榛子胚珠不同发育阶段circRNA的分析与鉴定

doi:10.16420/j.issn.0513-353x.2020-0653

摘要/Abstract

摘要：

在榛子胚珠不同发育时期筛选差异表达的circRNA,进一步鉴定其差异表达的靶mRNA,以期明确circRNA在胚珠中的表达特征及其对胚珠发育的调控作用。在榛子胚珠形成期（Ov1）、早期生长期（Ov2）、快速生长期（Ov3）和成熟期（Ov4）进行胚珠样品采集和Illumina高通量测序,结合已有的mRNA测序分析结果,鉴定参与胚珠发育的差异表达circRNA（DEcircRNA）及其靶差异表达mRNA（DEmRNA）。共鉴定到7 122条circRNA,59.54%的circRNA来自外显子区域。在成对比较Ov1-vs-Ov2、Ov1-vs-Ov3、Ov1-vs-Ov4、Ov2-vs-Ov3、Ov2-vs-Ov4和Ov3-vs-Ov4中,分别鉴定到123、162、187、85、105和21条DEcircRNA;DEcircRNA的表达有明显的发育时期特异性;有199条DEcircRNA的靶mRNA也发生差异表达,两者表达变化高度同步。GO（Gene Ontology）分类结果表明DEcircRNA参与胚珠发育过程中代谢的调控。随机选择了4个差异表达circRNA及靶mRNA,用qRT-PCR分析其在4个胚珠发育阶段表达量的变化,证实qRT-PCR与Illumina高通量测序结果相一致。

关键词: 榛子, 转录组, 胚珠, 环状RNA

Abstract:

The differentially expressed circRNA（DEcircRNA）of hazel ovule were screened at different developmental stages,and their target differentially expressed mRNA（DEmRNA）was further identified to clarify the regulatory effect of circRNAs on ovule development. Samples were collected from ovule formation stage（Ov1）,early growth stage（Ov2）,rapid growth stage（Ov3）and maturation stage（Ov4）of hazel. The DEcircRNA involved in ovule development was identified based on the available results of mRNA sequencing analysis at four developmental stages. A total of 7 122 circRNAs were identified,and 59.54% of them came from exon region. In pairwise comparisons of Ov1-vs-Ov2,Ov1-vs-Ov3,Ov1-vs-Ov4,Ov2-vs-Ov3,Ov2-vs-Ov4 and Ov3-vs-Ov4,123,162,187,85,105 and 21 DEcircRNAs were identified respectively. The expression pattern of DEcircRNAs was developmental stage specific. Correlation analysis showed that 199 target mRNAs of DEcircRNA were also differentially expressed. The expression changes of DEcircRNA and their responding DEmRNA were highly synchronized. The results of GO（Gene Ontology）classification indicated that DEcircRNAs were involved in the regulation of metabolism during ovule development. Four DEcircRNAs and their target differentially expressed mRNAs at four ovule development stages were randomly selected and subjected to qRT-PCR analysis,and their expression levels changes were consistent with the results of high-throughput sequencing of microRNAs.

Key words: hazel, transcriptome, ovule, circular RNA

中图分类号:

S664.4

刘剑锋, 孙莹, 魏珩, 贺红利, 张兴政, 程云清. 榛子胚珠不同发育阶段circRNA的分析与鉴定[J]. 园艺学报, 2021, 48(6): 1053-1066.

LIU Jianfeng, SUN Ying, WEI Heng, HE Hongli, ZHANG Xingzheng, CHENG Yunqing. Analysis and Identification of circRNAs of Hazel Ovule at Different Developmental Stages[J]. Acta Horticulturae Sinica, 2021, 48(6): 1053-1066.

