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
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摘要
【目的】探究miR403的分子进化特性及其在龙眼体胚发生早期中的表达模式。【方法】根据龙眼转录组数据库结合miRBase数据库下载得到植物miR403成熟体(miRNA,MicroRNA)和前体(pre-miRNA,Precursor MicroRNA)序列,对其构建系统发育树、序列多重比对、前体二级结构分析;并克隆龙眼miR403 5’末端cDNA序列,比较miR403启动子顺式作用元件。利用qPCR(Quantitative real time-PCR)探究龙眼体胚发生早期miR403和pre-miR403的表达模式。【结果】龙眼及其他17个物种中miR403的36个前体和42成熟体序列的系统发育树以及序列多重比对显示,植物miR403高度保守且主要来源于pre-miR403的3臂,而pre-miR403分歧较大,序列仅在miRNA生成区域保守;不同物种之间miR403成员数量差异不大;pre-miR403最小折叠自由能为-35.70～-55.50 kal·mol~(-1)。克隆获得龙眼primiR403(miR403初级体)5’末端cDNA序列,确认其第一个碱基A为转录起始位点(TSS,transcription start site)。龙眼等5种植物MIR403启动子中具有大量光、激素和逆境胁迫等元件。龙眼早期体胚发生过程中miR403和pre-miR403整体为显著下调趋势。【结论】龙眼miR403与其他植物miR403具有高度的保守性以及物种之间的特异性,miR403和pre-miR403下调表达可能有利于促进龙眼早期体胚发生。
【Objective】MiR403, a miRNA family unique to dicotyledonous plants, plays an important role in plant disease resistance, growth and development. Previous studies showed that miR403 guided cleavage AGO2 mRNA at transcription level, which played an significant role in defensing virus. In addition, miR403 could response to stress and express specifically in different organization. Besides,when over-expressed miR403, tomato showed slow growth, dwarfism, delayed flowering, and poor fertility. Dimocarpous longan, a member of the Sapindaceae family, was an important economic fruit tree in the subtropics. According to the previous researches, miRNA is important to longan somatic embryogenesis(SE). However, plants miR403 evolutionary characteristics and its function in plants somatic embryos have not been reported yet. Therefore, we investigated the molecular evolution characteristics of plant miR403 and its expression profile during longan early SE.【Methods】Plant miR403 matures(miRNA) and precursors(pre-miRNA) sequences were obtained from longan transcriptome database and miRBase database. The secondary structure analysis of plant pre-miR403 sequences were performed by Mfold online software. The phylogenetic tree of plant miR403 and pre-miR403 were performed by MEGA 7.0 through NJ method(Neighbor-joining) with 1000 bootstrap. In addition, the sequences multiple alignment were carried out by iTOL and WebLogo softwares. RLM-RACE(RNA ligase-mediated amplification of cDNA ends) was used to obtain longan miR403 5'c DNA(coding DNA) end sequence. The cDNA of RLM-RACE was synthesized using Gene Racer kit. DNAMAN 6.0 was used to design the primers of RLM-RACE. Based on the longan miR403 transcription start site(TSS), the promoter sequence was extracted of 2 800 bp upstream from the TSS. The promoter of athmiR403 was collected from NCBI according to the pri-ath-miR403(primary-ath-miR403) loading in NCBI(https://www.ncbi.nlm.nih.gov/). Then, the TSS of citrus, soybean and grape miR403 members were predicted by BDGP(http://www.fruitfly.org/seq_tools/promoter.html) and the TSSP(www.softbreey.com) software and promoter sequences were extracted form their genome. Next, five plant miR403 promoters cis-acting elements were predicted using PlantCARE database. Finally, the expression patterns of miR403 and pre-miR403 during longan early SE were verified by SYBR Prumix EX TaqTMII kit, Tip-mix kit and Roche LightCycler 480 real-time fluorescence quantitative PCR. The expression of miR403 was corrected by U6 SnRNA, miR408-5 p1 and ath5.8 S*, and the expression of premiR403 was corrected by FSD, EF-1α and EIF-4α. The expressions profiles of miR403 and premiR403 were obtained by 2-?Ctmethod. The statistical analysis miR403 and pre-miR403 relative expression were performed through SPSS19.0 software.【Results】In this study, phylogenetic tree of 36 premiR403 s and 42 miR403 s sequences from longan and other 17 species showed that plant miR403 was a highly conserved miRNA family, and there was no significant difference in the number of miR403 among different species. MiR403 was mainly derived from the 3-terminal arm of pre-miR403, and only a few was derived from the 5-terminal arm of pre-miR403. The minimum folding free energy of plant pre-miR403 s was from-35.70 kal·mol~(-1) to-55.50 kal·mol~(-1). After amplifying the 5'end c DNA sequence of longan miR403 by RLM-RACE technology, a 154 bp cDNA fragment without 3'end sequence was obtained, and it was confirmed that the first base A was the TSS. The TSS of soybean, citrus and grape were predicted by the BDGP and the TSSP softwares. Then, we extracted MIR403 promoter from longan, Arabidopsis thaliana, soybean, citrus and grape genome databases for analyzing the csiacting elements. The results showed that the promoters had a large number of conserved elements such as light responsiveness elements, hormone responsiveness elements and stress responsiveness elements,implying that the regulatory factors of miR403 was common in different species. qPCR analysis showed that the expression levels of miR403 and pre-miR403 was significant decreased in longan early SE, suggesting that down-regulation of miR403 and pre-miR403 would be possible to mediate longan early SE.【Conclusion】Our researches showed that plant miR403 s was highly conserved. In addition,the down regulation of pre-miR403 and miR403 might be promotive to longan early SE.
