园艺学报 ›› 2022, Vol. 49 ›› Issue (1): 100-116.doi: 10.16420/j.issn.0513-353x.2020-0850
谢思艺1,3, 周承哲1,2,3, 朱晨1,2,3, 詹冬梅1,3, 陈兰1,3, 吴祖春1,3, 赖钟雄1,2, 郭玉琼1,3,*()
收稿日期:
2021-04-23
修回日期:
2021-07-26
出版日期:
2022-01-25
发布日期:
2022-01-24
通讯作者:
郭玉琼
E-mail:guoyq828@163.com
基金资助:
XIE Siyi1,3, ZHOU Chengzhe1,2,3, ZHU Chen1,2,3, ZHAN Dongmei1,3, CHEN Lan1,3, WU Zuchun1,3, LAI Zhongxiong1,2, GUO Yuqiong1,3,*()
Received:
2021-04-23
Revised:
2021-07-26
Online:
2022-01-25
Published:
2022-01-24
Contact:
GUO Yuqiong
E-mail:guoyq828@163.com
摘要:
TIFY转录因子是陆地植物特有的转录因子,按照结构域特征共分为4个亚家族TIFY、PPD、JAZ、ZML,调控植物的生长发育过程。以茶树TIFY转录因子为研究对象,从茶树基因组数据库中共鉴定了22个CsTIFY转录因子家族成员,都包含了TIFY结构域。基因结构分析发现,大部分CsTIFY家族成员外显子数在5到12之间,同一亚家族所含Motif和外显子数相似。系统进化树表明,6个物种122个TIFY家族基因分为4个组,其中茶树与葡萄和杨树亲缘关系更近。启动子顺式作用元件分析发现CsTIFY家族含有大量响应SA、ABA、低温和干旱等的元件。亚细胞定位试验结果表明CsTIFY1、CsJAZ1和CsJAZ5定位于细胞核。不同茶树品种的组织表达量分析发现,大部分CsTIFY在成熟叶、根和果实中相对表达量较高。在外源ABA、SA和MeJA处理下,大部分CsJAZ上调表达,而CsZML的表达量下降;低温和干旱处理下,CsZML1表达量都极显著上升。
中图分类号:
谢思艺, 周承哲, 朱晨, 詹冬梅, 陈兰, 吴祖春, 赖钟雄, 郭玉琼. 茶树CsTIFY家族全基因组鉴定及非生物胁迫和激素处理中主要基因表达分析[J]. 园艺学报, 2022, 49(1): 100-116.
XIE Siyi, ZHOU Chengzhe, ZHU Chen, ZHAN Dongmei, CHEN Lan, WU Zuchun, LAI Zhongxiong, GUO Yuqiong. Genome-wide Identification and Expression Analysis of CsTIFY Transcription Factor Family Under Abiotic Stress and Hormone Treatments in Camellia sinensis[J]. Acta Horticulturae Sinica, 2022, 49(1): 100-116.
用途 Purpose | 基因 Gene | 上游引物序列(5′-3′) Forward primer | 下游引物序列(5′-3′) Reverse primer |
---|---|---|---|
qRT-PCR | CsTIFY1 | TGAGTAACCGATTCGCTGG | GCTTGTGGAAACCCATCTCTG |
CsTIFY2 | TCAACGAGGAAGAGGCTATGG | GGGTCCTCATCCTCAACCATT | |
CsTIFY3 | GCCTCTGTTGGTGGTTCTTC | CGGTTATTCATCTCAGTCCCT | |
CsTIFY4 | TCTCCGCCACTTCTGATCTG | GCGAATCGGTTACTCATCTCAG | |
CsPPD1 | CCTACCAGATGTGCGAGAATG | TCGCCTTCATCCAAGCTGAT | |
CsPPD2 | GCAAATTCTCCATTGCTCGC | TCCACGACGGTCTTCTCATC | |
CsJAZ1 | TCCTCCAAACCTCTTCCCAAT | CACAGAGCCAGAAGCAATCAC | |
CsJAZ2 | TCATTGATCGCCGAAGAAGC | GAATAGAGAGGCAAAGCAGG | |
CsJAZ4 | TGACCCACTTGCTGCTTCT | GAACGCCACCTTGTCCAT | |
CsJAZ5 | TGTCGAGTTCGTCTGGTTCT | CCGAATGTGCCTTTCTTCTC | |
CsJAZ6 | CGGTGGCTGTCAAAGAAGGTATT | CTCTAATCGCCATTTCAGTCCCT | |
CsJAZ7 | TCATTGATCGCCGAAGAAGC | GAATAGAGAGGCAAAGCAGGAG | |
CsJAZ8 | CGGATTACAACCACCACCAC | TCGTTAGCGGCTGTTCCTGT | |
CsZML1 | GGAGGGAGGAATCTTTGTTTGG | CTGCCAGTTTCTCCTGTTACAAC | |
CsZML2 | CTAAACGTTCCCAACCACTTCAC | GTCGTTGAGAGAGTGAGTTTCGT | |
