尼泊尔黄堇Aux/IAA基因家族的鉴定与UVB处理下表达模式分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0401

摘要/Abstract

摘要：

【目的】鉴定尼泊尔黄堇Aux/IAA基因家族，为尼泊尔黄堇Aux/IAA基因功能研究及抵御逆境胁迫提供理论基础。【方法】基于尼泊尔黄堇全长转录组数据，鉴定和分析尼泊尔黄堇Aux/IAA（ChAux-IAA）基因家族成员，并对其蛋白质理化性质、系统进化树、基因结构、蛋白质三级结构及蛋白质互作关系、启动子顺式作用元件进行分析。此外，对尼泊尔黄堇进行不同时间（0.5 h、48 h）UV-B处理，检测ChAux/IAA基因家族的表达模式。【结果】尼泊尔黄堇ChAux/IAA基因家族共有20个成员，大多数位于细胞核中，少数位于叶绿体、线粒体和细胞壁中。ChAux/IAA基因家族编码蛋白属于亲水性蛋白。ChAux/IAA基因家族的顺式作用元件中存在光响应元件，表明该基因家族可能参与植物的生长发育和抵御紫外胁迫。通过RT-qPCR分析发现，在UV-B处理下，20个ChAux/IAA的表达量均较对照组增加，其中ChIAA2、ChIAA3、ChIAA4、ChIAA10、ChIAA13和ChIAA21表达量增加更加显著，表明ChAux/IAA基因家族在其抵御紫外线胁迫方面具有重要功能。【结论】共鉴定出20个ChAux/IAA基因家族成员，ChAux/IAA基因家族在UV-B胁迫中发挥重要作用。

关键词: 尼泊尔黄堇, Aux/IAA基因家族, UV-B胁迫, 全长转录组, 生物信息学, 基因表达分析

Abstract:

【Objective】 To identify the Aux/IAA gene family in Corydalis hendersonii, and to provide a theoretical basis for the study of the function of C. hendersonii Aux/IAA genes and their resistances to adverse stresses.【Method】Based on the full-length transcriptome data of C. hendersonii, the members of the Aux/IAA（ChAux-IAA）gene family were identified and analyzed, and their proteins'physicochemical properties, phylogenetic tree, gene structure, protein tertiary structure, and protein interaction relationships were analyzed. In addition, C. Hendersonii was treated with UV-B at different times（0.5 h and 48 h）to detect the expression patterns of the ChAux/IAA gene family. 【Result】The ChAux/IAA gene family of C. hendersonii comprises a total of 20 members, most of which are located in the cell nucleus, with a minority found in chloroplasts, mitochondria, and cell walls. The proteins encoded by the ChAux/IAA gene family are hydrophilic. The presence of light-responsive elements in the cis-acting elements of the ChAux/IAA gene family suggests that this gene family may be involved in plant growth and development and in resisting ultraviolet stress. Analysis via RT-qPCR revealed that under UVB treatment, the expressions of all 20 ChAux/IAA members increased compared to the control group, with ChIAA2, ChIAA3, ChIAA4, ChIAA10, ChIAA13 and ChIAA21 showing a more significant increase in expression. This indicates that the ChAux/IAA gene family plays an important role in the plant's resistance to ultraviolet stress. 【Conclusion】A total of 20 ChAux/IAA gene family members are identified, and the ChAux/IAA gene family plays an important role in UV-B stress.

Key words: Corydalis hendersonii Hemsl., Aux/IAA gene family, UV-B stress, transcriptome, bioinformatics, analysis on gene expression

梁佳林, 赵爽, 李幸儿, 赵成周, 李萍. 尼泊尔黄堇Aux/IAA基因家族的鉴定与UVB处理下表达模式分析[J]. 生物技术通报, 2024, 40(12): 182-192.

LIANG Jia-lin, ZHAO Shuang, LI Xing-er, ZHAO Cheng-zhou, LI Ping. Identification of Aux/IAA Gene Family in Corydalis hendersonii Hemsl. and Analysis on Their Expression Pattern under UVB Treatment[J]. Biotechnology Bulletin, 2024, 40(12): 182-192.

