芒果2个不同花芽分化时期转录组分析
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摘要
【目的】分析芒果不同花芽分化时期的转录组,了解成花过程相关基因的表达情况,为芒果开花时间的分子调控机制研究提供理论参考。【方法】以芒果品种南逗迈4号为材料,采用Illumina高通量测序技术,分别对其新梢停长期(I期)和基部膨大期(II期)2个不同花芽分化时期顶芽进行转录组测序,并利用基因功能注释、差异表达基因筛选等生物信息学方法对测序获得的高质量序列进行分析。通过实时荧光定量PCR(q PCR)检测差异表达基因的表达情况以验证转录组测序结果的可信度。【结果】共获得85691条Unigenes,平均长度为870 bp,N50为1077 bp,与Uni Prot数据库比对,发现48589条Unigenes有同源信息,注释比例为56.7%,广泛涉及细胞组分、生物过程和分子功能三大类,共55个小类,其中参与生物过程的Unigene数量最多(有21个小类),以参与分子功能的Unigene数量最少(有14个小类)。以Fold-Change≥2为条件,筛选出花芽分化I和II期转录组间的2031个差异表达基因,其中1073个基因表达上调,958个基因表达下调,涉及247条代谢通路,富集的KEGG通路为内质网蛋白加工、次生物质的生物合成和积累、糖代谢和光合作用等,其中注释为内质网蛋白加工的基因数量最多。从2个不同时期顶芽的转录组数据中获得了春化、光周期、赤霉素(GA)、成花抑制、自主和年龄等途径的开花相关基因。将Mi SOC1、Mi VIN3、Mi Dof和Mi MADS1的q PCR检测结果与其转录组测序结果进行比对,发现这4个基因虽然在II期的表达量比I期上调差异倍数上存在一定差异,但表达量变化趋势一致,说明转录组测序结果的可信度较高。【结论】芒果花芽分化过程与内质网蛋白加工、次生代谢生物合成、植物激素信号转导、淀粉和蔗糖代谢等途径密切相关,可为芒果花期人工调控和产期调节提供参考。
【Objective】The transcriptome of mango(Mangifera indica L.) in two different bud differentiation periods was sequencing to comprehend the expression of genes related to mango flowering process. It laid the foundation for research on the molecular regulation mechanism of flowering time of mango.【Method】Illumina high-throughput sequencing technology was applied to detect the transcriptome sequencing data of mango variety Nandoumai 4 in two different flower bud differentiation stages,namely the withhold growing period of the shoots(stage I)and the base expanding period(stage II). High quality sequences in the results were analyzed by bioinformatics methods,such as gene function annotation and differentially expressed genes screening. Real time fluorescent quantitative PCR(q PCR)was used for the determination of differentially expressed genes to check the reliability of transcriptome sequencing.【Result】A total of 85691 Unigenes were obtained with average length of 870 bp. Their N50 was 1077 bp. Comparing with Uni Prot database,48589 Unigenes were recorded in it with homologous information,with the annotation proportion of 56.7%. They were involved in 55 subcategories,3 categories(cellular component,biological process and molecular function). The largest amount of unigenes(21 subcategories)participated in biological process,and the smallest amount of unigenes(14 subcategories)in molecular function. Screening with Fold-Change≥2 as the condition,there were 2031 genes expressed differentially between transcriptome in stageⅠand stageⅡ,among which 1073 were up regulated and 958 were down regulated. A total of247 metabolic pathways were involved. The enrichment KEGG pathways functioned in endoplasmic reticulum protein processing,synthesis and accumulation of secondary metabolites,glycometabolism and photosynthesis. Genes with annotation of endoplasmic reticulum protein processing were more than others. Through the analysis of transcriptome of mango flower buds in two different developing stages,genes related to flowering were found in vernalization pathway,photoperiod pathway,gibberellin(GA)pathway,repression pathway,autonomous pathway and aging pathway. The results of the transcriptome and q RT-PCR of four genes(Mi SOC1,Mi VIN3,Mi Dof and Mi MADS1)showed the same change trend in gene expression,suggested the transcriptome result was credible,although there were some differences in expression changing multiple among the four genes at stage Ⅱ compared with stageⅠ.【Conclusion】It is identified that Endoplasmic reticulum protein processing,synthesis of secondary metabolites,hormone signal transduction,metabolic pathways of starch and sucrose were closely related to differentiation of mango flower bud. This study provides theoretical foundation for artificial regulation of flowering time and fruit maturity of mango.