图/表 9

参考文献 50

[1]	Ambros V R. 2004. The functions of animal microRNAs. Nature, 431:350-355. doi: 10.1038/nature02871 URL
[2]	An Lin-jun, Luan Jia-yu, Ren Li, Li Hui-yu, Xia De-an. 2019. Bioinformatics and expression characteristics analysis of BpTCP8 in Betula platyphylla Suk. Journal of Nanjing Forestry Univeristy(Natural Sciences Edition), 43:67-73. (in Chinese)
	安琳君, 栾嘉豫, 任丽, 李慧玉, 夏德安. 2019. 白桦 BpTCP8基因生物信息学及表达特性分析. 南京林业大学学报(自然科学版), 43:67-73.
[3]	Apweiler R, Bairoch A M, Wu C H, Barker W C, Boeckmann B, Ferro S, Gasteiger E, Huang H, Lopez R, Magrane M. 2004. UniProt:the universal protein knowledgebase. Nucleic Acids Research, 32:115-119. doi: 10.1093/nar/gkh151 URL
[4]	Ashburner M, Ball C A, Blake J A, Botstein D, Butler H, Cherry J M, Davis A P, Dolinski K, Dwight S S, Eppig J T, Harris M A, Hill D P, Issel-Tarver L, Kasarskis A, Lewis S, Matese J C, Richardson J E, Ringwald M, Rubin G M, Sherlock G. 2000. Gene ontology:tool for the unification of biology. Nature Genetics, 25:25-29. doi: 10.1038/75556 URL
[5]	Ashwalfluss R, Meyer M, Pamudurti N R, Ivanov A, Bartok O, Hanan M, Evantal N, Memczak S, Rajewsky N, Kadener S. 2014. circRNA biogenesis competes with pre-mRNA splicing. Molecular Cell, 56:55-66. doi: 10.1016/j.molcel.2014.08.019 URL
[6]	Bartel D P. 2004. microRNAs:genomics,biogenesis,mechanism,and function. Cell, 116:281-297. pmid: 14744438
[7]	Bartel D P. 2009. microRNAs:target recognition and regulatory functions. Cell, 136:215-233. doi: 10.1016/j.cell.2009.01.002 pmid: 19167326
[8]	Bartel D P. 2018. Metazoan microRNAs. Cell, 173:20-51. doi: S0092-8674(18)30286-1 pmid: 29570994
[9]	Cao H P, Zhang L, Tan X F, Long H X, Shockey J M. 2014. Identification,classification and differential expression of oleosin genes in tung tree (Vernicia fordii). PLoS ONE, 9:e88409. doi: 10.1371/journal.pone.0088409 URL
[10]	Cao M J, Wang Z, Wirtz M, Hell R, Oliver D J, Xiang C B. 2013. SULTR3;1 is a chloroplast-localized sulfate transporter in Arabidopsis thaliana. Plant Journal, 73:607-616. doi: 10.1111/tpj.2013.73.issue-4 URL
[11]	Cheng Yun-qing, Zhang Li-na, Zhao Yong-bin, Liu Jian-feng. 2018. Analysis of SSR markers information and primer selection from transcriptome sequence of hybrid hazelnut Corylus heterophylla × C. avellana. Acta Horticulturae Sinica, 45 (1):139-148. (in Chinese)
	程云清, 张丽娜, 赵永斌, 刘剑锋. 2018. 平欧杂交榛转录组中SSR信息分析和引物筛选. 园艺学报, 45 (1):139-148.
[12]	Deng Yang-yang, Li Jian-qi, Wu Song-feng, Zhu Yun-ping, Chen Yao-wen, He Fu-chu. 2006. Integrated nr database in protein annotation system and its localization. Computer Engineering, 32:71-74. (in Chinese)
	邓泱泱, 荔建琦, 吴松锋, 朱云平, 陈耀文, 贺福初. 2006. nr数据库分析及其本地化. 计算机工程, 32:71-73.
[13]	Eddy S R. 1998. Profile hidden Markov models. Bioinformatics, 14:755-763. pmid: 9918945
[14]	Gao Zhen, Luo Meng, Wang Lei, Song Shiren, Zhao Liping, Xu Wenping, Zhang Caixi, Wang Shiping, Ma Chao. 2019. The research advance of plant circular RNA. Acta Horticulturae Sinica, 46 (1):171-181.
	高振, 骆萌, 王磊, 宋士任, 赵丽萍, 许文平, 张才喜, 王世平, 马超. 2019. 植物环状RNA研究进展. 园艺学报, 46 (1):171-181.
[15]	Hansen T B, Jensen T I, Clausen B H, Bramsen J B, Finsen B, Damgaard C K, Kjems J. 2013. Natural RNA circles function as efficient microRNA sponges. Nature, 495:384-388. doi: 10.1038/nature11993 URL
[16]	Hu Xiao-yi, Tan Xiao-feng, Tian Xiao-ming, Liu Qiao, Luo Qian, Chen Hong-peng, Hu Fang-min. 2008. Identification and analysis of an aquaporin (CoPIP1-1)in the seeds of Camellia oleifer. Scientia Silvae Sinicae, 44:48-56. (in Chinese)
	胡孝义, 谭晓风, 田晓明, 刘巧, 罗茜, 陈鸿鹏, 胡芳名. 2008. 油茶种子水通道蛋白CoPIP1-1的鉴定与分析. 林业科学, 44:51-59.
[17]	Jeck W R, Sharpless N E. 2014. Detecting and characterizing circular RNAs. Nature Biotechnology, 32:453-461. doi: 10.1038/nbt.2890 URL
[18]	Kanehisa M, Goto S, Kawashima S, Okuno Y, Hattori M. 2004. The KEGG resource for deciphering the genome. Nucleic Acids Research, 32:277-280.
