植物学报 ›› 2016, Vol. 51 ›› Issue (2): 202-209.DOI: 10.11983/CBB15088 cstr: 32102.14.CBB15088
李冬梅1, 王路雅1, 张澜玥2, 帖子阳2, 毛惠平1,*(
)
收稿日期:2015-05-18
接受日期:2015-09-04
出版日期:2016-03-01
发布日期:2016-03-31
通讯作者:
E-mail: 基金资助:
Dongmei Li1, Luya Wang1, Lanyue Zhang2, Ziyang Tie2, Huiping Mao1,*(
)
Received:2015-05-18
Accepted:2015-09-04
Online:2016-03-01
Published:2016-03-31
Contact:
E-mail: 摘要: AtPROPEP是拟南芥(Arabidopsis thaliana)具有7个成员的基因家族, 编码内源短肽激素。AtPROPEP基因家族编码的蛋白质C端23个氨基酸短肽能够被2个同源激酶受体AtPEPR1和AtPEPR2识别并结合, 引起下游反应。然而, 对于该家族成员AtPROPEP2,3−6的表达对茉莉酸(JA)和水杨酸(SA)的响应以及在根生长中的作用并不清楚。GUS染色和定量RT-PCR分析结果表明, AtPROPEP2-6的表达对于JA和SA的响应不同, 暗示着它们可能通过不同的方式参与植物的先天免疫反应。AtPROPEP3和AtPROPEP4过表达植株的表型分析表明, AtPROPEP3和AtPROPEP4促进拟南芥根的生长。
李冬梅, 王路雅, 张澜玥, 帖子阳, 毛惠平. 拟南芥短肽激素PROPEP基因家族在根生长中的作用机理. 植物学报, 2016, 51(2): 202-209.
Dongmei Li, Luya Wang, Lanyue Zhang, Ziyang Tie, Huiping Mao. Mechanism of Arabidopsis Short Peptide Hormones PROPEP Gene Family in the Root Growth. Chinese Bulletin of Botany, 2016, 51(2): 202-209.
| Primer name | Sequences (5'-3') | Function |
|---|---|---|
| AtPROPEP1_PF-HindIII AtPROPEP1_PR-BamHI AtPROPEP2_PF-HindIII AtPROPEP2_PR-BamHI AtPROPEP3_PF-BamHI AtPROPEP3_PR-NotI AtPROPEP4_PF-HindIII AtPROPEP4_PR-EcoRI AtPROPEP5_PF-HindIII AtPROPEP5_PR-BamHI AtPROPEP6_PF-XhoI AtPROPEP6_PR-EcoRI | CCCAAGCTTGTAAATTATAGTGAAAGGTACGG GCGGATCCTGAGATCTGATAAGACAGAGG CCCAAGCTTCGCATTCGCTTTTTTCTTTTTG GCGGATCCTGAAATCCAATAGTTTGTGAG GCGGATCCTATTTTAACAGTCAACAGCTATTTGG TTGCGGCCGCCGTTGACTTCTTAATCTTTTTTTG CCCAAGCTTAATAAGGATGAATAAAAAGTTTGGG CCGGAATTCGTTTTTCTTCAATTCTGCTTCGTG CCCAAGCTTTACTTAATTTCTTGTGAGAAACTTG GCGGATCCCTTCGCTATCTTCTAAGTTCCTC CCCTCGAGTGATATCTAAGTCCAACTTGGTG CCGGAATTCGTTTTTTGTTTTCTTTCTCTTCTT | AtPROPEP1 promoter clone AtPROPEP2 promoter clone AtPROPEP3 promoter clone AtPROPEP4 promoter clone AtPROPEP5 promoter clone AtPROPEP6 promoter clone |
表1 AtPROPEPs启动子克隆引物序列
Table 1 Sequences of AtPROPEPs promoter clone primers
| Primer name | Sequences (5'-3') | Function |
|---|---|---|
| AtPROPEP1_PF-HindIII AtPROPEP1_PR-BamHI AtPROPEP2_PF-HindIII AtPROPEP2_PR-BamHI AtPROPEP3_PF-BamHI AtPROPEP3_PR-NotI AtPROPEP4_PF-HindIII AtPROPEP4_PR-EcoRI AtPROPEP5_PF-HindIII AtPROPEP5_PR-BamHI AtPROPEP6_PF-XhoI AtPROPEP6_PR-EcoRI | CCCAAGCTTGTAAATTATAGTGAAAGGTACGG GCGGATCCTGAGATCTGATAAGACAGAGG CCCAAGCTTCGCATTCGCTTTTTTCTTTTTG GCGGATCCTGAAATCCAATAGTTTGTGAG GCGGATCCTATTTTAACAGTCAACAGCTATTTGG TTGCGGCCGCCGTTGACTTCTTAATCTTTTTTTG CCCAAGCTTAATAAGGATGAATAAAAAGTTTGGG CCGGAATTCGTTTTTCTTCAATTCTGCTTCGTG CCCAAGCTTTACTTAATTTCTTGTGAGAAACTTG GCGGATCCCTTCGCTATCTTCTAAGTTCCTC CCCTCGAGTGATATCTAAGTCCAACTTGGTG CCGGAATTCGTTTTTTGTTTTCTTTCTCTTCTT | AtPROPEP1 promoter clone AtPROPEP2 promoter clone AtPROPEP3 promoter clone AtPROPEP4 promoter clone AtPROPEP5 promoter clone AtPROPEP6 promoter clone |
