矮秆小粒水稻潇湘矮的形态学与分子遗传学分析
收稿日期: 2016-11-24
修回日期: 2017-02-06
网络出版日期: 2017-05-10
基金资助
国家自然科学基金资助项目(31471472);国家重点研发计划资助项目(2016YFD0101801);浙江省自然科学基金资助项目(LY14C30009)
Morphological and Molecular Genetic Analysis of a Dwarf and Small Grain Rice Variety Xiaoxiang’ai
Received date: 2016-11-24
Revised date: 2017-02-06
Online published: 2017-05-10
【目的】研究揭示潇湘矮矮秆小粒的遗传机制,为潇湘矮的育种利用提供理论基础。【方法】利用水稻育种过程中自然突变而得到的稳定遗传的矮秆小粒水稻材料潇湘矮,对其进行农艺性状考查、赤霉素(GA3)和油菜素内酯(BR)敏感性分析、遗传学分析,并利用潇湘矮与其近等基因系NIL(NIP)衍生的F2群体对控制矮秆小粒的基因xxa进行图位克隆,最终利用转基因互补试验验证候选基因。【结果】潇湘矮除表现为矮秆小粒外,其穗长变短,穗型紧凑,千粒重极显著下降。不同浓度梯度的赤霉素(GA3)和油菜素内酯(BR)处理,发现潇湘矮对GA3部分敏感,而对BR不敏感。遗传分析发现其符合孟德尔3:1分离规律。图位克隆将xxa基因定位于第5染色体InDel标记F81和F82之间约70 kb区间的物理距离内。该区间包含8个开放阅读框(ORF)。其中,第5个ORF(LOC_Os05g26890)被注释为水稻株高基因D1。序列分析发现,潇湘矮的D1基因在第5和12外显子分别有1个碱基的无义替换和3个碱基的缺失,其中第12外显子3个碱基的缺失导致1个赖氨酸的缺失。转基因互补显示D1可以恢复潇湘矮的表型。【结论】潇湘矮控制株高的途径可能与GA3代谢有关;潇湘矮矮秆小粒表型符合单隐性核基因控制的遗传规律;潇湘矮矮秆小粒性状是由D1基因突变所致。
吕育松, 谢耘丰, 圣忠华, 邬亚文, 唐绍清, 胡培松, 魏祥进 . 矮秆小粒水稻潇湘矮的形态学与分子遗传学分析[J]. 中国水稻科学, 2017 , 31(3) : 238 -246 . DOI: 10.16819/j.1001-7216.2017.6156 238
【Objective】To uncover the genetic mechanism of a dwarf and small grain variety Xiaoxiang’ai (XXA), and lay an important theoretical foundation in rice dwarfing breeding.【Method】Rice variety XXA characterized as a dwarf and small grain phenotype was derived from natural mutation under breeding process. The agronomic traits of XXA were investigated. Its sensibility to phytohormone was analyzed under GA3 and BR treatments at various concentrations. Genetic analysis of XXA with different varieties and fine mapping of xxa gene from the F2 population derived from the cross between near-isogenic lines NIL(NIP) and XXA were done. Furthermore, complementary assay was performed to confirm the candidate gene.【Result】The XXA shows short spikes and compact panicles, with significant decrease in 1000-grain weight under GA3 and BR treatments, we found that XXA is partially sensitive to GA3, whereas insensitive to BR. The segregation behavior in each of the derived F2 populations between XXA and either varieties was consistent with the Mendelian monogenic ratio of 3:1. The xxa gene was finally narrowed down to a 70 kb physical region between the InDel markers F81 and F82 on chromosome 5. Within this region, eight open reading frames (ORFs) were predicted, of which, ORF5 (LOC_Os05g26890) has been annotated as a Dwarf1 gene (D1). Furthermore, sequence analysis of all the region revealed that there are only two mutation sites including one 1-bp nonsense mutation in the 5th exon and a 3-bp deletion that resulted in a lysine deletion in the 12th exon of D1. The complementary assay showed that the abnormal phenotype of XXA can be recovered.【Conclusion】The dwarf and small grain phenotype is probably associated with GA3 metabolism. The abnormal phenotype is mainly controlled by a single recessive gene. The D1 indisputably corresponds to mutation phenotype of XXA.
