Chinese Journal OF Rice Science >
Genetic Mapping and Proteomic Analysis of the Dwarf and Low-tillering Mutant dlt3 in Rice
Received date: 2018-01-07
Revised date: 2018-02-10
Online published: 2018-11-10
【Objective】Plant height is an important agronomic trait for crops because dwarf cultivars are more resistant to lodging. Many factors are responsible for dwarfism in plants, but gibberellin (GA) and brassinosteroid (BR) are the most intensely studied factors, since their molecular mechanism in regulating plant height is important for both basic and applied researches of plant breeding. 【Method】In the present study, we reported the characterization and genetic mapping of a dwarf and low-tillering mutant, dlt3, gained through gamma ray mutagenesis of O. sativa indica 9311. The agronomic traits of the dlt3 mutant including plant height, tiller number, lamina inclination, leaf length and width, and seed-setting rate were analyzed by morphological observation and statistics. The responses to GA and BR were detected based on the changes of leaf angle by exogenous application of BR and GA-induced α-amylase activity. The F2 population and molecular markers were used for genetic mapping of the dlt3 gene. Furthermore, isobaric tags for the relative and absolute quantitation (iTRAQ)-based proteomic method were applied to determine the proteomics of the dlt3 mutant as compared with wild type. 【Result】The plant height, tiller number, leaf angle, leaf length and seed-setting rate were significantly reduced in dlt3. The mutant also showed increased width of leaf blades, dark green and crinkled leaves. The mutant showed normal response to GA, but no response to exogenous BR. Genetic analysis revealed that the dlt3 phenotype was controlled by a single recessive nuclear gene. The dlt3 gene was mapped on the short arm of chromosome 6 between molecular markers RM2615 and R6M14. The iTRAQ-based proteomic analysis revealed that a total of 330 proteins were differentially expressed in the dlt3 mutant, including 222 up-regulated proteins and 108 down-regulated proteins. It showed that four proteins were directly involved in BR signaling pathway and several proteins involved in plant height and developmental regulation were differentially expressed in the mutant. Furthermore, many proteins especially some kinase and phosphatase related proteins, Ca2+ binding related proteins and zinc finger containing proteins were significantly enriched in the mutant. 【Conclusion】These results indicate that the dlt3 is a BR-insensitive dwarf and low-tillering mutant. The DLT3 locus, whose mutation results in abnormal BR signaling, could play important roles in regulating both plant height and other aspects of rice growth and development.
Key words： rice; dwarf and low-tillering; dlt3; brassinosteroid (BR); genetic mapping; proteomics
Minjuan ZHANG, Shuaijun LI, Qiongqiong CHEN, Xiuqing JING, Kunming CHEN, Chunhai SHI, Wenqiang LI . Genetic Mapping and Proteomic Analysis of the Dwarf and Low-tillering Mutant dlt3 in Rice[J]. Chinese Journal OF Rice Science, 2018 , 32(6) : 529 -537 . DOI: 10.16819/j.1001-7216.2018.8001
| [1] | Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush G S, Kitano H, Matsuoka M.Green revolution: A mutant gibberellin-synthesis gene in rice.Nature, 2002, 416(6882): 701-702. |
| [2] | Hedden P.The genes of the green revolution.Trends Genet, 2003, 19(1): 5-9. |
| [3] | Smith S M, Li C, Li J.1-Hormone function in plants.Horm Metabol Signal Plants, 2017: 1-38. |
| [4] | Santner A, Calderon-Villalobos L I A,Estelle M. Plant hormones are versatile chemical regulators of plant growth.Nat Chem Biol, 2009, 5(5): 301-307. |
| [5] | Hedden P, Phillips A L.Gibberellin metabolism: New insights revealed by the genes.Trends Plant Sci, 2000, 5(12): 523-530. |
| [6] | Clouse S D, Sasse J M.Brassinosteroids: Essential regulators of plant growth and development.Annu Rev Plant Physiol Plant Mol Biol, 1998, 49(49): 427-451. |
