落叶松体胚发育中12个针叶树特异microRNAs表达分析

张俊红; 吴涛; 韩素英; 齐力旺; 张守攻

落叶松体胚发育中12个针叶树特异microRNAs表达分析

1.
中国林业科学研究院林业研究所,北京 100091
2.
中国林业科学研究院森林生态环境与保护研究所,北京 100091
基金项目:
国家"973"计划项目(2009CB119100)和国家自然基金(30830086)
中图分类号: S791.22

Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

1.
Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
2.
Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China
CLC number: S791.22

摘要: 以落叶松体细胞胚胎发育8个阶段的材料构建小RNA文库,进行了Solexa测序、miRNA鉴定、靶基因预测、qRT-PCR定量分析等研究。结果表明:发现的87个miRNAs中,29个为针叶树特异的,来自12个家族,分别为miR946、miR947、miR950、miR951、miR1311、miR1312、miR1313、miR1314、miR1316、miR3701、miR3702和miR3704,其中25个长度为22nt,且在小RNA文库中的最低拷贝数为18(miR1311),最高拷贝数为33 009(miR950)。发现12个miRNA家族的34个靶基因可能参与生长发育、抗病、AGO反馈调节等多方面遗传调控;qRT-PCR分析表明:72% miRNAs表达次高峰在后期单胚或早期子叶胚,在中期子叶胚最低,最高峰在后期子叶胚,分别与子叶形成、胚胎后熟、胚胎休眠的生理活动相吻合,而miRNA前体的表达模式不同于miRNA,前体变化幅度小于成熟体。
- 日本落叶松
- / 体细胞胚胎
- / microRNA
- / 针叶树
- / 表达分析
Abstract: In order to identify miRNA and investigate it’s expression profile during somatic embryogenesis (SE) of larch, a small RNA library was constructed and sequenced by Solexa technique, and expression analysis of miRNAs and four precursors during eight developmental stages of SE was evaluated by qRT-PCR. The results demonstrated that a total of 87 miRNAs was identified, of which 29 miRNAs from 12 families were specific to conifers, including miR946, miR947, miR950, miR951, miR1311, miR1312, miR1313, miR1314, miR1316, miR3701, miR3702 and miR3704, which could be used for further analysis and next experiment. Of these, 25 miRNAs were 22nt in length, while 4 miRNAs were 21nt. The abundance of miRNAs varied largely in the small RNA library, such as miR1311 was detected by 18 copies, while miR950 with 33 009 copies. 34 potential target genes were predicted for 12 miRNA families, which were linked to many respects of regulation, including plant growth and development, disease resistance, AGO protein feedback regulation. qRT-PCR analysis showed that, for 72% miRNAs, the minor expression peak was at late single embryo or early cotyledonary embryo, the lowest level was at middle cotyledonary embryo, while the major peak was at late cotyledonary embryo, which was consistent with the physiology events of cotyledon formation, embryo pre-mature, and embryo dormancy. However, the expression model of miRNA precursor was differed from that of miRNA, and precursor expression with much smaller extent than that of miRNA. The discovery of miRNAs specific to conifers and their targets using embryogenic tissue could provide references for the study on the growth and development of gymnosperms.
- Larix leptolepis
- / somatic embryo
- / microRNA
- / conifer
- / expression analysis

