共查询到17条相似文献，搜索用时 531 毫秒
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《果树学报》2015,(6)
【目的】系统研究‘库尔勒香梨’脱萼组和宿萼组花器官的转录组表达情况,筛选2者之间的差异表达基因,为进一步阐明‘库尔勒香梨’萼片脱落与宿存分子机理奠定基础。【方法】采用Illumina高通量测序技术对‘库尔勒香梨’脱萼组和宿萼组代表样品进行测序,利用生物信息学方法筛选2者的差异表达基因和进行功能基因预测。【结果】Trinity法拼接后得到48 894条unigenes序列,从中选取了2组样品中共表达和单独表达的SPL转录因子进行了功能探究,发现3条unigenes可能与萼片发育有关。筛选的差异表达基因获得了Gene Ontology和KEGG数据库的注释,涉及植物激素信号转导、光合代谢、苯丙氨酸代谢、芳香族化合物合成、细胞发育等与萼片发育相关过程。【结论】筛出了2组样品的差异表达基因,获得了与萼片发育相关的基因及其对应的功能注释,为今后进一步研究‘库尔勒香梨’萼片脱落与宿存分子机理奠定了良好基础。 相似文献
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《果树学报》2016,(5)
【目的】筛选‘库尔勒香梨’萼片脱落与宿存相关基因,研究其在脱萼组和宿萼组样品中的表达情况。【方法】以‘库尔勒香梨’脱萼和宿萼花器官转录组测序数据为基础,从Unigenes中选出一条转录组和数字表达谱测序结果共有的与萼片脱落相关的具有NAC结构域的差异基因,命名为kfpNAC,对其进行生物信息学分析,并通过实时荧光定量PCR分析其在花期不同阶段不同器官中的表达量。【结果】kfpNAC的序列长度是1 612 bp,包含一个编码308个氨基酸的长度为927 bp的开放阅读框(ORF),一个294 bp的5’端非编码区和一个391 bp的3’端非编码区,并且在3’端非编码区有一个poly A结构。‘库尔勒香梨’kfpNAC核酸和氨基酸序列与其他植物的NAC基因具有高度的同源性。实时荧光定量结果显示其在盛花期脱萼组全花样品中的表达量显著高于宿萼组全花和子房,并且在盛花期子房中的表达量显著高于萼片。【结论】克隆和鉴定了一个新的‘库尔勒香梨’kfpNAC基因,属于NAC基因家族的NAM亚群,该基因与‘库尔勒香梨’萼片脱落密切相关。 相似文献
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利用新一代高通量测序技术,对猕猴桃雌花和雄花中表达的小RNA进行了测序,分别得到雌花18 408 610条序列和雄花11 191 469条序列。通过生物信息学分析,共鉴定和预测得到39个保守miRNA家族和400个新的miRNA家族,其中有170个miRNA家族在雌、雄花样品间显著差异表达。对差异表达miRNA进行靶基因的预测及注释,结果显示,靶基因产物具有包含核苷三磷酸水解酶的磷酸环结构域的miRNA数量最多。在猕猴桃25号连锁群（Chr25）上共预测得到3个miRNA,其中novel-ach-miR362的靶基因Achn298021可能与猕猴桃花的性别发育有关。 相似文献
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《中国瓜菜》2015,(4):6-12
研究辣椒花药中sRNA分布,筛选育性相关miRNA,通过miRNA及其靶基因的表达分析探讨miRNA对雄性育性的调控。利用高通量测序技术对辣椒细胞核雄性不育两用系处于小孢子单核靠边期的花药进行sRNA测序,同时分析两材料间miRNA的表达差异。通过qRT-PCR技术验证差异分析结果,并对miRNA及其靶基因在小孢子发育不同时期的表达模式进行分析。在构建的不育株和可育株两个文库中共发掘出857个可信度高的miRNA;筛选得到42个表达差异显著的miRNA;通过靶基因预测与注释,预测出的差异表达miRNA的靶基因中与繁殖有关的基因有152个,与生殖过程有关的基因有151个。通过qRT-PCR验证了选出的9条miRNA的存在,其表达差异与测序分析结果基本一致;单核靠边期多数miRNA负调控其靶基因,不同发育时期其调控模式会发生变化。本研究为揭示辣椒miRNA与雄性育性的关系提供了重要信息。 相似文献
