水稻OsENO2-2基因过表达对水稻抽穗期的影响
收稿日期: 2017-05-18
修回日期: 2017-09-15
网络出版日期: 2018-01-10
基金资助
国家自然科学基金资助项目(31600981);江汉大学科研启动项目(1019-06210002)
Overexpression of OsENO2-2 Affects Heading Date in Rice
Received date: 2017-05-18
Revised date: 2017-09-15
Online published: 2018-01-10
【目的】 OsENO2-2是OsENO2通过可变剪切产生的短转录本,其cDNA序列从粳稻中花11中分离获得。本研究的主要目的是初步分析OsENO2-2在水稻抽穗期调控中的作用。【方法】 构建了OsENO2-2的超量表达载体,获得转基因植株,通过对表型的观察和统计,分析目的基因在过量表达条件下的功能,并利用反向遗传学方法验证该基因的功能。【结果】 OsENO2-2过量表达导致水稻在长日照条件下的抽穗期推迟,而短日照条件下抽穗期无明显变化。通过qRT-PCR方法对水稻开花关键基因的检测发现,在长日照条件下,RFT1的表达量在超表达材料中显著下调,其他开花重要基因表达量在野生型和超表达材料中没有显著变化;而在短日照条件下,所有检测基因的表达量在野生型和超表达材料中均没有显著变化。【结论】 在长日照条件下,OsENO2-2主要通过调控RFT1基因的表达来调控水稻抽穗期。
刘小云, 李晓, 李腾飞, 苏鲁方 . 水稻OsENO2-2基因过表达对水稻抽穗期的影响[J]. 中国水稻科学, 2018 , 32(1) : 12 -22 . DOI: 10.16819/j.1001-7216.2018.7055
【Objective】 OsENO2-2 was isolated from japonica cultivar Zhonghua 11 as a short transcript produced by alternative splicing of OsENO2 which is homologous to AtENO2. To reveal the function of OsENO2-2,【Method】 we constructed an overexpression vector of OsENO2-2 and obtained the transgenetic plants. Based on the observation and statistics of phenotype in overexpressed plant and wild-type, we validated the function of OsENO2-2 using reverse genetics method. 【Result】 Overexpression of OsENO2-2 transgenetic plants delayed heading date only under long-day conditions but not short-day conditions. qRT-PCR expression analysis of the critical flowering time genes showed that in long-day conditions, the expression of RFT1 in overexpressed plants was downregulated, and the expression levels of the other tested genes did not change significantly in wild-type and overexpressed plants; but in short-day conditions, the expression of all the tested genes in wild-type and over-expression plants did not change significantly. 【Conclusion】 OsENO2-2 might be involved in the photoperiodic pathways to regulate the heading date in rice by regulating the expression of RFT1 gene.
Key words： rice; overexpression; OsENO2-2; enolase gene; heading date
| [1] | 徐铨,奥本裕,王晓雪.水稻开花期调控分子机理研究进展.植物遗传资源学报,2014,15(1):129-136. |
| [1] | Xu Q,Okumoto Y,Wang X E.Research progress on regulatory molecular mechanisms of flowering time in riceJ Plant Genet Resour,2014,15(1):129-136. (in Chinese with English abstract) |
| [2] | Imaizumi T,Kay S.Photoperiodic control of flowering: not only by coincidence.Trends Plant Sci,2006,11(11):550-558. |
| [3] | Itoh H,Nonoue Y,Yano M,Izawa T.A pair of floral regulators sets critical day length forHd3a florigen expression in rice. Nat Genet,2010,42(7):635-638. |
| [4] | Tsuji H,Taoka K I,Shimamoto K.Regulation of flowering in rice: Two florigen genes, a complex gene network, and natural variation.Curr Opin Plant Biol,2011,14(1):45-52. |
| [5] | Yano M,Katayose Y,Ashikari M,Yamanouchi U,Monna L,Fuse T,Baba T,Yamamoto K,Umehara Y,Nagamura Y,Sasaki T. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS. Plant Cell,2000,12(12):2473-2484. |
| [6] | Tamaki S,Matsuo S,Wong H L,Yokoi S,Shimamoto K.Hd3a protein is a mobile flowering signal in rice.Science,2007,316(5827):1033-1036. |
| [7] | Hayama R,Coupland G.The molecular basis of diversity in the photoperiodic flowering responses ofArabidopsis and rice. Plant Physiol,2004,135(2):677-684. |
