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[1]刘炜婳,林争春,刘彦英,等.野生蕉miR395a前体克隆与进化特性及启动子分析[J].应用与环境生物学报,2018,24(01):89-96.[doi:10.19675/j.cnki.1006-687x.2017.09042]
 LIU Weihua,LIN Zhengchun,LIU Yanying,et al.Cloning and evolution characteristics of pre-miR395a and promoter analysis in wild banana ($Musa itinerans$)[J].Chinese Journal of Applied & Environmental Biology,2018,24(01):89-96.[doi:10.19675/j.cnki.1006-687x.2017.09042]
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野生蕉miR395a前体克隆与进化特性及启动子分析()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
24卷
期数:
2018年01期
页码:
89-96
栏目:
研究论文
出版日期:
2018-02-09

文章信息/Info

Title:
Cloning and evolution characteristics of pre-miR395a and promoter analysis in wild banana ($Musa itinerans$)
作者:
刘炜婳林争春刘彦英李汉生倪珊珊林玉玲赖钟雄?
福建农林大学园艺植物生物工程研究所 福州 350002
Author(s):
LIU Weihua LIN Zhengchun LIU Yanying LI Hansheng NI Shanshan LIN YulingLAI Zhongxiong**
Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
关键词:
野生蕉pre-miR395a克隆进化特性启动子分析qPCR
Keywords:
wild banana (Musa itinerans) pre-miR395a cloning evolution characteristic promoter analysis qPCR
分类号:
S668.101 : Q78
DOI:
10.19675/j.cnki.1006-687x.2017.09042
摘要:
植物miR395参与硫代谢调控,并在应答逆境胁迫等方面起重要调控作用. 为了解香蕉miR395的分子特性和进化规律,在参考香蕉基因组信息的基础上,结合RT-PCR技术,从三明野生蕉(Musa itinerans)叶片中获得81 nt miR395a前体序列,包含21 nt miR395a成熟体序列. 进化特性分析表明,植物miR395a前体存在于27个物种中,野生蕉pre-miR395a序列与双子叶耧斗菜(Aquilegia viridiflora Pall.)最为接近,与单子叶的水稻、玉米、高粱最远,表明野生蕉pre-miR395a的起源比较特殊;前体序列长度差异较大(57-226 nt);miR395a成熟体序列分析发现,来自5p臂上的miR395a序列特异性较大. 野生蕉miR395a前体能形成典型的发卡结构,miR395a成熟体位于前体的3p臂上. 转录起始位点分析表明,野生蕉pri-miR395a转录起始区域位于-87 bp - -38 bp,A为转录起始位点. 启动子顺式作用元件预测分析表明,野生蕉miR395a启动子包含多个激素、胁迫、生物钟及光响应元件,可能与响应胁迫密切相关. qPCR检测结果也发现miR395a的表达量在低温胁迫下极显著下调,表明响应低温胁迫. 综上表明三明野生蕉在响应低温胁迫过程中miR395a 起着重要调控作用,这对于进一步研究野生蕉miR395a在低温胁迫中的作用机制具有参考价值. (图7 表3 参41)
Abstract:
MiR395 participates in sulfur metabolism control and plays an important regulatory role in stress response. To explore its structure, molecular characteristics, evolution, and promoter sequence characteristics in wild banana (Musa itinerans) from Sanming in Fujian Province, we referred to the data of the banana genome and performed RT-PCR to sequence pre-miR395a, which contained 84 nt, including a 21-nt mature sequence. The evolution analysis showed that miR395a was distributed in 27 plant species, among which the length of pre-miR395a varied remarkably (between 57 and 226 nt), and miR395a-5p was considerably specific; miR395a was close to aqc-miR395a, but very far from osa-miR395a, zma-miR395a, and abi-miR395a, suggesting the possible specific origin of miR395a in wild banana. Mfold predicted that pre-miR395a spontaneously formed a typical, stable hairpin secondary structure, and the mature miR395a sequence was located in the 3p arm. Transcriptional start site (TSS) predicted that the TSS region of pri-miR395a was likely between -87 bp and -38 bp with TSS A. The promoter sequence analysis indicated that the miR395a promoter included many cis-elements such as hormone, light, stress responses, and circadian control. The qPCR assay further confirmed that miR395a played a crucial role in cold stress response in wild banana. The 84 nt sequence of pre-miR395a was obtained in wild banana, which was close to the dicot plant aqc-MIR395a as per evolution analysis. Thus, pri-miR395a might be associated with stress responses in plants. The qPCR assay indicated that the expression levels of miR395a decreased very significantly during low temperature stress in wild banana. Therefore, miR395a was inferred to play essential regulating roles in response to low temperature stress in wild banana, which was a valuable reference for further exploring the functional mechanism of miR395a during low temperature stress in wild banana.

