miR168家族进化特性及其在砂梨休眠期的表达模式分析

doi:10.16420/j.issn.0513-353x.2021-0103

园艺学报 ›› 2022, Vol. 49 ›› Issue (5): 958-972.doi: 10.16420/j.issn.0513-353x.2021-0103

miR168家族进化特性及其在砂梨休眠期的表达模式分析

梁沁¹^,², 张延晖³, 康开权⁴, 刘瑾航¹, 李亮¹, 冯宇¹, 王超¹, 杨超¹^,^*(), 李永裕¹^,^*()

¹福建农林大学园艺学院,福州 350002
²广安市广安区现代农业园区服务中心,四川广安 638500
³福建省永泰县农业农村局经济作物站,福州 350700
⁴福建省连江市国有林场,福州 350500

收稿日期:2022-01-11 修回日期:2022-04-01 出版日期:2022-05-25 发布日期:2022-05-25
通讯作者: 杨超,李永裕 E-mail:yycc32@126.com;lilin3182@163.com
基金资助:
国家自然科学基金项目(31501694);国家自然科学基金项目(31902067);高等学校博士学科点专项科研基金项目(2011351511011)

Molecular Evolution of MiR168 Family and Their Expression Profiling During Dormancy of Pyrus pyrifolia

LIANG Qin¹^,², ZHANG Yanhui³, KANG Kaiquan⁴, LIU Jinhang¹, LI Liang¹, FENG Yu¹, WANG Chao¹, YANG Chao¹^,^*(), LI Yongyu¹^,^*()

¹College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China
²Guang’an Modern Agricultural Park Service Center,Guang’an,Sichuan 638500,China
³Economic Crop Station,Agricultural and Rural Bureau of Yongtai County,Fuzhou 350700,China
⁴Lianjiang State-Owned Forest Farm in Fujian Province,Fuzhou 350500,China

Received:2022-01-11 Revised:2022-04-01 Online:2022-05-25 Published:2022-05-25
Contact: YANG Chao,LI Yongyu E-mail:yycc32@126.com;lilin3182@163.com

摘要/Abstract

摘要：

为了探究miR168家族的进化特性及其在梨休眠进程中的表达模式,通过检索实验室前期构建的砂梨（‘黄花’梨）miRNA文库、转录组数据库和miRBase数据库,得到植物miR168家族成员的成熟体和前体序列。从‘黄花’梨花芽中克隆了两条miR168前体,发现梨miR168家族包含2条前体序列和4条成熟体序列。采用生物信息分析和实时荧光定量PCR（qRT-PCR）,对梨miR168家族的成熟体序列及前体（pre-miR168）序列进行分析,与已知植物miR168构建系统发育进化树,预测其靶基因,分析其在梨休眠发生进程中的表达情况。结果表明,梨pre-miR168序列均能形成稳定的发夹结构;植物pre-miR168两端序列较保守,而中间序列不保守;mi168成熟体保守性较高,进化树分析表明梨pre-miR168与苹果的亲缘关系最近,miR168成熟体主要分为两大类。梨miR168调控的潜在靶基因主要包含Argonaute1蛋白基因（protein argonaute 1,AGO1）和胼胝质合成酶3基因（callose synthase 3,CalS3）共21条mRNA,都是以裂解靶标的方式进行调控。qRT-PCR分析结果表明,梨miR168a相对表达量在整个休眠期间呈先上升后下降的单峰动态变化;靶基因AGO1的表达呈上升趋势,靶基因CalS3的表达呈下降趋势;AGO1的表达趋势与梨花芽的萌芽率呈正相关关系,随着萌芽率的升高而升高,而CalS3表达趋势与梨花芽的萌芽率呈负相关关系,随萌芽率升高而降低。梨miR168与其他植物miR168的序列具有高度保守性,miR168与AGO1可能通过ABA来调控梨花芽的冬季休眠,通过上调表达梨miR168a而抑制CalS3表达的方式促进梨花芽冬季休眠的解除。

关键词: 梨, miR168, 进化特性, 休眠, 表达模式

Abstract:

