施钙量对白菜生长和钙转运关键基因家族ACA表达的影响

Effects of calcium application rate on the growth and calcium transport key gene family ACA expression in Chinese cabbage

  • 摘要: 【目的】探究施钙量对白菜生长和钙转运关键基因家族ACA表达的影响,了解白菜钙吸收、转运和富集的分子调控机制,为进一步开展富钙白菜新品种选育及栽培中钙肥合理施用提供理论基础。【方法】采用黑叶五月慢和上海六月慢2个白菜品种进行盆栽试验,设0、0.2、0.4、0.6和0.8 g/kg施钙量处理,以0 g/kg为对照,测定白菜地上部和根系生物量、钙含量及钙积累量,选取对照和0.8 g/kg施钙量处理白菜叶片进行转录组测序,筛选差异表达基因(DEGs)并进行GO功能注释和KEGG信号通路富集分析,通过实时荧光定量PCR验证施钙量对ACA基因家族表达的影响。【结果】随着施钙量的增加,黑叶五月慢和上海六月慢地上部和根系干重、钙含量均呈先升高后降低的变化趋势,2个白菜品种地上部和根系干重、钙含量分别在0.4、0.6 g/kg施钙量处理下达最大值且显著高于对照(P<0.05,下同);2个白菜品种地上部钙积累量随着施钙量的增加均呈先升高后降低的变化趋势,而根系钙积累量整体呈升高趋势。0.8 g/kg施钙量处理与对照黑叶五月慢间存在87个DEGs,0.8 g/kg施钙量处理与对照上海六月慢间存在176个DEGs,DEGs主要注释到细胞过程、代谢过程、结合、催化活性等GO功能条目,显著富集到酪氨酸代谢、硫代葡萄糖苷生物合成、植物激素信号传导等KEGG信号通路。从DEGs中鉴定出25个ACA基因家族成员。实时荧光定量PCR检测结果表明,0.6和0.8 g/kg施钙量处理下2个白菜品种ACA4-4ACA10-1基因相对表达量均较对照显著上调,黑叶五月慢ACA11-2基因相对表达量在0.6 g/kg施钙量处理下较对照显著上调。对白菜ACA基因相对表达量与钙含量进行相关分析发现,ACA4-4ACA10-1ACA11-2基因相对表达量与白菜钙含量均呈正相关,其中ACA4-4基因相对表达量与白菜钙含量相关性达显著水平。【结论】适量施钙可显著提高白菜地上部和根系干重及钙含量。筛选出钙转运关键基因ACA4-4ACA10-1ACA11-2,推测ACA基因家族成员在白菜钙转运和积累中具有重要作用。

     

    Abstract: 【Abstract】To explore the effects of calcium application rate on expression of the key gene family ACA of growth and calcium transport in Chinese cabbage, and to study the molecular regulation mechanism of calcium absorption, transport and enrichment in cabbage, which could provide theoretical basis for further breeding of new calcium-rich Chinese cabbage and the rational application of calcium fertilizer in cultivation. 【Method】A pot experiment was conducted on 2 varieties of Chinese cabbage, Heiyewuyueman and Shanghailiuyueman, with 0, 0.2, 0.4, 0.6 and 0.8 g/kg calcium application rates to determine aboveground and root biomass, calcium content and calcium accumulation of Chinese cabbage, and 0 g/kg was as control. Control and 0.8 g/kg Chinese cabbage leaves were selected for transcriptome sequencing, differential expression genes(DEGs) were screened, GO function annotation and KEGG signaling pathway enrichment analysis were performed, and the effects of calcium application on ACA gene family expression were verified by real-time fluorescence quantitative PCR. 【Result】With the increase of calcium application rate, the aboveground dry weight, root dry weight and calcium content of Heiyewuyueman and Shanghailiuyueman increased first and then decreased, calcium content in shoots and roots of the 2 varieties reached the peak at 0.4 and 0.6 g/kg calcium application rates and were significantly higher than control(P<0.05, the same below). The above-ground calcium accumulation of the 2 varieties showed a trend of increasing first and then decreasing with the increase of calcium application rate. The accumulation of calcium in root system showed an increasing trend. There were 87 DEGs between 0.8 g/kg calcium application rate treatment and control in Heiyewuyueman, and 176 DEGs between 0.8 g/kg calcium application rate treatment and control in Shanghailiuyueman. DEGs was mainly annotated on GO function items such as cellular process, metabolic process, binding and catalytic activity. KEGG signaling pathways such as tyrosine metabolism, glucoside biosynthesis and plant hormone signaling transduction were significantly enriched. Twenty-five ACA gene family members were identified from DEGs. The results of real-time fluorescence quantitative PCR showed that the relative expression levels of ACA4-4 and ACA10-1 genes in 2 Chinese cabbage varieties treated with 0.6 and 0.8 g/kg calcium application rate were significantly up-regulated compared with the control, relative expression level of ACA11-2 gene in Heiyewuyueman treated with 0.6 g/kg calcium application rate was significantly up-regulated compared with the control. The correlation analysis between ACA gene expression and calcium content in Chinese cabbage showed that the relative expression levels of ACA4-4,ACA10-1 and ACA11-2 genes were positively correlated with calcium content in Chinese cabbage, and the relative expression levels of ACA4-4 genes were significantly correlated with calcium content in Chinese cabbage.【Conclusion】The dry weight and calcium content of aboveground and roots of Chinese cabbage are significantly increased by applying proper amount of calcium. Key calcium transport genes ACA4-4, ACA10-1 and ACA11-2 are screened, suggesting that ACA gene family members play an important role in calcium transport and accumulation in Chinese cabbage.

     

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