Key words： rice; dwarf and small grain; gene mapping; mechanized propagation
| [1] | Khush G S.Green revolution: the way forward.Nat Rev Genet, 2001, 2(10): 815-822. |
| [2] | Hu C H.Evaluation of breeding semidwarf rice by induced mutation and hybridization.Euphytica, 1973, 22: 562-574. |
| [3] | 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: 701-702. |
| [4] | Peng J, Richards D E, Hartley N M, Murphy G P, Devos K M, Flintham J E, Beales J, Fish L J, Worland A J, Pelica F, Sudhakar D, Christou P, Snape J W, Gale M D, Harberd N P.“Green revolution” genes encode mutant gibberellin response modulators.Nature, 1999, 400: 256-261. |
| [5] | 张云辉, 张所兵, 林静, 汪迎节, 方先文. 水稻株高基因克隆及功能分析的研究进展. 中国农学通报, 2014, 30: 1-7. |
| [5] | Zhang Y H, Zhang S B, Fang X W, Lin J, Wang Y J, Fang X W.Research progress on cloning and functional analysis of plant height genes in rice(Oryza sativa L.).Chin Agric Sci Bull, 2014, 30: 1-7.(in Chinese with English abstract) |
| [6] | Fujioka S, Yokota T.Biosynthesis and metabolism of brassinosteroids.Annu Rev Plant Biol, 2003, 54: 137-164. |
| [7] | Itoh H, Ueguchi-Tanaka M, Sato Y, Ashikari M, Matsuoka M.The gibberellin signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei.Plant Cell, 2002, 14: 57-70. |
| [8] | Hirano K1, Asano K, Tsuji H, Kawamura M, Mori H, Kitano H, Ueguchi-Tanaka M, Matsuoka M. Characterization of the molecular mechanism underlying gibberellin perception complex formation in rice.Plant Cell, 2010, 22: 2680-2696. |
| [9] | Smith S M, Waters M T.Strigolactones: Destruction- dependent perception?Curr Biol, 2012, 22: 924-927. |
| [10] | Monna L1, Kitazawa N, Yoshino R, Suzuki J, Masuda H, Maehara Y, Tanji M, Sato M, Nasu S, Minobe Y. Positional cloning of rice semidwarfing gene, sd-1: Rice “green revolution gene” encodes a mutant enzyme involved in gibberellin synthesis.DNA Res, 2002, 9: 11-17. |
| [11] | 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. |
| [12] | Ueguchi-Tanaka M, Ashikari M, Nakajima M, Itoh H, Katoh E, Kobayashi M, Chow T Y, Hsing Y I, Kitano H, Yamaguchi I, Matsuoka M.GIBBERELLIN INSENSITIVE DWARF1 encodes a soluble receptor for gibberellin. Nature, 2005, 437: 693-698. |
| [13] | Izawa Y, Takayanagi Y, Inaba N, Abe Y, Minami M, Fujisawa Y, Kato H, Ohki S, Kitano H, Iwasaki Y.Function and expression pattern of the alpha subunit of the heterotrimeric G protein in rice.Plant Cell Physiol, 2010, 51: 271-281. |
| [14] | Kamei T, Matozaki T, Sakisaka T.Coendocytosis of cadherin and c-Met coupled to disruption of cell-cell adhesion in MDCK cells-regulation by Rho, Rac and Rab small G proteins.Oncogene, 1999, 18: 6776-6784. |
| [15] | Wu Y, Fu Y, Zhao S, Gu P, Zhu Z, Sun C, Tan L.CLUSTERED PRIMARY BRANCH 1, a new allele of DWARF11, controls panicle architecture and seed size in rice.Plant Biotechnol J, 2016, 14: 377-386. |
| [16] | Tong H, Liu L, Jin Y, Du L, Yin Y, Qian Q, Zhu L, Chu 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: 2562-2577. |
| [17] | Shi Z.Characterization and cloning of SMALL GRAIN 4, a novel DWARF11 allele that affects brassinosteroid biosynthesis in rice.Chin Sci Bull, 2015, 60: 905-915. |