[19]	Koonin E V, Fedorova N D, Jackson J D, Jacobs A R, Krylov D M, Makarova K S, Mazumder R, Mekhedov S L, Nikolskaya A N, Rao B S. 2004. A comprehensive evolutionary classification of proteins encoded in complete eukaryotic genomes. Genome Biology, 5:1-28.
[20]	Kristensen L S, Andersen M S, Stagsted L V W, Ebbesen K K, Hansen T B, Kjems J. 2019. The biogenesis,biology and characterization of circular RNAs. Nature Reviews Genetics, 20:675-691. doi: 10.1038/s41576-019-0158-7
[21]	Le Hir R, Sorin C, Chakraborti D, Moritz T, Schaller H, Tellier F, Robert S, Morin H, Bako L, Bellini C. 2013. ABCG9,ABCG11 and ABCG14 ABC transporters are required for vascular development in Arabidopsis. Plant Journal, 76:811-824. doi: 10.1111/tpj.12334 URL
[22]	Li B, Ruotti V, Stewart R M, Thomson J A, Dewey C N. 2010. RNA-Seq gene expression estimation with read mapping uncertainty. Bioinformatics, 26:493-500. doi: 10.1093/bioinformatics/btp692 URL
[23]	Li J Q, Yang J, Zhou P, Le Y P, Zhou C W, Wang S M, Xu D Z, Lin H K, Gong Z H. 2015a. Circular RNAs in cancer:novel insights into origins,properties,functions and implications. American Journal of Cancer Research, 5:472-480.
[24]	Li Y B, Fan C C, Xing Y Z, Jiang Y H, Luo L J, Sun L, Shao D, Xu C J, Li X H, Xiao J H. 2011. Natural variation in GS5 plays an important role in regulating grain size and yield in rice. Nature Genetics, 43:1266-1269. doi: 10.1038/ng.977 URL
[25]	Li Z Y, Huang C, Bao C, Chen L, Lin M, Wang X L, Zhong G L, Yu B, Hu W C, Dai L M. 2015b. Exon-intron circular RNAs regulate transcription in the nucleus. Nature Structural & Molecular Biology, 22:256-264. doi: 10.1038/nsmb.2959 URL
[26]	Liu J F, Cheng Y Q, Yan K, Liu Q, Wang Z W. 2012. The relationship between reproductive growth and blank fruit formation in Corylus heterophylla Fisch. Scientia Horticulturae, 136:128-134. doi: 10.1016/j.scienta.2012.01.008 URL
[27]	Liu J F, Luo Q Z, Zhang X Z, Zhang Q, Cheng Y Q. 2020. Identification of vital candidate microRNA/mRNA pairs regulating ovule development using high-throughput sequencing in hazel. BMC Developmental Biology, 20:13. doi: 10.1186/s12861-020-00219-z URL
[28]	Liu J F, Zhang H D, Cheng YQ, Kafkas S, Guney M. 2014. Pistillate flower development and pollen tube growth mode during the delayed fertilization stage in Corylus heterophylla Fisch. Plant Reproduction, 27:145-152. doi: 10.1007/s00497-014-0248-9 URL
[29]	Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT method. Methods, 25:402-408. pmid: 11846609
[30]	Lü Dong-lin, Guo Yi-wen, Han Rui, Jiang Jing. 2018. Characterization of gene expression in anthocyanin synthesisand salt tolerance of Betula pendula 'Purple Rain'. Journal of Nanjing Forestry Univeristy(Natural Sciences Edition), 42:25-32. (in Chinese)
	吕东林, 林琳, 郭译文, 韩锐, 姜静. 2018. 紫雨桦耐盐性及花色苷合成相关基因的表达特性. 南京林业大学学报(自然科学版), 42:28-35.
[31]	Ma L, Li T, Hao C Y, Wang Y Q, Chen X H, Zhang X Y. 2016. TaGS5-3A,a grain size gene selected during wheat improvement for larger kernel and yield. Plant Biotechnology Journal, 14:1269-1280. doi: 10.1111/pbi.2016.14.issue-5 URL
[32]	Memczak S, Jens M, Elefsinioti A, Torti F, Krueger J, Rybak A, Maier L, Mackowiak S D, Gregersen L H, Munschauer M. 2013. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature, 495:333-338. doi: 10.1038/nature11928 URL
[33]	Nanjo Y, Oka H, Ikarashi N, Kaneko K, Kitajima A, Mitsui T, Munoz F, Rodriguezlopez M, Barojafernandez E, Pozuetaromero J. 2006. Rice plastidial N-glycosylated nucleotide pyrophosphatase/phosphodiesterase is transported from the ER-golgi to the chloroplast through the secretory pathway. Plant Cell, 18:2582-2592. doi: 10.1105/tpc.105.039891 URL
[34]	Philips A, Nowis K, Stelmaszczuk M, Podkowiński J, Handschuh L, Jackowiak P, Figlerowicz M. 2020. Arabidopsis thaliana cbp80, c2h2,and flk knockout mutants accumulate increased amounts of circular RNAs. Cells, 9:1937. doi: 10.3390/cells9091937 URL