To explore the evolutionary characteristics of miR168 gene family and its expression pattern in dormancy process of pear,the mature and precursor sequences of all miR168 family members in plants were obtained from miRBase 22.0 and miRNA library of pear（‘Huanghua’）created by our laboratory. We cloned two precursors of miR168 from the pear flower buds,which contained two mature sequences. Bioinformatics analysis and real-time quantitative PCR（qRT-PCR）were used to analyze the mature sequences and the pre-miR168 sequences of the miR168 family of pear. The phylogenetic tree was constructed with known plant miR168,and based on target genes prediction,the expression of miR168 and target genes during dormancy development in pear was analyzed. RNAfold prediction analysis showed that the pre-miR168 sequence of pear could form stable hairpin structure. Further analysis showed that the sequences of both ends of plant pre-miR168 were conservative,while the intermediate sequences were not. The mature miR168 was highly conserved,and the evolutionary tree analysis was mainly divided into two categories. Phylogenetic analysis found that the affinity of pre-miR168 of pear was closely related to apple,which showed that species affinity is the main factor for the evolutionary characteristics of the precursor of miR168 family. PsRNA target prediction showed that a total of 21 potential mRNAs of target genes were regulated by miR168 by means of cleavage,which mainly contained protein argonaute 1 （AGO1）and callose synthase 3（CalS3）. qRT-PCR analysis showed that a dynamic change of “rise-decline expression pattern of miR168a was observed during the dormancy process of pear bud. In addition,the expression of target gene AGO1 showed an increase trend,while the expression of target gene CalS3 showed a decrease trend. Furthermore,the expression of CalS3 showed a negative correlation with germination rate of pear bud,declined with the increase of germination rate. Taken together,the work also showed that the sequence of plant miR168s was highly conserved,and may regulate flower bud dormancy together with AGO1 through ABA. The up-regulated expression of miR168a in pear may inhibit the expression of CalS3 and thus promote the release of dormancy in pear during dormancy in winter.

Key words: pear, microRNA168, evolutionary characteristics, dormancy, expression pattern

中图分类号:

S661.2

梁沁, 张延晖, 康开权, 刘瑾航, 李亮, 冯宇, 王超, 杨超, 李永裕. miR168家族进化特性及其在砂梨休眠期的表达模式分析[J]. 园艺学报, 2022, 49(5): 958-972.

LIANG Qin, ZHANG Yanhui, KANG Kaiquan, LIU Jinhang, LI Liang, FENG Yu, WANG Chao, YANG Chao, LI Yongyu. Molecular Evolution of MiR168 Family and Their Expression Profiling During Dormancy of Pyrus pyrifolia[J]. Acta Horticulturae Sinica, 2022, 49(5): 958-972.

