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作物学报 ›› 2017, Vol. 43 ›› Issue (10): 1489-1498.doi: 10.3724/SP.J.1006.2017.01489

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

活性炭促进玉米幼胚离体培养幼苗生长与发育的转录组分析

王金萍1,2,孙果忠2,*,王海波2,*   

  1. 1河北农业大学农学院, 河北保定 071001；2河北省农林科学院遗传生理研究所/河北省植物转基因中心, 河北石家庄 050051
  • 收稿日期:2017-03-21 修回日期:2017-04-20 出版日期:2017-10-12 网络出版日期:2017-05-08
  • 通讯作者: 孙果忠, E-mail: 13933023804@163.com; 王海波, E-mail: nkywanghb@163.com
  • 基金资助:

    本研究由河北省财政专项(2009055001, F15R25)资助。

Transcriptome Analysis of Promotive Effects of Active Carbon on Growth and Development of Maize Seedlings from in vitro Cultured Immature Embryos

WANG Jin-Ping1,2,SUN Guo-Zhong2,*,WANG Hai-Bo2,*   

  1. 1 College of Agriculture, Agricultural University of Hebei, Baoding 071001, China; 2 Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences / Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
  • Received:2017-03-21 Revised:2017-04-20 Published:2017-10-12 Published online:2017-05-08
  • Contact: Sun Guozhong, E-mail: 13933023804@163.com;Wang Haibo, E-mail: nkywanghb@163.com
  • Supported by:

    This study was supported by the Financial Fund Program of Hebei Province (2009055001, F15R25).

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摘要：

以玉米自交系昌7-2为试材, 比较了14 d龄幼胚在MS和MSA(MS加活性炭)培养基上离体培养9 d的幼苗生长情况, 并利用转录组测序技术分析了2种培养基上的玉米幼苗地上部和地下部的基因表达差异。结果表明, 活性炭可以显著促进玉米幼苗的生长与发育。活性炭主要影响地下部基因表达。加入活性炭后, 幼苗地上部表达上调的基因有1612个, 下调的基因有530个；幼苗地下部表达上调的基因有69个, 下调的基因有78个。GO功能显著性分析表明, 地下部差异表达基因(DEGs)主要涉及DNA包装、DNA包装复合体和水解酶活性, 地上部DEGs主要涉及脂类代谢、胞外区和过氧化物酶活性。KEGG富集分析表明, 地下部DEGs主要涉及能量、碳水化合物、脂类和氨基酸代谢, 以及细胞周期和植物激素转导途径；地上部DEGs主要涉及泛醌和其他萜-醌类物质合成途径。活性炭促进细胞周期途径中的关键基因(CYC、CDH1、MCM3、PCNA2和BUBR1)、生长素信号传导基因(Aux/IAA)以及细胞色素基因(CYP450)显著上调表达。利用实时荧光定量PCR (qRT-PCR)验证了10个DEGs的表达模式与转录组测序结果一致, 证实了转录组数据与分析的可靠性。

关键词: 玉米, 幼胚, 活性炭, 转录组

Abstract:

Immature embryos from the maize inbred line Chang 7-2 were collected at 14 days after pollination, and cultured on MS or MSA medium (MS medium plus active carbon) for nine days at 24°C. Active carbon significantly accelerated the growth and development of maize seedlings from cultured immature embryos. Using RNA-seq technique, the genes involved in the growth promotive effects of active carbon were analyzed. The presence of active carbon in the medium affected the gene expression in seedlings. Number of up- and down-regulated genes was 1612 and 530 in roots, as well as 69 and 78 in shoots, respectively, indicating that active carbon mainly affects gene expression in roots. GO enrichment analysis showed that differentially expressed genes (DEGs) in roots were mainly involved in DNA packaging, DNA packaging complex and hydrolase activity; the DEGs in shoots were mainly involved in lipid metabolic process, extracellular region and peroxidase activity. The KEGG enrichment analysis showed that the DEGs in roots were significantly associated with energy metabolism, carbohydrate metabolism, lipid metabolism, amino acid metabolism, cell cycle and plant hormone signal transduction. The DEGs in shoots were significantly associated with biosynthesis of ubiquinone and other terpenoid-quinone compounds. Several key genes involved in the cell cycle pathway (i.e., CYC, CDH1, MCM3, PCNA2, and BUB1), signal transduction of auxin (Aux/IAA) and cytochrome function (CYP450 oxidase) were significantly up-regulated by active carbon. Ten DEGs were confirmed by Real-time quantitative PCR assay, suggesting that our data and analysis of transcriptome sequencing are reliable.

Key words: Maize, Immature embryo, Active carbon, Transcriptome

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