【Objective】MiR403, a miRNA family unique to dicotyledonous plants, plays an important role in plant disease resistance, growth and development. Previous studies showed that miR403 guided cleavage AGO2 mRNA at transcription level, which played an significant role in defensing virus. In addition, miR403 could response to stress and express specifically in different organization. Besides,when over-expressed miR403, tomato showed slow growth, dwarfism, delayed flowering, and poor fertility. Dimocarpous longan, a member of the Sapindaceae family, was an important economic fruit tree in the subtropics. According to the previous researches, miRNA is important to longan somatic embryogenesis(SE). However, plants miR403 evolutionary characteristics and its function in plants somatic embryos have not been reported yet. Therefore, we investigated the molecular evolution characteristics of plant miR403 and its expression profile during longan early SE.【Methods】Plant miR403 matures(miRNA) and precursors(pre-miRNA) sequences were obtained from longan transcriptome database and miRBase database. The secondary structure analysis of plant pre-miR403 sequences were performed by Mfold online software. The phylogenetic tree of plant miR403 and pre-miR403 were performed by MEGA 7.0 through NJ method(Neighbor-joining) with 1000 bootstrap. In addition, the sequences multiple alignment were carried out by iTOL and WebLogo softwares. RLM-RACE(RNA ligase-mediated amplification of cDNA ends) was used to obtain longan miR403 5'c DNA(coding DNA) end sequence. The cDNA of RLM-RACE was synthesized using Gene Racer kit. DNAMAN 6.0 was used to design the primers of RLM-RACE. Based on the longan miR403 transcription start site(TSS), the promoter sequence was extracted of 2 800 bp upstream from the TSS. The promoter of athmiR403 was collected from NCBI according to the pri-ath-miR403(primary-ath-miR403) loading in NCBI(https://www.ncbi.nlm.nih.gov/). Then, the TSS of citrus, soybean and grape miR403 members were predicted by BDGP(http://www.fruitfly.org/seq_tools/promoter.html) and the TSSP(www.softbreey.com) software and promoter sequences were extracted form their genome. Next, five plant miR403 promoters cis-acting elements were predicted using PlantCARE database. Finally, the expression patterns of miR403 and pre-miR403 during longan early SE were verified by SYBR Prumix EX TaqTMII kit, Tip-mix kit and Roche LightCycler 480 real-time fluorescence quantitative PCR. The expression of miR403 was corrected by U6 SnRNA, miR408-5 p1 and ath5.8 S*, and the expression of premiR403 was corrected by FSD, EF-1α and EIF-4α. The expressions profiles of miR403 and premiR403 were obtained by 2-?Ctmethod. The statistical analysis miR403 and pre-miR403 relative expression were performed through SPSS19.0 software.【Results】In this study, phylogenetic tree of 36 premiR403 s and 42 miR403 s sequences from longan and other 17 species showed that plant miR403 was a highly conserved miRNA family, and there was no significant difference in the number of miR403 among different species. MiR403 was mainly derived from the 3-terminal arm of pre-miR403, and only a few was derived from the 5-terminal arm of pre-miR403. The minimum folding free energy of plant pre-miR403 s was from-35.70 kal·mol~(-1) to-55.50 kal·mol~(-1). After amplifying the 5'end c DNA sequence of longan miR403 by RLM-RACE technology, a 154 bp cDNA fragment without 3'end sequence was obtained, and it was confirmed that the first base A was the TSS. The TSS of soybean, citrus and grape were predicted by the BDGP and the TSSP softwares. Then, we extracted MIR403 promoter from longan, Arabidopsis thaliana, soybean, citrus and grape genome databases for analyzing the csiacting elements. The results showed that the promoters had a large number of conserved elements such as light responsiveness elements, hormone responsiveness elements and stress responsiveness elements,implying that the regulatory factors of miR403 was common in different species. qPCR analysis showed that the expression levels of miR403 and pre-miR403 was significant decreased in longan early SE, suggesting that down-regulation of miR403 and pre-miR403 would be possible to mediate longan early SE.【Conclusion】Our researches showed that plant miR403 s was highly conserved. In addition,the down regulation of pre-miR403 and miR403 might be promotive to longan early SE.