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【目的】了解‘库尔勒香梨’miR159家族成员的进化特征及其在萼片脱落与宿存中的作用。【方法】以‘库尔勒香梨’Small RNA高通量测序中获得的miR159家族前体序列和成熟体序列为基础进行系统发育树构建、前体和成熟体的序列比对、前体二级结构预测、WebLogo保守性分析、靶基因预测和表达量分析。【结果】‘库尔勒香梨’miR159家族存在2个前体成员(psi-MIR159a、psi-MIR159c)和2个成熟体成员(psi-miR159a、psi-miR159c)。2个前体成员的序列均能形成典型稳定的茎环结构,且序列完全一致,它们的最小折叠自由能(dG)都为-93.95 kal·mol~(-1)。系统发育进化树显示,‘库尔勒香梨’和苹果MIR159家族亲缘关系很近。WebLogo分析表明,miR159家族成熟体的保守性很高,其保守序列为5'-UUUGGAUUGAAGGGAGCUCUAUU-3',但‘库尔勒香梨’的碱基保守性较低。靶基因预测显示,‘库尔勒香梨’miR159基因家族靶基因为转录因子bHLH91、BES1/BZR1同源蛋白4、热激70 ku蛋白质、Trihelix转录因子GT-1、G型凝集素S-受体类丝氨酸/苏氨酸蛋白激酶RLK1等。实时荧光定量PCR(qPCR)显示,‘库尔勒香梨’miR159家族的2个成员在初花期、盛花期和末花期3个时期萼片脱落组和宿存组的表达量有显著差异。【结论】‘库尔勒香梨’miR159家族在进化过程中既有保守性又有多样性,可能在萼片脱落过程中起作用。 相似文献
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《果树学报》2019,(11)
【目的】番木瓜是典型的呼吸跃变型果实,外源乙烯处理使番木瓜呼吸跃变提前,促进果实成熟。分离番木瓜果实成熟相关miRNA,为深入了解呼吸跃变型果实的成熟分子机制奠定基础。【方法】利用高通量测序技术对乙烯(ETH)、1-MCP和清水对照(CG)处理的番木瓜果实进行miRNA和转录组高通量测序,然后对测序获得数据进行生物信息学分析,进行miRNA鉴定和靶基因预测,并与转录组测序结果进行关联分析。【结果】乙烯、1-MCP和对照处理分别获得10 734 196、16 486 803和16 067 290条纯净序列,共鉴定出523个miRNA。其中,已知miRNA个数为1-MCP(303)、CG(214)和ETH(239),新miRNA个数为1-MCP(184)、CG(188)和ETH(114)。与对照相比,在乙烯处理中上调和下调表达的miRNA分别是123和72条。靶基因预测共获得5 053个靶基因,KEGG功能富集分析显示它们参与了戊糖、葡萄糖醛酸转换、淀粉和蔗糖代谢、卟啉和叶绿素代谢、类胡萝卜素合成等代谢途径。筛选出的番木瓜果实成熟衰老相关候选miRNA,包含4个果实软化调控相关miRNA(miR167-y、miR4993-x、miR3946-x和miR5059-x)、3个果实颜色调控相关miRNA(miR4993-x、miR815-y和miR7810-x)、3个激素调控相关miRNA(miR4993-x、miR8004-x和miR9722-x)和4个转录因子调控相关miRNA(miR5641-y、miR9722-x、miR838-y和miR319-y)。【结论】筛选的番木瓜果实成熟衰老相关miRNA为今后果实成熟衰老调控网络研究提供了可能的线索。 相似文献
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番茄茉莉酸缺失突变体灰霉菌侵染响应miRNA及其表达分析 总被引：1，自引：0，他引：1
为揭示miRNA对番茄灰霉菌胁迫的响应机制，以番茄茉莉酸（JA）缺失突变体def1、spr2及其野生型（CM）为试验材料，构建了3个材料接种灰霉菌前后（0 h、48 h）2个时期的miRNA文库，并采用Illumina平台测序，对测序数据进行生物信息分析，结合实时荧光定量检测目的miRNA及其预测的靶基因表达情况。结果表明灰霉菌侵染番茄后，JA缺失突变体def1和spr2的病情指数显著高于CM，H2O2含量低于CM。高通量测序鉴定了130个已知miRNA和811个新miRNA。进一步筛选出8个保守miRNA（Sly-miR156e-3p、Sly-miR166c-5p、Sly-miR171f、Sly-miR172b、Sly-miR319a、Sly-miR390b-5p、Sly-miR399b、Sly-miR482d-5p），其在6个样本中表达模式各异。预测差异miRNA靶基因122个，结合qRT-PCR技术分析了8个miRNA和8个靶基因的表达情况，与高通量测序结果基本一致。推测Sly-miR156e-3p、Sly-miR390b-5p、Sly-miR399b、Sly-miR482d-5p通过依赖JA信号途径参与番茄对灰霉病的抗性。 相似文献
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通过生物信息分析发现，马铃薯miR319基因家族（Stu-miR319）共有3个前体基因，可生成5种成熟序列。序列比对发现，miR319成熟体序列在不同物种间具有高度的保守性，保守位点的分布差异较大。通过构建miR319前体基因的系统演化树发现，Stu-miR319的3个前体基因分属不同分支，且不同物种间的演化关系存在差异。组织特异性表达分析结果表明，Stu-miR319的5种成熟体在叶片中均有高表达。通过构建Stu-miR319前体基因过量表达转基因植株以及转录组测序分析发现，Stu-miR319-5p、Stu-miR319-3p、Stu-miR319a-5p和Stu-miR319b各有4个候选靶基因，Stu-miR319a-3p有2个候选靶基因。通过透射电镜分析发现，在Stu-miR319过量表达的植株中，导管细胞的细胞壁均有降解趋势，说明Stu-miR319基因家族对调控导管细胞细胞壁的完整性具有重要作用。 相似文献
