一个水稻“斑马叶”叶色突变体基因zebraleaf2(zl2)的图位克隆

doi:10.3969/j.issn.10017216.2013.03.002

中国水稻科学 ›› 2013, Vol. 27 ›› Issue (3): 231-239.DOI: 10.3969/j.issn.10017216.2013.03.002

一个水稻“斑马叶”叶色突变体基因zebraleaf2(zl2)的图位克隆

刘胜^{1, 2, #} ,魏祥进^{1, #} ,邵高能¹ ,唐绍清^{1, *} ,胡培松^{1, *}

¹中国水稻研究所国家水稻改良中心水稻生物学国家重点实验室, 浙江杭州 310006; ²杭州师范大学生命与环境科学学院, 浙江杭州 310036;

收稿日期:2012-11-20 修回日期:2013-01-25 出版日期:2013-05-10 发布日期:2013-05-10
通讯作者: 唐绍清1, * ,胡培松1, *
基金资助:
国家自然科学基金资助项目(31201193, 31201195); 浙江省自然科学基金资助项目(Y12C130012); 国家863计划资助项目(2011AA10A101); 中央级公益性科研院所基本科研业务费专项(2012RG0021)。

Mapbased Cloning of A ‘Zebra’ Leaf Mutant Gene zl2 in Rice

LIU Sheng ^{1, 2, #} , WEI Xiangjin ^{1, #} , SHAO Gaoneng¹, TANG Shaoqing ^{1, *} , HU Peisong ^{1, *}

¹State Key Laboratory of Rice Biology, Key Laboratory of Rice Biology and Breeding of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China; ²College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China;

Received:2012-11-20 Revised:2013-01-25 Online:2013-05-10 Published:2013-05-10
Contact: TANG Shaoqing1, *, HU Peisong1, *

摘要/Abstract

摘要： 从粳稻品种Asominori组培后代中获得一个稳定遗传的黄绿相间叶色突变体（zebra leaf 2， zl2）。该突变体在苗期表现为黄绿相间的斑马状，分蘖后期斑马叶性状逐渐减弱，到抽穗期叶片逐渐变为淡黄色。与野生型相比，zl2 在3叶期、分蘖盛期、抽穗期及成熟期叶片的叶绿素、类胡萝卜素含量显著降低，成熟后其结实率、千粒重、株高也显著下降。电镜观察结果显示，苗期zl2叶片黄色部分叶肉细胞中叶绿体显微结构发生了明显的异常，而绿色部分与野生型基本一致。遗传分析结果表明，zl2突变性状受一对隐性核基因控制。从zl2与籼稻品种南京11衍生的F2群体中挑选1607株表现为突变性状的分离单株，最终将该突变基因定位于第11染色体约164.3 kb的区域内。基因预测表明该区域内存在13个ORFs，其中ORF12编码一个类胡萝卜素异构酶，序列分析表明突变体中的该基因第10个内含子与第11外显子的交界处碱基A突变为T，导致cDNA发生错误剪切，缺失4个碱基，产生移码突变，并于第395个氨基酸处提前终止。RTPCR分析表明，相对野生型在突变体中ZL2的表达量显著下降，同时叶色相关基因PORA、RbcL、RbcS、 Cab1、Cab2、 psaA、psbA、OsDVR表达量也显著下降，而HEMA1、YGL1、V1、V2、SPP、OsPPR的表达量显著上升。结果表明ZL2在水稻叶绿素合成及叶绿体发育中起着重要作用。

