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. 2021 Jun 13;10(6):1204.
doi: 10.3390/plants10061204.

Identification and Expression Profiling of Nonphosphorus Glycerolipid Synthase Genes in Response to Abiotic Stresses in Dendrobium catenatum

Xinqiao Zhan  1   2 , Yichun Qian  2 , Bizeng Mao  2
Affiliations

Identification and Expression Profiling of Nonphosphorus Glycerolipid Synthase Genes in Response to Abiotic Stresses in Dendrobium catenatum

Xinqiao Zhan et al. Plants (Basel). .

Abstract

Dendrobium catenatum, a valuable Chinese herb, frequently experiences abiotic stresses, such as cold and drought, under natural conditions. Nonphosphorus glycerolipid synthase (NGLS) genes are closely linked to the homeostasis of membrane lipids under abiotic stress in plants. However, there is limited information on NGLS genes in D. catenatum. In this study, a total of eight DcaNGLS genes were identified from the D. catenatum genome; these included three monogalactosyldiacylglycerol synthase (DcaMGD1, 2, 3) genes, two digalactosyldiacylglycerol synthase (DcaDGD1, 2) genes, and three sulfoquinovosyldiacylglycerol synthase (DcaSQD1, 2.1, 2.2) genes. The gene structures and conserved motifs in the DcaNGLSs showed a high conservation during their evolution. Gene expression profiling showed that the DcaNGLSs were highly expressed in specific tissues and during rapid growth stages. Furthermore, most DcaNGLSs were strongly induced by freezing and post-freezing recovery. DcaMGD1 and DcaSQDs were greatly induced by salt stress in leaves, while DcaDGDs were primarily induced by salt stress in roots. Under drought stress, most DcaNGLSs were regulated by circadian rhythms, and DcaSQD2 was closely associated with drought recovery. Transcriptome analysis also revealed that MYB might be regulated by circadian rhythm and co-expressed with DcaNGLSs under drought stress. These results provide insight for the further functional investigation of NGLS and the regulation of nonphosphorus glycerolipid biosynthesis in Dendrobium.

Keywords: Dendrobium; abiotic stress; circadian regulation; nonphosphorus glycerolipid synthase.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Phylogenetic analysis of NGLSs in D. catenatum, rice, and Arabidopsis. A total of eight NGLSs domain-containing proteins from D. catenatum, nine from rice, and six from Arabidopsis were used to construct the unrooted neighbor-joining (NJ) tree with a bootstrap of 1000 replicates. Four sucrose synthases (SS) and five UDP-glucose epimerases (UGE) were divided from the SQD1 and SQD2 clusters.
Figure 2
Figure 2
Domain organization and gene structure of DcaNGLSs. (a,b) The conserved motifs of DcaNGLSs are predicted by MEME. (c) The DcaNGLSs structures are constructed by GSDS 2.0.
Figure 3
Figure 3
Predicted cis-elements in DcaNGLS promoters. Promoter sequences (−2000 bp) of 8 DcaNGLSs are analyzed by PlantCARE. The upstream length to the translation start site can be inferred according to the scale at the bottom.
Figure 4
Figure 4
Expression patterns of DcaNGLSs in different tissues. (a) DcaNGLSs are highly expressed in the specific tissues. (b) KEGG enrichment analysis in green root tip vs. white root comparison. (c) KEGG enrichment analysis in leaf vs. flower buds comparison. (d) The Sankey diagram shows the regulatory relationships between transcription factors and DcaNGLSs leaf vs. flower buds. The color scale represents the color code for the log2 fold-change of gene expression in a.
Figure 5
Figure 5
Expression patterns of DcaNGLSs in four stages. (a) DcaNGLSs are highly expressed in the S2. (b) KEGG enrichment analysis in S2 vs. S3 comparison. (c) Expression patterns of glycerolipid metabolism-related genes. (d) The expression levels of transcription factors in S2 vs. S3 comparison. (e). The expression distribution of ERF in S2 vs. S3 comparison. S1: plant experiences vegetative growth with few polysaccharides (4 months after sprouting); S2: plant accumulates polysaccharides rapidly (10 months after sprouting); S3: plant develops into a mature stage with the highest polysaccharide content (12 months after sprouting); S4: plant begins to die and the polysaccharide content decreases rapidly (16 months after sprouting). The color scale represents the color code for the log2 fold-change in gene expression in (a,c,d).
Figure 6
Figure 6
Expression of DcaNGLSs in response to abiotic stresses. (a) Expression of DcaNGLSs in response to salinity treatment. The seedlings were supplied with 250 mM NaCl, and the leaf and root sample were harvested at 0, 4, and 12 h. (b) Expression of DcaNGLSs in response to cold treatment. The 2-year-old plants were treated with cold acclimation (CA), freezing (FT), and post-freezing recovery (FR). (c) Expression of DcaNGLSs in response to drought. The seedlings were watered on the 1st day, dried from the 2nd to the 7th day, and re-watered on the 8th day. Leaves were collected at different times; DR5/DR8, DR6/DR10, and DR7/DR15 indicate sampling at 06:30 and 18:30 on the 2nd, 7th, and 9th days, respectively, and DR11 indicates sampling at 18:30 on the 8th day. (d). Effect of the time of day on DcaNGLSs expression induced by drought stress. The experiment was repeated three times with similar results. Color scale represents the color code for the log2 fold-change in gene expression. CK, control group.
Figure 7
Figure 7
Co-expression analysis DcaNGLSs and MYB in response to drought. (a) Co-expression analysis of DcaNGLSs and MYB under drought stress. (b) The expression levels of MYB under drought stress. The color scale represents the color code for the log2 fold-change in gene expression.
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