引文
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[5] LIN Y,LIN L,LAI R,LIU W,CHEN Y,ZHANG Z,XU X,LAI Z. MicroRNA390-directed TAS3 cleavage leads to the production of tasiRNA-ARF3/4 during somatic embryogenesis in Dimocarpus longan Lour.[J]. Frontiers in Plant Science,2015,6(127):1119.
[6] LI W,ZHANG S,HAN S,WU T,ZHANG J,QI L. Regulation of LaMYB33 by miR159 during maintenance of embryogenic potential and somatic embryo maturation in Larix kaempferi(Lamb.)Carr[J]. Plant Cell Tissue&Organ Culture,2013,113(1):131-136.
[7]苏立遥,张帅,陈旭,徐小萍,赖钟雄,林玉玲. miR171家族成员分子特性及miR171b调控靶标在龙眼体胚发生早期的表达分析[J].果树学报,2018,35(11):1324-1334.SU Liyao,ZHANG Shuai,CHEN Xu,XU Xiaoping,LAI Zhongxiong,LIN Yuling. Molecular characteristics of miR171family members and analysis of the expression pattern of miR171b regulatory targets during early somatic embryogenesis in longan[J]. Journal of Fruit Science,2018,35(11):1324-1334.
[8] LI H,ZHANG J,YANG Y,JIA N,WANG C,SUN H. MiR171and its target gene SCL6,contribute to embryogenic callus induction and torpedo-shaped embryo formation during somatic embryogenesis in two lily species[J]. Plant Cell Tissue&Organ Culture,2017,130(3):591-600.
[9] SZYRAJEW K,BIELEWICZ D,DOLATA J,WóJCIK A M,NOWAK K,SZCZYGIE?-SOMMER A,SZWEYKOWSKAKULINSKA Z,JARMOLOWSKI A,GAJ M D. MicroRNAs are intensively regulated during induction of somatic embryogenesis in Arabidopsis[J]. Frontiers in Plant Science,2017,8:18.
[10] LIU Y,HAN S,DING X,LI M,ZHANG L,LI W,XU H,LI Z,QI L. Transcriptome analysis of m RNA and miRNA in somatic embryos of Larix leptolepis subjected to hydrogen treatment[J].International Journal of Molecular Sciences,2016,17(11):1951.
[11] LIN Y,LAI Z. Comparative analysis reveals dynamic changes in miRNAs and their targets and expression during somatic embryogenesis in longan(Dimocarpus longan Lour.)[J]. Plos One,2013,8(4):e60337.
[12] SUNKAR R,JAGADEESWARAN G. In silico identification of conserved microRNAs in large number of diverse plant species[J]. Bmc Plant Biology,2008,8(1):37.
[13] JAGTAP S,SHIVAPRASAD P V. Diversity,expression and mRNA targeting abilities of Argonaute-targeting miRNAs among selected vascular plants[J]. BMC Genomics,2014,15(1):1049.
[14] BAO D,GANBAATAR O,CUI X,YU R,BAO W,FALK B W,WURIYANGHAN H. Down-regulation of genes coding for core RNAi components and disease resistance proteins via corresponding microRNAs might be correlated with successful Soybean mosaic virus infection in soybean[J]. Molecular Plant Pathology,2018,19(4):948-960.
