Identification and Expression Analysis of GRAS Gene Family in Saccharum spontaneum
HUANG Cui-Lin1, LIN Ping-Ping2, ZHAO Xin-Wang2, ZHANG Mu-Qing1,*
1 Guangxi Key Laboratory of Sugarcane Biology/State Key Laboratory of Unitization and Conservation for Subtropical Agri-Biological Resources, Guangxi University, Nanning 530005, China; 2 National Sugarcane Engineering Technology Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Abstract：Sugarcane (Saccharum officinarum) is an important sugar and biofuel crop, and its yield is affected by various biotic and abiotic stresses. S. spontaneum is one of the important parents in sugarcane cultivation and breeding. GRAS (GAI-RGA-SCR) transcription factors play important roles in the regulation of plant photomorphogenesis, seed germination, root growth and defense responses. In this study, 173 members of the GRAS gene family were identified from the genome of Saccharum spontaneum using the Hidden Markov Model (HMM) configuration file of the GRAS domain, which were distributed on 30 chromosomes of S. spontaneum. Phylogenetic analysis by maximum likelihood method showed that 173 GRAS genes of S. spontaneum could be further divided into 8 subfamilies. Motif and gene structure analysis showed that 173 GRAS genes contained at least one conserved GRAS domain at the C-terminal, and 61.2% of GRAS genes contained at least one intron. The analysis of cis-acting elements in the 2 000 bp sequence promoter upstream of 173 GRAS genes found that all GRAS promoter region had cis-acting elements in response to biotic and abiotic stresses, indicating that they might be involved in the regulation process of a variety of stress. The spatiotemporal transcriptional expression analysis found that the members of the GRAS family from S. spontaneum significantly expressed at different stages of stem and leaf development. qRT-PCR verified the expression of 9 genes in mature stem and leaf tissues, and the results were consistent with transcriptome data, suggesting that their family members played different roles in the growth and development regulation of S. spontaneum. The expression of SsPAT1.8-1 and SsDELLA6-2 were induced by the pathogen of sugarcane pokkah boeng disease; SsPAT1.8-2, SsPAT1.9, SsPAT1.10, SsDELLA2, SsSCR2, SsDELLA1-2, SsSCL3.1-2 and SsHAM9 were induced by Sugarcane mosaic virus (ScMV); SsSCL3.2-1 was induced by drought stress. qRT-PCR results showed that after ScMV infection, SsSCR2 was up-regulated in leaf +1 of resistant variety 'B48', SsHAM9 was up-regulated in leaf +1 and leaf -3, and SsLISCL11-1 was down-regulated in leaf -3. It is inferred that these GRAS family genes might play an important regulatory role in sugarcane response to different stresses. These results lay a foundation for further analyzing the function and regulation mechanism of GRAS family genes in sugarcane, and provide valuable gene resources for stress resistance breeding of sugarcane.
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