Phenotypical Analysis and Fine Mapping of a Semi-dominant Dwarfism Gene Si-dd1 in Rice

doi:10.16819/j.1001-7216.2016.5113

Abstract

Abstract:

A semi-dominant dwarf rice mutant (termed as Si-dd1) was obtained from japonica rice variety Nipponbare by tissue culture mutagenesis. Morphological analysis showed that Si-dd1(AA) and Si-dd1(Aa) exhibited shorter plant height, decreased seed-setting rate, delayed heading, increased number of primary rachis branch and secondary rachis branch compared to the wild type (WT,Nipponbare). Physiological test results showed Si-dd1 had similar response to 24-Epibrassinosteroid to the WT, while it was insensitive to Gibberellin at 10^-4μmol/L. Moreover, Western blot results also confirmed GID2 expression level in Si-dd1 differed greatly from WT. The cytological analysis showed that Si-dd1 had smaller stomas in the leaf midrib, increased mesophyll cells and stem vascular bundles compared to WT. Genetic analysis and map-based clone showed that the Si-dd1 is controlled by a single semi-dominant gene, and locates in a 244 kb region on chromosome 6.

Key words: rice, semi-dominant, dwarf, genetic analysis, map-based clone

摘要：

以粳稻品种日本晴在组织培养过程中分离出来的一个半显性矮秆突变体Si-dd1为研究对象,通过形态学分析,发现与野生型日本晴相比,矮秆Si-dd1(AA)和半矮秆Si-dd1(Aa)植株的株高降低。结实率下降,生育期延长,一次枝梗和二次枝梗增加。激素处理结果表明突变体Si-dd1(AA)与野生型对油菜素内酯反应基本相同,而在高浓度赤霉素处理下,突变体Si-dd1(AA)表现为一定程度上的钝感。Western blot对GID2表达量分析也确定这一结果。组织切片实验表明,突变体Si-dd1(AA)相对于野生型叶片主脉气孔变小,叶肉细胞增多,茎维管束数目增加。遗传分析结果表明该突变体基因受一对核基因控制。进一步利用分子标记将该基因定位在水稻第6染色体约244 kb区间内,目前该区段并未发现已报道的矮秆相关基因。

关键词: 水稻, 半显性, 矮秆, 遗传分析, 图位克隆

CLC Number:

Q755
S511.01

Yong-tao CUI, Li-wen WU, Shi-kai HU, De-yong REN, Chang-wei GE, Wei-jun YE, Guo-jun DONG, Long-biao GUO, Xing-ming HU. Phenotypical Analysis and Fine Mapping of a Semi-dominant Dwarfism Gene Si-dd1 in Rice[J]. Chinese Journal OF Rice Science, 2016, 30(2): 152-160.

崔永涛, 吴立文, 胡时开, 任德勇, 葛常伟, 叶卫军, 董国军, 郭龙彪, 胡兴明. 水稻半显性矮秆基因Si-dd1的表型分析和精细定位[J]. 中国水稻科学, 2016, 30(2): 152-160.

