狗尾草ARF基因家族的鉴定与表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0318

摘要/Abstract

摘要：

【目的】 生长素响应因子ARF（auxin response factor）可以调控生长素响应基因的表达，在植物生长发育中发挥着重要作用。从狗尾草全基因组范围内鉴定出SvARF基因家族成员，分析相关基因表达特征，为后续深入研究其与狗尾草生长发育的关系提供支撑。【方法】 利用生物信息学方法从狗尾草基因组中鉴定了ARF转录因子成员，分析其系统进化、染色体定位和基因结构等情况。利用RT-qPCR（quantitative real-time polymerase chain reaction）分析该家族成员在生长素类除草剂2,4-D异辛酯处理后的表达响应情况。【结果】 共鉴定到24个狗尾草SvARF基因家族成员，可分成4个亚家族，同一亚家族SvARF基因的结构相似且基因表达模式相近。不同SvARF基因的表达模式存在差异，且有组织特异性，III亚家族的SvARF基因在根茎分蘖处的表达量均较高。2,4-D异辛酯处理后，有9个SvARF基因的相对表达量发生显著变化，其中SvARF5、SvARF6、SvARF9、SvARF10、SvARF17和SvARF18表达量显著上调，SvARF14、SvARF22和SvARF24表达量显著下调。【结论】 狗尾草SvARF基因家族有24个成员。生长素类除草剂影响SvARF基因的表达，暗示SvARF基因在抑制狗尾草植株分蘖中发挥重要作用。

关键词: ARF转录因子, 狗尾草, 系统进化, 结构特征, 表达分析, 分蘖, 2,4-D异辛酯

Abstract:

【Objective】 The auxin response factor（ARF）plays an important role in plant growth and development through regulating the expression of auxin response genes. The SvARF gene family members were identified from the Setaria viridis genome, and the expression characteristics of related genes were analyzed, providing support for further research on the relationship between SvARF and the growth and development of S. viridis. 【Method】 The ARF transcription factors were identified from the S. viridis genome by bioinformatics. The phylogenetic evolution, chromosome location, and gene structure were also analyzed. Real-time quantitative polymerase chain reaction（RT-qPCR）were used to analyze the expression responses of SvARF gene to 2,4-D isooctyl esters. 【Result】 A total of 24 SvARF genes were identified. The members of SvARF gene family can be divided into 4 subfamilies. SvARF genes in the same subfamily shared similar expression pattern and genes in subfamily III expressed specifically in the tillers. After 2,4-D isooctyl ester treatment, the relative expressions of nine SvARF genes changed significantly. SvARF5, SvARF6, SvARF9, SvARF10, SvARF17 and SvARF18 were up-regulated, while SvARF14, SvARF22, and SvARF24 were down-regulated. 【Conclusion】 There are 24 members in the SvARF gene family. Auxin pesticide affects the expressions of SvARF genes, which may decrease the plant tiller number.

Key words: ARF transcription factor, Setaria viridis, phylogenetic evolution, structure characteristics, gene expression, tillering, 2,4-D isooctyl ester

王其昕, 张凌馨, 钟妮娜, 熊王丹. 狗尾草ARF基因家族的鉴定与表达分析[J]. 生物技术通报, 2024, 40(11): 227-235.

WANG Qi-xin, ZHANG Ling-xin, ZHONG Ni-na, XIONG Wang-dan. Identification and Expression Analysis of ARF Gene Family in Setaria viridis[J]. Biotechnology Bulletin, 2024, 40(11): 227-235.

图/表 9

表1 荧光定量 PCR 引物

Table 1 Primer sequences for quantitative real time PCR

基因名称Gene name	正向引物 Forword primer（5'-3'）	反向引物 Reverse primer（5'-3'）
SvARF5	GCAGTTCCGTCACATCTACCG	CGAGGAGTCATCCCAGTCAA
SvARF6	ACCCGTGGGAGTCATTTGTG	GATAGGTAGCGTGGATCGTTTC
SvARF9	AATTGGCTCGTTCGATTACG	ACTTGCCCTTGGCTGCTGT
SvARF10	TATTTGGGTTTCCTCTTGGTAG	TTCTGAACATTCTGGGTAGCCT
SvARF14	CCCTTCTTGCTCAACATCACC	CTTCGGCAGTTCCTTAGTCG
SvARF17	TTCTCCAAAGCCTGAGCAAT	CAAATCTTGGAACCCCGAAG
SvARF18	TGATGGTGAACTAAGACTGGGTG	AGACTGGACGGTTGAGGCTC
SvARF22	CCTGACCTCTGGCACCCTGAT	TCCCAAGCACAAGCACCCAC
SvARF24	GGTTTCTCATCTCAAGGTCGTG	AGCCTCCGTGCTACTTTCAT
SvActin	CAGTGGACGCACAACAGGTAT	AGCAAGGTCAAGACGGAGAAT

图1 SvARF的染色体分布

Fig. 1 Chromosomes distribution of SvARF genes

图2 狗尾草和水稻的ARF蛋白质的系统进化分析

Fig. 2 Phylogenetic analysis of ARF protein in S. viridis and rice（Oryza sativa）

图3 狗尾草SvARF基因家族之间的共线性分析

Fig. 3 Collinearity analysis among the SvARF gene family in S. viridis

图4 狗尾草SvARF基因结构分析

Fig. 4 Analysis of SvARF gene structure in S. viridis

图5 狗尾草ARF蛋白质三级结构预测

Fig. 5 Prediction of the tertiary structures of ARF proteins in S. viridis

图6 SvARF在狗尾草不同组织中表达模式

Fig. 6 Expression pattern of SvARF genes in different tissues of S. viridis

图7 喷施2,4-D异辛酯后SvARF基因在狗尾草分蘖中的相对表达情况以SvARF基因在对照T-0时的表达量作为1，相应地计算处理时该基因表达水平。T-1和T-2分别表示2,4-D异辛酯处理浓度为1 050和4 200 g a.i./ha。**表示P<0.01，下同

Fig. 7 Relative expression analysis of SvARF genes in the tillers of S. viridis after spraying 2,4-D isooctyl ester The expression of SvARF genes under control T0-1 was defined as 1, and the relative expression of SvARF gene under treatment was determined by the relative fold changes between treatment and control. T-1 and T-2 refer to the concentration of 2,4-D isooctyl ester at 1 050 and 4 200 g a.i./ha, respectively. ** indicates P<0.01. The same below

图8 喷施2,4-D异辛酯后狗尾草分蘖中的IAA含量 T-0表示2,4-D异辛酯处理浓度为0 g a.i./ha

Fig. 8 Content of IAA in the tillers of S. viridis after spraying 2,4-D isooctyl ester T-0 refers to the concentration of 2,4-D isooctyl ester at 0 g a.i./ha

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