图/表 12

参考文献 53

[1]	Arora R, Rowland L, Tanino K. 2003. Induction and release of bud dormancy in woody perennials:a science comes of age. HortScience, 38 (5):911-921. doi: 10.21273/HORTSCI.38.5.911 URL
[2]	Bartel D P. 2004. MicroRNAs:genomics,biogenesis,mechanism,and function. Cell, 116 (2):281-297. doi: 10.1016/s0092-8674(04)00045-5 pmid: 14744438
[3]	Budak H, Akpinar B A. 2015. Plant miRNAs:biogenesis,organization and origins. Functional & Integrative Genomics, 15 (5):523-531.
[4]	Chen X. 2005. microRNA biogenesis and function in plants. Febs Letters, 579 (26):5923-5931. doi: 10.1016/j.febslet.2005.07.071 URL
[5]	Cui Hai-Fang, Zhang Fan, Yin Jun-long, Guo Ying-qi, Yue Yan-ling. 2017. Callose deposition and pollen development. Journal of Yunnan Agricultural University, 32 (3):551-557. (in Chinese)
	崔海芳, 张凡, 尹俊龙, 郭瑛琪, 岳艳玲. 2017. 胼胝质沉积与花粉发育. 云南农业大学学报, 32 (3):551-557.
[6]	Ding Q, Zeng J, He X-Q. 2016. MiR169 and its target PagHAP2-6 regulated by ABA are involved in poplar cambium dormancy. Journal of Plant Physiology, 198:1-9. doi: 10.1016/j.jplph.2016.03.017 pmid: 27111502
[7]	Fazli W, Adeeb S, Taous K, Young K Y. 2010. MicroRNAs:synthesis,mechanism,function,and recent clinical trials. Biochimica et Biophysica Acta, 1803 (11):1231-243. doi: 10.1016/j.bbamcr.2010.06.013 pmid: 20619301
[8]	Gao J, Ni X, Li H, Hayat F, Shi T, Gao Z. 2021. miR169 and PmRGL2 synergistically regulate the NF-Y complex to activate dormancy release in Japanese apricot(Prunus mume Siebet Zucc.). Plant Molecular Biology, 105:83-87. doi: 10.1007/s11103-020-01070-3 URL
[9]	Hammond S M, Bernstein E, Beach D, Hannon G J. 2000. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells of science. Nature:International Weekly Journal of Science, 404 (6775):293-296.
[10]	Hemery G E, Savill P S, Pyror S N. 2005. Applications of the crown diameter-stem diameter relationship for different species of broadleaved trees. Forest Ecology Management, 215:285-294. doi: 10.1016/j.foreco.2005.05.016 URL
[11]	Jian Ling-cheng, Wang Hong. 2004. Plasmodesmatal dynamics in both woody poplar and herbaceous winter wheat under controlled short day and in field winter period. Acta Botanica Sinica, 46 (2):230-235. (in Chinese)
	简令成, 王红. 2004. 杨树和冬小麦在短日照和寒冬中的胞间连丝动态. 植物学报, 46 (2):230-235.
[12]	Jiang Weibing, Han Haozhang, Wang Liangju, Ma Kai. 2003. Advance in research of chilling requirement and the mechanism of deciduous fruit crops. Journal of Fruit Science, 20 (5):364-368. (in Chinese)
	姜卫兵, 韩浩章, 汪良驹, 马凯. 2003. 落叶果树需冷量及其机理研究进展. 果树学报, 20 (5):364-368.
[13]	Mathieu J, Levi J, Yant, Felix Mürdter, Frank Küttner, Markus Schmid. 2009. Repression of flowering by the miR172 target SMZ. PLoS Biology, 7 (7):e1000148. doi: 10.1371/journal.pbio.1000148 URL
[14]	Ko J H, Prassinos C, Han K H. 2006. Developmental and seasonal expression of PtaHB1,a Populus gene encoding a class III HD-Zip protein,is closely associated with secondary growth and inversely correlated with the level of microRNA(miR166). New Phytologist, 169 (3):469-478. doi: 10.1111/j.1469-8137.2005.01623.x URL
[15]	Lang G A, Early J D, Martin G C, Darnell R L. 1987. Endo-,para-,and ecodormancy:physiological terminology and classification for dormancy research. HortScience, 22 (3):371-377.
[16]	Li Su-fen, Zhao Yan, Ji Lu-sha. 2014. Influences of AGOl to the development of the Arabidopsis leaves with the serration margin. Journal of Hebei University of Science and Technology, 35 (1):51-57. (in Chinese)
	李素芬, 赵燕, 冀芦沙. 2014. AGOl基因对拟南芥叶边缘锯齿状发育的影响. 河北科技大学学报, 35 (1):51-57.
[17]	Li W, Cui X, Meng Z, Huang X, Xie Q, Wu H, Jin H, Zhang D, Liang W. 2012. Transcriptional regulation of Arabidopsis MIR168a and ARGONAUTE1 homeostasis in abscisic acid and abiotic stress responses. Plant Physiology, 158 (3):1279-1292. doi: 10.1104/pp.111.188789 URL
[18]	Li Xiao-dong, Lu Guo-yi, Yu Yan 1996. Effect of endogenous hormones on the dormancy of garlic(Allium sativum L.). Acta Horticulturae Sinica. 23 (2):150-154 (in Chinese)
	李晓东, 陆帼一, 于燕. 1996. 休眠‘蔡家坡’蒜内源激素水平的变化规律. 园艺学报, 23 (2):150-154.
[19]	Liu C, Xin Y, Xu L, Cai Z, Xue Y, Liu Y, Xie D, Liu Y, Qi Y. 2018. Arabidopsis ARGONAUTE 1 binds chromatin to promote gene transcription in response to hormones and stresses. Developmental Cell, 44 (3):348-361. doi: 10.1016/j.devcel.2017.12.002 URL