[35]	Ranocha P, Dima O, Nagy R, Felten J, Corratgé-Faillie C, Novák O, Morreel K, Lacombe B, Martinez Y, Pfrunder S, Jin X, Renou J P, Thibaud J B, Ljung K, Fischer U, Martinoia E, Boerjan W, Goffner D. 2013. Arabidopsis WAT 1 is a vacuolar auxin transport facilitator required for auxin homoeostasis. Nature Communications, 4:2625. doi: 10.1038/ncomms3625 URL
[36]	Shi Tian-pei, Wang Xin-yue, Hou Hao-bin, Zhao Zhi-da, Shang Ming-yu, Zhang Li. 2020. Analysis and identification of circRNAs of skeletal muscle at different stages of sheep embryos based on whole transcriptome sequencing. Scientia Agricultura Sinica, 53:642-657. (in Chinese)
	石田培, 王欣悦, 侯浩宾, 赵志达, 尚明玉, 张莉. 2020. 基于全转录组测序的绵羊胚胎不同发育阶段骨骼肌circRNA的分析与鉴定. 中国农业科学, 53:642-657.
[37]	Tan J J, Zhou Z J, Niu Y J, Sun X Y, Deng Z P. 2017. Identification and functional characterization of tomato circRNAs derived from genes involved in fruit pigment accumulation. Scientific Reports, 7:8594. doi: 10.1038/s41598-017-08806-0 URL
[38]	Tatusov R L, Galperin M Y, Natale D A, Koonin E V. 2000. The COG database:a tool for genome-scale analysis of protein functions and evolution. Nucleic Acids Research, 28:33-36.
[39]	Tay Y, Rinn J L, Pandolfi P P. 2014. The multilayered complexity of ceRNA crosstalk and competition. Nature, 505:344-352. doi: 10.1038/nature12986 URL
[40]	Tian Xue-yao, Zhou Jie, Wang Bao-song, He Kai-yue, He Xu-dong. 2020. Cloning and expression pattern analysis of NAC genes in Salix. Journal of Nanjing Forestry Univeristy(Natural Sciences Edition), 44:123-128. (in Chinese)
	田雪瑶, 周洁, 王保松, 何开跃, 何旭东. 2020. 柳树 NAC基因的克隆与表达模式分析. 南京林业大学学报(自然科学版), 44:123-128.
[41]	Wang H L, Xiao Y, Wu L, Ma D C. 2018. Comprehensive circular RNA profiling reveals the regulatory role of the circRNA-000911/miR-449a pathway in breast carcinogenesis. International Journal of Oncology, 52:743-754.
[42]	Wang Y, Xiong Z Y, Li Q, Sun Y Y, Jin J, Chen H, Zou Y, Huang X G, Ding Y. 2019. Circular RNA profiling of the rice photo-thermosensitive genic male sterile line Wuxiang S reveals circRNA involved in the fertility transition. BMC Plant Biology, 19:340. doi: 10.1186/s12870-019-1944-2 URL
[43]	Wang Y X, Wang Q, Gao L P, Zhu B Z, Luo Y B, Deng Z P, Zuo J H. 2017a. Integrative analysis of circRNAs acting as ceRNAs involved in ethylene pathway in tomato. Physiologia Plantarum, 161:311-321. doi: 10.1111/ppl.2017.161.issue-3 URL
[44]	Wang Z P, Liu Y F, Li D W, Li L, Zhang Q, Wang S B, Huang H W. 2017b. Identification of circular RNAs in kiwifruit and their species-specific response to bacterial canker pathogen invasion. Frontiers in Plant Science, 8:413.
[45]	Xiong H, Shekhar S, Tan P N, Kumar V. 2004. Exploiting a support-based upper bound of Pearson's correlation coefficient for efficiently identifying strongly correlated pairs//Proceedings of the tenth ACM SIGKDD international conference on knowledge discovery and data mining. Seattle:Association for Computing Machinery:334-343.
[46]	Xu C J, Liu Y, Li Y B, Xu X D, Xu C G, Li X H, Xiao J H, Zhang Q F. 2015. Differential expression of GS5 regulates grain size in rice. Journal of Experimental Botany, 66:2611-2623. doi: 10.1093/jxb/erv058 URL
[47]	Yao J T, Zhao S H, Liu Q P, Lü M Q, Zhou D X, Liao Z J, Nan K J. 2017. Over-expression of CircRNA_100876 in non-small cell lung cancer and its prognostic value. Pathology-Research and Practice, 213:453-456. doi: 10.1016/j.prp.2017.02.011 URL
[48]	Zhang Y, Zhang X O, Chen T, Xiang J F, Yin Q F, Xing Y H, Zhu S S, Yang L, Chen L L. 2013. Circular intronic long noncoding RNAs. Molecular Cell, 51:792-806. doi: 10.1016/j.molcel.2013.08.017 URL
[49]	Zhao W, Cheng Y H, Zhang C, You Q B, Shen X J, Guo W, Jiao Y Q. 2017. Genome-wide identification and characterization of circular RNAs by high throughput sequencing in soybean. Scientific Reports, 7:5636. doi: 10.1038/s41598-017-05922-9 URL
[50]	Zhou R, Yu X Q, Ottosen C O, Zhao T M. 2020. High throughput sequencing of CircRNAs in tomato leaves responding to multiple stresses of drought and heat. Horticultural Plant Journal, 6 (1):34-38. doi: 10.1016/j.hpj.2019.12.004 URL