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梨花芽休眠相关miRNA的鉴定和差异表达分析 总被引：1，自引：0，他引：1
为了探究梨花芽休眠进程中miRNA的表达模式和靶基因,利用Solexa测序技术、生物信息学分析和实时荧光定量PCR（qPCR）技术,对内休眠、内休眠解除和生态休眠解除3个时期梨花芽的miRNA进行高通量测序、筛选和鉴定。结果表明,内休眠、内休眠解除和生态休眠解除时期3个样本文库中分别有12 276 226、10 135 952、11 453 981条Unique序列。miRNA主要分布在21 ~ 24 nt之间,其中长度为24 nt的数量最多。共检测到151个已知的miRNA,属于39个不同的家族,并利用生物信息学软件预测到了209个新的miRNAs。比较分析从内休眠进入到生态休眠解除的整个休眠转换时期差异表达的miRNA,筛选出8个miRNA（ahy-miR156b-5p、cpa-miR319、aly-miR172c-3p、aau-miR396、mdm-miR858、aly-miR171b-3p、bdi-miR160f和hbr-miR166a）,其靶基因主要参与转录调控、信号传导等过程。利用qPCR验证了8个miRNA及其8个靶基因的表达。 相似文献
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Li Ma Chenjing Li Maosong Pei Fujun Cao Shaowen Quan 《The Journal of Horticultural Science and Biotechnology》2019,94(3):305-316
The objective of the study was to learn more about some newly identified miRNAs related to calyx persistence in Korla fragrant pear (Pyrus sinkiangensis Yu). Small RNAs were subjected to high-throughput sequencing after extraction from the ovaries and sepals of flowers with either a deciduous or a persistent calyx. Differentially expressed miRNAs were screened, and 73 new miRNAs were obtained. Twenty of these new miRNAs were selected to further validate all of the new miRNAs. Their mature miRNAs were cloned and identified, the secondary structures of the precursor miRNAs (pre-miRNAs) were analysed, and then qRT-PCR analysis was conducted. The results showed that the mature sequences of nine new miRNAs (novel_miRNA) in different samples were consistent with the results of high-throughput sequencing. Overall, this study improved current methods for the molecular cloning and identification of fruit tree miRNA. Nine new miRNA types were identified. This study laid a good foundation for elucidating the biological functions of these new miRNAs. 相似文献
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Differentially expressed microRNAs during solid endosperm development in coconut (Cocos nucifera L.)