[20]	Liu Chang. 2017. Function of nuclear Argonaute 1 in response to various stimuli in Arabidopsis[Ph. D. Dissertation]. Beijing: China Agricultural University.
	刘畅. 2017. 拟南芥核内Argonaute 1在响应各类刺激中的功能研究[博士论文]. 北京: 中国农业大学. (in Chinese)
[21]	Liu G, Li W, Zheng P, Xu T, Chen L, Liu D, Hussain S, Teng Y. 2012. Transcriptomic analysis of‘Suli’pear(Pyrus pyrifolia white pear group) buds during the dormancy by RNA-Seq. BMC Genomics, 13 (1): 10.1186/1471-2164-13-700. doi: 10.1186/1471-2164-13-700
[22]	Liu Guo-qin, Zheng Peng-hua, Sayed Hussain, Teng Yuan-wen. 2013. Characteristics of two dormancy-associated MADS-box genes in pear and their expression analysis during the dormancy. Acta Horticulturae Sinica, 40 (4):724-732. (in Chinese)
	刘国琴, 郑鹏华, Hussain S, 滕元文. 2013. 梨两个休眠相关MADS-box基因特征及在其休眠过程中的表达分析. 园艺学报, 40 (4):724-732.
[23]	Liu Lin, Quan Xian-qing, Zhao Xiao-mei, Huang Li-hua, Feng Shang-cai, Huang Kun-yan, Zhou Xiao-yan, Su Wen-ting. 2013. Arabidopsis callose synthase gene GSL8 is required for cell wall formation and establishment and maintenance of quiescent center. Chinese Bulletin of Botany, 48 (4):389-397. (in Chinese) doi: 10.3724/SP.J.1259.2013.00389 URL
	刘林, 全先庆, 赵小梅, 黄力华, 冯尚彩, 黄坤艳, 周晓燕, 粟文婷. 2013. 拟南芥胼胝质合酶基因GSL8参与细胞壁形成和根端静止中心建立与维持. 植物学报, 48 (4):389-397. doi: 10.3724/SP.J.1259.2013.00389
[24]	Ma Xin-rui, Li Liang, Liu Jin-hang, Yang Meng-jie, Chen Jie, Liang Qin, Wu Shao-hua, LI Yong-yu. 2018. Identification and differentially expressed analysis of microRNA associated with dormancy of pear flower buds. Acta Horticulturae Sinica, 45 (11):2089-2105. (in Chinese)
	马鑫瑞, 李亮, 刘瑾航, 杨梦洁, 陈洁, 梁沁, 吴少华, 李永裕. 2018. 梨花芽休眠相关miRNA的鉴定和差异表达分析. 园艺学报, 45 (11):2089-2105.
[25]	Miao C, Wang Z, Zhang L, Yao J, Zhu J K. 2019. The grain yield modulator miR156 regulates seed dormancy through the gibberellin pathway in rice. Nature Communications, 10 (1):3822. doi: 10.1038/s41467-019-11830-5 URL
[26]	Muthamilarasan M, Manoj Prasad. 2013. Cutting-edge research on plant miRNAs. Current Science, 104 (3):287-289.
[27]	Niu Q, Li J, Cai D, Qian M, Jia H, Bai S, Sayed H, Liu G, Teng Y, Zheng X. 2016. Dormancy-associated MADS-box genes and microRNAs jointly control dormancy transition in pear(Pyrus pyrifolia white pear group)flower bud. Journal of Experimental Botany, 67 (1):239-257. doi: 10.1093/jxb/erv454 URL
[28]	Niu Qing-feng. 2016. The molecular mechanism and molecular network of pear flower bud dormancy transition[Ph. D. Dissertation]. Hangzhou: Zhejiang University. (in Chinese)
	牛庆丰. 2016. 梨花芽休眠转换的分子网络及调控机制[博士论文]. 杭州: 浙江大学.
[29]	Olsen J E. 2010. Light and temperature sensing and signaling in induction of bud dormancy in woody plants. Plant Molecular Biology, 73 (1-2):37-47. doi: 10.1007/s11103-010-9620-9 URL
[30]	Pacey-Miller T, Scott K, Ablett E, Tingey S, Ching A, Henry R. 2003. Genes associated with the end of dormancy in grapes. Functional Integrative Genomics, 3 (4):144-152. doi: 10.1007/s10142-003-0094-6 URL
[31]	Peng S, Liang D, Zhang Z, Yin W, Xia X. 2013. Identification of drought-responsive and novel Populus trichocarpa microRNAs by high-throughput sequencing and their targets using degradome analysis. BMC Genomics, 14 (1):233. doi: 10.1186/1471-2164-14-233 URL
[32]	Rajesh Kumar Singh, Pal Miskolczi, Jay P Maurya, Rishikesh P Bhalerao. 2019. A tree ortholog of SHORT VEGETATIVE PHASE floral repressor mediates photoperiodic control of bud dormancy. Current Biology, 29 (1):128-133. doi: S0960-9822(18)31472-6 pmid: 30554900
[33]	Rhoades M, Reinhart B, Lim L, Burge C, Bartel B, Bartel D. 2002. Prediction of plant microRNA targets. Cell, 110 (4):513-520. doi: 10.1016/S0092-8674(02)00863-2 URL
[34]	Rinne P L, Kaikuranta P M, Schoot C V D. 2001. The shoot apical meristem restores its symplasmic organization during chilling-induced release from dormancy. Plant Journal, 26 (3):249-264. pmid: 11439114
[35]	Rinne P L, Schoot C V D. 2003. Plasmodesmata at the crossroads between development,dormancy,and defense. Canadian Journal of Botany, 276 (1-2):133-137.
[36]	Rohde A, Bhalerao R. 2007. Plant dormancy in the perennial context. Trends in Plant Science, 12 (5):217-223. doi: 10.1016/j.tplants.2007.03.012 URL
