收稿日期: 2014-09-23
修回日期: 2014-12-10
网络出版日期: 2015-03-10
基金资助
上海市水稻产业技术体系建设项目[沪农科产字(2014)第3号];上海市科委重点科技攻关项目(10391900602);国家自然科学基金资助项目(31101131);国家科技支撑计划重大项目(2011BAD16B03);江苏省农业自主创新资金资助项目[CX(12)1003]
Physiological Characterization and Gene Identification of a Yellow-Green Leaf Mutant ygl11(t) in Rice
Received date: 2014-09-23
Revised date: 2014-12-10
Online published: 2015-03-10
在水稻品种南粳41中发现了一个黄绿叶自然突变体,经过多代连续自交形成了稳定的突变系,命名为ygl11(t),ygl11(t)整个生育期叶片都表现为黄绿色。对苗期、分蘖盛期、齐穗期突变体和野生型的叶绿素含量进行测定,ygl11(t) 的叶绿素含量是野生型的45.7%~74.7%,叶绿素a 含量是野生型的55.2%~87.5%,叶绿素b含量是野生型的12.5%~25.3%,ygl11(t) 的类胡萝卜素的含量是野生型的62.3%~97.0%。ygl11(t)在分蘖盛期的净光合速率显著高于野生型,花后10 d,ygl11(t)的净光合速率比野生型略低。对突变体叶片中叶绿体的超微结构进行观察,发现突变体叶绿体内的类囊体基粒片层数目减少且严重扭曲变形。遗传分析表明,ygl11(t)叶色性状受1对隐性核基因控制。利用SSR分子标记将YGL11(t)初步定位在水稻第10染色体的长臂上,进一步利用新开发的InDel和CAPS标记将YGL11(t)定位在58.1 kb的物理距离内。对该区段内存在的开放阅读框进行序列分析,发现突变体ygl11(t) 中编码叶绿素a氧化酶(chlorophyll a oxygenase 1)基因(OsCAO 1) 的第9个外显子存在2个碱基缺失,从而导致提前出现终止密码子,初步分析OsCAO1即为YGL11(t)的候选基因。
吴书俊, 杨杰, 闫影, 张丽霞, 范方军, 朱金燕, 李文奇, 仲维功, 曹黎明, 王军 . 水稻黄绿叶突变体ygl11(t)的生理特性和基因克隆[J]. 中国水稻科学, 2015 , 29(2) : 111 -118 . DOI: 10.3969/j.issn.1001-7216.2015.02.001
A spontaneous mutant showing yellow-green leaf throughout the growth period, designated as ygl11(t), was screened and identified from rice cultivar Nanjing 41. This mutant was grown for several generations and showed stable mutation phenotype. The ygl11(t) mutant has reduced chlorophyll accumulation and abnormal chloroplast development. The average contents of total chlorophyll, chlorophyll a, chlorophyll b, and carotene in the ygl11(t) mutant were 45.7% to 74.7%, 55.2% to 87.5%, 12.5% to 25.3%, and 62.3% to 97.0% of these in wild type, respectively, at the stages of seedling, tillering and heading. The leaf net photosynthetic rate of the ygl11(t) was significantly higher at maximum tillering stage and slightly lower at 10 days after flowering than that of wild type. Ultrastructure analysis revealed that the grana lamellae of chloroplast thylakoid were less in number and abnormally developed in the ygl11(t) mutant. Genetic analysis suggested that the phenotype of ygl11(t) was controlled by a recessive nuclear gene. YGL11(t)was prelimarily mapped on the long arm of rice chromosome 10, and was finally narrowed to a 58.1 kb region containing six open reading frames (ORFs). All the six ORFs were amplified and sequenced. A 2-bp deletion was found in the 9th exon of the OsCAO1gene, encoding a chlorophyll a oxygenase 1, which results in a premature stop codon and protein truncation in the ygl11(t) mutant. These results suggest that the OsCAO1 gene is the candidate of YGL11(t).