关键词: 叶色突变体;遗传分析, 图位克隆, 表达分析, 水稻

Abstract: A yellow and green zebra leaf mutant zebraleaf 2 (zl2) displayed a distinctive zebra leaf at seedling stage, and the leaves of the mutant gradually turned yellow after heading. Compared with wild type, the chlorophyll and carotenoid contents of zl2 were significantly reduced at trileaf stage, tillering stage, heading stage and mature stage. The seed setting rate, grain weight and plant height of zl2 also decreased significantly. The ultrastructures of chloroplasts in the mesophyll cells of the yellow part of seedling of zl2 have obvious differences from that of wild type. Genetic analysis showed that the phenotype of zl2 was controlled by a single recessive nuclear gene. Total 1607 zebra leaf plants selected from the F2 population of zl2 / Nanjing 11 were used to finemap the mutant gene. Finally, ZL2 gene was narrowed to the 164.3 kb region on chromosome 11. There are 13 ORFs in this region, and a single base mutation was found in the ORF12 which encode a carotenoid isomerase. And as a result, four bases were missed in cDNA of ORF12 of zl2, leading to frameshift and the amino acid premature termination at the 395th. Compared with the wild type, the expression level of ZL2 and the genes involved in the control of leaf color and photosynthesis, such as PORA, RbcL, RbcS， Cab1， Cab2， psaA, psbA and OsDVR decreased significantly in the mutant, while those of HEMA1, YGL1, V1, V2, SPP and OsPPR increase significantly. The results suggested that ZL2 act a very important role in chlorophyll biosynthesis, chloroplast development and photosynthesis in rice.

Key words: leaf mutant, genetic analysis, mapbased cloning, express analysis, rice

中图分类号:

刘胜1, 2, # ,魏祥进1, # ,邵高能1 ,唐绍清1, * ,胡培松1, *. 一个水稻“斑马叶”叶色突变体基因zebraleaf2(zl2)的图位克隆[J]. 中国水稻科学, 2013, 27(3): 231-239.

LIU Sheng1, 2, #, WEI Xiangjin1, #, SHAO Gaoneng1, TANG Shaoqing1, *, HU Peisong1, *. Mapbased Cloning of A ‘Zebra’ Leaf Mutant Gene zl2 in Rice[J]. Chinese Journal of Rice Science, 2013, 27(3): 231-239.

参考文献

\[1\]刘聪利, 魏祥进, 邵高能, 等. 水稻叶色突变分子机制的研究进展. 中国稻米， 2012, 18(4): 1521.

\[2\]何冰, 刘玲珑, 张文伟, 等. 植物叶色突变体. 植物生理学通讯, 2006, 42(1): 19.

\[3\]邢才, 王贵学, 黄俊丽, 等. 植物叶绿素突变体及其分子机理的研究进展. 生物技术通报, 2008(5): 1012.

\[4\]Gustafsson A. The plastid development in various types of chlorophyllmutations. Hereditas, 1942, 28: 483492.

\[5\]Iwata N, Omura T, Satoh H. Linkage studies in rice(Oryza sativa L.)on some mutants for physiological leaf spots. J Fac Agric, Kyushu Univ, 1978, 22: 243251.

\[6\]Khush G S, Kinoshita T. Rice karyotype, marker genes and linkage groups //Khush G S, Torenniessen G H. Rice Biotechnology. Wallingford Oxon, UK: CAB Internantional, 1991: 83108.

\[7\]钱前, 朱旭东, 曾大力, 等. 细胞质基因控制的新特异材料白绿苗的研究. 作物品种资源, 1996（4）: 1112.

\[8\]李国权,李梦,吴殿武，等. 辐射诱发大豆叶绿素缺失突变及其遗传分析. 吉林农业大学学报, 1987, 9(3): 4651.

\[9\]谢戎, 邵继荣, 孙敬三, 等. 水稻温敏叶片间断失绿过程中的氨基酸组分变化. 乐山师范学学报, 2001(4): 2830.

\[10\]陶华, 薛庆中, 田振涛. 淡绿叶标记光温敏核不育水稻叶色及育性的遗传分析. 作物学报, 2005, 31(4): 438433.

\[11\]Chen G, Bi Y R, Li N. EGY1 encodes a membraneassociated and ATPindependent metalloprotease that is required for chloroplast development. Plant J, 2005, 41: 364375.

\[12\]Wu Z M, Zhang X, He B, et al. A chlorophylldeficient rice mutant with impaired chlorophyllide esterification in chlorophyll biosynthesis. Plant Physiol, 2007, 145(1): 2940.

\[13\]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.

