欢迎访问作物学报,今天是

作物学报 ›› 2014, Vol. 40 ›› Issue (04): 581-590.doi: 10.3724/SP.J.1006.2014.00581

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

两个水稻品种镉积累相关基因表达及其分子调控机制

黄志熊1,2,王飞娟2,蒋晗2,李志兰3,丁艳菲2,江琼2,陶月良4,朱诚1,2,*   

  1. 1 浙江大学生命科学学院 / 生理生化国家重点实验室, 浙江杭州310058; 2 中国计量学院生命科学学院 / 浙江省生物计量及检验检疫技术重点实验室, 浙江杭州310018; 3 浙江省自然科学基金委员会, 浙江杭州310012; 4 温州大学生命与环境科学学院, 浙江温州325035
  • 收稿日期:2013-09-09 修回日期:2014-01-12 出版日期:2014-04-12 网络出版日期:2014-02-17
  • 通讯作者: 朱诚,E-mail: pzhch@cjlu.edu.cn; Tel: 0571-86914510
  • 基金资助:

    本研究由国家“十二五”科技支撑计划项目(2012BAK17B03)和浙江省自然科学基金项目(Y3110334, LY13C020002, Y2090945)资助。

A Comparison of Cadmium-Accumulation-Associated Genes Expression and Molecular Regulation Mechanism between Two Rice Cultivars (Oryza sativa L. subspecies japonica)

HUANG Zhi-Xiong1,2,WANG Fei-Juan2,JIANG Han2,LI Zhi-Lan3,DING Yan-Fei2,JIANG Qiong2,TAO Yue-Liang4,ZHU Cheng1,2,*   

  1. 1 State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, China; 2 Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China JiLiang University, Hangzhou 310018, China; 3 Nature Science Foundation Committee of Zhejiang Province, Hangzhou 310012, China; 4 College of Life and Environment Sciences, Wenzhou University, Wenzhou 325035, China
  • Received:2013-09-09 Revised:2014-01-12 Published:2014-04-12 Published online:2014-02-17
  • Contact: 朱诚,E-mail: pzhch@cjlu.edu.cn; Tel: 0571-86914510

摘要:

内源小干扰RNAs (small interfering RNAs, siRNAs)DNA甲基化在植物生长发育和适应环境胁迫中调控基因的表达。对于植物来说, (Cadmium, Cd)是一种非必需且具有毒性的元素。为研究DNA甲基化和siRNAs在水稻(Oryza sativa L.) Cd积累相关基因表达调控方面的作用, 比较了Cd高积累品种(秀水110)Cd低积累品种(秀水11)Cd积累相关基因的表达情况。结果表明, 秀水110秀水11叶片中, 除植物Cd抗性蛋白(plant cadmium resistance protein, PCR)基因家族成员OsPCR1的表达水平呈现出显著的差异外, 其他Cd积累相关基因的表达水平不存在显著的差异。说明OsPCR1基因可能参与调控水稻体内Cd的积累。利用荧光实时定量PCR (qRT-PCR)技术研究了水稻叶片中siRNA表达水平在水稻5个不同生长发育期中的变化情况。数据显示水稻叶片中与OsPCR1基因外显子2匹配的siRNA的丰度和OsPCR1基因的表达水平呈负相关。进一步利用McrBC-qRT-PCR技术研究OsPCR1基因外显子2甲基化水平在水稻5个不同生长发育期中的变化情况表明, Cd处理条件下水稻叶片中OsPCR1基因外显子2甲基化水平和该基因的表达水平也呈负相关。说明, 在水稻体内Cd积累的过程中, siRNA和OsPCR1基因外显子2甲基化可能参与调控OsPCR1基因的表达。这些结果对于研究OsPCR1基因的功能和培育Cd低积累水稻品种具有重要的理论指导意义。

关键词: siRNA, DNA甲基化, OsPCR1, 水稻, Cd积累

Abstract:

