Identification and Expressions under Low-temp/Draught Stress of Polyamine Synthesis Genes in Oil Palms
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摘要:
目的 研究多胺合成酶基因在油棕生长发育及非生物胁迫应答中的作用，为油棕抗性品种培育提供理论基础。 方法 从油棕基因组中鉴定出15个编码多胺合成酶的基因，并对其进行生物信息学分析和表达模式分析。 结果 多胺合成酶的氨基酸长度为318～720，分子量为36.00～46.43 kDa，等电点为4.78～6.31，蛋白不稳定指数为29.26～48.73，脂溶指数为76.46～94.41，总平均亲水性为−0.315～0.082，外显子数目为1～11个，具有特定的蛋白保守基序和保守结构域。进化分析发现油棕多胺合成酶基因可以分为3个家族，EgADCs与水稻的亲缘关系较近，EgACLs、EgSPDSs与椰枣的亲缘关系较近，EgSPMSs与玉米的亲缘关系较近。启动子上鉴定出大量植物激素响应、逆境胁迫响应和光响应的顺式作用元件。组织表达分析发现多胺合成酶基因在油棕不同组织中均有表达；多胺合成酶基因受低温和脱水胁迫的诱导。 结论 多胺合成酶基因参与油棕对低温和脱水胁迫的应答，其中EgSPDS1、EgSPDS2、EgSPMS1、EgADC1、EgADC2、EgACL5-2、EgACL5-3、EgSAMDC3-1、EgSAMDC4-1、EgSAMDC4-2和EgACL5-1受低温胁迫的诱导，EgSPDS1、EgSPDS2、EgADC1、EgADC2、EgSAMDC4-1和Eg SAMDC4-2、EgACL5-1受脱水胁迫的诱导。 Abstract:Objective Genetic information and resource of low-temp/draught resistant oil palms were studied with respect to the stress responses of polyamines synthase gene in the plants for breeding purposes. Method Bioinformatics of polyamines synthase genes in oil palm were gathered from the genome bank, and their expressions under abiotic stresses analyzed. Result Fifteen polyamines synthase genes were identified from the oil palm genome. There were between 318 and 720 amino acids in the genes with a molecular weight ranging from 36.00 kDa to 46.43 kDa and an isoelectric point between 4.78 and 6.31. The instability indices of the genes were found in between 29.26 and 48.73, the aliphatic indices in between 76.46 and 94.41, the gravy in between −0.315 and 0.082, and the exon number in between 1 and 11. They contained both specific conserved protein motifs and conserved domain. A phylogenetic analysis divided them into 3 families, i.e., EgADCs closely related to rice, EgACLs and EgSPDSs closely related to data palm, and EgSPMSs closely related to maize. Many cis-acting elements that response to phytohormone, stresses, and light were found in the promoters. The expressions of the polyamine synthase genes existed in different tissues of the plant and could be induced by either low temperature or draught stress. Specifically, EgSPDS1, EgSPDS2, EgSPMS1, EgADC1, EgADC2, EgACL5-2, EgACL5-3, EgSAMDC3-1, EgSAMDC4-1, EgSAMDC4-2, and EgACL5-1 were found to be upregulated by the imposed cold stress; while EgSPDS1, EgSPDS2, EgADC1, EgADC2, EgSAMDC4-1 EgSAMDC4-2, and EgACL5-1 by dehydration. Conclusion The polyamine synthase genes were significantly expressed in tissues of the oil palms that could be induced by either low temperature or draught stress. Those responded to the abiotic stresses were identified in this study. -
Key words:
- Oil palm /
- polyamine synthase /
- cold /
- dehydration /
- gene expression
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图 1 油棕多胺合成酶基因在染色体上的分布
Figure 1. Chromosome location of polyamine synthase genes in oil palm
图 2 油棕多胺合成酶的进化关系、保守蛋白基序、结构域和基因结构分析
Figure 2. Phylogenetic relationships, architecture of conserved protein motifs and conserved domain, and gene structure of polyamine synthase genes in oil palm
图 3 油棕和其他植物多胺合成酶基因家族的进化树
Figure 3. Phylogenetic tree of polyamine synthase genes in oil palms and other plants
图 4 油棕多胺合成酶启动子顺式作用元件分析
Figure 4. Cis-acting elements in promoter of polyamine synthase in oil palms
图 5 油棕多胺合成酶基因在不同组织中的表达模式
Figure 5. Expressions of polyamine synthase genes in tissues of oil palms
图 6 多胺合成酶基因在低温胁迫下表达模式
注：柱形图上方不同小写字母表示在0.05水平差异显著（下同）。
Figure 6. Expressions of polyamine synthase genes under low-temp stress
Note：Different lowercase letters at the top of the bar chart indicate significant differences at the 0.05 level (the same as below).