RNA	基因名称 Gene ID	引物序列（5′-3′） Primer sequence	产物长度/bp Amplicon size
DEcircRNA	00262:4702\|5542	F：GTTCTGTTCCCTGCCTCCAAT;R：CCATTTTCCCAACATTCCTTAGT	129
	03593:4109\|4340	F：AAACAAGAGGCCGAAACTAG;R：ATCCATTGTCTGCAAGTGTT	115
	05162:7223\|7649	F：CAGTTCAGGGTAGTTGTCG;R：GTTTGCTAGGCTTAGATACTTC	103
	05531:13219\|13378	F：TGTGCCCTTCATGTTCTCCTT;R：CACGAGGAGGATTGGAGGAAG	133
DEmRNA	g3238	F：CGGTAATAAGAGCCCAGGTGC;R：CACCTGTGCCGTGAATTTGTC	142
	g18070	F：CGCTCTTGCTCAGGCTTGTC;R：GAGAAGGCATGGATACCCGTC	87
	g21729	F：CTGCAAAGAACTCCCTGTGGTAA;R：TTGGCAGGGCTCAGGCATT	129
	g22409	F：GCGGTATCAAGAACCAAAGGG;R：GAGCGTCAAACCAGCAAGTG	110
内参基因 Reference gene	Beta actin	F：CCCTCACAATTTCACGCTCG;R：ATGAGGGTTATGCCCTCCCA	135