| [8] | Komiya R,Yokoi S,Shimamoto K.A gene network for long-day flowering activates RFT1 encoding a mobile flowering signal in rice.Development,2009,136(20):3443-3450. |
| [9] | Hayama R,Yokoi S,Tamaki S,Yano M,Shimamoto K.Adaptation of photoperiodic control pathways produces short-day flowering in rice.Nature,2003,422: 719-722. |
| [10] | Kim S L,Lee S,Kim H J,Nam H G,An G.OsMADS51 is a short-day flowering promoter that functions upstream ofEhd1, OsMADS14, and Hd3a. Plant Physiol,2007,145(4):1484-1494. |
| [11] | Xue W,Xing Y,Weng X,Zhao Y,Tang W,Wang L,Zhou H,Yu S,Xu C,Li X,Zhang Q.Natural variation inGhd7 is an important regulator of heading date and yield potential in rice. Nat Genet,2008,40(6):761-767. |
| [12] | Wu C,You C,Li C,Long T,Chen G,Byrne M E,Zhang Q.RID1, encoding a Cys2/His2-type zinc finger transcription factor, acts as a master switch from vegetative to floral development in rice.Proc Natl Acad Sci USA,2008,105(35):12915-12920. |
| [13] | Matsubara K,Yamanouchi U,Wang Z X,Minobe Y,Izawa T,Yano M.Ehd2, a rice ortholog of the maize INDETERMINATE1 gene, promotes flowering by up-regulating Ehd1. Plant Physiol,2008,148(3):1425-1435. |
| [14] | Park S J,Kim S L,Lee S,Je B I,Piao H L,Park S H,Kim C M,Ryu C H,Xuan Y H,Colasanti J,An G,Han C D.RiceIndeterminate 1 (OsId1) is necessary for the expression of Ehd1 (Early heading date 1) regardless of photoperiod. Plant J,2008,56(6):1018-1029. |
| [15] | Lu S J,Wei H,Wang Y,Wang H M,Yang R F,Zhang X B,Tu J M.Overexpression of a transcription factorOsMADS15 modifies plant architecture and flowering time in rice(Oryza sativa L.). Plant Mol Biol Rep,2012,30(6):1461-1469. |
| [16] | Arora R,Agarwal P,Ray S,Singh A,Singh V,Tyagi A K,Kapoor S.MADS-box gene family in rice: Genome-wide identification, organization and expression profiling during reproductive development and stress.BMC Genom,2007,8(1):242. |
| [17] | Pancholi V.Multifunctional α-enolase: Its role in diseases.Cell Mol Life Sci,2001,58: 902-920. |
| [18] | Àngels D R,Anna R B,Ana G M,Roser L A.α-enolase, a multifunctional protein: Its role on pathophysiological situations. J Biomed Biotechnol, 2012(7):1-12. |
| [19] | Kang M,Abdelmageed H,Lee S,Reichert A,Mysore K S,Allen R D.AtMBP-1, an alternative translation product of LOS2, affects abscisic acid responses and is modulated by the E3 ubiquitin ligase AtSAP5.Plant J,2013,76(3):481-493. |
| [20] | Eremina M,Rozhon W,Yang S,Poppenberger B.ENO2 activity is required for the development and reproductive success of plants, and is feedback-repressed by AtMBP-1.Plant J,2015,81(6):895-906. |
| [21] | Sun X,Cao Y,Yang Z,Xu C,Li X,Wang S.Xa26, a gene conferring resistance to Xanthomonas oryzae pv. oryzae in rice, encodes an LRR receptor kinase-like protein. Plant J,2004,37: 517-527. |
| [22] | 卢扬江,郑康乐.提取水稻DNA 的一种简易方法.中国水稻科学,1992,6(1):47-48. |
| [22] | Lu Y J,Zheng K L.A simple method for isolation of rice DNA.Chin J Rice Sci,1992,6(1):47-48. (in Chinese with English abstract) |
| [23] | Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2[-Delta Delta C(T)] method.Methods,2001,25(4):402-408. |
| [24] | Hiei Y,Ohta S,Komari T,Kumashiro T.Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.Plant J,1994,6(2):271-282. |
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