图 7 多胺合成酶基因在脱水胁迫下表达模式
Figure 7. Expressions of polyamine synthase genes under draught stress
表 1 油棕多胺合成酶基因qRT-PCR引物序列
Table 1. Sequence of qRT-PCR primer of polyamine synthase genes in oil palms
基因 Gene 正向引物 Forward primer 反向引物 Reverse primer 产物长度 Product length EgSPDS1 CCAGCAGGAGTCTTTACGCA CAGCCCCAAGTGTCAGCATA 122 EgSPDS1L TGTCTGCCGCCAAACCTTTA ATGAAACCAATCACCCCGCT 84 EgSPMS1 CCGATTGGTCCAGCTCAAGA AAGCCACATGCTCTCTGCTT 108 EgSPMS2 TGCGCCTGAAGGGATGTATG GGCCTTTCAACAAGTTCCCG 81 EgADCL GAAGGGCAAGTTCGGTCTGA AGCCGATGTGAAAGTGGAGG 107 EgADC GTGATGTTCGAGGGGCTCAA GCACCAGATAGGGCATGGAA 112 EgACL5-1 ACTTGAACGGCGAGTTCCTT AAGAACAACATGCCCTCCCA 135 EgACL5-2 CATGGGGATGGGTTATGGCA ACAAGGAACTCGCCGTTCAA 118 EgACL5-3 CTATGCTGACACTTGGGGCT CGGCAGATGCAAAGGTCTTG 137 EgACL5-4 CCAGCAGGAGTCTTTACGCA CAGCCCCAAGTGTCAGCATA 122 EgSAMDC2 TCAGCGGACGATCCACTCTA TGCTGAGTTGTGCTGGTTCT 96 EgSAMDC3-1 CAGCCTTCTCCCCATCGTAG GATGATGCCGGATCGCCTAT 110 EgSAMDC3-2 TGAAGCCATGGGTCTCAACC CCTTCTTCGACCATGTGCCT 143 EgSAMDC3-3 AGTACTCCCGTGGGACCTTT AAGCCTTACCACCCGAACTG 120 EgSAMDC4-1 GGAGATGACCGAGTTGACGG AATCGTCGAGTATCGGTCGC 121 EgSAMDC4-2 TTACTCGATGAACGGCCTCG CCGGAATACGTTCACGACCT 142 β-actin CTCAACCCCAAGGCGAAC GTAACACCATCTCCCGAGTCAA 100 
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表 2 油棕多胺合酶基因的理化性质分析
Table 2. Physicochemical properties of polyamine synthase genes in oil palms
基因名
Symbol基因登录号
Gene accession基因座登录号
Gene locus氨基酸长度
Number of amino acids分子量（kDa）
Molecular weight等电点
Theoretical pI蛋白不稳定指数
Instability index脂溶指数
Aliphatic index总平均亲水性
GravyEgSPDS1 XM_010943910.3 LOC105060266 346 37.92 4.80 48.15 85.29 −0.094 EgSPDS2 XM_010930791.3 LOC105050680 342 37.52 4.78 48.1 83.16 −0.137 EgSPMS1 XM_010915332.3 LOC105039242 389 42.70 5.31 45.35 81.93 −0.236 EgSPMS2 XM_019849278.2 LOC105040703 363 40.12 5.61 42.47 85.62 −0.131 EgACL5-1 XM_010934133.2 LOC105053105 332 37.74 5.36 30.61 82.20 −0.307 EgACL5-2 XM_010936715.1 LOC105055022 334 37.42 5.91 30.65 78.53 −0.315 EgACL5-3 XM_029263566.1 LOC105041819 318 36.00 5.10 29.26 83.71 −0.291 EgADC1 XM_010910687.3 LOC105035222 720 76.58 5.14 42.79 92.82 0.082 EgADC2 XM_010920850.3 LOC105043339 696 74.08 5.73 42.71 94.41 0.028 EgSAMDC1 XM_019846142.1 LOC105031981 356 38.93 5.47 50.07 85.51 −0.008 EgSAMDC2 XM_010906317.3 LOC105032002 423 46.43 6.05 46.75 80.21 −0.152 EgSAMDC3-1 XM_010930896.3 LOC105050758 402 43.89 4.96 48.73 80.27 −0.083 EgSAMDC3-2 XM_010915747.2 LOC105039560 382 41.89 5.01 44.15 86.05 0.000 EgSAMDC4-1 XM_010923664.3 LOC105045398 340 37.34 6.31 46.89 81.18 −0.082 EgSAMDC4-2 XM_010933625.3 LOC105052707 333 36.27 6.17 46.26 76.46 −0.158
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