RNA	基因名称 Gene ID	引物序列（5′-3′） Primer sequence	产物长度/bp Amplicon size
DEcircRNA	00262:4702\|5542	F：GTTCTGTTCCCTGCCTCCAAT;R：CCATTTTCCCAACATTCCTTAGT	129
	03593:4109\|4340	F：AAACAAGAGGCCGAAACTAG;R：ATCCATTGTCTGCAAGTGTT	115
	05162:7223\|7649	F：CAGTTCAGGGTAGTTGTCG;R：GTTTGCTAGGCTTAGATACTTC	103
	05531:13219\|13378	F：TGTGCCCTTCATGTTCTCCTT;R：CACGAGGAGGATTGGAGGAAG	133
DEmRNA	g3238	F：CGGTAATAAGAGCCCAGGTGC;R：CACCTGTGCCGTGAATTTGTC	142
	g18070	F：CGCTCTTGCTCAGGCTTGTC;R：GAGAAGGCATGGATACCCGTC	87
	g21729	F：CTGCAAAGAACTCCCTGTGGTAA;R：TTGGCAGGGCTCAGGCATT	129
	g22409	F：GCGGTATCAAGAACCAAAGGG;R：GAGCGTCAAACCAGCAAGTG	110
内参基因 Reference gene	Beta actin	F：CCCTCACAATTTCACGCTCG;R：ATGAGGGTTATGCCCTCCCA	135

注释数据库Annotation database	注释数量Annotated number	基因长度Gene length
注释数据库Annotation database	注释数量Annotated number	（300 ~ 999 bp）	（> = 1 000 bp）
COG	1 440	103	1 335
GO	2 398	263	2 130
KEGG	1 741	188	1 548
KOG	2 377	241	2 133
Pfam	2 859	243	2 616
Swissprot	2 793	280	2 509
eggNOG	3 440	367	3 068
NR	3 509	391	3 113
总数 All annotated	3 511	391	3 115