【Objective】The transcriptome of mango(Mangifera indica L.) in two different bud differentiation periods was sequencing to comprehend the expression of genes related to mango flowering process. It laid the foundation for research on the molecular regulation mechanism of flowering time of mango.【Method】Illumina high-throughput sequencing technology was applied to detect the transcriptome sequencing data of mango variety Nandoumai 4 in two different flower bud differentiation stages,namely the withhold growing period of the shoots(stage I)and the base expanding period(stage II). High quality sequences in the results were analyzed by bioinformatics methods,such as gene function annotation and differentially expressed genes screening. Real time fluorescent quantitative PCR(q PCR)was used for the determination of differentially expressed genes to check the reliability of transcriptome sequencing.【Result】A total of 85691 Unigenes were obtained with average length of 870 bp. Their N50 was 1077 bp. Comparing with Uni Prot database,48589 Unigenes were recorded in it with homologous information,with the annotation proportion of 56.7%. They were involved in 55 subcategories,3 categories(cellular component,biological process and molecular function). The largest amount of unigenes(21 subcategories)participated in biological process,and the smallest amount of unigenes(14 subcategories)in molecular function. Screening with Fold-Change≥2 as the condition,there were 2031 genes expressed differentially between transcriptome in stageⅠand stageⅡ,among which 1073 were up regulated and 958 were down regulated. A total of247 metabolic pathways were involved. The enrichment KEGG pathways functioned in endoplasmic reticulum protein processing,synthesis and accumulation of secondary metabolites,glycometabolism and photosynthesis. Genes with annotation of endoplasmic reticulum protein processing were more than others. Through the analysis of transcriptome of mango flower buds in two different developing stages,genes related to flowering were found in vernalization pathway,photoperiod pathway,gibberellin(GA)pathway,repression pathway,autonomous pathway and aging pathway. The results of the transcriptome and q RT-PCR of four genes(Mi SOC1,Mi VIN3,Mi Dof and Mi MADS1)showed the same change trend in gene expression,suggested the transcriptome result was credible,although there were some differences in expression changing multiple among the four genes at stage Ⅱ compared with stageⅠ.【Conclusion】It is identified that Endoplasmic reticulum protein processing,synthesis of secondary metabolites,hormone signal transduction,metabolic pathways of starch and sucrose were closely related to differentiation of mango flower bud. This study provides theoretical foundation for artificial regulation of flowering time and fruit maturity of mango.
引文
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孟晓庆,侯智霞,张兰.2013.高等植物成花转变分子调控机制研究进展[J].生物技术进展,3(1):1-6.[Meng X Q,Hou Z X,Zhang L.2013.Progress on molecular and genetic mechanisms of floral control in higher plants[J].Current Biotechnology,3(1):1-6.]
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彭磊,高小俊,龙雯虹,吴兴恩,周玲,董广平,王莹.2011.短截后芒果花芽分化期间ABA含量的变化[J].云南农业大学学报,26(3):434-436.[Peng L,Gao X J,Long WH,Wu X E,Zhou L,Dong G P,Wang Y.2011.Changes of ABA contents in mango during floral differentiation after heading-back[J].Journal of Yunnan Agricultural University,26(3):434-436.]
孙俪,徐启江.2009.高等植物开花诱导途径信号整合的分子机制[J].生物技术通报,20(6):885-890.[Sun L,Xu QJ.2009.Molecular mechanisms of the signals integration of floral inductive pathways in high plants[J].Letters inBiotechnology,20(6):885-890.]
孙颖,谭晓风,罗敏,李建安.2014.油桐花芽2个不同发育时期转录组分析[J].林业科学,50(5):70-74.[Sun Y,Tan X F,Luo M,Li J A.2014.The sequencing analysis of transcriptome of Vernicia fordii flower buds at two development stages[J].Scientia Silvae Sinicae,50(5):70-74.]
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