草业学报 ›› 2022, Vol. 31 ›› Issue (3): 47-59.DOI: 10.11686/cyxb2020593
高莉娟1(), 张正社1, 文裕2, 宗西方1, 闫启1, 卢丽燕1, 易显凤3, 张吉宇1()
收稿日期:
2020-12-29
修回日期:
2021-02-09
出版日期:
2022-03-20
发布日期:
2022-01-15
通讯作者:
张吉宇
作者简介:
Corresponding author. E-mail: zhangjy@lzu.edu.cn基金资助:
Li-juan GAO1(), Zheng-she ZHANG1, Yu WEN2, Xi-fang ZONG1, Qi YAN1, Li-yan LU1, Xian-feng YI3, Ji-yu ZHANG1()
Received:
2020-12-29
Revised:
2021-02-09
Online:
2022-03-20
Published:
2022-01-15
Contact:
Ji-yu ZHANG
摘要:
bHLH转录因子家族不仅参与了植物的生长发育,而且在植物响应逆境胁迫和次生代谢方面发挥着关键作用。在全基因组水平对重要饲草及能源植物象草的bHLH转录因子家族进行了鉴定及分析,并利用转录组数据及定量RT-PCR分析了象草bHLH转录因子对赤霉素(GA3)和多效唑(PAC)的响应。结果显示:在象草中共鉴定出229个具有完整保守结构域的bHLH基因家族成员(CpbHLH001~CpbHLH229),不均匀地分布于14条染色体上;系统进化分析结果表明,229个CpbHLHs可被分为18个亚类,其中C亚类的成员数量最多,为41个;此外,相同亚家族中的大多数基因具有相似的基因结构和保守基序;基于转录组数据的表达谱分析结果发现,多数bHLH基因在象草茎尖组织中均对GA3和PAC有响应。随机挑选9个表达量较高的基因进一步通过qPCR进行验证。结果显示,经外源GA3和PAC处理之后,这9个基因因不同处理而差异表达,表明这9个基因可能与GA3和PAC介导的信号通路有关。综上所述,本研究为象草bHLH转录因子家族的生物学功能奠定了基础。
高莉娟, 张正社, 文裕, 宗西方, 闫启, 卢丽燕, 易显凤, 张吉宇. 象草全基因组bHLH转录因子家族鉴定及表达分析[J]. 草业学报, 2022, 31(3): 47-59.
Li-juan GAO, Zheng-she ZHANG, Yu WEN, Xi-fang ZONG, Qi YAN, Li-yan LU, Xian-feng YI, Ji-yu ZHANG. Genome-wide identification and expression analysis of the bHLH transcription factor family in Cenchrus purpureus[J]. Acta Prataculturae Sinica, 2022, 31(3): 47-59.
基因名称Gene name | 上游引物Forward primer (5'-3') | 下游引物Reverse primer (5'-3') |
---|---|---|
CpbHLH002 | CAAGATCAGCAAGATGGACAGG | GGGTGACTCAAGCTCATTCTG |
CpbHLH013 | ACAAGCAAACCAACACTTCTG | GCTCTCCTTTAACACCTTAACCT |
CpbHLH017 | AGCAAGTTGAGTTCTTGTCC | TCTTTGTGCAGGAGTGTAGAC |
CpbHLH019 | TGCGGAAAGGTTGAGAAGG | TGATCTCGTCAAGCATCACC |
CpbHLH025 | ACAAACAAGACAGATAGGGCAG | GAGAGTGGAATATCAGCAACCAG |
CpbHLH030 | ACAAGCAAACCAACACTTCAG | CTCGTTCAGCACCTTGACC |
CpbHLH055 | GTCAAGGTACTAAGCATGAGCA | TCTTCTTCCATCAGCTTTGTG |
CpbHLH156 | AGCAACAGCGAGAATGGC | CAGCAGCTTCATCCTCTGG |
CpbHLH158 | AGCTGGTTGCTGATATTCCAC | TCTTCCATCAGCTTTGCAACC |
CpEE1a | ACTACGAGCAAGAGTTGGAAAC | TGGATGGCTGGAAGAGGAC |
表1 实时定量PCR引物序列
Table 1 Primer sequences used in real-time quantitative PCR
基因名称Gene name | 上游引物Forward primer (5'-3') | 下游引物Reverse primer (5'-3') |
---|---|---|
CpbHLH002 | CAAGATCAGCAAGATGGACAGG | GGGTGACTCAAGCTCATTCTG |
CpbHLH013 | ACAAGCAAACCAACACTTCTG | GCTCTCCTTTAACACCTTAACCT |
CpbHLH017 | AGCAAGTTGAGTTCTTGTCC | TCTTTGTGCAGGAGTGTAGAC |
CpbHLH019 | TGCGGAAAGGTTGAGAAGG | TGATCTCGTCAAGCATCACC |
CpbHLH025 | ACAAACAAGACAGATAGGGCAG | GAGAGTGGAATATCAGCAACCAG |
CpbHLH030 | ACAAGCAAACCAACACTTCAG | CTCGTTCAGCACCTTGACC |
CpbHLH055 | GTCAAGGTACTAAGCATGAGCA | TCTTCTTCCATCAGCTTTGTG |
CpbHLH156 | AGCAACAGCGAGAATGGC | CAGCAGCTTCATCCTCTGG |
CpbHLH158 | AGCTGGTTGCTGATATTCCAC | TCTTCCATCAGCTTTGCAACC |
CpEE1a | ACTACGAGCAAGAGTTGGAAAC | TGGATGGCTGGAAGAGGAC |
图2 象草CpbHLH转录因子的保守基序分析a: CpbHLH基因家族的系统进化树Phylogenetic tree of the CpbHLH family; b: CpbHLH基因家族的基因结构Gene structure of the CpbHLH family; 绿色框、黑色线条分别代表外显子和内含子The green boxes, blacklines represent the exon and intron, respectively; c: CpbHLH基因家族的保守基序Conserved motifs of the CpbHLH family.