参考文献/References:

1 张锐, 赖钟雄, 刘生财, 张梓浩, 吴金寿. 福建野生蕉(Musa spp., AB group) 3个自然居群遗传多样性的RAPD分析[J]. 福建农林大学学报(自然科学版), 2013, 42 (3): 246-251 [Zhang R, Lai ZX, Liu SC, Zhang ZH, Wu JS. Ananlysis of genetic diversity of 3 natural populations of the wild banana (Musa spp., AB group) in Fujian by RAPD markers [J]. J Fujian Agric For Univ (Nat Sci Ed), 2013, 42 (3): 246-251]
2 陈红俊, 赖钟雄, 刘炜婳, 赖恭梯, 张梓浩, 叶炜, 刘生财, 林玉玲. 福建东南部野生蕉7个自然居群289份样品的ISSR分析[J]. 热带作物学报, 2013, 33 (12): 2115-2124 [Chen HJ, Lai ZX, Liu WH, Lai GT, Zhang ZH, Ye W, Liu SC, Lin YL. ISSR analysis of 289 samples from 7 wild banana groups distributed in southeastern Fujian Province [J]. Chin J Trop Crops, 2013, 33 (12): 2115-2124]
3 赖恭梯, 赖钟雄, 刘炜婳, 叶炜, 林玉玲, 刘生财, 陈裕坤, 张梓浩, 吴高杰. 福建中部3个野生蕉自然居群基于NTSYS和STRUCTURE软件的ISSR分析[J]. 热带作物学报, 2014, 35 (2): 223-231 [Lai GT, Lai ZX, Liu WH, Ye W, Lin YL, Liu SC, Chen YK, Zhang ZH, Wu GJ. ISSR analysis of 3 natural populations of the wild banana distributed in the middle of Fujian Province based on NTSYS and STRUCTURE [J]. Chin J Trop Crops, 2014, 35 (2): 223-231]
4 张妙霞. 野生香蕉(Musa spp., AB group)抗寒相关基因的克隆与表达分析[D]. 福州: 福建农林大学, 2010 [Zhang MX. Cloning and expression analysis of cold resistance relative genes of the wild banana (Musa spp., AB group) [D]. Fuzhou: Fujian Agriculture and Forestry University, 2010]
5 赖恭梯. 福建香蕉种质资源试管保存及野生蕉ISSR与抗寒性分析[D]. 福州: 福建农林大学, 2014 [Lai GT. In vitro conservation of Musa spp. germplasm resources and ISSR and cold-resistant analyses of the wild banana in Fujian Province [D]. Fuzhou: Fujian Agriculture and Forestry University, 2014]
6 Liu WH, Cheng CZ, Lai GT , Lin YL, Lai ZX. Molecular cloning and expression analysis of KIN10 and cold-acclimation related genes in wild banana ‘Huanxi’ (Musa itinerans) [J]. SpringerPlus, 2015, 4 (1): 1-7
7 刘炜婳. 福建野生蕉离体繁殖及其抗性基因克隆与定量表达分析[D]. 福州: 福建农林大学, 2013 [Liu WH. Micropropagation and cloning and quantitative expression of resistant genes of the wild bananas in Fujian Province [D]. Fuzhou: Fujian Agriculture and Forestry University, 2013]
8 杨洋. 三明野生蕉和天宝蕉WHIRLY基因克隆及其定量表达分析[D]. 福州: 福建农林大学, 2014 [Yang Y. Cloning and expression of WHIRLY in the wild banana from Sanming City and Musa spp. cv. Tianbaojiao [D]. Fuzhou: Fujian Agriculture and Forestry University, 2014]
9 Feng X, Lai Z, Lin Y, Lai G, Lian C. Genome-wide identification and characterization of the superoxide dismutase gene family in Musa acuminata cv. Tianbaojiao (AAA group) [J]. BMC Genom, 2015, 16 (1): 823.