| [18] | Zhou F, Lin Q, Zhu L, Ren Y, Zhou K, Shabek N, Wu F, Mao H, Dong W, Gan L, Ma W, Gao H, Chen J, Yang C, Wang D, Tan J, Zhang X, Guo X, Wang J, Jiang L, Liu X, Chen W, Chu J, Yan C, Ueno K, Ito S, Asami T, Cheng Z, Wang J, Lei C, Zhai H, Wu C, Wang H, Zheng N, Wan J.D14-SCF(D3)-dependent degradation of D53 regulates strigolactone signalling.Nature, 2013, 504(7480): 406-410. |
| [19] | Jiang L, Liu X, Xiong G, Liu H, Chen F, Wang L, Meng X, Liu G, Yu H, Yuan Y, Yi W, Zhao L, Ma H, He Y, Wu Z, Melcher K, Qian Q, Xu H E, Wang Y, Li J.DWARF 53 acts as a repressor of strigolactone signalling in rice.Nature, 2013, 504: 401-405. |
| [20] | Huang N, Courtois B, Khush G S.Association of quantitative trait loci for plant height with major dwarfing genes in rice.Heredity, 1996, 77: 130-137. |
| [21] | Moncada P, Martínez C P, Borrero J.Quantitative trait loci for yield and yield components in an Oryza sativa × Oryza rufipogon BC2F2 population evaluated in an upland environment.Theor Appl Genet, 2002, 102: 41-52. |
| [22] | 罗炬, 邵高能, 魏祥进, 陈明亮, 唐绍清, 焦桂爱, 谢黎虹, 胡培松. 一个控制水稻株高QTL qPH3的遗传分析. 中国水稻科学, 2012, 26(4): 417-422. |
| [22] | Luo J, Shao G N, Wei X J, Chen M L, Tang S Q, Jiao Q A, Xie L H, Hu P S.Genetic analysis of a QTL qPH3 for plant height in rice.Chin J Rice Sci, 2012, 26(4): 417-422. (in Chinese with English abstract) |
| [23] | 余应弘, 吴云天, 曾翔, 袁隆平. 水稻矮秆小粒突变体潇湘矮的特征特性及其遗传鉴定. 杂交水稻, 2007, 22(6): 67-70. |
| [23] | Yu Y H, Wu Y T, Zeng X, Yuan L P.Characterization and genetic studies on dwarf rice mutant Xiaoxiangai with small grains.Hybrid Rice, 2007, 22(6): 67-70. (in Chinese with English abstract) |
| [24] | 郑贵朝, 胡事君. 提高水稻愈伤组织植株再生能力几种方法的评价. 杂交水稻, 2005, 20(2): 54-57. |
| [24] | Zheng C G, Hu S J.Evaluation of some culture methods for enhancing plant regeneration ability of rice callus.Hybrid Rice, 2005, 20(2): 54-57. (in Chinese with English abstract) |
| [25] | Ashikari M, Wu J, Yano M, Sasaki T, Yoshimura A.Rice gibberellin-insensitive dwarf mutant gene Dwarf 1 encodes the alpha-subunit of GTP-binding protein.Proc Natl Acad Sci USA, 1999, 96: 10284-10289. |
| [26] | Ueguchi-Tanaka M, Fujisawa Y, Kobayashi M.Rice dwarf mutant d1, which is defective in the alpha subunit of the heterotrimeric G protein, affects gibberellin signal transduction. Proc Natl Acad Sci USA, 2000, 97: 11638-11643. |
| [27] | Hartweck L M, Olszewski N E.Rice GIBBERELLIN INSENSITIVE DWARF1 is a gibberellin receptor that illuminates and raises questions about GA signaling.Plant Cell, 2006, 18: 278-282. |
| [28] | Choe S, Dilkes B P, Gregory B D, Ross A S, Yuan H, Noguchi T, Fujioka S, Takatsuto S, Tanaka A, Yoshida S, Tax F E, Feldmann K A.The Arabidopsis dwarf 1 mutant is defective in the conversion of 24-methylenecholesterol to campesterol in brassinosteroid biosynthesis.Plant Physiol, 1999, 119: 897-907. |
| [29] | Ferrero-Serrano A, Assmann S M.The alpha-subunit of the rice heterotrimeric G protein, RGA1, regulates drought tolerance during the vegetative phase in the dwarf rice mutant d1.J Exp Bot, 2016, 67(11): 3433-3443. |
/
| 〈 |
|
〉 |