Figures/Tables 10

References 36

[1]	Spielmeyer W, Ellis M H, Chandler P M.Semidwarf (sd-1), “green revolution” rice, contains a defective gibberellin 20-oxidase gene.Proc Natl Acad Sci USA, 2002, 99: 9043-9048.
[2]	Yin H F, Gao P, Liu C W, et al.SUI-family genes encode phosphatidylserine synthases and regulate stem development in rice.Planta, 2013, 237: 15-27.
[3]	Komorisono M, Ueguchi-Tanaka M, Aichi I, et al.Analysis of the rice mutant dwarf and gladius leaf 1 , aberrant katanin-mediated microtubule organization causes up-regulation of gibberellin biosynthetic genes independently of gibberellin signaling.Plant Physiol, 2005, 138: 1982-1993.
[4]	Iwamoto M, Kiyota S, Hanada A, et al.The multiple contributions of phytochromes to the control of internode elongation in rice.Plant Physiol, 2011, 157: 1187-1195.
[5]	Toyomasu T, Kagahara T, Hirose Y, et al.Cloning and characterization of cDNAs encoding ent-copalyl diphosphate synthases in wheat: Insight into the evolution of rice phytoalexin biosynthetic genes.Biosc Biotech Biochem, 2009, 73: 772-775.
[6]	Hirano K, Asano K, Tsuji H, et al.Characterization of the molecular mechanism underlying gibberellin perception complex formation in rice.Plant Cell, 2010, 22: 2680-2696.
[7]	Ueguchi-Tanaka M, Nakajima M, Katoh E, et al.Molecular interactions of a soluble gibberellin receptor, GID1, with a rice DELLA protein, SLR1, and gibberellin.Plant Cell, 2007, 19: 2140-2155.
[8]	Oikawa T, Koshioka M, Kojima K, et al.A role of OsGA20ox1, encoding an isoform of gibberellin 20-oxidase, for regulation of plant stature in rice.Plant Mol Biol, 2004, 55: 687-700.
[9]	Hirano K, Kouketu E, Katoh H, et al.The suppressive function of the rice DELLA protein SLR1 is dependent on its transcriptional activation activity.Plant J, 2012, 71: 443-453.
[10]	Yang M F, Qi W W, Sun F, et al.Overexpression of rice LRK1 restricts internode elongation by down-regulating OsKO2.Biotech Lett, 2013, 35: 121-128.
[11]	Ueguchi-Tanaka M, Fujisawa Y, Kobayashi M, et al.Rice dwarf mutant d1, which is defective in the α subunit of the heterotrimeric G protein, affects gibberellin signal transduction.Proc Natl Acad Sci USA, 2000, 97: 11638-11643.
[12]	Wang L, Xu Y, Ma Q, et al.Heterotrimeric G protein α subunit is involved in rice brassinosteroid response.Cell Res, 2006, 16: 916-922.
[13]	Hu X, Qian Q, Xu T, et al.The U-box E3 ubiquitin ligase TUD1 functions with a heterotrimeric G α subunit to regulate brassinosteroid-mediated growth in rice.PLoS Gene, 2013, 9: e1003391.
[14]	Sakamoto T, Morinaka Y, Inukai Y, et al.Auxin signal transcription factor regulates expression of the brassinosteroid receptor gene in rice.Plant J, 2013, 73: 676-688.
[15]	Hong Z, Ueguchi-Tanaka M, Umemura K, et al.A rice brassinosteroid-deficient mutant, ebisu dwarf (d2), is caused by a loss of function of a new member of cytochrome P450.Plant Cell, 2003, 15: 2900-2910.
[16]	Zhi H, Miyako U, Shozo F, et al.The rice brassinosteroid-deficient dwarf2 Mutant, defective in the rice homolog of Arabidopsis DIMINUTO/DWARF1, is rescued by the endogenously accumulated alternative bioactive brassinosteroid, dolichosterone.Plant Cell, 2005, 17(8): 2243-2254.
[17]	Thirumurugan T, Ito Y, Kubo T, et al.Identification, characterization and interaction of HAP family genes in rice.Mol Genet Genom, 2008, 279: 279-289.
[18]	Hong Z, Ueguchi-Tanaka M, Shimizu-Sato S, et al.Loss-of-function of a rice brassinosteroid biosynthetic enzyme, C-6 oxidase, prevents the organized arrangement and polar elongation of cells in the leaves and stem.Plant J, 2002, 32: 495-508.
[19]	Tanabe S, Ashikari M, Fujioka S, et al.A novel cytochrome P450 is implicated in brassinosteroid biosynthesis via the characterization of a rice dwarf mutant, dwarf11, with reduced seed length.Plant Cell, 2005, 17: 776-790.
[20]	Bai M Y, Zhang L Y, Gampala S S, et al.Functions of OsBZR1 and 14-3-3 proteins in brassinosteroid signaling in rice.Proc Natl Acad Sci USA, 2007, 104: 13839-13844.
[21]	Sang D J, Chen D Q, Liu G F, et al.Strigolactones regulate rice tiller angle by attenuating shoot gravitropism through inhibiting auxin biosynthesis.Proc Natl Acad Sci USA, 2014, 111: 11199-11204.
[22]	Ito S, Kitahata N, Umehara M, et al.A new lead chemical for strigolactone biosynthesis inhibitors.Plant Cell Physiol, 2010, 51: 1143-1150.
[23]	Arite T, Umehara M, Ishikawa S, et al.d14, a strigolactone-insensitive mutant of rice, shows an accelerated outgrowth of tillers.Plant Cell Physiol, 2009, 50: 1416-1424.
[24]	Zou J H, Zhang S Y, Zhang W P, et al.The rice HIGH-TILLERING DWARF1 encoding an ortholog of Arabidopsis MAX3 is required for negative regulation of the outgrowth of axillary buds.Plant J, 2006, 48: 687-696.
[25]	Jiang L, Liu X, Xiong G S, et al.DWARF 53 acts as a repressor of strigolactone signalling in rice.Nature, 2014, 506: 401-405.
[26]	Liu W, Kohlen W, Lillo A, et al.Strigolactone biosynthesis in medicago truncatula and rice requires the symbiotic GRAS-type transcription factors NSP1 and NSP2.Plant Cell, 2011, 23: 3853-3865.
[27]	Guo S Y, Xu Y Y, Liu H H, et al.The interaction between OsMADS57 and OsTB1 modulates rice tillering via DWARF14.Nat Comm, 2013, 4(3): 1566.
[28]	Du H, Wu N, Fu J, et al.A GH3 family member, OsGH3-2, modulates auxin and abscisic acid levels and differentially affects drought and cold tolerance in rice.J Exp Bot, 2012, 63: 6467-6480.
[29]	Sazuka T, Kamiya N, Nishimura T, et al.A rice tryptophan deficient dwarf mutant, tdd1, contains a reduced level of indole acetic acid and develops abnormal flowers and organless embryos.Plant J, 2009, 60: 227-241.
[30]	Sato-Izawa K, Nakaba S, Tamura K, et al.DWARF50 (D50), a rice (Oryza sativa L.) gene encoding inositol polyphosphate 5-phosphatase, is required for proper development of intercalary meristem.Plant Cell Environ, 2012, 35: 2031-2044.
[31]	Zhang B C, Liu X L, Qian Q, et al.Golgi nucleotide sugar transporter modulates cell wall biosynthesis and plant growth in rice.Proc Natl Acad Sci USA, 2011, 108: 5110-5115.
[32]	Luan W J, Liu Y Q, Zhang F X, et al.OsCD1 encodes a putative member of the cellulose synthase-like D sub-family and is essential for rice plant architecture and growth.Plant Biotech J, 2011, 9: 513-524.
[33]	Rogers S O, Bendich A J.Extraction of DNA from milligram amountsof fresh, herbarium and mummified plant tissues.Plant Mol Biol, 1985, 5: 69-76.
[34]	Huang L M, Sun Q W, Qin F J, et al.Down-regulation of a SILENT INFORMATION REGULATOR2-related histone deacetylase gene, OsSRT1, induces DNA fragmentation and cell death in rice.Plant Physiol, 2007, 144: 1508-1519.
[35]	Zhang Q, Xu J, Li Y, et al.Morphological, anatomical and genetic analysis for a rice mutant with abnormal hull.J Genet Genom, 2007, 34: 519-526.
[36]	Yang D L, Li Q, Deng Y W, et al.Altered disease development in the eui mutants and Eui overexpressors indicates that gibberellins negatively regulate rice basal disease resistane.Mol Plant, 2008, 1: 528-537.