[1]	Juarez M T, Kui J S, Thomas J, et al. microRNA-mediated repression of rolled leaf1 specifies maize leaf polarity[J]. Nature, 2004, 428(6978):84-88
[2]	Parry G, Calderon-Villalobos L I, Prigge M et al. Complex regulation of the TIR1/AFB family of auxin receptors[J]. Proc Natl Acad Sci U S A, 2009, 106(52):22540-22545
[3]	Liu H H, Tian X, Li Y J, et al. Microarray-based analysis of stress-regulated microRNAs in Arabidopsis thaliana[J]. RNA, 2008, 14(5):836-843
[4]	Jones-Rhoades M W, Bartel D P, Bartel B. MicroRNAs and their regulatory roles in plants[J]. Annu Rev Plant Biol, 2006, 57:19-53
[5]	Lu S f, Sun Y H, Amerson H, et al. MicroRNAs in loblolly pine (Pinus taeda L.) and their association with fusiform rust gall development [J]. The Plant Journal,2007, 51:1077-1098
[6]	Yakovlev I A, Fossdal C G, Johnsen O. MicroRNAs, the epigenetic memory and climatic adaptation in Norway spruce[J]. New Phytol, 2010, 187(4):1154-1169
[7]	孙晓梅,张守攻,齐力旺,等.日本落叶松自由授粉家系纸浆材材性遗传变异的研究[J].林业科学研究,2003,16( 5): 515-522
[8]	Bowe L M, Gwe'nae¨le C, dePamphilis C W. Phylogeny of seed plants based on all three genomic compartments: Extant gymnosperms are monophyletic and Gnetales’ closest relatives are conifers [J]. Proc Natl Acad Sci U S A, 2000, 97:4092-4097
[9]	Filonova L H, Bozhkov P V, von Arnold S. Developmental pathway of somatic embryogenesis in Picea abies as revealed by time-lapse tracking[J]. J Exp Bot, 2000, 51(343):249-264
[10]	Cairney J, Pullman G S. The cellular and molecular biology of conifer embryogenesis[J]. New Phytol, 2007, 176(3):511-536
[11]	Kirst M, Johnson A F, Baucom C, et al. Apparent homology of expressed genes from wood-forming tissues of loblolly pine (Pinus taeda) with Arabidopsis thaliana[J]. Proc Natl Acad Sci U S A, 2003, 100(12):7383-7388
[12]	Lorenz W W, Sun F, Liang C, et al. Water stress-responsive genes in loblolly pine (Pinus taeda) roots identified by analyses of expressed sequence tag libraries[J]. Tree Physiol, 2006, 26(1):1-16
[13]	Cairney J, Zheng L, Cowels A, et al. Expressed sequence tags from loblolly pine embryos reveal similarities with angiosperm embryogenesis[J]. Plant Mol Biol, 2006, 62(4-5):485-501
[14]	Ahuja M R, Neale a D B. Evolution of Genome Size in Conifers[J]. Silvae Genetica, 2005, 54(3):126-137
[15]	Li M, Xia Y, Gu Y, et al. MicroRNAome of porcine pre- and postnatal development[J]. PLoS One, 2010, 5(7):e11541
[16]	Kozomara A, Griffiths-Jones S. miRBase: integrating microRNA annotation and deep-sequencing data[J]. Nucleic Acids Res, 2011, 39(Database issue):D152-157
[17]	Dai X, Zhao a P X. psRNATarget: a plant small RNA target analysis server[J]. Nucleic Acids Research, 2011, 39(Web Server issue):W155-159
[18]	Vaucheret H, Vazquez F, Crete P, et al. The action of ARGONAUTE1 in the miRNA pathway and its regulation by the miRNA pathway are crucial for plant development[J]. Genes Dev, 2004, 18(10):1187-1197
[19]	Yang H, Matsubayashi Y, Nakamura K, et al. Oryza sativa PSK gene encodes a precursor of phytosulfokine-alpha, a sulfated peptide growth factor found in plants[J]. Proc Natl Acad Sci U S A, 1999, 96(23):13560-13565
[20]	Lindsey K, Casson S, Chilley P. Peptides:new signaling molecules in plants[J]. Trends Plant Sci, 2002, 7(2): 78-83
[21]	Allen E, Xie Z X, Gustafson A M, et al. microRNA-directed phasing during trans-acting siRNA biogenesis in plants [J]. Cell, 2005, 121(2):207-221
[22]	Howell M D, Fahlgren N, Chapman E J, et al. Genome-wide analysis of the RNA-DEPENDENT RNA POLYMERASE6/DICER-LIKE4 pathway in Arabidopsis reveals dependency on miRNA- and tasiRNA-directed targeting [J]. Plant Cell, 2007, 19(3):926-942
[23]	Allen E, Xie Z X, Gustafson A M, et al. Evolution of microRNA genes by inverted duplication of target gene sequences in Arabidopsis thaliana[J]. Nature Genetics, 2004, 36(12):1282-1290
[24]	Vazquez F, Blevins T, Ailhas J, et al. Evolution of Arabidopsis MIR genes generates novel microRNA classes[J]. Nucleic Acids Res, 2008, 36(20):6429-6438
[25]	Braybrook S A, Stone S L, Park S, et al. Genes directly regulated by LEAFY COTYLEDON2 provide insight into the control of embryo maturation and somatic embryogenesis[J]. Proc Natl Acad Sci U S A, 2006, 103(9):3468-3473
[26]	Holdsworth M J, Bentsink L, Soppe W J. Molecular networks regulating Arabidopsis seed maturation, after-ripening, dormancy and germination[J]. New Phytol, 2008, 179(1):33-54