In plants, as in other eukaryotes, endogenous small interfering RNAs (siRNAs), a class of small non-coding RNAs, andDNA methylation regulate gene expression in developmental processes and adaptating to environmental stresses, including Cd stress. Cadmium (Cd) is a non-essential heavy metal and highly toxic to plants. To investigate the regulatory role of siRNAs and DNA methylation on genes involved in heavy metals transport, we compared these genes’ expression profiles between a high Cd-accumulating rice (Oryza sativa L. subspecies japonica) cultivar (Xiushui 11) and a low Cd-accumulating rice cultivar (Xiushui110). At five rice development stages investigated, the difference of these genes expression level between the two rice cultivars was not significant except OsPCR1, indicating OsPCR1 may be important in Cd transport in rice. Furthermore, quantitative real time PCR (qRT-PCR) was performed to examine the expression level of a siRNA matched OsPCR1 second exon. Results indicated that the expression level of the siRNA negatively correlated with OsPCR1 expression level at the five stages. In addition, McrBC-qRT-PCR technology was used to determine DNA methylation level, showing that OsPCR1 expression level also negatively correlated with OsPCR1 second exon methylation level. These results of regulatory roles of siRNA and DNA methylation on OsPCR1 expression will contribute to the studies on OsPCR1 function and rice breeding for low Cd accumulation.

Key words: siRNA, DNA methylationOsPCR1, Rice (Oryza sativa L.), Cadmium accumulation

[1]Uraguchi S, Kamiya T, Sakamoto T, Kssai K, Sato Y, Nagamura Y, Yoshida A, Kyozuka J, Ishikawa S, Fujiwara T. Low-affinity cation transporter (OsLCT1) regulates cadmium transport into rice grains. Proc Natl Acad Sci USA, 2011, 108: 20959–20964



[2]Ueno D, Koyama E, Yamaji N, Ma J F. Physiological, genetic, and molecular characterization of a high-Cd-accumulating rice cultivars, Jarjan. J Exp Bot, 2011, 62: 2262–2272



[3]Uraguchi S, Mori S, Kuramata M, Kawasaki A, Arao T, Ishikawa S. Root-to-shoot Cd translocation via the xylem is the major process determining shoot and grain cadmium accumulation in rice. J Exp Bot, 2009, 60: 2677–2688



[4]Takahashi R, Ishimaru Y, Senoura T, Shimo H, Ishikawa S, Arao T, Nakanishi H, Nishizawa N K. The OsNRAMP1 iron transporter is involved in Cd accumulation in rice. J Exp Bot, 2011, 62: 4843–4850



[5]Kuramata M, Masuya S, Takahashi Y, Kitagawa E, Inoue C, Ishikawa S, Youssefian S, Kusano T. Novel cysteine-rich peptides from Digitaria ciliaris and Oryza sativa enhance tolerance to cadmium by limiting its cellular accumulation. Plant Cell Physiol, 2009, 50: 106–117



[6]Shen G M, Zhu C, Du Q Z. Genome-wide identification of PHYTOCHELATIN and PHYTOCH_SYNTH domain-containing phytochelatin family from rice. Electronic J Biol, 2010, 6:73–79



[7]Shimo H, Ishimaru Y, An G, Yamakawa T, Nakanishi H, Nishizawa N K. Low cadmium (LCD), a novel gene related to cadmium tolerance and accumulation in rice. J Exp Bot, 2011, 62: 5727–5734



[8]Song W Y, Choi K S, Alexis D A, Martinoia E, Lee Y. Brassica juncea plant cadmium resistance 1 protein (BjPCR1) facilitates the radial transport of calcium in the root. Proc Natl Acad Sci USA, 2010, 108: 1908–19813



[9]Song W Y, Choi K S, Kim D Y, Geisler M, Park J, Vincenzetti V, Schellenberg M, Kim S H, Lim Y P, Noh E W, Lee Y, Martinoia E. Arabidopsis PCR2 is a zinc exporter involved in both zinc extrusion and long-distance zinc transport. Plant Cell, 2010, 22: 2237–2252



[10]Cobbett C, Goldsbrough P. Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostasis. Annu Rev Plant Biol, 2002, 53: 159–182



[11]Song W Y, Hörtensteiner S, Tomioka R, Lee Y, Martinoia E. Common functions or only phylogenetically related? The large family of PLAC8 motif-containing/PCR genes. Mol Cells, 2011, 31: 1–7