10 Jonesrhoades MW, Bartel DP. Computational identification of plant microRNAs and their targets, including a stress-induced miRNA [J]. Mol Cell, 2004, 14 (6): 787-799
11 Gang L, Yang F, Yu D. MicroRNA395 mediates regulation of sulfate accumulation and allocation in Arabidopsis thaliana [J]. Plant J, 2010, 62 (6): 1046-1057
12 Liang G, Yu D. Reciprocal regulation among miR395, aps and sultr2;1 in Arabidopsis thaliana [J]. Plant Signal Behav, 2010, 5 (10): 1257-1259
13 Takahashi H, Watanabetakahashi A, Smith FW, Blakekalff M, Hawkesford MJ, Saito K. The roles of three functional sulphate transporters involved in uptake and translocation of sulphate in Arabidopsis thaliana [J]. Plant J For Cell Mol Biol, 2000, 23 (2): 171-182
14 Takahashi H, Yamazaki M, Sasakura N, Watanabe A, Leustek T, Engler JA, Engler G, Montagu MV, Saito K. Regulation of sulfur assimilation in higher plants: a sulfate transporter induced in sulfate-starved roots plays, a central role in Arabidopsis thaliana [J]. PNAS, 1997, 94 (20): 11102-11107
15 Jonesrhoades MW, Bartel DP. Computational identification of plant microRNAs and their targets, including a stress-induced miRNA [J]. Mol Cell, 2004, 14 (6): 787-799
16 黄思齐. 镉胁迫下水稻和油菜中microRNA的克隆、鉴定与miR395功能分析[D]. 南京: 南京农业大学, 2010 [Huang SQ. Isolation and identification of microRNA in Oryza sativa and Brassica napus undercd stress and functional characterization of MIR395 [D]. Nanjing: Nanjing Agricultural University, 2010]
17 Kawashima CG, Yoshimoto N, Maruyamanakashita A, Tsuchiya YN, Saito K, Takahashi H, Dalmay T. Sulphur starvation induces the expression of microRNA-395 and one of its target genes but in different cell types [J]. Plant J For Cell Mol Biol, 2009, 57 (2): 313-321
18 Lin LL, Wu CC, Huang HC, Chen HJ, Hsu-Liang H, Hsueh-Fen J. Identification of microRNA 395a in 24-epibrassinolide-regulated root growth of Arabidopsis thaliana using microRNA arrays [J]. Int J Mol Sci, 2013, 14 (7): 14270-14286
19 曾长英, 周玉飞, 彭明. MiR395abcd与其靶基因ATRX在不同耐寒木薯品种中对低温响应的差异性分析[J]. 基因组学与应用生物学, 2015, 34 (11): 2441-2447 [Zeng CY, Zhou YF, Peng M. The differential analysis of miR395abcd and its target gene ATRX response to low temperature in different cold tolerance cassava varieties [J]. Genom Appl Biol, 2015, 34 (11): 2441-2447]
20 刘炜婳, 林争春, 冯新, 赖钟雄.天宝蕉HOS1基因启动子克隆及生物信息学分析 [J]. 福建农业学报, 2016, 31 (8): 820-825 [Liu WH, Lin ZC, Feng X, Lai ZX. Cloning and bioinformatic analysis of HOS1 promoter from the ‘Tianbao’ banana (Musa acuminate, AAA group) [J]. Fujian J Agric Sci, 2016, 31 (8): 820-825]
21 刘炜婳, 林玉玲, 赖钟雄. 福州宦溪野生蕉CBF7的cDNA与启动子克隆及在不同温度下的表达特性 [J]. 应用与环境生物学报, 2017, 23 (2): 200-208 [Liu WH, Lin YL, Lai ZX. Cloning and expression profile of cDNA and promoter of CBF7 in wild banana (Musa itinerans) from Huanxi of Fuzhou city [J]. Chin J Appl Environ Biol, 2017, 23 (2): 200-208]
22 Nawrocki EP, Burge SW, Bateman A, Daub J, Eberhardt RY, Eddy SR, Floden EW, Gardner PP, Jones TA, Tate J, Finn RD. Rfam 12.0: updates to the RNA families database [J]. Nucleic Acids Res, 2015, 43: 130-137
23 Scott IM, Clarke SM, Wood JE, Mur LA. Salicylate accumulation inhibits growth at chilling temperature in Arabidopsis [J]. Plant Physiol, 2004, 135 (2): 1040-1049
24 Kosová K, Prá?il IT, Vítámvás P, Dobrev P, Motyka V, Floková K, Novák O, Ture?ková V, Rol?ik J, Pe?ek B, Trávni?ková A, Gaudinová A, Galiba G, Janda T, Vlasáková E, Prá?ilová P, Vanková R. Complex phytohormone responses during the cold acclimation of two wheat cultivars differing in cold tolerance, winter samanta and spring Sandra [J]. J Plant Physiol, 2012, 169 (6): 567-576