图/表 9

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引物名称Primer name	上游引物Forward primer（5'-3'）	下游引物Reverse primer（5'-3'）
ChIAA1	GCGAGCGAGATTCTGAGTGAGAG	GCATCCAGTCCCCATCCTTGTC
ChIAA2	GATGTGCCGTGGGAGATGTTTATTG	TGGTGAGTGACTGGAGACAAGTTC
ChIAA3	AGGATGCTGGTTGGAGATGATGTC	GTGTCATTGCTTGCTTGCTGTTTC
ChIAA4	GATGGGTTGCGTGAGGAACAAAG	TGATGAGGGAAACAAAGAGGAAGGG
ChIAA5	GATGGGTTGCGTGAGGAACAAAG	TGATGAGGGAAACAAAGAGGAAGGG
ChIAA6	TAGAGGTGGAAATGGTTGTGGGAAG	CGCCTTGGGAGACATAGGAGTAAC
ChIAA7	ATGCTATGAAACCCTTGCCCAAAC	AACCACATGCTCCTCTCCACTTG
ChIAA8	GCGAGCGAGATTCTGAGTGAGAG	GCATCCAGTCCCCATCCTTGTC
ChIAA9	TAGAGGTGGAAATGGTTGTGGGAAG	CGCCTTGGGAGACATAGGAGTAAC
ChIAA10	AGGATGCTGGTTGGAGATGATGTC	GTGTCATTGCTTGCTTGCTGTTTC
ChIAA11	CCATCGGCATCCAAAGCACAAG	CCTTCTCCTCATCGGCGTTACTC
ChIAA12	AAGGAGACAGGATGTTGGTTGGAG	TGTCTTTCCTTGCTTGTTGCTTCC
ChIAA13	GGAAGGAAGGAAGGAAGGAAGGAAG	AGTTGTAGTCGTCTTTGTCGTCTCC
ChIAA14	GGAAGGAAGGAAGGAAGGAAGGAAG	AGTTGTAGTCGTCTTTGTCGTCTCC
ChIAA16	AAGGAGACAGGATGTTGGTTGGAG	TGTCTTTCCTTGCTTGTTGCTTCC
ChIAA18	GATGGGTTGCGTGAGGAACAAAG	TGATGAGGGAAACAAAGAGGAAGGG
ChIAA19	TACGACGACGAAGACGAAGAACTC	CAAATCTTTCCTCCTCCGTTTTCCG
ChIAA21	TAGAGGTGGAAATGGTTGTGGGAAG	CGCCTTGGGAGACATAGGAGTAAC
ChIAA23	AGGATGCTGGTTGGAGATGATGTC	GTGTCATTGCTTGCTTGCTGTTTC
ChIAA24	TTCTGATGCTTGTGACGATGTTGAC	GTTCTAACCGGCGGCCATCC
GAPDH	AGTGGACTTAACTTGCCGACTTGAG	GAACTGAGACCTATGCCAGCCTTG