[12]Song W Y, Martinoia E, Lee J, Kim D, Kim D Y, Vogt E, Shim D, Choi K S, Hwang I, Lee Y. A novel family of cys-rich membrane proteins mediates cadmium resistance in Arabidopsis. Plant Physiol, 2004, 135: 1027–1039



[13]Borsani O, Zhu J, Verslues P E, Sunkar R, Zhu J K. Endogenous siRNAs derived from a pair of natural cis-antisense transcripts regulate salt tolerance in Arabidopsis. Cell, 2005, 123: 1279–1291



[14]Carthew R W, Sontheimer E J. Origins and mechanisms of miRNA and siRNAs. Cell, 2009, 136: 642–655



[15]Moldovan D, Spriggs A, Yang J, Pogson B J, Dennis E S, Wilson I W. Hypoxia-responsive microRNAs and trans-acting small interfering RNAs in Arabidopsis. J Exp Bot, 2010, 61: 165–177



[16]Yan Y, Zhang Y, Sun Z, Fu Y, Chen X, Fang R. Small RNAs from MITE-derived stem-loop precursors regulate abscisic acid signaling and abiotic stress responses in rice. Plant J, 2011, 65: 820–828



[17]Yao Y, Ni Z, Peng H, Sun F, Xin M, Sunkar R, Zhu J K, Sun Q. Non-coding small RNAs responsive to abiotic stress in wheat (Triticum aestivum L. ). Funct Integr Genomic, 2010, 10: 187–190



[18]Kim V N, Han J, Siomi M C. Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol, 2009, 10: 126–139



[19]Hannon G J. RNA interference. Nature, 2002, 418: 244–251



[20]Song J J, Smith S K, Hannon G J, Joshua-Tor L. Crystal structure of Argonaute and its implications for RISC slicer activity. Science, 2004, 305: 1434–1437



[21]Law J A, Jacobsen S E. Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat Rev Genet, 2010, 11: 204–220



[22]Chan S W L, Henderson I R, Jacobsen S E. Gardening the genome: DNA methylation in Arabidopsis thaliana. Nat Rev Genet, 2005, 6: 351–360



[23]Boyko A, Blevins T, Yao Y, Golubov A, Bilichak A, Ilnytskyy Y, Hollander J. Transgenerational adaptation of Arabidopsis to stress requires DNA methylation and the function of dicer-like proteins. PLoS One, 2010, 5: e9514



[24]Choi C S, Sano H. Abiotic-stress induces demethylation and transcriptional activation of a gene encoding a glycerophosphodiesterase-like protein in tobacco plants. Mol Genet Genomics, 2007, 277: 589–600



[25]Greco M, Chiappetta A, Bruno L, Bitonti M B. In Posidonia oceanica cadmium induces changes in DNA methylation and chromatin patterning. J Exp Bot, 2012, 63: 695–709



[26]Verhoeven K J F, Jansen J J, Dijk P J, Biere A. Stress-induced DNA methylation changes and their heritability in asexual dandelions. New Phytol, 2010, 185: 1108–1118



[27]Raj S, Brautigen K, Hamanishi E T, Wilkins O, Thomas B R, Schroeder W, Mansfield S D, Plant A L, Campbell M M. Clone history shapes populous drought response. Proc Natl Acad Sci USA, 2011, 108: 12521–12526



[28]Ball M P, Li J B, Gao Y, Lee J H, LeProust E M, Park I H, Xie B, Daley G Q, Church G M. Targeted and genome-scale strategies reveal gene-body methylation signatures in human cells. Nat Biotechnol, 2009, 27: 361–368



[29]Hellman A, Chess A. Gene Body-specific methylation on the active X chromosome. Science, 2007, 315: 1141–1143



[30]Lister R, Pelizzola M, Dowen R H, Hawkins R D, Hon G, Tonti-Filippini J, Nery J R, Lee L, Ye Z, Ngo Q M, Edsall L, Antosiewicz-Bourget J, Stewart R, Ruotti V, Millar A H, Thomson J A, Ren B, Ecker J R. Human DNA methylomes at base resolution show widespread epigenomic differences. Nature, 2009, 462: 315–322