25 Dong CJ, Li L, Shang QM, Liu XY, Zhang ZG. Endogenous salicylic acid accumulation is required for chilling tolerance in cucumber (cucumis sativus, l.) seedlings [J]. Planta, 2014, 240 (4): 687-700
26 Kang HM, Saltveit ME. Chilling tolerance of maize, cucumber and rice seedling leaves and roots are differentially affected by salicylic acid [J]. Physiol Plantarum, 2002, 115 (4): 571-576
27 Senaratna T, Touchell D, Bunn E, Dixon K. Acetyl salicylic acid (aspirin) and salicylic acid induce multiple stress tolerance in bean and tomato plants [J]. Plant Growth Reg, 2000, 30 (2): 157-161
28 Ta?gín E, ?kke? A, Nalbanto?lu B. Effects of salicylic acid and cold on freezing tolerance in winter wheat leaves [J]. Plant Growth Regul, 2003, 41 (3): 231-236
29 Horvath E, Szalai MP, Paldi E, Janda T. Exogenous 4-hydroxybenzoic acid and salicylic acid modulate the effect of short-term drought and freezing stress on wheat plants [J]. Biol Plantarum, 2007, 51 (3): 480-487
30 Huang X, Chen MH, Yang LT, Li YR, Wu JM. Effects of exogenous abscisic acid on cell membrane and endogenous hormone contents in leaves of sugarcane seedlings under cold stress [J]. Sugar Technol, 2015, 17 (1): 59-64
31 Shinkawa R, Morishita A, Amikura K, Machida R, Murakawa H, Kuchitsu K, Ishikawa M. Abscisic acid induced freezing tolerance in chilling-sensitive suspension cultures and seedlings of rice [J]. BMC Res Notes, 2013, 6 (1): 351-365
32 Pál M, Ja T, Szalai G. Abscisic acid may alter the salicylic acid-related abiotic stress response in maize [J]. J Agron Crop Sci, 2011, 197 (5): 368-377
33 Furukawa J, Abe Y, Mizuno H, Matsuki K, Sagawa K, Kojima M, Sakakibara H, Iwai H, Satoh S. Seasonal fluctuation of organic and inorganic components in xylem sap of populus nigra [J]. Plant Root, 2011, 5 (5): 56-62
34 Lee SJ, Kang JY, Park HJ, Kim MD, Bae MS, Choi HI, Kim SY. DREB2C interacts with ABF2, a bZIP protein regulating abscisic acid-responsive gene expression, and its overexpression affects abscisic acid sensitivity [J]. Plant Physiol, 2010, 153 (2): 716-727
35 韦弟, 李杨瑞, 邸南南, 卜朝阳, 闭志强. 乙烯利提高香蕉幼苗抗寒性的生理效应[J]. 热带作物学报, 2009, 30 (10): 1447-1451 [Wei D, Li YR, Di NN, Bu ZY, Bi ZQ. Effect of ethephon on cold resistance of young plants of banana (Musa AAA Cavendish subgroup) [J]. Chin J Trop Crop, 2009, 30 (10): 1447-1451]
36 Shi Y, Tian S, Hou L, Huang X, Zhang X, Guo H, Yang S. Ethylene signaling negatively regulates freezing tolerance by repressing expression of CBF and type-a ARR genes in Arabidopsis [J]. Plant Cell, 2012, 24 (6): 2578-2595
37 金鹏, 吕慕雯, 孙萃萃, 郑永华, 孙明. Meja与低温预贮对枇杷冷害和活性氧代谢的影响[J]. 园艺学报, 2012, 39 (3): 461-468 [Jin P, Lü MW, Sun CC, Zheng YH, Sun M. Effects of methyl jasmonate in combination with low temperature conditioning on chilling injury and active oxygen metabolism in loquat fruits [J]. Acta Horticult Sin, 2012, 39 (3): 461-468]
38 Hu Y, Jiang L, Wang F, Yu D. Jasmonate regulates the inducer of CBF expression-c-repeat binding factor/DRE binding factor1 cascade and freezing tolerance in Arabidopsis [J]. Plant Cell, 2013, 25 (8): 2907-2924
39 Achard P, Gong F, Cheminant S, Alioua M, Hedden P, Genschik P. The cold-inducible CBF1 factor: dependent signaling pathway modulates the accumulation of the growth-repressing della proteins via its effect on gibberellin metabolism [J]. Plant Cell, 2008, 20 (8): 2117-2129
40 李静, 崔继哲, 弭晓菊. 生长素与植物逆境胁迫关系的研究进展[J].生物技术通报, 2012 (6): 13-17 [Li J, Cui JZ, Mi XJ. Progress of studies on relationship between auxin and plant response to abiotic stress [J]. Biotechnol Bull, 2012 (6): 13-17]
41 Daniel X, Sugano S, Tobin EM. Ck2 phosphorylation of CCA1 is necessary for its circadian oscillator function in Arabidopsis [J]. PNAS, 2004, 101 (9): 3292-3297

更新日期/Last Update: 2018-02-09