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[17] ZHANG C,XIAN Z,HUANG W,LI Z. Evidence for the biological function of miR403 in tomato development[J]. Scientia Horticulturae,2015,197:619-626.
[18] KHAKSEFIDI R E,MIRLOHI S,KHALAJI F,FAKHARI Z,SHIRAN B,FALLAHI H,RAFIEI F,BUDAK H,EBRAHIMIE E. Differential expression of seven conserved microRNAs in response to abiotic stress and their regulatory network in Helianthus annuus[J]. Frontiers in Plant Science,2015,6:741.
[19] PUZEY J R,KARGER A,AXTELL M,KRAMER E M. Deep annotation of Populus trichocarpa microRNAs from diverse tissue sets[J]. Plos One,2012,7(3):e33034.
[20] XU Q,LIU Y,ZHU A,WU X,YE J,YU K,GUO W,DENG X.Discovery and comparative profiling of microRNAs in a sweet orange red-flesh mutant and its wild type[J]. Bmc Genomics,2010,11(1):246.
[21] MICA E,PICCOLO V,DELLEDONNE M,FERRARINI A,PEZZOTTI D,CASATI C,FABBRO C D,VALLE G,POLICRITI A,MORGANTE M,PESOLE G,PèM E,HORNER D S. Erratum to:High throughput approaches reveal splicing of primary microRNA transcripts and tissue specific expression of mature microRNAs in Vitis vinifera[J]. Bmc Genomics,2010,11(1):109.
[22] KULCHESKI F R,OLIVEIRA L F D,MOLINA L G,ALMER?O M P,RODRIGUES F A,MARCOLINO J,BARBOSA J F,STOLF-MOREIRA R,NEPOMUCENO A L,MARCELINOGUIMAR?ES F C,ABDELNOOR R V,NASCIMENTO L C,CARAZZOLLE M F,PEREIRA G A,MARGIS R. Identification of novel soybean microRNAs involved in abiotic and biotic stresses[J]. BMC Genomics,2011,12(1):307.
[23] LIN Y,MIN J,LAI R,WU Z,CHEN Y,YU L,CHENG C,JIN Y,TIAN Q,LIU Q,LIU W,ZHANG C,LIN L,HU Y,ZHANG D,THU M,ZHANG Z,LIU S,ZHONG C,FANG X,WANG J,YANG H,VARSHNEY R K,YIN Y,LAI Z. Genome-wide sequencing of longan(Dimocarpus longan Lour.)provides insights into molecular basis of its polyphenol-rich characteristics[J]. Gigascience,2017,6(5):1-14.
[24] LIN Y,LAI Z. Reference gene selection for qPCR analysis during somatic embryogenesis in longan tree[J]. Plant Science,2010,178(4):359-365.
[25] ZHAO X,CHEN S,ZHAO L,ZHANG X,MA P,GUO Z. Evolution of MIR166 Gene family in land plants[J]. Plant Diversity&Resources,2014,36(3):331-341.
[26] CLARKE J T,WARNOCK R C M,DONOGHUE P C J. Establishing a time-scale for plant evolution[J]. New Phytologist,2011,192(1):266-301.
[27] SMITH S A,BEAULIEU J M,DONOGHUE M J. An uncorrelated relaxed-clock analysis suggests an earlier origin for flowering plants[J]. Proceedings of the National Academy of Sciences of the United States of America,2010,107(13):5897-5902.
[28] MOREA E G O,SILVA E M D,SILVA G F F E,VALENTE G T,ROJAS C H B,VINCENTZ M,NOGUEIRA F T S. Erratum to:Functional and evolutionary analyses of the miR156 and miR529families in land plants[J]. Bmc Plant Biology,2016,16(1):1.
[29] WANG R,XU L,ZHU X,ZHAI L,WANG Y,YU R,GONG Y,LIMERA C,LIU L. Transcriptome-wide characterization of novel and heat-stress-responsive microRNAs in Radish(Raphanus Sativus L.)using next-generation sequencing[J]. Plant Molecular Biology Reporter,2014,33(4):867-880.
[30] SUN X,XU L,WANG Y,YU R,ZHU X,LUO X,GONG Y,WANG R,LIMERA C,ZHANG K. Identification of novel and salt-responsive miRNAs to explore miRNA-mediated regulatory network of salt stress response in radish(Raphanus sativus L.)[J]. BMC Genomics,2015,16(1):1-16.