Dongdong Li Yusheng Zheng Li Wan Xiaoming Zhu Zhekui Wang 《Scientia Horticulturae》2009,122(4):666-669
MicroRNAs (miRNAs) are a class of 20–24 nt, endogenously expressed, non-coding RNAs that play important regulatory roles in plants and animals. To identify miRNAs potentially involved in tissue development and compound anabolism, we studied miRNA expression profiles in endosperm of coconut at different developmental stages. Based on the annotation in miRBase (release 10.1), we measured a total of 179 miRNAs in immature (95 expressed miRNAs) and mature tissues (176 expressed miRNAs) using microarrays, respectively. The comparative analyses on miRNA expression profiles between these two groups of tissues showed that 23 miRNAs were up-regulated and nine miRNAs were down-regulated in matured endosperm. We further confirmed the increased expression of four miRNAs and decreased expression of a miRNA in immature endosperm using real-time PCR. Moreover, we computationally predicted the target genes of 32 miRNAs with differential expression (p < 0.01), and identified the lowest-score targets of six miRNAs. Finally, we discussed the potential functional relevance of several differentially expressed miRNAs. 相似文献
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ZHANG Si-yi LU Zhong-ming SONG Xin-han ZHANG Hong-bin CHEN Liang-si LUO Xiao-ning CHEN Shao-hua WU Yi-long 《园艺学报》2013,29(1):86-92
AIM：To investigate the differential microRNA expression profiles between laryngeal cancer and adjacent normal laryngeal mucosa. METHODS：Forty two pairs of laryngeal cancer tissue and adjacent normal laryngeal mucosa tissue were collected. Ten pairs of samples were used for determining microRNA expression by the method of miRNA microarray chip. Data analysis was performed to find out the significant differential microRNA expression profile in laryngeal cancer, and the difference was verified by quantitative real-time PCR (qRT-PCR) analysis on another 32 pairs of samples. Methyl thiazolyl tetrazolium (MTT) assay and colony-forming assay were used to analyze the proliferation of Hep2 cells induced by miR-125a-5p. RESULTS：Both miRNA microarray and qRT-PCR showed that the expression of let-7f-5p, miR-10a-5p, miR-125a-5p, miR-144-3p, miR-195-5p and miR-203 was down-regulated in laryngeal cancer tissues. miR-125a-5p suppressed the proliferation of Hep2 cells. CONCLUSION：The results of microarray are accordant with those of qRT-PCR. Significant difference of miRNA expression profiles between laryngeal cancer and adjacent normal laryngeal mucosa indicates that miRNAs may play a role in carcinogenesis and progression of laryngeal cancer. miR-125a-5p inhibits the proliferation of Hep2 cell, indicating a novel therapeutic target against laryngeal cancer. 相似文献
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Ghunichigul ABAQ Maynur DAWUT Yimit RAHMAN Aynur YUSUP Wureyatiguli KEWEIER Marjangul ABLITIP 《园艺学报》2017,33(7):1317-1322
AIM:To explore the effect of pinobanksin-3-acetate (PB3A) on microRNA (miRNA) expression profile of human colon cancer cells for providing new methods of treatment of colon cancer and development of targeted drug.METHODS:The method of miRNA expression profiling was used to observe the miRNA differential expression in human colon cancer SW480 cells after treated with PB3A.The expression of miRNA-198 and miRNA-296-5p in the SW480 cells was detected by RT-qPCR.The network databases of miRWalk,MicroT,miRanda and so on were used to predict the target genes regulated by these miRNAs,and pathway significant enrichment analysis was performed.RESULTS:miRNA microarray analysis showed that after treated with propolis flavonoid PB3A for 24 h,267 miRNAs with differential expression twice or more in the SW480 cells were observed.Among them,there were 30 miRNAs with 10-fold or more differential expression,in which 28 were up-regulated and 2 were down-regulated.The results of RT-qPCR showed that the expression levels of miRNA-198 and miRNA-296-5p were consistent with the results of miRNA microarray analysis,and the difference was statistically significant (P<0.05).Bioinformatic analysis revealed that miRNA-198 has 859 target genes,and miRNA-296-5p has 906 target genes.The target genes of miRNA-198 were clustered in pathways in cancer,axon guidance,Wnt signaling pathway,regulation of actin cytoskeleton,insulin signaling pathway and MAPK signaling pathway,while the target genes of miRNA-296-5p were clustered in axon guidance,Wnt signaling pathway,MAPK signaling pathway,endocytosis,melanogenesis,insulin signaling pathway and calcium signaling pathway.CONCLUSION:Propolis flavonoid PB3A affects the expression of miRNA in colon cancer SW480 cells.The abnormal expression of miRNA-198 and miRNA-296-5p may be involved in the inhibitory effect of PB3A on colon cancer. 相似文献
