转录组学对转基因杨树表型变化的代谢调控机制解析
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国家自然科学基金(32071486)


Transcriptomics Analysis of the Metabolic Regulation Mechanism of Phenotypic Changes in Transgenic Poplar
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    摘要:

    D型细胞周期蛋白(Dtype cyclin)调控着细胞周期G1/S的转变,在植物生长发育过程中发挥重要作用。转基因杨树 PtoCYCD2;1(OEPtoCYCD2;1)植株出现明显的表型变化,株高降低,茎粗变细且叶片发生卷曲。该研究以转基因杨树 OEPtoCYCD2;1为研究材料,通过转录组学测序和生理指标变化并结合植株表型特征分析 PtoCYCD2;1在植物生长发育中的功能,为研究木本植物D型细胞周期蛋白功能提供理论基础。结果表明:(1)在 OEPtoCYCD2;1中共鉴定得到1 269个差异表达基因,其中有700个上调表达,569个下调表达。分析发现,有26个属于AP2/ERF转录因子的基因上调表达;有8个下调的差异表达基因富集在木质部合成通路中;在碳代谢通路中共富集27个下调差异表达基因,其中有8个基因富集到卡尔文循环通路中。(2)qRTPCR实验结果显示,9个差异表达基因的qRTPCR结果与RNAseq测定的表达水平变化趋势一致,表明所用RNAseq结果可靠。(3)生理指标分析发现,与野生型(WT)相比,转基因杨树 OEPtoCYCD2;1的幼叶和成熟叶的总叶绿素含量分别增加57.36%和78.22%;成熟叶的可溶性糖含量下降了12.72%;幼叶和成熟叶中的木质素含量分别下降了4.48%和8.03%;幼茎和成熟茎中的木质素含量分别下降了20.03%和31.63%。研究认为,转基因杨树 OEPtoCYCD2;1通过影响杨树碳代谢和木质素合成过程中相关基因的表达,从而造成转基因植株相应代谢物含量减少,最终导致植株表型改变,总体生物量降低。

    Abstract:

    Dtype cyclin regulates the G1/S transition of the cell cycle and plays an important role in the growth and development of plants. Transgenic poplar PtoCYCD2;1 (OEPtoCYCD2;1) plants showed obvious phenotypic changes, with reduced plant height, thinner stems and curled leaves. Using transgenic poplar OEPtoCYCD2;1 as the research material, the study analyzed the function of PtoCYCD2;1 in plant growth and development by linking plant phenotypes to transcriptomics sequencing and physiological index changes. This study provides a theoretical basis for the functional study of Dtype cyclins in woody plants. Studies have shown that: (1) a total of 1 269 differentially expressed genes were identified in OEPtoCYCD2;1, of which 700 were upregulated and 569 were downregulated. Through analysis, it was found that 26 genes belonging to AP2/ERF transcription factors were upregulated; 8 downregulated differentially expressed genes were enriched in the xylem synthesis pathway; a total of 27 downregulated differentially expressed genes were enriched in the carbon metabolism pathway. In the carbon metabolism pathway, 8 genes were enriched in the Calvin cycle pathway. (2) The results of qRTPCR experiments showed that the qRTPCR results of 9 differentially expressed genes were consistent with the trend of expression levels determined by RNAseq, indicating the reliability of the RNAseq results. (3) Physiological index analysis showed that compared with wild type (WT), the total chlorophyll content of young and mature leaves of transgenic poplar OEPtoCYCD2;1 increased by 57.36% and 78.22%, respectively; and the soluble sugar content of mature leaves decreased by 12.72%; and the lignin content in young and mature leaves decreased by 4.48% and 8.03%, respectively and the lignin content in young and mature stems decreased by 20.03% and 31.63%, respectively. This study indicated that transgenic poplar OEPtoCYCD2;1 affects the expression of related genes in the carbon metabolism and lignin synthesis process of poplar, thereby reducing the contents of corresponding metabolites in transgenic plants, eventually leading to changes in plant phenotype and overall biomass reduction.

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和成成,吴照群,许 楠,等.转录组学对转基因杨树表型变化的代谢调控机制解析[J].西北植物学报,2022,42(5):751-759

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  • 在线发布日期: 2022-06-16
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