[37]	Rui Shi, Vincent L Ching. 2005. Facile means for quantifying microRNA expression by real-time PCR. Biotechniques, 39 (4):519-524. doi: 10.2144/000112010 pmid: 16235564
[38]	Ruttink T, Arend M, Morreel K, Storme V, Rombauts S, Fromm J, Bhalerao R P, Boerjan W, Rohde A. 2007. A molecular timetable for apical bud formation and dormancy induction in poplar. Plant Cell Reports, 19 (8):2370-2390.
[39]	Shi Ting, Gao Zhi-hong, Zhang Zhen, Zhuang Wei-bing. 2010. Advance of research on microRNA in flower development regulation. Chinese Agricultural Science Bulletin, 26 (13):267-271. (in Chinese)
	侍婷, 高志红, 章镇, 庄维兵. 2010. MicroRNA参与植物花发育调控的研究进展. 中国农学通报, 26 (13):267-271.
[40]	Siré C, Moreno A B, Garcia-Chapa M, López-Moya J J, Segundo B S. 2009. Diurnal oscillation in the accumulation of Arabidopsis microRNAs,miR167,miR168,miR171 and miR398. Febs Letters, 583 (6):1039-1044. doi: 10.1016/j.febslet.2009.02.024 URL
[41]	Su Li-Yao, Jiang Meng-Qi, Huang Shu-Qi, Xu Xiao-Ping, Li Xue, Chen Xu, Lai Zhong-Xiong, Lin Yu-Ling. 2019. Analysis of the molecular evolution characteristics of miR403 and its expression pattern during analysis of the molecular evolution characteristics of miR403 and its expression pattern during early somatic embryogenesis in Dimocarpus longan. Journal of Fruit Science, 36 (7):846-856. (in Chinese)
	苏立遥, 蒋梦琦, 黄倏祺, 徐小萍, 厉雪, 陈旭, 赖钟雄, 林玉玲. 2019. miR403分子进化特性及其在龙眼体胚发生早期的表达模式分析. 果树学报, 36 (7):846-856.
[42]	Sun L, Ai X, Li W, Guo W, Deng X, Hu C, Zhang J, Wu K. 2012. Identification and comparative profiling of miRNAs in an early flowering mutant of trifoliate orange and its wild type by genome-wide deep sequencing. PLoS ONE, 7 (8):doi: 10.1371/journal.pone.0043760. doi: 10.1371/journal.pone.0043760 URL
[43]	Várallyay Éva, Válóczi Anna, Ágyi Ákos, Burgyán József, Havelda Zoltán. 2017. Plant virus-mediated induction of miR168 is associated with repression of ARGONAUTE1 accumulation. The EMBO Journal, 36 (7):846-856.
[44]	Wang J W, Park M, Wang L J, Koo Y, Chen X Y, Weigel D, Poethig R, Sederoff R. 2011. MiRNA control of vegetative phase change in trees. PLoS Genetics, 7 (2):doi: 10.1371/journal.pgen.1002012. doi: 10.1371/journal.pgen.1002012 URL
[45]	Wang Xin-chao, Ma Chun-lei, Yang Ya-jun. 2011. Progress in research on bud dormancy and its regulation mechanisms in perennial plants. Chinese Journal of Applied & Environmental Biology, 17 (4):589-595. (in Chinese)
	王新超, 马春雷, 杨亚军. 2011. 多年生植物的芽休眠及调控机理研究进展. 应用与环境生物学报, 17 (4):589-595.
[46]	Wang Yan-fang, Zhou Rui-lian, Zhao Yan-hong. 2015. Molecular evolution of miR-171 gene family and prediction of their target genes. Life Science Research, 19 (6):479-483. (in Chinese)
	王艳芳, 周瑞莲, 赵彦宏. 2015. miR-171基因家族进化分析及靶基因预测. 生命科学研究, 19 (6):479-483.
[47]	Welling A, Palva E T. 2010. Molecular control of cold acclimation in trees. Physiologia Plantarum, 127 (2):167-181. doi: 10.1111/j.1399-3054.2006.00672.x URL
[48]	Wu G, Park M Y, Conway R. S, Wang J-W, Weigel D, Poethig R S. 2009. The sequential action of miR156 and miR172 regulates developmental timing in Arabidopsis. Cell, 138 (4):750-759. doi: 10.1016/j.cell.2009.06.031 URL
[49]	Xian Zhi-qiang. 2014. Isolation and characterisation of SlAGO genes and study on biological function of miR168 and AGO1 in tomato[Ph. D. Dissertation]. Chongqing: Chongqing University. (in Chinese)
	先志强. 2014. 番茄AGO基因的分离鉴定及SlmiR168,AGO1在番茄生长发育过程中的功能研究[博士论文]. 重庆: 重庆大学.
[50]	Yadav S R, Yan D, Sevilem I, Ykä Helariutta. 2014. Plasmodesmata-mediated intercellular signaling during plant growth and development. Frontiers in Plant Science, 5,doi: 10.3389/fpls.2014.00044. doi: 10.3389/fpls.2014.00044 URL
[51]	Yamane H, Ooka T, Jotatsu H, Hosaka Y, Sasaki R, Tao R. 2011. Expressional regulation of PpDAM5 and PpDAM6,peach(Prunus persica) dormancy-associated MADS-box genes,by low temperature and dormancy-breaking reagent treatment. Journal of Experimental Botany, 62 (10):3481-3488. doi: 10.1093/jxb/err028 URL
[52]	Zhang Y, Wang Y, Gao X, Liu C, Gai S. 2018. Identification and characterization of microRNAs in tree peony during chilling induced dormancy release by high-throughput sequencing. Scientific Reports, 8 (1):4537. doi: 10.1038/s41598-018-22415-5 pmid: 29540706
[53]	Zhao X, Chen S, Zhao L, Zhang X, Ma P, Gou Z. 2014. Evolution of MIR166 gene family in land plants. Plant Diversity and Resources, 36 (3):331-341.