[31]Kawanabe T, Fujimoto R, Sasaki T, Taylor J M, Dennis E S. A comparison of transcriptome and epigenetic status between closely related species in the genus Arabidopsis. Gene, 2012, 506: 301–309



[32]Zilberman D, Gehring M, Tran R K, Ballinger T, Henikoff S. Genome-wide analysis of Arabidopsis thaliana DNA methylation uncovers an interdependence between methylation and transcription. Nat Genet, 2007, 39: 61–69



[33]Suzuki M M, Bird A. DNA methylation landscapes: provocative insights from epigenomics. Nat Rev Genet, 2008, 9: 465–476



[34]He J, Zhu C, Ren Y, Yan Y, Jiang D. Genotypic variation in grain cadmium concentration of lowland rice. J Plant Nutr Soil Sci, 2006, 169: 711–716



[35]何俊瑜,任艳芳,朱诚,蒋德安. 镉胁迫对不同水稻品种种子萌发、幼苗生长和淀粉酶活性的影响. 中国水稻科学, 2008, 22: 399–404



He J Y, Ren Y F, Zhu C, Jiang D A. Effects of cadmium stress on seed germination, seedling growth, and amylase activities in rice. Chin J Rice Sci, 2008, 22: 399–404 (in Chinese with English abstract)



[36]Ding Y, Chen Z, Zhu C. Microarray-based analysis of cadmium-responsive microRNAs in rice (Oryza sativa). J Exp Bot, 2011, 62: 3563–3573



[37]Pfaffl M W. A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res, 2001, 29: e45



[38]Serra I A, Procaccini G, Intrieri M C, Migliaccio M, Mazzuca S, Innocenti A M. Comparison of ISSR and SSR markers for analysis of genetic diversity in the seagrass Posidonia oceanica. Marine Ecology Progress Series, 2007, 338: 71–79



[39]Teixeira F K, Heredia F, Sarazin A, Roudier F, Boccara M, Ciaudo C, Cruaud C, Poulain J, Berdasco M, Fraga M F, Voinnet O, Wincker P, Esteller M, Colot V. A role for RNAi in the selective correction of DNA methylation defects. Science, 2009, 323: 1600–1604



[40]Johnson C, Bowman L, Adai A T, Vance V, Sundaresan V. CSRDB: a small RNA integrated database and browser resource for cereals. Nucleic Acids Res, 2007, 35: D829–D833



[41]Brandeis M, Ariel M, Cedar H. Dynamics of DNA methylation during development. Bioessays, 1993, 15: 709–713



[42]赵嵘, 胡丽玲, 孔繁强, 左爱军. PXR基因外显子3甲基化与肠癌细胞对5氟尿嘧啶的耐药性相关. 中国生物化学与分子生物学报, 2013, 29: 63–69



Zhao R, Hu L L, Kong F Q, Zuo A J. Association between pregnane X receptor gene exon3 methylation with 5-fluorouracil resistance of the colon cancer cells. Chin J Biochem Mol Biol, 2013, 29: 63–69 (in Chinese with English abstract)



[43]Hohn T, Corsten S, Rieke S, Muller M, Rothnie H. Methylation of coding region alone inhibits gene expression in plant protoplasts. Proc Natl Acad Sci USA, 1996, 93: 8334–8339



[44]Rountree M R, Selker E U. DNA methylation inhibits elongation but not initiation of transcription in Neurospora crassa. Genes Dev, 1997, 11: 2383–2395



[45]Xiao Z, Wang C, Mo D, Li J, Chen Y, Zhang Z, Cong P. Promoter CpG methylation status in porcine Lyn is associated with its expression levels. Gene, 2012, 511: 73–78



[46]Foret S, Kucharski R, Pellegrini M, Feng S, Jacobsen S E, Robinson G E, Maleszka R. DNA methylation dynamics, metabolic fluxes, genesplicing, and alternative phenotypes in honey bees. Proc Natl Acad Sci USA, 2012, 109: 4968–4973