[1]	李龙 , 张立峰 , 齐力旺 , 韩素英 . 日本落叶松体细胞胚胎发生相关基因LaSERK1的克隆与表达分析. 林业科学研究, 2013, 26(6): 673-680.
[2]	范艳如 , 兰倩 , 韩素英 , 齐力旺 , 张立峰 . 日本落叶松LaSPL2和LaSPL3在体细胞胚发育中的表达分析. 林业科学研究, 2021, 34(5): 79-87. doi: 10.13275/j.cnki.lykxyj.2021.005.009
[3]	李丹蕾 , 张瑞芝 , 王峰 , 陈俏丽 , 牛春阳 , 零雅茗 , 郝昕 . MicroRNA转录后调控欧美杨R2R3-MYBs抗锈菌表达. 林业科学研究, 2017, 30(2): 254-259. doi: 10.13275/j.cnki.lykxyj.2017.02.010
[4]	朱建峰 , 李万峰 , 杨文华 , 韩素英 , 齐力旺 . 中间锦鸡儿生长发育过程中5个miRNAs及其靶基因的表达模式分析. 林业科学研究, 2013, 26(S1): 45-51.
[5]	肖霞 , 张立峰 , 齐力旺 , 韩素英 . miR396在落叶松体细胞胚胎中的功能研究. 林业科学研究, 2016, 29(2): 227-233.
[6]	王菁 , 李爱 , 王春国 , 宋文芹 , 陈成彬 . 日本落叶松 UDPGDH基因的cDNA克隆和表达分析. 林业科学研究, 2013, 26(S1): 76-81.
[7]	卢孟柱 , 王晓茹 , Alfred E. Szmidt . PCR-SSCP用于针叶树种遗传分析的可行性. 林业科学研究, 2000, 13(4): 349-354.
[8]	孙海涛 , 杨玲 , 齐力旺 , 李万峰 . 日本落叶松胚状体干化处理对萌发的影响. 林业科学研究, 2024, 37(2): 90-95. doi: 10.12403/j.1001-1498.20230311
[9]	王楠 , 胡宝全 , 王春国 , 宋文芹 , 陈成彬 . 黑杨三倍体与二倍体叶片中miRNA表达差异研究. 林业科学研究, 2013, 26(S1): 33-38.
[10]	吴涛 , 张俊红 , 韩素英 , 杨文华 , 李万峰 , 齐力旺 . 日本落叶松小RNA文库构建及其microRNA鉴定. 林业科学研究, 2012, 25(6): 677-684.
[11]	吴涛 , 韩素英 , 张俊红 , 李万峰 , 杨文华 , 齐力旺 . 日本落叶松种子萌发过程中18个miRNAs的表达变化. 林业科学研究, 2013, 26(S1): 9-17.
[12]	刘超 , 张力鹏 , 王春国 , 宋文芹 , 陈成彬 . 日本落叶松EST-SSR标记挖掘及特征分析. 林业科学研究, 2013, 26(S1): 60-68.
[13]	董雷鸣 , 张守攻 , 孙晓梅 . 日本落叶松全双列交配生长性状的遗传分析. 林业科学研究, 2019, 32(4): 11-18. doi: 10.13275/j.cnki.lykxyj.2019.04.002
[14]	周亚楠 , 李爱 , 陈成彬 , 王春国 , 宋文芹 . 日本落叶松杂种及亲本 4CL基因的克隆和SNP多态性分析. 林业科学研究, 2013, 26(S1): 18-24.
[15]	易敏 , 张守攻 , 谢允慧 , 孙晓梅 . 日本落叶松咖啡酸-O-甲基转移酶基因 LkCOMT的克隆及单核苷酸多态性分析. 林业科学研究, 2013, 26(S1): 52-59.
[16]	易敏 , 张守攻 , 谢允慧 , 孙晓梅 . 日本落叶松纤维素合酶基因片段的克隆及单核苷酸多态性分析. 林业科学研究, 2015, 28(3): 303-310.
[17]	张婷 , 丁贵杰 , 文晓鹏 . 马尾松紫色酸性磷酸酶基因PmPAP1的克隆与表达模式分析. 林业科学研究, 2016, 29(6): 797-806.
[18]	李正红 , 孙振元 , 刘秀贤 , 彭镇华 . 地锦体细胞胚胎发生研究. 林业科学研究, 2005, 18(1): 36-40.
[19]	侯成林 , 王有智 . 针叶树斑痣盘菌科真菌病原调查*. 林业科学研究, 1995, 8(4): 426-428.
[20]	张俊红 , 张守攻 , 童再康 , 李万峰 , 韩素英 , 齐力旺 . 植物中MicroRNA及其它非编码小RNA的保守性和进化历程. 林业科学研究, 2013, 26(S1): 103-108.

点击查看大图

计量

文章访问数: 3126
HTML全文浏览量: 184
PDF下载量: 1659
被引次数: 0

姓名
邮箱
手机号码
标题
留言内容
验证码

留言板

落叶松体胚发育中12个针叶树特异microRNAs表达分析

Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

计量

落叶松体胚发育中12个针叶树特异microRNAs表达分析

English Abstract

Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

全文HTML

目录

留言板

落叶松体胚发育中12个针叶树特异microRNAs表达分析

Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

计量

出版历程

落叶松体胚发育中12个针叶树特异microRNAs表达分析

English Abstract

Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

全文HTML

目录