用途Purpose	引物名称Primer name	引物序列（5′-3′）Primer sequence
克隆Clone	ppy-miR168a-F	ATAGAGGCGTAGGCCTGTGT
	ppy-miR168a-R	TGATGGTGGGAATTGGGATAT
克隆Clone	ppy-miR168b-F	TGTCTGTTTGGGCGTTGATT
	ppy-miR168b-R	AAACTATTGTAGATCAGACAACAACGC
qRT-PCR	Ppy-miR168a	TCGCTTGGTGCAGGTCG
	AGO1-F	CACCAGCACCACAACAATA
	AGO1-R	ACCACCACCATAGCCTCT
	CalS3-F	AGAGGCGGATAACTCG
	CalS3-R	GCAACACGAAGAATGG
	Actin-F	CCATCCAGGCTGTTCTCTC
	Actin-R	GCAAGGTCCAGACGAAGG
	5SrRNA-F	TAGGCGTAGAGGAACCACACCAAT
	5SrRNA-R	ACAGTATCGTCACCGCAGTAGAGT

用途Purpose	引物名称Primer name	引物序列（5′-3′）Primer sequence
克隆Clone	ppy-miR168a-F	ATAGAGGCGTAGGCCTGTGT
	ppy-miR168a-R	TGATGGTGGGAATTGGGATAT
克隆Clone	ppy-miR168b-F	TGTCTGTTTGGGCGTTGATT
	ppy-miR168b-R	AAACTATTGTAGATCAGACAACAACGC
qRT-PCR	Ppy-miR168a	TCGCTTGGTGCAGGTCG
	AGO1-F	CACCAGCACCACAACAATA
	AGO1-R	ACCACCACCATAGCCTCT
	CalS3-F	AGAGGCGGATAACTCG
	CalS3-R	GCAACACGAAGAATGG
	Actin-F	CCATCCAGGCTGTTCTCTC
	Actin-R	GCAAGGTCCAGACGAAGG
	5SrRNA-F	TAGGCGTAGAGGAACCACACCAAT
	5SrRNA-R	ACAGTATCGTCACCGCAGTAGAGT

科 Family	物种 Species	简写 Logogram	miR168数量 Quantity of miR168	pre-miR168数量 Quantity of pre-miR168
十字花科Brassicaceae	拟南芥Arabidopsis thaliana	ath	4	2
	油菜Brassica napus	bna	2	2
	琴叶拟南芥Arabidopsis lyrata	aly	4	2
	芜菁Brassica rapa	bra	6	3
	亚麻荠Camelina sativa	csa	1	1
禾本科Poaceae	青稞Hordeum vulgare	hvu	2	1
	二穗短柄草Brachypodium distachyon	bdi	2	1
	甘蔗属Saccharum sp	ssp	1	1
	甘蔗Saccharum officinarum	sof	2	2
	水稻Oryza sativa	osa	3	2
	高粱Sorghum bicolor	sbi	1	1
	玉米Zea mays	zma	4	2
	山羊草Aegilops tauschii	ata	1	1
豆科Leguminosae	大豆Glycine max	gma	2	2
	蒺藜苜蓿Medicago truncatula	mtr	4	3
	大叶相思Acacia auriculiformis	aau	1	1
	豇豆Vigna unguiculata	vun	1	1
	百脉根Lotus japonicus	lja	1	1
科 Family	物种 Species	简写 Logogram	miR168数量 Quantity of miR168	pre-miR168数量 Quantity of pre-miR168
杨柳科Salicaceae Mirb	毛果杨Populus trichocarpa	ptc	4	2
葡萄科Vitaceae Juss	葡萄Vitis vinifera	vvi	1	1
毛茛科Ranunculaceae Juss	耧斗菜Aquilegia viridiflora	aqc	1	1
芸香科Rutaceae Juss	脐橙Citrus sinensis	csi	2	1
	柑橘Citrus reticulata	cre	1	1
	克萊门柚Citrus clementina	ccl	1	1
大戟科Rosaceae Juss	木薯Manihot esculenta	mes	1	1
	三叶橡胶树 Hevea brasiliensis	hbr	1	1
	蓖麻 Ricinus communis	rco	1	1
葫芦科Cucurbitaceae	甜瓜Cucumis melo	cme	1	1
菊科Asteraceae Bercht	刺苞菜蓟Cynara cardunculus	cca	1	1
亚麻科Linaceae	亚麻Linum usitatissimum	lus	2	2
茄科Solanaceae Juss	烟草Nicotiana tabacum	nta	5	5
茄科Solanaceae Juss	番茄Lycopersicon esculentum	sly	4	2
蔷薇科Rosaceae Juss	苹果Malus domestica	mdm	2	2
	碧桃Prunus persica	ppe	1	1
	森林草莓Fragaria vesca	fve	1	1
凤梨科Bromeliaceae Juss	莺歌凤梨Vriesea carinata	vcr	4	2
松科Pinaceae Lindl	挪威云杉Picea abies	pab	2	2
百合科Liliaceae Juss	芦笋Asparagus officinalis	aof	2	2
梧桐科Sterculiaceae	可可Theobroma cacao	tcc	1	1