| [7] | Lanahan M B, Ho T H (1988) Slender barley: A constitutive gibberellin-response mutant.Planta, 1988, 175(1): 107-114. |
| [8] | Tong H N, Jin Y, Liu W B, Li F, Fang J, Yin Y H, Qian Q, Zhu L H, Chu C C.DWARF AND LOW-TILLERING, a new member of the GRAS family, plays positive roles in brassinosteroid signaling in rice. Plant J, 2009, 58(5): 803-816. |
| [9] | Murray M G, Thompson W F.Rapid isolation of high molecular weight plant DNA.Nucl Acids Res, 1980, 8(19): 4321-4325. |
| [10] | Wang Z Q, Xu X Y, Gong Q Q, Xie C, Fan W, Yang J L, Lin Q S, Zheng S J.Root proteome of rice studied by iTRAQ provides integrated insight into aluminum stress tolerance mechanisms in plants.J Proteomics, 2014, 98(4): 189-205. |
| [11] | 李磊, 薛芗, 左示敏, 陈宗祥, 张亚芳, 李前前, 朱俊凯, 马玉银, 潘学彪, 潘存红. 抑制OsAGO1a基因的表达导致水稻叶片近轴面卷曲. 中国水稻科学, 2013, 27(3): 223-230. |
| [11] | Li L, Xue X, Zuo S M, Chen Z X, Zhang Y F, Li Q Q, Zhu J K, Ma Y Y, Pan X B, Pan C H.Suppressed expression ofOsAGO1a leads to adaxial leaf rolling in rice. Chin J Rice Sci, 2013, 27(3): 223-230. |
| [12] | Wu S Y, Xie Y R, Zhang J J, Ren Y L, Zhang X, Wang J L, Guo X P, Wu F Q, Sheng P K, Wang J, Wu C, Wang H, Huang S, Wan [J]. VLN2 regulates plant architecture by affecting microfilament dynamics and polar auxin transport in rice. Plant Cell, 2015, 27(10): 2829-2845. |
| [13] | Sunohara H, Kawai T, Shimizu-Sato S, Sato Y, Sato K, Kitano H.A dominant mutation of TWISTED DWARF 1 encoding an alpha-tubulin protein causes severe dwarfism and right helical growth in rice. Genes Genet Syst, 2009, 84(3): 209-218. |
| [14] | Li W, Wu J, Weng S, Zhang Y, Zhang D, Shi C.Identification and characterization of dwarf 62, a loss-of-function mutation in DLT/OsGRAS-32 affecting gibberellin metabolism in rice. Planta, 2010, 232(6): 1383-1396. |
| [15] | Tong H N, Liu L C, Jin Y, Du L, Yin Y H, Qian Q, Zhu L H, Chu C C.DWARF AND LOW-TILLERING acts as a direct downstream target of a GSK3/SHAGGY-like kinase to mediate brassinosteroid responses in rice. Plant Cell, 2012, 24(6): 2562-2577. |
| [16] | Hirano K, Yoshida H, Aya K, Kawamura M, Hayashi M, Hobo T, Sato-Izawa K, Kitano H, Ueguchi-Tanaka M, Matsuoka M.SMALL ORGAN SIZE 1 and SMALL ORGAN SIZE 2/DWARF AND LOW-TILLERING form a complex to integrate auxin and brassinosteroid signaling in rice. Mol Plant, 2017, 10(4): 590-604. |
| [17] | Sun L, Li X, Fu Y, Zhu Z, Tan L, Liu F, Sun X, Sun X, Sun C.GS6, a member of the GRAS gene family, negatively regulates grain size in rice. J Integr Plant Biol, 2013, 55(10): 938-949. |
| [18] | Li D, Wang L, Wang M, Xu Y Y, Luo W, Liu Y J, Xu Z H, Li J, Chong K.EngineeringOsBAK1 gene as a molecular tool to improve rice architecture for high yield. Plant Biotechnol J, 2009, 7(8): 791-806. |
| [19] | Tanaka A, Nakagawa H, Tomita C, Shimatani Z, Ohtake M, Nomura T, Jiang C, Dubouzet J G, Kikuchi S, Sekimoto H, Yokota T, Asami T, Kamakura T, Mori M.BRASSINOSTEROID UPREGULATED1, encoding a helix-loop-helix protein, is a novel gene involved in brassinosteroid signaling and controls bending of the lamina joint in rice. Plant Physiol, 2009, 151(2): 669-680. |
| [20] | Sakamoto T, Morinaka Y, Inukai Y, Kitano H, Fujioka S.Auxin signal transcription factor regulates expression of the brassinosteroid receptor gene in rice. Plant J, 2013, 73(4): 676-688. |
| [21] | Purwestri Y A, Ogaki Y, Tamaki S, Tsuji H, Shimamoto K.The 14-3-3 protein GF14c acts as a negative regulator of flowering in rice by interacting with the florigen Hd3a.Plant Cell Physiol, 2009, 50(3): 429-438. |
| [22] | Bai M Y, Zhang L Y, Gampala S S, Zhu S W, Song W Y, Chong K, Wang Z Y.Functions of OsBZR1 and 14-3-3 proteins in brassinosteroid signaling in rice.Proc Natl Acad Sci USA, 2007, 104(34): 13839-13844. |
| [23] | Boonburapong B, Buaboocha T.Genome-wide identification and analyses of the rice calmodulin and related potential calcium sensor proteins. .BMC Plant Biol, 2007, 7(1): 4. |
/
| 〈 |
|
〉 |