Physiobiochemical Changes and Gene Expressions of Polygonatum cyrtonema Hua Seeds undergone Post-ripening Treatment
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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 Physiobiochemical changes and differentially expressed genes in Polygonatumcyrtonema Hua seeds after a post-ripening treatment were studied. Method Contents of superoxide dismutase, α-amylase, β-amylase, soluble protein, crude fat, and soluble sugar at 6 seed development stages were monitored. Differentially expressed genes were analyzed with high-throughput sequencing followed by a qRT-PCR verification. Result (1) The post-ripening treatment raised the soluble sugar content in the seeds gradually in the initial 4 stages and slightly declined in the last two stages, while induced a declining trend on the crude fat and protein. Meanwhile, the activities of α-amylase and β-amylase increased. (2) Significantly enriched differentially expressed genes were observed in the sucrose and starch metabolic pathways as well as the hormone signal transduction pathway during the process. Conclusion The post-ripening treatment on the P. cyrtonema seeds brought about the conversion and metabolism of starch and other nutrients as well as endogenous hormones interactions promoting the physiological changes and dormancy release. -
图 1 多花黄精种子生理后熟过程的表型分析
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 on physiological post-ripening process of P. cyrtonema Hua seeds
T0: post-mature 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 on nutrients and enzymes in P. cyrtonema Hua seeds during post-ripening treatment
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的共有、上调和下调差异基因韦恩图。
Figure 3. Transcriptome sequencing on physiological post-ripening process of P. cyrtonema Hua seeds
a: number of differentially expressed genes; b: principal component analysis; c–e: shared, upregulated, and downregulated differential genes of 5 comparison groups (T0 vs. T1, T0 vs. T2, T0 vs. T3, T0 vs. T4, and T0 vs. T5), respectively.
图 5 植物激素信号转导通路基因表达模式
红色方框表示基因的上调，绿色方框表示基因的下调，黄色方框表示同时存在上调和下调基因。
Figure 5. Gene expressions of plant hormone signaling pathways
Red, green, yellow boxes indicate up-regulated, down-regulated and both up-regulated and down-regulated genes.
表 1 荧光定量PCR引物序列
Table 1. Sequences of qPCR primers
基因名称
Gene name正向引物
Forward primer (5'-3')反向引物
Reverse prime (5'-3')TGA10 TGGTCGCCAATAATGCCTCC GAAGGCTTGCGCAGAATAGG IAA18 TTGCTGCTGGGAATCAGAGG ATGCGGTCTCCTTCATCGTC IAA30 TGCGGTACATCTTGAGGTCG GAGGAGTCCAGCACCAATCC TIFY 10 TTTCGGATTCGGGTAAGCCC CTGAATTCGGGTTCGGCTCT GAPDH GCCTGTGGTTATCACTGCCT CGGATCGTCTGCTCCACTAC 
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