科 Family	物种 Species	简写 Logogram	miR168数量 Quantity of miR168	pre-miR168数量 Quantity of pre-miR168
十字花科Brassicaceae	拟南芥Arabidopsis thaliana	ath	4	2
	油菜Brassica napus	bna	2	2
	琴叶拟南芥Arabidopsis lyrata	aly	4	2
	芜菁Brassica rapa	bra	6	3
	亚麻荠Camelina sativa	csa	1	1
禾本科Poaceae	青稞Hordeum vulgare	hvu	2	1
	二穗短柄草Brachypodium distachyon	bdi	2	1
	甘蔗属Saccharum sp	ssp	1	1
	甘蔗Saccharum officinarum	sof	2	2
	水稻Oryza sativa	osa	3	2
	高粱Sorghum bicolor	sbi	1	1
	玉米Zea mays	zma	4	2
	山羊草Aegilops tauschii	ata	1	1
豆科Leguminosae	大豆Glycine max	gma	2	2
	蒺藜苜蓿Medicago truncatula	mtr	4	3
	大叶相思Acacia auriculiformis	aau	1	1
	豇豆Vigna unguiculata	vun	1	1
	百脉根Lotus japonicus	lja	1	1
科 Family	物种 Species	简写 Logogram	miR168数量 Quantity of miR168	pre-miR168数量 Quantity of pre-miR168
杨柳科Salicaceae Mirb	毛果杨Populus trichocarpa	ptc	4	2
葡萄科Vitaceae Juss	葡萄Vitis vinifera	vvi	1	1
毛茛科Ranunculaceae Juss	耧斗菜Aquilegia viridiflora	aqc	1	1
芸香科Rutaceae Juss	脐橙Citrus sinensis	csi	2	1
	柑橘Citrus reticulata	cre	1	1
	克萊门柚Citrus clementina	ccl	1	1
大戟科Rosaceae Juss	木薯Manihot esculenta	mes	1	1
	三叶橡胶树 Hevea brasiliensis	hbr	1	1
	蓖麻 Ricinus communis	rco	1	1
葫芦科Cucurbitaceae	甜瓜Cucumis melo	cme	1	1
菊科Asteraceae Bercht	刺苞菜蓟Cynara cardunculus	cca	1	1
亚麻科Linaceae	亚麻Linum usitatissimum	lus	2	2
茄科Solanaceae Juss	烟草Nicotiana tabacum	nta	5	5
茄科Solanaceae Juss	番茄Lycopersicon esculentum	sly	4	2
蔷薇科Rosaceae Juss	苹果Malus domestica	mdm	2	2
	碧桃Prunus persica	ppe	1	1
	森林草莓Fragaria vesca	fve	1	1
凤梨科Bromeliaceae Juss	莺歌凤梨Vriesea carinata	vcr	4	2
松科Pinaceae Lindl	挪威云杉Picea abies	pab	2	2
百合科Liliaceae Juss	芦笋Asparagus officinalis	aof	2	2
梧桐科Sterculiaceae	可可Theobroma cacao	tcc	1	1

靶基因注释 Target gene annotation	靶基因编号 Target accession	期望值 Expection
AGO1蛋白 Protein argonaute 1-like transcript variant	XM_018645622.1	0
	XM_018647395.1	0
	XM_009349800.2	0
	XM_018645623.1	0
	XM_018647396.1	0
	XM_009357682.2	0
胼胝质合成酶3（CalS3） Callose synthase 3（CalS3）transcript variant	XM_018649103.1	2.5
	XM_009376827.2	2.5
	XM_018651590.1	2.5
	XM_018651591.1	2.5
	XM_009365800.2	2.5
	XM_018649104.1	2.5
	XM_009365801.2	2.5
	XM_009365802.2	2.5
	XM_009365803.2	2.5
AGO1蛋白Protein argonaute 1-like transcript variant	XM_009359066.2	2.5
	XM_009349798.2	2.5
	XM_009359065.2	2.5
	XM_009349797.2	2.5
	XM_009357684.2	2.5
	XM_009357683.2	2.5

miR168家族进化特性及其在砂梨休眠期的表达模式分析

Molecular Evolution of MiR168 Family and Their Expression Profiling During Dormancy of Pyrus pyrifolia