CsZML3 | TGCAGCACATGAGAAATGG | GAACACATAAACCTGACCCTGG | |
CsZML4 | CCAGAGACGAATCACCAATCC | CTCACCACCCACAACTTCATC | |
CsZML5 | CCGATTCCACTTCAAGCAAG | TCCTCCTCCTCCGTCTTCAT | |
CsZML6 | TCGATGAAGACGGAGGAGG | GGGTTGTTATTCACAGGGAGG | |
CsZML7 | CGAATCCACAGCCACTTCAAG | TCCGCCTCATCCATAGCATC | |
CsZML8 | CTGAGACCAACCACGAAGCT | TCGAATCGGATCTGAGGGTT | |
内参基因 | β-ACTNI | GCCATCTTTGATTGGAATGG | GGTGCCACAACCTTGATCTT |
Reference gene | SAND | CCAATTGCCCCCTTAATGAC | GCAATCATTTCCTTCGTGGAG |
亚细胞定位 | CsTIFY1-1302 | ACTCTTGACCATGGTAGATCTATGATAGATAGACAGGTTGCAAATCAT | ATCTGATCCACCTTTACTAGTCTAAAAAATTGCCTTGTTCGGG |
Subcellular | |||
localization | CsJAZ1-1302 | ACTCTTGACCATGGTAGATCTATGGCATCGAGATCAGCTGTTG | ATCTGATCCACCTTTACTAGTCTAATCTTCATATCTTGATGTATTCTTCCC |
CsJAZ5-1302 | ACTCTTGACCATGGTAGATCTATGTCGAGTTCGTCTGGTTCTGC | ATCTGATCCACCTTTACTAGTCTACAAATGGTGCTCAAACTGCA |
表1 茶树CsTIFY基因家族引物序列
Table 1 CsTIFY gene family primer sequences
用途 Purpose | 基因 Gene | 上游引物序列(5′-3′) Forward primer | 下游引物序列(5′-3′) Reverse primer |
---|---|---|---|
qRT-PCR | CsTIFY1 | TGAGTAACCGATTCGCTGG | GCTTGTGGAAACCCATCTCTG |
CsTIFY2 | TCAACGAGGAAGAGGCTATGG | GGGTCCTCATCCTCAACCATT | |
CsTIFY3 | GCCTCTGTTGGTGGTTCTTC | CGGTTATTCATCTCAGTCCCT | |
CsTIFY4 | TCTCCGCCACTTCTGATCTG | GCGAATCGGTTACTCATCTCAG | |
CsPPD1 | CCTACCAGATGTGCGAGAATG | TCGCCTTCATCCAAGCTGAT | |
CsPPD2 | GCAAATTCTCCATTGCTCGC | TCCACGACGGTCTTCTCATC | |
CsJAZ1 | TCCTCCAAACCTCTTCCCAAT | CACAGAGCCAGAAGCAATCAC | |
CsJAZ2 | TCATTGATCGCCGAAGAAGC | GAATAGAGAGGCAAAGCAGG | |
CsJAZ4 | TGACCCACTTGCTGCTTCT | GAACGCCACCTTGTCCAT | |
CsJAZ5 | TGTCGAGTTCGTCTGGTTCT | CCGAATGTGCCTTTCTTCTC | |
CsJAZ6 | CGGTGGCTGTCAAAGAAGGTATT | CTCTAATCGCCATTTCAGTCCCT | |
CsJAZ7 | TCATTGATCGCCGAAGAAGC | GAATAGAGAGGCAAAGCAGGAG | |
CsJAZ8 | CGGATTACAACCACCACCAC | TCGTTAGCGGCTGTTCCTGT | |
CsZML1 | GGAGGGAGGAATCTTTGTTTGG | CTGCCAGTTTCTCCTGTTACAAC | |
CsZML2 | CTAAACGTTCCCAACCACTTCAC | GTCGTTGAGAGAGTGAGTTTCGT | |
CsZML3 | TGCAGCACATGAGAAATGG | GAACACATAAACCTGACCCTGG | |
CsZML4 | CCAGAGACGAATCACCAATCC | CTCACCACCCACAACTTCATC | |
CsZML5 | CCGATTCCACTTCAAGCAAG | TCCTCCTCCTCCGTCTTCAT | |
CsZML6 | TCGATGAAGACGGAGGAGG | GGGTTGTTATTCACAGGGAGG | |
CsZML7 | CGAATCCACAGCCACTTCAAG | TCCGCCTCATCCATAGCATC | |
CsZML8 | CTGAGACCAACCACGAAGCT | TCGAATCGGATCTGAGGGTT | |
内参基因 | β-ACTNI | GCCATCTTTGATTGGAATGG | GGTGCCACAACCTTGATCTT |