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[2] ARMENTA-MEDINA A,LEPE-SOLTERO D,XIANG D,DATLA R,ABREU-GOODGER C,GILLMOR C S. Arabidopsis thaliana miRNAs promote embryo pattern formation beginning in the zygote[J]. Developmental Biology,2017,431(2):145-151.
[3] LIN Y,LAI Z,TIAN Q,LIN L,LAI R,YANG M,ZHANG D,CHEN Y,ZHANG Z. Endogenous target mimics down-regulate miR160 mediation of ARF10,-16,and-17 cleavage during somatic embryogenesis in Dimocarpus longan Lour.[J]. Frontiers in Plant Science,2015,6:956.
[4] LIN Y,LAI Z,LIN L,LAI R,TIAN Q,YE W,ZHANG D,YANG M,CHEN Y,ZHANG Z. Endogenous target mimics,microRNA167,and its targets ARF6,and ARF8,during somatic embryo development in Dimocarpus longan Lour.[J]. Molecular Breeding,2015,35(12):227.
[5] LIN Y,LIN L,LAI R,LIU W,CHEN Y,ZHANG Z,XU X,LAI Z. MicroRNA390-directed TAS3 cleavage leads to the production of tasiRNA-ARF3/4 during somatic embryogenesis in Dimocarpus longan Lour.[J]. Frontiers in Plant Science,2015,6(127):1119.
[6] LI W,ZHANG S,HAN S,WU T,ZHANG J,QI L. Regulation of LaMYB33 by miR159 during maintenance of embryogenic potential and somatic embryo maturation in Larix kaempferi(Lamb.)Carr[J]. Plant Cell Tissue&Organ Culture,2013,113(1):131-136.
[7]苏立遥,张帅,陈旭,徐小萍,赖钟雄,林玉玲. miR171家族成员分子特性及miR171b调控靶标在龙眼体胚发生早期的表达分析[J].果树学报,2018,35(11):1324-1334.SU Liyao,ZHANG Shuai,CHEN Xu,XU Xiaoping,LAI Zhongxiong,LIN Yuling. Molecular characteristics of miR171family members and analysis of the expression pattern of miR171b regulatory targets during early somatic embryogenesis in longan[J]. Journal of Fruit Science,2018,35(11):1324-1334.
[8] LI H,ZHANG J,YANG Y,JIA N,WANG C,SUN H. MiR171and its target gene SCL6,contribute to embryogenic callus induction and torpedo-shaped embryo formation during somatic embryogenesis in two lily species[J]. Plant Cell Tissue&Organ Culture,2017,130(3):591-600.
[9] SZYRAJEW K,BIELEWICZ D,DOLATA J,WóJCIK A M,NOWAK K,SZCZYGIE?-SOMMER A,SZWEYKOWSKAKULINSKA Z,JARMOLOWSKI A,GAJ M D. MicroRNAs are intensively regulated during induction of somatic embryogenesis in Arabidopsis[J]. Frontiers in Plant Science,2017,8:18.
[10] LIU Y,HAN S,DING X,LI M,ZHANG L,LI W,XU H,LI Z,QI L. Transcriptome analysis of m RNA and miRNA in somatic embryos of Larix leptolepis subjected to hydrogen treatment[J].International Journal of Molecular Sciences,2016,17(11):1951.
[11] LIN Y,LAI Z. Comparative analysis reveals dynamic changes in miRNAs and their targets and expression during somatic embryogenesis in longan(Dimocarpus longan Lour.)[J]. Plos One,2013,8(4):e60337.
[12] SUNKAR R,JAGADEESWARAN G. In silico identification of conserved microRNAs in large number of diverse plant species[J]. Bmc Plant Biology,2008,8(1):37.
[13] JAGTAP S,SHIVAPRASAD P V. Diversity,expression and mRNA targeting abilities of Argonaute-targeting miRNAs among selected vascular plants[J]. BMC Genomics,2014,15(1):1049.
[14] BAO D,GANBAATAR O,CUI X,YU R,BAO W,FALK B W,WURIYANGHAN H. Down-regulation of genes coding for core RNAi components and disease resistance proteins via corresponding microRNAs might be correlated with successful Soybean mosaic virus infection in soybean[J]. Molecular Plant Pathology,2018,19(4):948-960.
[15] ALLEN E,XIE Z,GUSTAFSON A M,SUNG GH,SPATAFORA J W,CARRINGTON J C. Evolution of microRNA genes by inverted duplication of target gene sequences in Arabidopsis thaliana[J]. Nature Genetics,2004,36(12):1282-1290.
