Physiological changes and gene expression patterns of Polygonatum cyrtonema Hua seeds during physiological post-ripening process
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摘要:
目的 探明多花黄精种子生理后熟过程中生理生化变化规律及差异基因表达情况。 方法 以多花黄精种子生理后熟过程6个阶段的种子为材料，采用化学测定方法，分析每个阶段的超氧化物歧化酶、α淀粉酶、β淀粉酶、可溶性蛋白、粗脂肪、可溶性糖含量。采用高通量转录组测序技术，对6个阶段的差异表达基因进行分析，并用qRT-PCR法对关键差异基因的表达量进行验证。 结果 （1）多花黄精种子生理后熟过程中，可溶性糖含量在前4个时期逐渐增高后，在后两个时期稍有下降；粗脂肪、蛋白质整体呈下降趋势；α淀粉酶活性及β淀粉酶活性呈现增强趋势。（2）多花黄精种子生理后熟过程中，蔗糖和淀粉代谢途径、激素信号转导通路等显著富集差异表达基因。 结论 多花黄精种子生理后熟过程中，淀粉等营养物质的代谢、转化与内源激素的协调作用，促进多花黄精种子的生理后熟。 Abstract:Objective In order to study the physiological and biochemical changes and differential gene expression of Polygonatum cyrtonema Hua seeds during physiological post-ripening. Method the contents of superoxide dismutase, α-amylase, β-amylase, soluble protein, crude fat and soluble sugar at each stage were analyzed by chemical determination method. The differentially expressed genes in 6 stages were analyzed by high-throughput sequencing technology, and the expression levels of key differentially expressed genes were verified by qRT-PCR. Result (1) During the physiological post-ripening process of Polygonatum cyrtonema Hua seeds, the soluble sugar content increased gradually in the first four stages, and slightly decreased in the last two stages; The crude fat and protein showed a decreasing trend. The activity of α-amylase and β-amylase increased. (2) During physiological postripening of Polygonatum cyrtonema Hua seeds, differentially expressed genes were significantly enriched in sucrose and starch metabolic pathways and hormone signal transduction pathways. Conclusion During the physiological post-ripening process of Polygonatum cyrtonema Hua seeds, the conversion of starch and other nutrients and the coordination of endogenous hormones promoted the physiological post-ripening of Polygonatum cyrtonema Hua seeds. This study is of great significance for understanding the physiological and biochemical changes of Polygonatum cyrtonema Hua seeds during post-ripening and the molecular mechanism of dormancy release. -
图 1 多花黄精种子生理后熟过程的表型分析
注：T0，后熟休眠期；T1，初始萌发期；T2，完全萌发期；T3，小球茎形成期；T4，胚芽形成期；T5，初生根茎形成期。
Figure 1. Superficial analysis of physiological post-ripening process of Polygonatum cyrtonema Hua seeds
Note: T0, postmature dormancy; T1, initial germination stage; T2, complete germination stage; T3, corm formation stage; T4, germ formation stage; T5, primary rhizome formation stage.
图 2 多花黄精种子生理后熟过程中营养物质与酶的动态变化
Figure 2. Dynamic changes of nutrients and enzymes in Polygonatum cyrtonema Hua seeds during physiological afterripening
图 3 多花黄精种子生理后熟过程的转录组测序分析
a: 差异表达基因数量； b: 主成分分析；c-e: T0vsT1, T0vsT2, T0vsT3, T0vsT4和T0vsT5五个比较组的共有、上调和下调差异基因韦恩图
Figure 3. Transcriptome sequencing analysis of physiological post-ripening process of Polygonatum cyrtonema Hua seeds
a: Number of differentially expressed genes; b: Principal component analysis; c-e: Five comparison groups (T0vsT1, T0vsT2, T0vsT3, T0vsT4 and T0vsT5) shared, up-regulated and down-regulated differential genes
图 5 植物激素信号转导通路基因表达模式
Figure 5. Gene expression patterns of plant hormone signaling pathways
表 1 荧光定量PCR引物序列
Table 1. Sequence of qPCR primers
基因名称
Gene name正向引物(5'-3')
Forward primer (5'-3')反向引物(5'-3')
Reverse prime (5'-3')TGA10 TGGTCGCCAATAATGCCTCC GAAGGCTTGCGCAGAATAGG IAA18 TTGCTGCTGGGAATCAGAGG ATGCGGTCTCCTTCATCGTC IAA30 TGCGGTACATCTTGAGGTCG GAGGAGTCCAGCACCAATCC TIFY 10 TTTCGGATTCGGGTAAGCCC CTGAATTCGGGTTCGGCTCT 内参基因 reference gene GCCTGTGGTTATCACTGCCT CGGATCGTCTGCTCCACTAC 
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