Reference gene | SAND | CCAATTGCCCCCTTAATGAC | GCAATCATTTCCTTCGTGGAG |
亚细胞定位 | CsTIFY1-1302 | ACTCTTGACCATGGTAGATCTATGATAGATAGACAGGTTGCAAATCAT | ATCTGATCCACCTTTACTAGTCTAAAAAATTGCCTTGTTCGGG |
Subcellular | |||
localization | CsJAZ1-1302 | ACTCTTGACCATGGTAGATCTATGGCATCGAGATCAGCTGTTG | ATCTGATCCACCTTTACTAGTCTAATCTTCATATCTTGATGTATTCTTCCC |
CsJAZ5-1302 | ACTCTTGACCATGGTAGATCTATGTCGAGTTCGTCTGGTTCTGC | ATCTGATCCACCTTTACTAGTCTACAAATGGTGCTCAAACTGCA |
基因名称 Gene name | ID | 氨基酸 Amino acid | 分子量/kD Molecular weight | 等电点 Isoelectric point | 内含子 Intron | 外显子 Exons | 不稳定指数 Instability index | 脂肪族指数 Aliphatic index | 平均疏水性 GRAVY | 亚细胞定位 Subcellular localization |
---|---|---|---|---|---|---|---|---|---|---|
CsTIFY1 | TEA007552.1 | 250 | 26.77771 | 9.90 | 1 | 2 | 52.44 | 55.04 | -0.694 | 细胞核 |
Cell nucleus | ||||||||||
CsTTFY2 | TEA012041.1 | 934 | 103.1778 | 5.57 | 11 | 12 | 49.30 | 84.03 | -0.140 | 叶绿体 |
Chloroplast | ||||||||||
CsTIFY3 | TEA032457.1 | 352 | 38.90351 | 10.25 | 4 | 5 | 53.41 | 72.81 | -0.461 | 液泡 Vacuole |
CsTIFY4 | TEA009327.1 | 393 | 41.65471 | 9.40 | 5 | 6 | 46.36 | 65.29 | -0.437 | 细胞核 |
Cell nucleus | ||||||||||
CsPPD1 | TEA005947.1 | 1 820 | 204.7527 | 6.14 | 16 | 17 | 41.51 | 92.61 | -0.120 | 胞间连丝 |
Plasmodesma | ||||||||||
CsPPD2 | TEA011549.1 | 1 114 | 125.3216 | 8.23 | 12 | 13 | 50.60 | 88.93 | -0.299 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ1 | TEA027049.1 | 212 | 23.87791 | 9.15 | 5 | 6 | 55.42 | 67.08 | -0.632 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ2 | TEA030826.1 | 148 | 16.39844 | 7.84 | 0 | 1 | 48.12 | 58.65 | -0.564 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ3 | TEA001501.1 | 261 | 29.26615 | 9.17 | 4 | 5 | 57.65 | 70.69 | -0.666 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ4 | TEA001681.1 | 410 | 42.84663 | 9.34 | 7 | 8 | 35.36 | 66.44 | -0.310 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ5 | TEA030190.1 | 294 | 31.53145 | 9.46 | 4 | 5 | 55.85 | 70.37 | -0.395 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ6 | TEA001821.1 | 436 | 46.20707 | 9.58 | 7 | 8 | 48.77 | 71.40 | -0.343 | 细胞质 Cytoplasm |
CsJAZ7 | TEA032228.1 | 220 | 24.45353 | 9.18 | 5 | 6 | 55.48 | 67.36 | -0.557 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ8 | TEA004474.1 | 136 | 15.60498 | 9.93 | 2 | 3 | 84.54 | 75.29 | -0.721 | 细胞核 |