Key words： rice; yellow-green leaf mutant; fine mapping; gene; OsCAO1
| [1] | 黄晓群, 赵海新, 董春林, 等. 水稻叶绿素合成缺陷突变体及其生物学研究进展. 西北植物学报, 2005, 25(8): 1685-1691. |
| [2] | 邓晓娟, 张海清, 王悦, 等. 水稻叶色突变基因研究进展. 杂交水稻, 2012, 27(5): 9-14. |
| [3] | 陈青, 卢芙萍, 徐雪莲. 水稻叶色突变体研究进展. 热带生物学报, 2010, 1(3): 269-281. |
| [4] | 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. |
| [5] | 杜鹏. 水稻黄叶基因YGL3的遗传分析和精细定位[学位论文]. 重庆: 西南农业大学, 2011: 25-26. |
| [6] | 孙小秋, 王兵, 肖云华, 等. 水稻ygl98黄绿叶突变基因的精细定位与遗传分析. 作物学报,2011,37(6): 991-997. |
| [7] | Zhou Y, Gong Z Y, Yang Z F, et al.Mutation of the light-induced yellow leaf 1 gene, which encodes a geranylgeranyl reductase, affects chlorophyll biosynthesis and light sensitivity in rice.Plos One, 2013, 8(9): 1-14. |
| [8] | 王军, 王宝和, 周丽慧, 等. 一个水稻新黄绿叶突变体基因的分子定位. 中国水稻科学, 2006, 20(5):455-459. |
| [9] | 刘梦梦, 桑贤春, 凌英华, 等. 水稻黄绿叶基因YGL4的遗传分析和分子定位. 作物学报, 2009, 35(8):1405-1409. |
| [10] | 吕典华, 宗学凤, 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. |
| [11] | 何颖红. 两个水稻叶色突变体的鉴定和基因定位. 北京:中国农业科学院,2011: 38-39. |
| [12] | 高家旭. 水稻黄绿叶突变体ygl80的遗传分析及基因定位. 成都:四川农业大学, 2010: 29-30. |
| [13] | 李秀兰, 孙小秋, 王平荣, 等. 一个新的水稻黄绿叶突变体的遗传分析与基因定位. 作物学报, 2010, 36(6):1050-1054. |
| [14] | 杨海莲, 刘敏, 郭旻, 等. 一个水稻黄绿叶突变体ygl10的遗传分析和基因定位. 中国水稻科学, 2014, 28(1): 41-48. |
| [15] | 邓晓梅, 叶胜海, 修芬连, 等. 一个水稻黄绿叶突变性状的遗传分析及基因定位. 核农学报, 2012, 26(2): 203-209. |
| [16] | Lichtenthaler H K.Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes.Methods Enzymol, 1987, 148(34): 350-382. |
| [17] | 金磊, 李霞, 魏晓东, 等.不同氮效率粳稻生育后期产量形成的生理基础. 华北农学报, 2013, 28(5): 175-186. |
| [18] | Li X, Cao K, Wang C, et al.Variation of photosynthetic tolerance of rice cultivars(Oryza sativa L.) to chilling temperature in the light.Afric J Biotechnol, 2010, 9(9): 1325-1337. |
| [19] | 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. |
| [20] | Dellaporta S L, Wood J, Hicks J B.A plant DNA minipreparation: Version II.Plant Mol Biol Rep, 1983, 1(4): 19-21. |
| [21] | Shen Y J, Jiang H, Jin J P, et al.Development of genome-wide DNA Polymorphism database for map-based cloning of rice genes.Plant Physiol, 2004, 135(3): 1198-1205. |
| [22] | Michelmore R W, Paran I, Kesseli R V.Identification of markers linked to disease-resistance genes by bulked segregant analysis: A rapid method to detect markers in specific genomic regions by using segregating populations.Proc Natl Acad Sci USA, 1991, 88(21): 9828-9832. |
| [23] | Lander E S, Green P, Abrahamson J, et al.Mapmaker: An interactive computer package for constructing primary genetic linkage maps of experimental and natural populations.Genomics, 1987, 1(2): 174-181. |
| [24] | Fromme P, Melkozernov A, Jordan P, et al.Structure and function of photosystem. I: Interaction with its soluble electron carriers and external antenna systems.FEBS Let, 2003, 555(1):40-44. |
| [25] | Beale S I.Green genes gleaned.Trends Plant Sci, 2005, 10(7): 309-312. |
| [26] | 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. |
| [27] | 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. |
| [28] | 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. |
| [29] | Wang P R, Gao J X, Wan C M, et al.Divinyl chlorophyll(ide) a can be convertedto monovinyl chlorophyll(ide) a by a divinyl reductase in rice.Plant Physiol, 2010, 153(3): 994-1003. |
| [30] | 李燕群,蒲翔,李春梅,等. 水稻507ys黄绿叶突变体的遗传鉴定与候选基因分析. 中国农业科学, 2014,47(2):221-229. |
| [31] | 孔萌萌, 余庆波, 张慧绮, 等. 控制水稻叶绿体发育基因OsALB23的定位. 植物生理与分子生物学学报, 2006, 32(4): 433-437. |
/
| 〈 |
|
〉 |