\[14\]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.

\[15\]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.

\[16\]Wang P R, Gao J X, Wan C M, 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1003.

\[17\]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.

\[18\]Gothandam K M, Kim E S, Cho H, et al. OsPPR1, a pentatricopeptide repeat protein of rice is essential for the chloroplast biogenesis. Plant Mol Biol, 2005, 58(3): 421433.

\[19\]Yue R Q, Wang X F, Chen J Y, et al. A rice stromal processing peptidase regulates chloplast and root development. Plant Cell Physiol, 2010, 51(3): 475485.

\[20\]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.

\[21\]Zhang H T, Li J J, Yoo J H, et al. Rice Chlorina1 and Chlorina9 encode Ch1D and Ch1I subunits of Mgchelatase, a key enzyme for chlorophyll synthesis and chloroplast development. Plant Mol Biol, 2006, 62(3): 325337.

\[22\]Hu J , Reddy V S, Wessler S R. The rice R gene family: Two distinct subfamilies containing several miniature invertedrepeat transposable elements.Plant Mol Biol, 2000, 42(5): 667678.

\[23\]Hortensteiner S. Chlorophyll degradation during senescence.Ann Rev Plant Biol, 2006, 57: 5557.

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

\[25\]吕典华, 宗学风, 王三根, 等. 两个水稻叶色突变体的光合特性研究.作物学报, 2009, 35(12): 23042308.

\[26\]The mapbased sequence of the rice genome. Nature, 2005, 436(7052): 793800.

\[27\]Wu K S, Tanksley S D. Abundance, polymorphism and genetic mapping of microsatellites in rice.Mol Gen Genet, 1993, 241(1/2): 225235.

\[28\]Chai C L Fang J, Liu Y, et al. ZEBRA2, encoding a carotenoid isomerase, is involved in photoprotection in rice. Plant Mol Biol, 2011， 75(3): 211221.

\[29\]Iwata N, Satoh H, Omura T. The relationships between chromosomes identified cytologically and linkage groups. J Fac Agric, Kyushu Univ, 1984, 1: 128132.

\[30\]McCouch S R, Kochert G, Yu Z H, et al. Molecular mapping of rice chromosome. Theor Appl Genet, 1988, 76: 815829.

\[31\]Wang Q S, Sang X C, Ling Y H, et al. Genetic analysis and molecular mapping of a novel gene for zebra mutation in rice(Oryza sativa L.). J Gene Genom, 2009, 36(11): 679684.

\[32\]Abenes M L P, Tabien R E, Mcouch S R, et al. Orientation and integration of the classical and molecular genetic maps of chromosome 11 in rice. Euphytica, 1994, 76: 8187.

\[33\]Kusumi K, Komori H, Satoh H, et al. Characterization of a zebra mutant of rice with increased susceptibility to ligt stress. Plant Cell Physiol, 2000, 41(2): 158164.

\[34\]何瑞锋, 丁毅, 余金洪, 等. 水稻“斑马叶”叶绿素含量及几种酶活性的变化. 武汉大学学报, 2000, 46(6): 761765.

\[35\]邵继荣, 王玉忠，孙敬三, 等. 水稻温敏型突变体叶片间断失绿的超微结构.植物学报, 1999, 41(1): 2024.

\[36\]Zhang D Y, Gong Z Y, Ye J W, et al. Effects of zebra leaf 1a new variegation mutationon chloroplast development in rice. J Mol Cell Biol, 2008, 41(5): 417422.

\[37\]Li J J, Pandeya D, Nath K, et al. ZEBRANECROSIS, a thylakoidbound protein, is critical for the photoprotection of developing chloroplasts during early leaf development. Plant J, 2010， 62: 713725.

