中国农业科技导报 ›› 2023, Vol. 25 ›› Issue (6): 11-21.DOI: 10.13304/j.nykjdb.2021.0924
收稿日期:
2021-11-01
接受日期:
2022-01-18
出版日期:
2023-06-01
发布日期:
2023-07-28
通讯作者:
阎媛媛
作者简介:
陈丽婷 E-mail:clt1158433820@163.com;
基金资助:
Received:
2021-11-01
Accepted:
2022-01-18
Online:
2023-06-01
Published:
2023-07-28
Contact:
Yuanyuan YAN
摘要:
开花是植物产生有性生殖器官的重要阶段,开花整合子整合来自环境和植物体内的信号,进而激活花芽分化,使植物在适宜的时间产生足够多的后代,保障生殖繁衍。棉花中开花整合子基因(FT、SOC1和LFY)已被克隆,但它们在棉花中的进化关系尚不清楚。参考陆地棉基因组,明确了棉属开花整合子同源基因数目,FT和LFY在棉属中进化具有保守性,而SOC1基因序列具有多样性。不同于其他物种,棉花开花整合子基因均在顶端分生组织和叶片中表达量较高,且随着棉苗的生长发育其表达量逐步升高。克隆了GhLFY-2、GhFT-2和GhSOC1-5基因,发现其促进开花的功能是保守的,但开花时间调控机制存在物种特异性。陆地棉中既存在保守的GhFT-2/GhSOC1-5/GhLFY-2线性调控路径,又进化出GhFT-1和GhSOC1-1直接调控GhAP1的路径,表明棉花开花时间调控机制的多样性可能与其对环境的适应性有关。
中图分类号:
陈丽婷, 阎媛媛. 陆地棉开花整合子调控机制解析[J]. 中国农业科技导报, 2023, 25(6): 11-21.
Liting CHEN, Yuanyuan YAN. Investigation of Regulatory Mechanism of Floral Integrators in Upland Cotton[J]. Journal of Agricultural Science and Technology, 2023, 25(6): 11-21.
引物名称 Primer name | 引物序列 Primer sequence(5’-3’) | 用途 Purpose |
---|---|---|
GhLFY-2-F(PstI) | TT | 基因克隆 Gene cloning |
GhLFY-2-R(SmaI) | TT | |
GhSOC1-4-F(EcoRI) | CC | |
GhSOC1.4-R(BamHI) | TT | |
GhFT-2-F(EcoRI) | TT | |
GhFT-2-R(XmaI) | TT | |
GhLFY-2-RT-F | AATTCGAGGGTGGTGATGAT | qRT-PCR |
GhLFY-2-RT-R | CTCCGTTACGATGAAAGGGT | |
GhSOC1-5-RT-F | GGCGCATAGAGAACGATACA | |
GhSOC1-5-RT-R | TTTGTATGCCGCCTATAACG | |
GhFT-2-RT-F | GGGATGATCTGAGGACCTTCT | |
GhFT-2-RT-R | CACCAGTTGTGGCTGGAATA | |
AtTUB2-F | ATCCGTGAAGAGTACCCAGAT | |
AtTUB2-R | AAGAACCATGCACTCATCAGC |
表1 本研究中所用引物序列
Table 1 Primer sequences in the study
引物名称 Primer name | 引物序列 Primer sequence(5’-3’) | 用途 Purpose |
---|---|---|
GhLFY-2-F(PstI) | TT | 基因克隆 Gene cloning |
GhLFY-2-R(SmaI) | TT | |
GhSOC1-4-F(EcoRI) | CC | |
GhSOC1.4-R(BamHI) | TT | |
GhFT-2-F(EcoRI) | TT | |
GhFT-2-R(XmaI) | TT | |
GhLFY-2-RT-F | AATTCGAGGGTGGTGATGAT | qRT-PCR |
GhLFY-2-RT-R | CTCCGTTACGATGAAAGGGT | |
GhSOC1-5-RT-F | GGCGCATAGAGAACGATACA | |
GhSOC1-5-RT-R | TTTGTATGCCGCCTATAACG | |
GhFT-2-RT-F | GGGATGATCTGAGGACCTTCT | |
GhFT-2-RT-R | CACCAGTTGTGGCTGGAATA | |
AtTUB2-F | ATCCGTGAAGAGTACCCAGAT | |
AtTUB2-R | AAGAACCATGCACTCATCAGC |
图2 植物开花整合子基因的进化树注:Gh—陆地棉;Gb—海岛棉;Ga—亚洲棉;Gr—雷蒙德氏棉;At—拟南芥;Pt—杨树;Nt—烟草;Vv—葡萄。
Fig. 2 Phylogenetic analysis of plant floral integrator genesNote:Gh—Gossypium hirsutum; Gb—Gossypium barbadense; Ga—Gossypium arboreum; Gr—Gossypium raimondii; At—Arabidopsis thaliana; Pt—Populus tomentosa; Nt—Nicotiana tabacum; Vv—Vitis vinifera.
图3 开花整合子基因在陆地棉中表达A:棉花开花整合子基因组织特异性表达;B:棉花开花整合子基因随棉苗生长在叶片中表达变化;C:棉花开花整合子基因随棉苗生长在SAM中表达变化。R—根;S—茎;L—叶;TLS—真叶期
Fig. 3 Expression of floral integrators genes in Gossypium hirsutumA: Tissue-specific expression of flowering integrator of Gossypium hirsutum; B: Expression trends of flowering integrators in cotton leaves during seedling development; C: Expression trends of flowering integrators in cotton SAM during seedling development. R—Root; S—Stem; L—Leaf; TLS—True leaf stage
图4 棉花开花整合子促进开花A:转基因拟南芥早花表型;B:转基因拟南芥基因的表达量及莲座叶数目统计;C:野生型和转基因拟南芥的开花时间,**表示与野生型相比差异显著(P<0.01);D:GhLFY-2转基因植株莲座叶叶腋里伸出单生花
Fig. 4 Cotton flowering integrators promotes floweringA: Early flowering phenotype of transgenic plants; B: Comparison of gene expression and flowering time between WT and transgenic plants; C: Flowering time of WT and transgenic lines, ** indicates significant differences (P<0.01); D: Solitary flowers produced in GhLFY-2 transgenic plants
图5 拟南芥开花整合子基因转录水平注:A、B和C分别为开花整合子在GhFT-2、GhSOC1-5和GhLFY-2转基因株系中的表达量。
Fig. 5 Expression of floral integrator genes in transgenic Arabidopsis plantsNote:A, B and C shows relative expression of flowering integrators in GhFT-2, GhSOC1-5 and GhLFY-2 transgenic plants, respectively.
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