Cell nucleus | ||||||||||
CsZML1 | TEA033832.1 | 385 | 42.00050 | 4.88 | 10 | 11 | 42.88 | 70.08 | -0.625 | 细胞核 |
Cell nucleus | ||||||||||
CsZML2 | TEA033836.1 | 271 | 30.51872 | 4.57 | 4 | 5 | 46.48 | 78.04 | -0.601 | 细胞核 |
Cell nucleus | ||||||||||
CsZML3 | TEA013465.1 | 303 | 33.49678 | 6.19 | 6 | 7 | 31.87 | 60.79 | -0.916 | 细胞核 |
Cell nucleus | ||||||||||
CsZML4 | TEA014550.1 | 234 | 25.20185 | 5.00 | 3 | 4 | 50.61 | 71.24 | -0.550 | 细胞核 |
Cell nucleus | ||||||||||
CsZML5 | TEA014541.1 | 352 | 38.21245 | 4.85 | 10 | 11 | 50.11 | 72.30 | -0.574 | 细胞核 |
Cell nucleus | ||||||||||
CsZML6 | TEA011504.1 | 352 | 38.21245 | 4.85 | 10 | 11 | 50.11 | 72.30 | -0.574 | 细胞核 |
Cell nucleus | ||||||||||
CsZML7 | TEA002032.1 | 383 | 41.64484 | 4.76 | 10 | 11 | 54.89 | 63.92 | -0.795 | 细胞核 |
Cell nucleus | ||||||||||
CsZML8 | TEA001414.1 | 461 | 49.38947 | 5.75 | 11 | 12 | 41.39 | 73.19 | -0.375 | 细胞核 |
Cell nucleus |
表2 茶树CsTIFY家族蛋白的理化特性
Table 2 Physicochemical properties of CsTIFY family proteins of tea tree
基因名称 Gene name | ID | 氨基酸 Amino acid | 分子量/kD Molecular weight | 等电点 Isoelectric point | 内含子 Intron | 外显子 Exons | 不稳定指数 Instability index | 脂肪族指数 Aliphatic index | 平均疏水性 GRAVY | 亚细胞定位 Subcellular localization |
---|---|---|---|---|---|---|---|---|---|---|
CsTIFY1 | TEA007552.1 | 250 | 26.77771 | 9.90 | 1 | 2 | 52.44 | 55.04 | -0.694 | 细胞核 |
Cell nucleus | ||||||||||
CsTTFY2 | TEA012041.1 | 934 | 103.1778 | 5.57 | 11 | 12 | 49.30 | 84.03 | -0.140 | 叶绿体 |
Chloroplast | ||||||||||
CsTIFY3 | TEA032457.1 | 352 | 38.90351 | 10.25 | 4 | 5 | 53.41 | 72.81 | -0.461 | 液泡 Vacuole |
CsTIFY4 | TEA009327.1 | 393 | 41.65471 | 9.40 | 5 | 6 | 46.36 | 65.29 | -0.437 | 细胞核 |
Cell nucleus | ||||||||||
CsPPD1 | TEA005947.1 | 1 820 | 204.7527 | 6.14 | 16 | 17 | 41.51 | 92.61 | -0.120 | 胞间连丝 |
Plasmodesma | ||||||||||
CsPPD2 | TEA011549.1 | 1 114 | 125.3216 | 8.23 | 12 | 13 | 50.60 | 88.93 | -0.299 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ1 | TEA027049.1 | 212 | 23.87791 | 9.15 | 5 | 6 | 55.42 | 67.08 | -0.632 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ2 | TEA030826.1 | 148 | 16.39844 | 7.84 | 0 | 1 | 48.12 | 58.65 | -0.564 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ3 | TEA001501.1 | 261 | 29.26615 | 9.17 | 4 | 5 | 57.65 | 70.69 | -0.666 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ4 | TEA001681.1 | 410 | 42.84663 | 9.34 | 7 | 8 | 35.36 | 66.44 | -0.310 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ5 | TEA030190.1 | 294 | 31.53145 | 9.46 | 4 | 5 | 55.85 | 70.37 | -0.395 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ6 | TEA001821.1 | 436 | 46.20707 | 9.58 | 7 | 8 | 48.77 | 71.40 | -0.343 | 细胞质 Cytoplasm |
CsJAZ7 | TEA032228.1 | 220 | 24.45353 | 9.18 | 5 | 6 | 55.48 | 67.36 | -0.557 | 细胞核 |
Cell nucleus | ||||||||||
CsJAZ8 | TEA004474.1 | 136 | 15.60498 | 9.93 | 2 | 3 | 84.54 | 75.29 | -0.721 | 细胞核 |
Cell nucleus | ||||||||||
CsZML1 | TEA033832.1 | 385 | 42.00050 | 4.88 | 10 | 11 | 42.88 | 70.08 | -0.625 | 细胞核 |
Cell nucleus | ||||||||||
CsZML2 | TEA033836.1 | 271 | 30.51872 | 4.57 | 4 | 5 | 46.48 | 78.04 | -0.601 | 细胞核 |
Cell nucleus | ||||||||||
CsZML3 | TEA013465.1 | 303 | 33.49678 | 6.19 | 6 | 7 | 31.87 | 60.79 | -0.916 | 细胞核 |
Cell nucleus | ||||||||||
CsZML4 | TEA014550.1 | 234 | 25.20185 | 5.00 | 3 | 4 | 50.61 | 71.24 | -0.550 | 细胞核 |
Cell nucleus | ||||||||||
CsZML5 | TEA014541.1 | 352 | 38.21245 | 4.85 | 10 | 11 | 50.11 | 72.30 | -0.574 | 细胞核 |
Cell nucleus | ||||||||||
CsZML6 | TEA011504.1 | 352 | 38.21245 | 4.85 | 10 | 11 | 50.11 | 72.30 | -0.574 | 细胞核 |
Cell nucleus | ||||||||||
CsZML7 | TEA002032.1 | 383 | 41.64484 | 4.76 | 10 | 11 | 54.89 | 63.92 | -0.795 | 细胞核 |
Cell nucleus | ||||||||||
CsZML8 | TEA001414.1 | 461 | 49.38947 | 5.75 | 11 | 12 | 41.39 | 73.19 | -0.375 | 细胞核 |
Cell nucleus |
图1 茶树(Cs)、葡萄(Vv)、拟南芥(At)、杨树(Pt)、水稻(Os)和二穗短柄草(Bd)的TIFY蛋白进化树
Fig. 1 TIFY protein phylogenetic tree of tea plant(Cs),grape(Vv),Arabidopsis(At),poplar(Pt), rice(Os)and Brachypodium distachyon(Bd)
图5 外源SA、ABA、MeJA处理下部分茶树CsTIFY基因的表达 *表示同一时间点差异显著(α = 0.05),**为极显著(α = 0.01);不同小写字母表示不同时间点差异显著(P < 0.05)。下同。
Fig. 5 Expression of CsTIFY genes in tea plants under exogenous SA,ABA,MeJA stresses * indicates significant differences at the same time point(α = 0.05),** indicates extremely significant(α = 0.01);Different lowercase letters indicate significant differences at different points in time(P < 0.05). The same below.
图6 干旱和4 ℃低温处理下部分茶树CsTIFY的表达
Fig. 6 CsTIFY gene expression of tea under abiotic stress drought and 4 ℃ treatments CsTIFY gene expression of tea under abiotic stress drought and 4 ℃ treatments
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