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AIM: To screen the chemotherapy resistance-related microRNAs (miRNAs) of colon cancer using gene chip technique, and to explore the mechanism of miRNAs regulating chemotherapy resistance. METHODS: Gene chip technique was used to analyze the expression of miRNAs in colon cancer cell line HCT8 and vincristine-resistant cell line HCT8/v, and screen the miRNAs with significantly different expression. The results were verified by RT-qPCR. The target genes of these miRNAs were predicted, and the Gene Ontology (GO) analysis and the signaling pathway analysis of the predicted genes were carried out. RESULTS: Altogether 342 miRNAs with significantly differential expression were selected, in which 190 were up-regulated, and 152 were down-regulated. The verification results of RT-qPCR showed that the expression of miR-125-5p, miR-181c-5p and miR-153-3 was consistent with the results of chip detection. The expression of miR-130a-3p and miR-149-3p was not consistent with the results of chip detection. The results of GO analysis showed that the main pathway of chemotherapy resistance-related genes was RNA polymerase II regulatory region sequence-specific DNA binding. The chemotherapy resistance-related genes played roles mainly through positive regulation and are mainly located in intracellular membrane-bound organelles. The results of KEGG analysis showed that the pathways associated with the most enriched chemotherapy resistance-related genes were axon guidance pathway, insulin signaling pathway, and phospholipase D signaling pathway.CONCLUSION: miRNAs are closely related to chemotherapy resistance in colon cancer. Through the researches on miRNAs, we can have a deeper understanding of the mechanism of chemotherapy resistance and provide new ideas for reversing chemotherapy resistance in colon cancer. 相似文献
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AIM：To identify differentially expressed microRNAs (miRNAs) in ischemic myocardial tissues from the rats with acute myocardial infarction (AMI) by miRNA array technique, and to predict their targets and analyze their functions using bioinformatics. METHODS：The rat models of AMI (n=3) were prepared by ligaturing the left anterior descending coronary artery (LAD) of Wistar rats. Electrocardiogram and blood pressure were detected during the operation, and the myocardial infarct size was measured by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. Ischemic myocardial tissues were isolated from the infarct area 4 h after ischemia. The same procedure in sham group (n=3) was performed except for ligaturing LAD. Total RNA was extracted from ischemic and normal myocardial tissues. miRNA was isolated from total RNA, labeled with Cy3 and hybridized on miRNA array. Real-time PCR was applied to verify the reliability of miRNA array results. The targets of differentially expressed miRNAs were predicted and their functions were analyzed by bioinformatics. RESULTS：Rat model of AMI was successfully prepared and verified by electrocardiogram detection, blood pressure measurement and pathological observation. Compared with sham group, microarray screening showed that total 11 AMI-related miRNAs were selected, including 6 up-regulated and 5 down-regulated. Three of them (rno-miR-181c, rno-miR-146b and rno-miR-208) were related to the cardiovascular functions, while the functions of the others (rno-miR-672*, rno-miR-743b, rno-miR-128, rno-miR-138-1*, rno-miR-336, rno-miR-138-2*, rno-miR-325-3p and rno-miR-3572) were unknown and might be novel AMI-related biomarkers. Parts of the miRNA targets were also related to the cardiovascular functions. CONCLUSION：Differentially expressed miRNAs in AMI rats may serve as novel biomarkers for diagnosis of AMI and potential targets for treatment of AMI. 相似文献