| Primer name | Sequences (5'-3') | Function |
|---|---|---|
| AtPROPEP3_CDSF-SalI AtPROPEP3_CDSR-BamHI AtPROPEP4_CDSF-SalI AtPROPEP4_CDSR-EcoRI | GCGTCGACATGGAGAATCTCAGAAATGG CGGGATCCCTAATTGTGTTTGCCTCCTT GCGTCGACATGGAGAGAGGAGTTTCTTA CGGAATTCCTAAAACGGCTTCTTGTTGG | AtPROPEP3 coding sequence clone AtPROPEP4 coding sequence clone |
表2 AtPROPEP3和AtPROPEP4引物序列
Table 2 Sequences of AtPROPEP3 and AtPROPEP4 clone primers
| Primer name | Sequences (5'-3') | Function |
|---|---|---|
| AtPROPEP3_CDSF-SalI AtPROPEP3_CDSR-BamHI AtPROPEP4_CDSF-SalI AtPROPEP4_CDSR-EcoRI | GCGTCGACATGGAGAATCTCAGAAATGG CGGGATCCCTAATTGTGTTTGCCTCCTT GCGTCGACATGGAGAGAGGAGTTTCTTA CGGAATTCCTAAAACGGCTTCTTGTTGG | AtPROPEP3 coding sequence clone AtPROPEP4 coding sequence clone |
| Primer name | Sequences (5'-3') |
|---|---|
| AtPROPEP2F AtPROPEP2R AtPROPEP3F AtPROPEP3R AtPROPEP4F AtPROPEP4R AtPROPEP5F AtPROPEP5R AtPROPEP6F AtPROPEP6R ACTINF ACTINR | CTCGACCAAGCTCTCATAGCTG CACAACGACATCATCGTCTTTC TCTTCTTCTTGCGATCTTTCGTCAT CTGAACTTGGCGTAGGCTTAGTC CTCAAGCTTCTCGGTTTGCGATC ACTTTCTCTCGACTTCTTTAGTAC GAGATTGTTGCAAGCTCATGCCTC AGTTGAAGTTTCGATAGATGAAGGT TGAAGTGTCTTGGTCTTGAGTC TGGTCTCCTTCTTAACACTGCTG ACGGTAACATTGTGCTCAGTGGTG CTTGGAGATCCACATCTGCTGGA |
表3 实时荧光定量PCR引物
Table 3 Primers used for quantitative RT-PCR
| Primer name | Sequences (5'-3') |
|---|---|
| AtPROPEP2F AtPROPEP2R AtPROPEP3F AtPROPEP3R AtPROPEP4F AtPROPEP4R AtPROPEP5F AtPROPEP5R AtPROPEP6F AtPROPEP6R ACTINF ACTINR | CTCGACCAAGCTCTCATAGCTG CACAACGACATCATCGTCTTTC TCTTCTTCTTGCGATCTTTCGTCAT CTGAACTTGGCGTAGGCTTAGTC CTCAAGCTTCTCGGTTTGCGATC ACTTTCTCTCGACTTCTTTAGTAC GAGATTGTTGCAAGCTCATGCCTC AGTTGAAGTTTCGATAGATGAAGGT TGAAGTGTCTTGGTCTTGAGTC TGGTCTCCTTCTTAACACTGCTG ACGGTAACATTGTGCTCAGTGGTG CTTGGAGATCCACATCTGCTGGA |
图2 AtPROPEP2,3−6启动子对茉莉酸(JA)和水杨酸(SA)的响应 (A) AtPROPEP2,3−6::GUS转基因植株在250 µmol∙L−1 JA和100 µmol∙L−1 SA处理下的GUS染色; (B) 定量RT-PCR检测AtPROPEP2,3−6在JA和SA处理下的表达水平
Figure 2 Response of AtPROPEP2,3−6 promoter to jasmonic acid (JA) and salicylic acid (SA) (A) GUS staining of transgenic seedlings harboring AtPROPEP2,3−6::GUS construct under 250 µmol∙L−1 JA and 100 µmol∙L−1 SA; (B) Quantitative RT-PCR analysis of AtPROPEP2,3−6 expression level under JA and SA treatments
图3 定量RT-PCR检测拟南芥过表达植株 (A) 35S::AtPROPEP3; (B) 35S::AtPROPEP4
Figure 3 Quantitative RT-PCR analysis of Arabidopsis over- expressing lines (A) 35S::AtPROPEP3; (B) 35S::AtPROPEP4
图4 拟南芥过量表达AtPROPEP3和AtPROPEP4植株根长增加 (A) 野生型与过量表达突变体在CK培养基上的表型; (B) 野生型与过量表达突变体在CK培养基上的根长分析(**P<0.01; * P<0.05)
Figure 4 Arabidopsis plants overexpressing AtPROPEP3 and AtPROPEP4 exhibit longer root than that of wild type (A) Seedlings of Col-0 and transgenic seedlings grown on CK medium; (B) Measurement of root growth under CK medium (**P<0.01; * P<0.05)
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