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 12

参考文献 53

相关文章 15

编辑推荐

Metrics

本文评价

访问总数:　当日访问总数:　当前在线人数:

处理日期（M-D）Treatment date	ppy-miR168a	AGO1	CalS3
11-01	0.22	0	4.00
11-25	3.37	0.05	1.03
12-27	1.12	0.58	0.75
01-23	1.00	5.91	0.59
相关系数Correlation coefficient		-0.223	-0.503

[1]	宋健坤, 杨英杰, 李鼎立, 马春晖, 王彩虹, 王然. 梨新品种‘鲁秀’[J]. 园艺学报, 2022, 49(S2): 3-4.
[2]	董星光, 曹玉芬, 张莹, 田路明, 霍宏亮, 齐丹, 徐家玉, 刘超, 王立东. 抗寒脆肉梨新品种‘玉翠香’[J]. 园艺学报, 2022, 49(S2): 5-6.
[3]	欧春青, 姜淑苓, 王斐, 马力, 张艳杰, 刘振杰. 早熟梨新品种‘兴梨蜜水’[J]. 园艺学报, 2022, 49(S2): 7-8.
[4]	张艳杰, 王斐, 欧春青, 马力, 姜淑苓, 刘振杰. 梨新品种‘中梨玉脆3’[J]. 园艺学报, 2022, 49(S2): 9-10.
[5]	范净, 陈启亮, 张靖国, 杨晓平, 杜威, 田瑞, 周德平, 胡红菊, . 中熟红皮砂梨新品种‘金彤’[J]. 园艺学报, 2022, 49(S2): 11-12.
[6]	王苏珂, 李秀根, 杨健, 王龙, 苏艳丽, 张向展, 薛华柏. 红皮梨新品种‘丹霞红’[J]. 园艺学报, 2022, 49(S2): 13-14.
[7]	王斐, 欧春青, 张艳杰, 马力, 姜淑苓. 晚熟耐贮梨新品种‘华秋’[J]. 园艺学报, 2022, 49(S1): 9-10.
[8]	宋健坤, 李鼎立, 杨英杰, 马春晖, 王彩虹, 王然. 梨新品种‘琴岛红’[J]. 园艺学报, 2022, 49(S1): 11-12.
[9]	郭伟珍, 赵京献, 李莹. 中早熟梨新品种‘美玉’[J]. 园艺学报, 2022, 49(9): 2051-2052.
[10]	刘金明, 郭彩华, 袁星, 亢超, 全绍文, 牛建新. 梨Dof家族基因鉴定及其在宿存与脱落萼片中的表达分析[J]. 园艺学报, 2022, 49(8): 1637-1649.
[11]	陶鑫, 朱荣香, 贡鑫, 吴磊, 张绍铃, 赵建荣, 张虎平. 梨果糖激酶基因PpyFRK5在果实蔗糖积累中的作用[J]. 园艺学报, 2022, 49(7): 1429-1440.
[12]	张秋悦, 刘昌来, 于晓晶, 杨甲定, 封超年. 盐胁迫条件下杜梨叶片差异表达基因qRT-PCR内参基因筛选[J]. 园艺学报, 2022, 49(7): 1557-1570.
[13]	周徐子鑫, 杨威, 毛美琴, 薛彦斌, 马均. 金边红苞凤梨叶色突变体色素鉴定及类胡萝卜素合成限速基因筛选[J]. 园艺学报, 2022, 49(5): 1081-1091.
[14]	向妙莲, 吴帆, 李树成, 马巧利, 王印宝, 肖刘华, 陈金印, 陈明. 外源褪黑素调控活性氧代谢诱导梨果实抗采后黑斑病[J]. 园艺学报, 2022, 49(5): 1102-1110.
[15]	王宏, 杨王莉, 蔺经, 杨青松, 李晓刚, 盛宝龙, 常有宏. 早熟砂梨‘苏翠1号’与其亲本‘翠冠’‘华酥’成熟果实差异代谢产物及差异基因比较分析[J]. 园艺学报, 2022, 49(3): 493-508.

miR168家族进化特性及其在砂梨休眠期的表达模式分析

Molecular Evolution of MiR168 Family and Their Expression Profiling During Dormancy of Pyrus pyrifolia

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 12

参考文献 53

相关文章 15

编辑推荐

Metrics

本文评价

访问总数: 当日访问总数: 当前在线人数:

访问总数:　当日访问总数:　当前在线人数: