园艺学报 ›› 2023, Vol. 50 ›› Issue (5): 1025-1036.doi: 10.16420/j.issn.0513-353x.2022-0177
刘嘉琦1, 公菲菲1, 张颢2, 景维坤1,2, 瞿素萍2, 马男1, 高俊平1, 孙小明1,*()
收稿日期:
2023-02-17
修回日期:
2023-04-27
出版日期:
2023-05-25
发布日期:
2023-05-31
通讯作者:
*(E-mail:xmsun@cau.edu.cn)基金资助:
LIU Jiaqi1, GONG Feifei1, ZHANG Hao2, JING Weikun1,2, QU Suping2, MA Nan1, GAO Junping1, SUN Xiaoming1,*()
Received:
2023-02-17
Revised:
2023-04-27
Published:
2023-05-25
Online:
2023-05-31
摘要:
为了解茉莉酸甲酯(methyl jasmonate,MeJA)在花瓣衰老中的作用机制,以月季‘Samantha’为材料,在花瓣中克隆了MeJA生物合成关键酶茉莉酸羧基甲基转移酶(jasmonic acid carboxyl methyltrans-ferase,JMT)基因RhJMT。同源蛋白序列比对和系统进化树分析表明,RhJMT具有保守的Methylyransf_7超家族结构域,属于SABATH蛋白家族。实时荧光定量PCR结果表明,RhJMT的表达量在花瓣进入衰老期后显著升高。通过病毒诱导的基因沉默(virus induced gene silencing,VIGS)技术对RhJMT进行功能验证,结果表明沉默RhJMT显著延缓花瓣衰老,外源施加MeJA能够抵消沉默RhJMT对花瓣衰老的延缓作用。以上结果说明RhJMT能够促进花瓣衰老,可能是通过控制MeJA的含量来发挥作用。
中图分类号:
刘嘉琦, 公菲菲, 张颢, 景维坤, 瞿素萍, 马男, 高俊平, 孙小明. 月季茉莉酸羧基甲基转移酶基因RhJMT对花瓣衰老的调控[J]. 园艺学报, 2023, 50(5): 1025-1036.
LIU Jiaqi, GONG Feifei, ZHANG Hao, JING Weikun, QU Suping, MA Nan, GAO Junping, SUN Xiaoming. Jasmonic Acid Carboxy Methyltransferase Gene RhJMT Regulates Petal Senescence in Rosa hybrida[J]. Acta Horticulturae Sinica, 2023, 50(5): 1025-1036.
用途 Function | 引物名称 Primer name | 引物序列(5′-3′) Sequence |
---|---|---|
克隆基因 Gene clone | RhJMT-F | GAGATAATGGAGGTAGAGCAAGTTCTTC |
RhJMT-R | GCTTCATATTGTATACTTGCACG | |
病毒诱导的基因沉默 Virus induced gene silencing | TRV1-F | TTACAGGTTATTTGGGCTAG |
TRV1-R | CCGGGTTCAATTCCTTATC | |
TRV2-F | TGGGAGATGATACGCTGTT | |
TRV2-R | CCTAAAACTTCAGACACG | |
TRV2-RhJMT-F | AAGGTTACCGAATTCTCTAGAGGTACTGTCCACTGACATAAGAGC | |
TRV2-RhJMT-R | CTCGAGACGCGTGAGCTCGGTACCGCTTCATATTGTATACTTGCACG | |
实时荧光定量PCR | RhUBI2-F | GCCCTGGTGCGTTCCCAACTG |
qRT-PCR | RhUBI2-R | CCTGCGTGTCTGTCCGCATTG |
qRT-RhJMT-F | GTACTGTCCACTGACATAAGAGC | |
qRT-RhJMT-R | GCTTCATATTGTATACTTGCACG | |
qRT-RhSAG12-F | AGCGGAGAAGCCTTTCAGTC | |
qRT-RhSAG12-R | CAGCATGGTTCAGGCTGGTA | |
qRT-RhSAG113-F | TGTGTCCTCTGTAACCAACAAGT | |
qRT-RhSAG113-R | CTCCAACTAGACCGCAGCAA |
表1 本研究中所用的引物
Table 1 Primers used in this study
用途 Function | 引物名称 Primer name | 引物序列(5′-3′) Sequence |
---|---|---|
克隆基因 Gene clone | RhJMT-F | GAGATAATGGAGGTAGAGCAAGTTCTTC |
RhJMT-R | GCTTCATATTGTATACTTGCACG | |
病毒诱导的基因沉默 Virus induced gene silencing | TRV1-F | TTACAGGTTATTTGGGCTAG |
TRV1-R | CCGGGTTCAATTCCTTATC | |
TRV2-F | TGGGAGATGATACGCTGTT | |
TRV2-R | CCTAAAACTTCAGACACG | |
TRV2-RhJMT-F | AAGGTTACCGAATTCTCTAGAGGTACTGTCCACTGACATAAGAGC | |
TRV2-RhJMT-R | CTCGAGACGCGTGAGCTCGGTACCGCTTCATATTGTATACTTGCACG | |
实时荧光定量PCR | RhUBI2-F | GCCCTGGTGCGTTCCCAACTG |
qRT-PCR | RhUBI2-R | CCTGCGTGTCTGTCCGCATTG |
qRT-RhJMT-F | GTACTGTCCACTGACATAAGAGC | |
qRT-RhJMT-R | GCTTCATATTGTATACTTGCACG | |
qRT-RhSAG12-F | AGCGGAGAAGCCTTTCAGTC | |
qRT-RhSAG12-R | CAGCATGGTTCAGGCTGGTA | |
qRT-RhSAG113-F | TGTGTCCTCTGTAACCAACAAGT | |
qRT-RhSAG113-R | CTCCAACTAGACCGCAGCAA |
图1 不同物种JMT蛋白序列比对分析 框选位置为Methylyransf_7 superfamily的保守结构域。Rh:月季;In:牵牛花;Na:烟草;Fv:野草莓;Gm:大豆;St:马铃薯。
Fig. 1 Alignment and analysis of JMT protein sequences from various plant species The box is selected for Methylyransf_7 superfamily’s conservative domain. Rh:Rosa hybrida;In:Ipomoea nil;Na:Nicotiana attenuata;Fv:Fragaria vesca;Gm:Glycine max;St:Solanum tuberosum.
图3 ‘Samantha’月季不同器官(A)和不同开放时期花瓣(B)中RhJMT的表达量
Fig. 3 The relative expression level of RhJMT in different tissues(A)and in different opening stages(B)of rose flowers
图7 沉默RhJMT与外源MeJA处理对‘Bride’月季花瓣圆片变色面积的影响
Fig. 7 Effects of silencing RhJMT and exogenous MeJA treatment on area of disc discoloration in Rosa hybrida‘Bride’ *** P < 0.001.
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