[16] LOBBES D,RALLAPALLI G,SCHMIDT D D,MARTIN C,CLARKE J. Serrate:a new player on the plant microRNA scene[J]. Embo Reports,2006,7(10):1052-1058.
[17] ZHANG C,XIAN Z,HUANG W,LI Z. Evidence for the biological function of miR403 in tomato development[J]. Scientia Horticulturae,2015,197:619-626.
[18] KHAKSEFIDI R E,MIRLOHI S,KHALAJI F,FAKHARI Z,SHIRAN B,FALLAHI H,RAFIEI F,BUDAK H,EBRAHIMIE E. Differential expression of seven conserved microRNAs in response to abiotic stress and their regulatory network in Helianthus annuus[J]. Frontiers in Plant Science,2015,6:741.
[19] PUZEY J R,KARGER A,AXTELL M,KRAMER E M. Deep annotation of Populus trichocarpa microRNAs from diverse tissue sets[J]. Plos One,2012,7(3):e33034.
[20] XU Q,LIU Y,ZHU A,WU X,YE J,YU K,GUO W,DENG X.Discovery and comparative profiling of microRNAs in a sweet orange red-flesh mutant and its wild type[J]. Bmc Genomics,2010,11(1):246.
[21] MICA E,PICCOLO V,DELLEDONNE M,FERRARINI A,PEZZOTTI D,CASATI C,FABBRO C D,VALLE G,POLICRITI A,MORGANTE M,PESOLE G,PèM E,HORNER D S. Erratum to:High throughput approaches reveal splicing of primary microRNA transcripts and tissue specific expression of mature microRNAs in Vitis vinifera[J]. Bmc Genomics,2010,11(1):109.
[22] KULCHESKI F R,OLIVEIRA L F D,MOLINA L G,ALMER?O M P,RODRIGUES F A,MARCOLINO J,BARBOSA J F,STOLF-MOREIRA R,NEPOMUCENO A L,MARCELINOGUIMAR?ES F C,ABDELNOOR R V,NASCIMENTO L C,CARAZZOLLE M F,PEREIRA G A,MARGIS R. Identification of novel soybean microRNAs involved in abiotic and biotic stresses[J]. BMC Genomics,2011,12(1):307.
[23] LIN Y,MIN J,LAI R,WU Z,CHEN Y,YU L,CHENG C,JIN Y,TIAN Q,LIU Q,LIU W,ZHANG C,LIN L,HU Y,ZHANG D,THU M,ZHANG Z,LIU S,ZHONG C,FANG X,WANG J,YANG H,VARSHNEY R K,YIN Y,LAI Z. Genome-wide sequencing of longan(Dimocarpus longan Lour.)provides insights into molecular basis of its polyphenol-rich characteristics[J]. Gigascience,2017,6(5):1-14.
[24] LIN Y,LAI Z. Reference gene selection for qPCR analysis during somatic embryogenesis in longan tree[J]. Plant Science,2010,178(4):359-365.
[25] ZHAO X,CHEN S,ZHAO L,ZHANG X,MA P,GUO Z. Evolution of MIR166 Gene family in land plants[J]. Plant Diversity&Resources,2014,36(3):331-341.
[26] CLARKE J T,WARNOCK R C M,DONOGHUE P C J. Establishing a time-scale for plant evolution[J]. New Phytologist,2011,192(1):266-301.
[27] SMITH S A,BEAULIEU J M,DONOGHUE M J. An uncorrelated relaxed-clock analysis suggests an earlier origin for flowering plants[J]. Proceedings of the National Academy of Sciences of the United States of America,2010,107(13):5897-5902.
[28] MOREA E G O,SILVA E M D,SILVA G F F E,VALENTE G T,ROJAS C H B,VINCENTZ M,NOGUEIRA F T S. Erratum to:Functional and evolutionary analyses of the miR156 and miR529families in land plants[J]. Bmc Plant Biology,2016,16(1):1.
[29] WANG R,XU L,ZHU X,ZHAI L,WANG Y,YU R,GONG Y,LIMERA C,LIU L. Transcriptome-wide characterization of novel and heat-stress-responsive microRNAs in Radish(Raphanus Sativus L.)using next-generation sequencing[J]. Plant Molecular Biology Reporter,2014,33(4):867-880.
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