protein is required for editing of rpoB transcripts and the rapid development of chloroplasts during early growth. Plant J, 2009, 58(1): 8296.

\[46\]Ishizaki Y, Tsunoyama Y, Hatano K, et al. A nuclearencoded sigma factor, Arabidopsis SIG6, recognizes sigma70 type chloroplast promoters and regulates early chloroplast development in cotyledons. Plant J, 2005, 42(2): 133144.

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