Scientia Agricultura Sinica ›› 2018, Vol. 51 ›› Issue (8): 1431-1447.doi: 10.3864/j.issn.0578-1752.2018.08.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Gene Expression Profiling of Foxtail Millet (Setaria italica L.) Under Drought Stress During Germination

XU BingXia, YIN MeiQiang, WEN YinYuan, PEI ShuaiShuai, KE ZhenJin, ZHANG Bin, YUAN XiangYang   

  1. College of Agriculture, Shanxi Agricultural University, Taigu 030801, Shanxi
  • Received:2017-11-03 Online:2018-04-16 Published:2018-04-16

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Abstract: 【Objective】Foxtail millet (Setaria italica L.) is a drought tolerant crop. The objective of this research is to get a lot of differently expressed genes during germination in response to drought stress by high-throughput sequencing, then to obtain the key gene and the related molecular mechanism at seed germination stage in foxtail millet under drought stress.【Method】Seed of JinGu45 was treated with 18%PEG-6000（PEG-stress）and distilled water germination(control sample) at 1 h, 10 h and 18 h as a test material, and the activities of SOD, POD and CAT were measured, respectively. SOD activity was assayed by nitro blue tetrazolium (NBT) method. POD activity was determined by guaiacol method, and the activity of CAT was measured by colorimetric method. Sample of control and PEG-stress that germinated for 10h and 18h were used to construct cDNA library by gene expression profiling technology. We compared reads to the reference genome by using Bowtie and analysed the result by using RSEM. Differential expression analysis used DESeq. The functional annotation of differently expresse genes were obtained by using NR, Swiss-Prot, KEGG, COG and GO online databases. The key genes that regulate germination in foxtail millet was obtained through analyse DEGs. The reliability of sequencing results was comfirmed by qRT-PCR. 【Result】The SOD activity of PEG-stress sample was higher than that in the control sample, but the activity of POD and CAT were lower than that in the control group. With the time of germination changes, the activity of SOD was increased, but the activity of CAT and POD was gradually decreased. The sequences of gene expression profile was highly consistent with the selected reference genome sequence, and the gene expression was highly heterogeneous. Expression analysis showed a total of 35470 genes, and with the selection criteria of RPKM ≥0.01, there were 24030 and 24 486 genes in the control samples and 24 019 and 23 877 genes in the samples under PEG drought stress, respectively. 456 and 545 DEGs were screened out during millet germination at 10h and 18h under drought stress, in which 87 and 267 DEGs were up-regulated and 369 and 278 DEGs were down-regulated. GO enrichment analysis showed that these DEGs were mainly relating to metabolism process, cell stimulation and response process. The KEGG enrichment analysis showed that these DEGs were associated with phenylpropanoid metabolism and plant hormone signal transduction. The results obtained from five genes tested by RT-PCR agreed with the trend of regulation identified by gene expression profile. 【Conclusion】DEGs were widely involved in the metabolism of biomacromolecule such as sugar, protein, nucleic acid, secondary metabolism and energy metabolism. SnRK2 and PAL genes may regulate seed germination in foxtail millet under drought stress.

Key words: foxtail millet (Searia italic L.), drought stress, seed germination, gene expression profile

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