材料 Material	株高 Plant height /cm	分蘖数 Tiller number	一次枝梗 Number of primary rachis branches	二次枝梗 Number of secondary rachis branches	结实率 Seed-setting rate/%	总粒数 Total grain number
野生型WT	83.8 a	18.7	8.0	9.0 a	88.8 a	72.7 a
半矮秆Si-dd1(Aa)	60.6 b	16.7	8.0	16.3 b	61.3 b	90.7 b
矮秆Si-dd1(AA)	43.7 c	16.3	9.6	16.3 b	1.1 c	97.3 b

材料 Material	株高 Plant height /cm	分蘖数 Tiller number	一次枝梗 Number of primary rachis branches	二次枝梗 Number of secondary rachis branches	结实率 Seed-setting rate/%	总粒数 Total grain number
野生型WT	83.8 a	18.7	8.0	9.0 a	88.8 a	72.7 a
半矮秆Si-dd1(Aa)	60.6 b	16.7	8.0	16.3 b	61.3 b	90.7 b
矮秆Si-dd1(AA)	43.7 c	16.3	9.6	16.3 b	1.1 c	97.3 b

组合 Cross	F₁	F₂				χ²(1:2:1)
组合 Cross	F₁	总株数 Total of plants		正常高秆 Normal plants	半矮秆 Semi-dwarf	矮秆 Dwarf
Si-dd1(AA)/9311	半矮秆 Semi-dwarf	184	48	94	42	0.478
Si-dd1(AA)/NJ06	半矮秆 Semi-dwarf	243	62	122	59	0.078
NJ06/Si-dd1(AA)	半矮秆 Semi-dwarf	193	47	100	46	0.264
9311/Si-dd1(AA)	半矮秆 Semi-dwarf	290	72	150	68	0.455

组合 Cross	F₁	F₂				χ²(1:2:1)
组合 Cross	F₁	总株数 Total of plants		正常高秆 Normal plants	半矮秆 Semi-dwarf	矮秆 Dwarf
Si-dd1(AA)/9311	半矮秆 Semi-dwarf	184	48	94	42	0.478
Si-dd1(AA)/NJ06	半矮秆 Semi-dwarf	243	62	122	59	0.078
NJ06/Si-dd1(AA)	半矮秆 Semi-dwarf	193	47	100	46	0.264
9311/Si-dd1(AA)	半矮秆 Semi-dwarf	290	72	150	68	0.455

组合 Cross	野生型 Wild type	半矮秆 Semi-dwarf	总株数 Total	χ²(1:1)
Si-dd1(Aa)/ 9311	54	48	102	0.353
Si-dd1(Aa)/ NJ06	59	54	113	0.220
9311 /Si-dd1(Aa)	52	45	97	0.505
NJ06 /Si-dd1(Aa)	52	48	100	0.160