Fig.2 The conserved motifs analysis of C. purpureus bHLH transcription factors
图3 象草与水稻bHLH转录因子的系统进化分析分枝上的数字表示自展值Numbers on major branches indicate bootstrap estimates for 1000 replicate analyses. Cp: 象草C. purpureus; Os: 水稻O. sativa; A~U: 不同亚类Different subgroups.
Fig.3 Phylogenetic analysis of bHLH transcription factors in C. purpureus and O. sativa
图4 象草CpbHLH基因在GA3和PAC处理1和48 h之后的表达分析红-白-蓝3色代表基因表达的强度,红色越亮代表表达量越高,蓝色越亮代表表达量越低,白色代表表达量为0 The three colors of red-white-blue represent the intensity of gene expression, the brighter the red, the higher the expression, the brighter the blue, the lower the expression, and the white represents the expression is 0. CK: 对照Control; GA3-1: GA3处理1 h GA3 treatment for 1 hour; GA3-48: GA3处理48 h GA3 treatment for 48 hour; PAC-1: PAC处理1 h PAC treatment for 1 hour; PAC-48: PAC处理48 h PAC treatment for 48 hour; 下同The same below.
Fig.4 Expression analysis of bHLH gene in C. purpureus after GA3 and PAC treatments for 1 and 48 hour
图6 象草bHLH基因的GO分类ML: 代谢过程Metabolic process; CP: 细胞过程Cellular process; SP: 单一有机体过程Single-organism process; BR: 生物调节Biological regulation; RS: 对刺激的响应Response to stimulus; LO: 定位Localization; CB: 细胞成分组织或生物组分Cellular component organization or biogenesis; DP: 发育过程 Developmental process; MP: 多细胞有机体过程Multicellular organismal process; SI: 信号Signaling; RP: 生殖过程Reproductive process; MO: 多有机体过程Multi-organism process; RD: 繁殖Reproduction; GR: 生长Growth; IP: 免疫系统过程Immune system process; RY: 节律过程Rhythmic process; CK: 细胞凋零Cell killing; LM: 运动Locomotion; BA: 生物附着Biological adhesion; BP: 生物相 Biological phase; CE: 细胞Cell; CP: 细胞组分Cell part; OG: 细胞器Organelle; ME: 细胞膜Membrane; MC: 膜部分Membrane part; OP: 器官部分Organelle part; MP: 大分子复合物Macromolecular complex; ML: 膜封闭腔Membrane-enclosed lumen; ER: 细胞外区域Extracellular region; CJ: 细胞结Cell junction; SY: 共质体Symplast; EP: 细胞外区域部分Extracellular region part; VN: 病毒Virion; VP: 病毒部分 Virion part; ND: 核Nucleoid; BD: 结合Binding; CA: 催化活性Catalytic activity; TA: 运输器活性Transporter activity; NA: 核酸结合转录因子活性Nucleic acid binding transcription factor activity; MR: 结构分子活性Structural molecule activity; AA: 抗氧化活性Antioxidant activity; EA: 电子载流子活性Electron carrier activity; MT: 分子传感器活性Molecular transducer activity; SA: 信号转导活性Signal transducer activity; TP: 转录因子活性,蛋白结合Transcription factor activity, protein binding; NA: 营养积累活性Nutrient reservoir activity; PT: 蛋白质标签Protein tag.
Fig.6 GO classification of CpbHLH genes
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