引物名称Primer name	上游引物Forward primer（5'-3'）	下游引物Reverse primer（5'-3'）
ChIAA1	GCGAGCGAGATTCTGAGTGAGAG	GCATCCAGTCCCCATCCTTGTC
ChIAA2	GATGTGCCGTGGGAGATGTTTATTG	TGGTGAGTGACTGGAGACAAGTTC
ChIAA3	AGGATGCTGGTTGGAGATGATGTC	GTGTCATTGCTTGCTTGCTGTTTC
ChIAA4	GATGGGTTGCGTGAGGAACAAAG	TGATGAGGGAAACAAAGAGGAAGGG
ChIAA5	GATGGGTTGCGTGAGGAACAAAG	TGATGAGGGAAACAAAGAGGAAGGG
ChIAA6	TAGAGGTGGAAATGGTTGTGGGAAG	CGCCTTGGGAGACATAGGAGTAAC
ChIAA7	ATGCTATGAAACCCTTGCCCAAAC	AACCACATGCTCCTCTCCACTTG
ChIAA8	GCGAGCGAGATTCTGAGTGAGAG	GCATCCAGTCCCCATCCTTGTC
ChIAA9	TAGAGGTGGAAATGGTTGTGGGAAG	CGCCTTGGGAGACATAGGAGTAAC
ChIAA10	AGGATGCTGGTTGGAGATGATGTC	GTGTCATTGCTTGCTTGCTGTTTC
ChIAA11	CCATCGGCATCCAAAGCACAAG	CCTTCTCCTCATCGGCGTTACTC
ChIAA12	AAGGAGACAGGATGTTGGTTGGAG	TGTCTTTCCTTGCTTGTTGCTTCC
ChIAA13	GGAAGGAAGGAAGGAAGGAAGGAAG	AGTTGTAGTCGTCTTTGTCGTCTCC
ChIAA14	GGAAGGAAGGAAGGAAGGAAGGAAG	AGTTGTAGTCGTCTTTGTCGTCTCC
ChIAA16	AAGGAGACAGGATGTTGGTTGGAG	TGTCTTTCCTTGCTTGTTGCTTCC
ChIAA18	GATGGGTTGCGTGAGGAACAAAG	TGATGAGGGAAACAAAGAGGAAGGG
ChIAA19	TACGACGACGAAGACGAAGAACTC	CAAATCTTTCCTCCTCCGTTTTCCG
ChIAA21	TAGAGGTGGAAATGGTTGTGGGAAG	CGCCTTGGGAGACATAGGAGTAAC
ChIAA23	AGGATGCTGGTTGGAGATGATGTC	GTGTCATTGCTTGCTTGCTGTTTC
ChIAA24	TTCTGATGCTTGTGACGATGTTGAC	GTTCTAACCGGCGGCCATCC
GAPDH	AGTGGACTTAACTTGCCGACTTGAG	GAACTGAGACCTATGCCAGCCTTG

蛋白名称Protein name	α-螺旋α-helix/%	β-折叠β-angle/%	随机卷曲Random coil/%	延伸连 Extended strand/%
ChIAA1	19.85	3.31	59.03	17.81
ChIAA2	26.23	6.49	46.23	21.04
ChIAA3	18.67	4.8	61.87	14.67
ChIAA4	16.02	4.14	64.64	15.19
ChIAA5	18.01	2.77	63.71	15.51
ChIAA6	13.75	3.44	66.76	16.05
ChIAA7	22.84	5.25	53.09	18.83
ChIAA8	16.09	2.21	65.93	15.77
ChIAA9	18.59	2.56	61.86	16.99
ChIAA10	19.80	2.97	61.39	15.84
ChIAA11	23.21	3.07	52.22	21.50
ChIAA12	17.29	3.01	56.77	22.93
ChIAA13	26.52	3.03	55.68	14.77
ChIAA14	22.96	3.89	57.20	15.95
ChIAA16	17.97	3.69	56.68	21.66
ChIAA18	7.73	2.42	76.81	13.04
ChIAA19	20.77	3.86	58.45	16.91
ChIAA21	31.89	4.86	47.03	16.22
ChIAA23	10.30	1.21	70.91	17.58
ChIAA24	26.14	5.88	45.75	22.22

蛋白名称Protein name	α-螺旋α-helix/%	β-折叠β-angle/%	随机卷曲Random coil/%	延伸连 Extended strand/%
ChIAA1	19.85	3.31	59.03	17.81
ChIAA2	26.23	6.49	46.23	21.04
ChIAA3	18.67	4.8	61.87	14.67
ChIAA4	16.02	4.14	64.64	15.19
ChIAA5	18.01	2.77	63.71	15.51
ChIAA6	13.75	3.44	66.76	16.05
ChIAA7	22.84	5.25	53.09	18.83
ChIAA8	16.09	2.21	65.93	15.77
ChIAA9	18.59	2.56	61.86	16.99
ChIAA10	19.80	2.97	61.39	15.84
ChIAA11	23.21	3.07	52.22	21.50
ChIAA12	17.29	3.01	56.77	22.93
ChIAA13	26.52	3.03	55.68	14.77
ChIAA14	22.96	3.89	57.20	15.95
ChIAA16	17.97	3.69	56.68	21.66
ChIAA18	7.73	2.42	76.81	13.04
ChIAA19	20.77	3.86	58.45	16.91
ChIAA21	31.89	4.86	47.03	16.22
ChIAA23	10.30	1.21	70.91	17.58
ChIAA24	26.14	5.88	45.75	22.22