[1]	郭展, 张运波. 水稻对干旱胁迫的生理生化响应及分子调控研究进展[J]. 中国水稻科学, 2024, 38(4): 335-349.
[2]	韦还和, 马唯一, 左博源, 汪璐璐, 朱旺, 耿孝宇, 张翔, 孟天瑶, 陈英龙, 高平磊, 许轲, 霍中洋, 戴其根. 盐、干旱及其复合胁迫对水稻产量和品质形成影响的研究进展[J]. 中国水稻科学, 2024, 38(4): 350-363.
[3]	许丹洁, 林巧霞, 李正康, 庄小倩, 凌宇, 赖美玲, 陈晓婷, 鲁国东. OsOPR10正调控水稻对稻瘟病和白叶枯病的抗性[J]. 中国水稻科学, 2024, 38(4): 364-374.
[4]	候小琴, 王莹, 余贝, 符卫蒙, 奉保华, 沈煜潮, 谢杭军, 王焕然, 许用强, 武志海, 王建军, 陶龙兴, 符冠富. 黄腐酸钾提高水稻秧苗耐盐性的作用途径分析[J]. 中国水稻科学, 2024, 38(4): 409-421.
[5]	胡继杰, 胡志华, 张均华, 曹小闯, 金千瑜, 章志远, 朱练峰. 根际饱和溶解氧对水稻分蘖期光合及生长特性的影响[J]. 中国水稻科学, 2024, 38(4): 437-446.
[6]	刘福祥, 甄浩洋, 彭焕, 郑刘春, 彭德良, 文艳华. 广东省水稻孢囊线虫病调查与鉴定[J]. 中国水稻科学, 2024, 38(4): 456-461.
[7]	陈浩田, 秦缘, 钟笑涵, 林晨语, 秦竞航, 杨建昌, 张伟杨. 水稻根系和土壤性状与稻田甲烷排放关系的研究进展[J]. 中国水稻科学, 2024, 38(3): 233-245.
[8]	缪军, 冉金晖, 徐梦彬, 卜柳冰, 王平, 梁国华, 周勇. 过量表达异三聚体G蛋白γ亚基基因RGG2提高水稻抗旱性[J]. 中国水稻科学, 2024, 38(3): 246-255.
[9]	尹潇潇, 张芷菡, 颜绣莲, 廖蓉, 杨思葭, 郭岱铭, 樊晶, 赵志学, 王文明. 多个稻曲病菌效应因子的信号肽验证和表达分析[J]. 中国水稻科学, 2024, 38(3): 256-265.
[10]	朱裕敬, 桂金鑫, 龚成云, 罗新阳, 石居斌, 张海清, 贺记外. 全基因组关联分析定位水稻分蘖角度QTL[J]. 中国水稻科学, 2024, 38(3): 266-276.
[11]	魏倩倩, 汪玉磊, 孔海民, 徐青山, 颜玉莲, 潘林, 迟春欣, 孔亚丽, 田文昊, 朱练峰, 曹小闯, 张均华, 朱春权. 信号分子硫化氢参与硫肥缓解铝对水稻生长抑制作用的机制[J]. 中国水稻科学, 2024, 38(3): 290-302.
[12]	周甜, 吴少华, 康建宏, 吴宏亮, 杨生龙, 王星强, 李昱, 黄玉峰. 不同种植模式对水稻籽粒淀粉含量及淀粉关键酶活性的影响[J]. 中国水稻科学, 2024, 38(3): 303-315.
[13]	关雅琪, 鄂志国, 王磊, 申红芳. 影响中国水稻生产环节外包发展因素的实证研究：基于群体效应视角[J]. 中国水稻科学, 2024, 38(3): 324-334.
[14]	许用强, 姜宁, 奉保华, 肖晶晶, 陶龙兴, 符冠富. 水稻开花期高温热害响应机理及其调控技术研究进展[J]. 中国水稻科学, 2024, 38(2): 111-126.
[15]	吕海涛, 李建忠, 鲁艳辉, 徐红星, 郑许松, 吕仲贤. 稻田福寿螺的发生、危害及其防控技术研究进展[J]. 中国水稻科学, 2024, 38(2): 127-139.

一个水稻“斑马叶”叶色突变体基因zebraleaf2(zl2)的图位克隆

Mapbased Cloning of A ‘Zebra’ Leaf Mutant Gene zl2 in Rice

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics