水稻花器官发育基因DPS2的鉴定和精细定位
#共同第一作者
收稿日期: 2019-11-15
修回日期: 2019-12-27
网络出版日期: 2020-03-10
基金资助
国家自然科学基金资助项目(31971872, 31661143006);中国农业科学院科技创新工程项目
Identification and Fine Mapping of Defective Pistil and Stamens 2 in Rice
#These authors contributed equally to this work
Received date: 2019-11-15
Revised date: 2019-12-27
Online published: 2020-03-10
【目的】鉴定和克隆花器官发育相关基因,为进一步研究水稻花发育的分子机制奠定基础。【方法】在大田常规种植条件下比较了突变体dps2 (Defective pistil and stamens 2)和野生型春江06的主要农艺性状及花器官形态特征差异;扫描电镜及石蜡切片观察花药结构并用染色法观察花粉和胚囊的育性;利用图位克隆方法进行基因精细定位;qRT-PCR分析了花发育相关基因在野生型和突变体中的表达水平。【结果】dps2突变体抽穗期变长,不能正常扬花,雄蕊和雌蕊皱缩且花药和柱头数目增多;进一步研究发现,dps2突变体花药腔室塌陷,内无可见小孢子,即使部分花药形成腔室,花粉粒也无淀粉积累呈干瘪状。此外,突变体胚囊育性也受到影响;遗传分析表明该突变性状受一对隐性核基因控制,该基因位于第4染色体短臂上91.2 kb的区间内,区间内未见花器官发育相关基因的报道。qRT-PCR检测发现,水稻ABCDE模型中的B类、C类和E类基因的表达在突变体中显著升高。【结论】dps2突变体的雄蕊及雌蕊均发育异常,最终导致完全不育,推测DPS2可能在水稻第3轮雄蕊发育和第4轮雌蕊发育调控中发挥重要作用。
王昌健, 陈龙, 代丽萍, 路雪丽, 贺金立, 杨龙, 胡江, 朱丽, 董国军, 张光恒, 高振宇, 任德勇, 陈光, 沈兰, 张强, 郭龙彪, 钱前, 曾大力 . 水稻花器官发育基因DPS2的鉴定和精细定位[J]. 中国水稻科学, 2020 , 34(2) : 115 -124 . DOI: 10.16819/j.1001-7216.2020.9122
【Objective】 Identification and cloning of organ development-related genes can lay a foundation for further study on the molecular mechanism of rice flower development.【Method】The main agronomic traits and morphological characteristics of floral organs of wild type Chunjing 06 and dps2 (Defective pistil and stamens 2) mutant were compared under conventional planting conditions in field. The structure of anther was observed under a scanning electronic microscope by paraffin sectioning, and the fertility of pollen and embryo sac was observed by staining. Map-based cloning was used for the gene localization. Quantitative RT-PCR was performed to analyze the relative expression of genes associated with flower development in the wild type and the mutant. 【Result】 Longer heading date, abnormal flowering, shriveled pistils and stamens, increased anthers and stigmas were presented in the dps2 mutant. Further study revealed that the anther chamber of the dps2 mutant collapsed, and no microspores were found. Though some anthers had their chambers, there was no starch accumulation in the shriveled pollen grains, and the embryo sac fertility of dps2 was also affected. Genetic analysis indicated that the mutant’s phenotype of dps2 was controlled by a single recessive nuclear gene. The DPS2 gene was mapped to a 91.2 kb region on the short arm of chromosome 4. The expression levels of some genes associated with class B, C and E genes in ABCDE model were up-regulated in the mutant. 【Conclusion】 The abnormal development of stamens and pistils attributed to the complete infertility of the dps2 mutant, which indicated that DPS2 play an important role in third round of stamen development and the fourth round of pistil development.
Key words： rice; dps2; floral organ development; gene mapping
| [1] | Pelaz S, Ditta G S, Baumann E, Wisman E, Yanofsky M F.B and C floral organ identity functions require SEPALLATA MADS-box genes[J]. Nature, 2000, 405(6783): 200-203. |
| [2] | Coen E S, Meyerowitz E M.The war of the whorls genetic interactions controlling flower development[J]. Nature, 1991, 353(6339): 31-37. |
| [3] | Ditta G, Pinyopich A, Robles P, Pelaz S, Yanofsky M F.The SEP4 gene of Arabidopsis thaliana functions in floral organ and meristem identity[J]. Current Biology, 2004, 14(21): 1935-1940. |
| [4] | Causier B, Schwarz-Sommer Z, Davies B.Floral organ identity: 20 years of ABCs[J]. Seminars in Cell and Developmental Biology, 2010, 21(1): 73-79. |
| [5] | Kalika P, Usha V.Double-stranded RNA interference of a rice PI/GLO paralog, OsMADS2, uncovers its second-whorl-specific function in floral organ patterning[J]. Genetics，2003, 165(4): 2301-2305. |
| [6] | Shri R Y, Kalika P, Usha V.Divergent regulatoryOsMADS2 functions control size, shape and differentiation of the highly derived rice floret second-whorl organ. Genetics, 2007, 176(1): 283. |
| [7] | Shan G Y, Shinnosuke O, Mayumi K, Hitoshi Y.Unequal genetic redundancy of rice PISTILLATA orthologs, OsMADS2 and OsMADS4, in lodicule and stamen development[J]. Plant & Cell Physiology, 2008, 49(5): 853. |
| [8] | Nagasawa M, Miyoshi M, Sano Y, Satoh H, Hirano H, Sakai H, Nagato Y.SUPERWOMAN1 and DROOPING LEAF genes control floral organ identify in rice[J]. Development, 2003, (130): 708-718 |
| [9] | Yun D P, Liang W Q, Dreni L D, Yin C S, Zhou Z G, Kater M M, Zhang D B.OsMADS16 genetically interacts with OsMADS3and OsMADS58 in specifying floral patterning in rice[J]. Molecular Plant, 2013, 6(3): 743-756. |
| [10] | Hu Y, Liang W Q, Yin C S, Yang X L, Ping B Z, Li A X, Jia R, Chen M J, Luo Z J, Cai Q, Zhao X X, Zhang D B, Yuan Z.Interactions of OsMADS1 with floral homeotic genes in rice flower development[J]. Molecular Plant, 2015, 8(9): 1366-1384. |
| [11] | Hu L F, Liang W Q, Yin C S, Cui X, Zong J, Wang X, Hu J P, Zhang D B.Rice MADS3 regulates ROS homeostasis during late anther development[J]. Plant Cell, 2011, 23(2): 515-533. |
| [12] | Yamaguchi T, Nagasawa N, Kawasaki S, Matsuoka M, Nagato Y, Hirano H Y.The YABBY gene DROOPING LEAF regulates carpel specification and midrib development in Oryza sativa[J]. Plant Cell, 2004, 16(2): 500-509. |
| [13] | Lopez-Dee ZP, Wittich P, Enrico P M, Rigola D, Del B I, Gorla M S, Kater M M, Colombo L.OsMADS13, a novel rice MADS-box gene expressed during ovule development[J]. Developmental Genetics, 2015, 25(3): 237-244. |
| [14] | Dreni L, Jacchia S, Fornara F, Fornari M, Ouwerkerk P B, An G, Colombo L, Kater M M.The D-lineage MADS-box gene OsMADS13 controls ovule identity in rice[J]. The Plant Journal, 2007, 52(4): 690-699. |
| [15] | Prasad K, Sriram P, Kumar C S, Kushalappa K, Vijayraghavan U.Ectopic expression of rice OsMADS1 reveals a role in specifying the lemma and palea, grass floral organs analogous to sepals[J]. Development Genes & Evolution, 2001, 211(6): 281-290. |
| [16] | Li H F, Liang W Q, Rui D J, Yin C S, Zong J, Kong H Z, Zhang D B.The AGL6-like gene OsMADS6 regulates floral organ and meristem identities in rice[J]. Cell Research, 2010, 20(3): 299. |
| [17] | Cui R F, Han J K, Zhao S Z, Su K M, Feng W, Du X Q, Xu Q J, Kang C, Theissen G, Zheng M.Functional conservation and diversification of class E floral homeotic genes in rice (Oryza sativa)[J]. Plant Journal, 2010, 61(5): 767-781. |
| [18] | Suzaki T, Sato M, Ashikari M, Miyoshi M, Nagato Y, Hirano H Y.The gene FLORAL ORGAN NUMBER1 regulates floral meristern size in rice and encodes a leucine-rich repeat receptor kinase orthologous to Arabidopsis CLAVATA1[J]. Development, 2004, 131(22): 5649-5657. |
| [19] | Suzaki T, Toriba T, Fujimoto M, Tsutsumi N, Kitano H, Hirano H Y.Conservation and diversification of meristem maintenance mechanism in Oryza sativa: Function of the FLORAL ORGAN NUMBER2 gene[J]. Plant & Cell Physiology, 2006, 47(12): 1591-1602. |
| [20] | Li J, Zhang W L, Xia Z H, Jiang G H, Qian Q, Li A L, Cheng Z K, Zhu L H, Long M, Zhai W X.A paracentric inversion suppresses genetic recombination at the FON3 locus with breakpoints corresponding to sequence gaps on rice chromosome 11L[J]. Molecular Genetics & Genomics ,2007, 277(3): 263-272. |
| [21] | 张向前, 邹金松, 朱海涛, 李晓燕, 曾瑞珍. 水稻早熟多子房突变体fon5的遗传分析和基因定位[J]. 遗传, 2008, 30(10): 1349-1355. |
| [21] | Zhang X Q, Zou J S, Zhu H T, Li X Y, Zeng R Z.Genetic analysis and gene mapping of an early flowering and multi-ovary mutant in rice (Oryza sativa L.)[J]. Hereditas, 2008, 30(10): 1349-1355. |
| [22] | 赵福永,王洁雅, 黄显波, 邓则勤, 林成豹, 严寒, 田志宏. 水稻花器官数目突变体fon6的研究初报[J]. 杂交水稻, 2011, 26(2): 52-57. |
| [22] | Zhao F Y, Wang J Y, Huang X B, Deng Z Q, Lin C B, Han Y, Tian Z H.A preliminary study on the floral organ number mutant fon6 in rice[J]. Hybrid Rice, 2011, 26(2): 52-57. |
| [23] | Thomson B, Zheng B, Wellmer F.Floral organogenesis: When knowing your ABCs is not enough[J]. Plant Physiol, 2017, 173(1): 56-64. |
| [24] | Livak K J, Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 2001, 25(4): 402-408. |
| [25] | Zhang D B, Wilson Z A.Stamen specification and anther development in rice[J]. Chinese Science Bulletin, 2009, 54(14): 2342-2353. |
| [26] | Xing Y Z, Zhang Q F.Genetic and molecular bases of rice yield[J]. Annual Review of Plant Biology, 2010, 61(1): 421-442. |
| [27] | Ikeda K, Sunohara H, Nagato Y.Development course of inflorescence and spikelet in rice[J]. Breeding Science, 2004, 54: 147-156. |
| [28] | 吴华茂. 水稻穗粒数相关QTL分析及水稻花器官发育突变体dps1的鉴定[D]. 中国科学院大学,2018. |
| [28] | Wu H M.Studies on rice QTL of grain number per panicle and mutant dps1 defective in floral organ development[D]. University of Chinese Academy of Science, 2018. |
| [29] | Yamaguchi T, Lee D Y, Miyao A, Hirochika H, An G, Hirano H Y.Functional diversification of the two C-class MADS box genes OSMADS3 and OSMADS58 in Oryza sativa[J]. Plant Cell, 2006, 18(1): 15-28. |
| [30] | Moreau F, Thévenon E, Blanvillain R, Lopezvidriero I, Francozorrilla J M, Dumas R, Parcy F, Morel P, Trehin C, Carles C C.The Myb-domain protein ULTRAPETALA1 INTERACTING FACTOR 1 controls floral meristem activities in Arabidopsis[J]. Development, 2016, 143(7): 1108-1119. |
| [31] | Prunet N, Yang W B, Das P, Meyerowitz E M, Jack T P.SUPERMAN prevents class B gene expression and promotes stem cell termination in the fourth whorl of Arabidopsis thaliana flowers[J]. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(27): 7166. |
| [32] | Sharma N, Xin R J, Kim D H, Sung S, Lange T, Huq E.NO FLOWERING IN SHORT DAY (NFL) is a bHLH transcription factor that promotes flowering specifically under short-day conditions in Arabidopsis[J]. Development, 2016, 143(4): 682. |
| [33] | Hugouvieux V, Silva C S, Jourdain A, Stigliani A, Charras Q, Conn V, Conn S J, Carles C C, Parcy F, Zubieta C.Tetramerization of MADS family transcription factors SEPALLATA3 and AGAMOUS is required for floral meristem determinacy in Arabidopsis[J]. Nucleic Acids Research, 2018. |
| [34] | Sommer R J, Retzlaff M, Goerlich K, Sander K, Tautz D.Evolutionary conservation pattern of zinc-finger domains of Drosophila segmentation genes[J]. Proceedings of the National Academy of Sciences of the United States of America, 1992, 89(22): 10782-10786. |
| [35] | Hiratsu K, Ohta M, Matsui K, Ohme-Takagi M.The SUPERMAN protein is an active repressor whose carboxy-terminal repression domain is required for the development of normal flowers[J]. Febs Letters, 2002, 514(2): 351-354. |
| [36] | Sakai H, Medrano LJ, Meyerowitz E M.Role of SUPERMAN in maintaining Arabidopsis floral whorl boundaries[J]. Nature, 1995, 378(6553): 199-203. |
| [37] | Gaiser J C, Robinson-Beers K, Gasser C S.The Arabidopsis SUPERMAN gene mediates asymmetric growth of the outer integument of ovules[J]. Plant Cell, 1995, 7(3): 333-345. |
| [38] | Nibau C, Stilio V S D, Wu H M, Cheung A Y. Arabidopsis and Tobacco SUPERMAN regulate hormone signalling and mediate cell proliferation and differentiation[J]. Journal of Experimental Botany, 2011, 62(3): 949-961. |
| [39] | Uemura A, Yamaguchi N, Xu Y, Wee W Y, Ichihashi Y, Suzuki T, Shibata A, Shirasu K, Ito T.Regulation of floral meristem activity through the interaction of AGAMOUS, SUPERMAN, and CLAVATA3 in Arabidopsis[J]. Plant Reproduction, 2017, 31(1): 89-105. |
/
| 〈 |
|
〉 |