[47]Shukla S, Kavak E, Gregory M, Imashimizu M, Shutinoski B, Kashlev M, Oberdoerffer P, Sandberg R, Oberdoerffer S. CTCF-promoted RNA polymerase II pausing links DNA methylation to splicing. Nature, 2011, 479: 74–79
[1] 田甜, 陈丽娟, 何华勤. 基于Meta-QTL和RNA-seq的整合分析挖掘水稻抗稻瘟病候选基因[J]. 作物学报, 2022, 48(6): 1372-1388.
[2] 郑崇珂, 周冠华, 牛淑琳, 和亚男, 孙伟, 谢先芝. 水稻早衰突变体esl-H5的表型鉴定与基因定位[J]. 作物学报, 2022, 48(6): 1389-1400.
[3] 周文期, 强晓霞, 王森, 江静雯, 卫万荣. 水稻OsLPL2/PIR基因抗旱耐盐机制研究[J]. 作物学报, 2022, 48(6): 1401-1415.
[4] 郑小龙, 周菁清, 白杨, 邵雅芳, 章林平, 胡培松, 魏祥进. 粳稻不同穗部籽粒的淀粉与垩白品质差异及分子机制[J]. 作物学报, 2022, 48(6): 1425-1436.
[5] 颜佳倩, 顾逸彪, 薛张逸, 周天阳, 葛芊芊, 张耗, 刘立军, 王志琴, 顾骏飞, 杨建昌, 周振玲, 徐大勇. 耐盐性不同水稻品种对盐胁迫的响应差异及其机制[J]. 作物学报, 2022, 48(6): 1463-1475.
[6] 杨建昌, 李超卿, 江贻. 稻米氨基酸含量和组分及其调控[J]. 作物学报, 2022, 48(5): 1037-1050.
[7] 杨德卫, 王勋, 郑星星, 项信权, 崔海涛, 李生平, 唐定中. OsSAMS1在水稻稻瘟病抗性中的功能研究[J]. 作物学报, 2022, 48(5): 1119-1128.
[8] 朱峥, 王田幸子, 陈悦, 刘玉晴, 燕高伟, 徐珊, 马金姣, 窦世娟, 李莉云, 刘国振. 水稻转录因子WRKY68在Xa21介导的抗白叶枯病反应中发挥正调控作用[J]. 作物学报, 2022, 48(5): 1129-1140.
[9] 王小雷, 李炜星, 欧阳林娟, 徐杰, 陈小荣, 边建民, 胡丽芳, 彭小松, 贺晓鹏, 傅军如, 周大虎, 贺浩华, 孙晓棠, 朱昌兰. 基于染色体片段置换系群体检测水稻株型性状QTL[J]. 作物学报, 2022, 48(5): 1141-1151.
[10] 王泽, 周钦阳, 刘聪, 穆悦, 郭威, 丁艳锋, 二宫正士. 基于无人机和地面图像的田间水稻冠层参数估测与评价[J]. 作物学报, 2022, 48(5): 1248-1261.
[11] 陈悦, 孙明哲, 贾博为, 冷月, 孙晓丽. 水稻AP2/ERF转录因子参与逆境胁迫应答的分子机制研究进展[J]. 作物学报, 2022, 48(4): 781-790.
[12] 王吕, 崔月贞, 吴玉红, 郝兴顺, 张春辉, 王俊义, 刘怡欣, 李小刚, 秦宇航. 绿肥稻秆协同还田下氮肥减量的增产和培肥短期效应[J]. 作物学报, 2022, 48(4): 952-961.
[13] 巫燕飞, 胡琴, 周棋, 杜雪竹, 盛锋. 水稻延伸因子复合体家族基因鉴定及非生物胁迫诱导表达模式分析[J]. 作物学报, 2022, 48(3): 644-655.
[14] 陈云, 李思宇, 朱安, 刘昆, 张亚军, 张耗, 顾骏飞, 张伟杨, 刘立军, 杨建昌. 播种量和穗肥施氮量对优质食味直播水稻产量和品质的影响[J]. 作物学报, 2022, 48(3): 656-666.
[15] 王琰, 陈志雄, 姜大刚, 张灿奎, 查满荣. 增强叶片氮素输出对水稻分蘖和碳代谢的影响[J]. 作物学报, 2022, 48(3): 739-746.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!