注释数据库Annotation database	注释数量Annotated number	基因长度Gene length
注释数据库Annotation database	注释数量Annotated number	（300 ~ 999 bp）	（> = 1 000 bp）
COG	1 440	103	1 335
GO	2 398	263	2 130
KEGG	1 741	188	1 548
KOG	2 377	241	2 133
Pfam	2 859	243	2 616
Swissprot	2 793	280	2 509
eggNOG	3 440	367	3 068
NR	3 509	391	3 113
总数 All annotated	3 511	391	3 115

成对比较 Paired comparison	差异表达circRNA DEcircRNA	log₂FC （circRNA）	差异表达mRNA DEmRNA	log₂FC （mRNA）	注释 Annotation
Ov1-vs-Ov4	19296:1590\|1950	-∞	Corylus_avellana_newGene_15264	-11.80	NPR4
Ov1-vs-Ov2	19296:1590\|1950	-∞	Corylus_avellana_newGene_15264	-7.74	NPR4
Ov1-vs-Ov3	19296:1590\|1950	-∞	Corylus_avellana_newGene_15264	-7.73	NPR4
Ov1-vs-Ov4	00262:4702\|5542	-∞	g3238	-6.19	NPP
Ov1-vs-Ov4	00262:4702\|5735	-∞	g3238	-6.19	NPP
Ov1-vs-Ov4	00262:5280\|5910	-∞	g3238	-6.19	NPP
Ov1-vs-Ov3	01486:5071\|5547	-∞	g10760	-4.39	ABCG9
Ov1-vs-Ov3	09164:4340\|5637	-∞	g27362	-4.32	SULTR3;1
Ov1-vs-Ov3	05982:1271\|3603	-∞	Corylus_avellana_newGene_58963	-3.97	SCPL18
Ov1-vs-Ov4	09164:4340\|5637	-∞	g27362	-3.93	SULTR3;1
Ov1-vs-Ov4	05162:7223\|7649	-4.21	g21729	-3.90	SGR
Ov1-vs-Ov4	05982:1271\|3603	-∞	Corylus_avellana_newGene_58963	-3.64	SCPL18
Ov1-vs-Ov3	05162:7223\|7649	-3.48	g21729	-3.08	SGR
Ov1-vs-Ov2	00262:4702\|5542	-∞	g3238	-3.03	NPP
Ov1-vs-Ov2	00262:4702\|5735	-4.55	g3238	-3.03	NPP
Ov2-vs-Ov4	05312:1137\|1462	∞	Corylus_avellana_newGene_56641	6.62	NHX4
Ov2-vs-Ov3	14244:2582\|2731	∞	g31169	6.87	OLE1
Ov2-vs-Ov4	01490:2219\|2537	∞	g10779	6.92	COR2
Ov2-vs-Ov3	03593:4109\|4340	∞	g18070	7.31	TIP3-2
Ov2-vs-Ov4	03593:4109\|4340	∞	g18070	7.63	TIP3-2
成对比较 Paired comparison	差异表达circRNA DEcircRNA	log₂FC （circRNA）	差异表达mRNA DEmRNA	log₂FC （mRNA）	注释 Annotation
Ov2-vs-Ov4	03974:6066\|6673	∞	g19072	7.80	SBP
Ov2-vs-Ov4	14244:2582\|2731	∞	g31169	7.82	OLE1
Ov1-vs-Ov2	04715:14790\|15271	∞	g20792	8.37	WAT1
Ov1-vs-Ov3	03593:4109\|4340	∞	g18070	9.34	TIP3-2
Ov1-vs-Ov3	14244:2582\|2731	∞	g31169	9.55	OLE1
Ov1-vs-Ov4	03593:4109\|4340	∞	g18070	9.59	TIP3-2
Ov1-vs-Ov3	05531:13219\|13378	∞	g22409	9.69	OLE5
Ov1-vs-Ov4	05531:13219\|13378	∞	g22409	9.99	OLE5
Ov1-vs-Ov4	05531:13219\|13433	∞	g22409	9.99	OLE5
Ov1-vs-Ov4	14244:2582\|2731	∞	g31169	10.35	OLE1