基金项目:北京市自然科学基金(6192018),国家重点研发计划课题?019YFD1001001(/div>
详细信息
杨淑敏。研究方向:园林植物与观赏园艺研究。Email9a href="//www.inggristalk.com/j/article/doi/10.12171/mailto:2638562727@qq.com">2638562727@qq.com 地址?00083 北京市海淀区清华东?5号北京林业大学园林学陡/p>
潘会堂,教授。研究方向:园林植物与观赏园艺研究。Email9a href="//www.inggristalk.com/j/article/doi/10.12171/mailto:htpan@bjfu.edu.cn">htpan@bjfu.edu.cn 地址同上
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出版历程
- 收稿日期:2021-06-30
- 录用日期:2023-03-07
- 修回日期:2022-01-10
- 网络出版日期:2023-03-10
Selection and validation of appropriate reference genes and expression analysis of jasmonic acid-related genes responding toMarssonina rosaeinRosaspecies and cultivars
Beijing Forestry University, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, National Engineering Research Center for Floriculture, College of Landscape Architecture, Beijing 100083, China
摘要:
目的为研究蔷薇属植物响应黑斑病侵染的稳定表达的最适内参基因,解析茉莉酸在蔷薇属植物响应蔷薇盘二孢侵染过程中的作用、/sec>
方法以黑斑病病原菌蔷薇盘二孢侵染?个蔷薇属?品种不同时间的离体叶片为材料,利用qRT-PCR技术及geNorm、NormFinder、BestKeeper软件?个候选内参基因(
ACT?i>GAPDH?i>PP2
A?i>Rcl2?i>SAND?i>TIP?i>TUA?i>TUB?i>UBC)的表达量进行测定和分析,利用筛选出的内参基因,对蔷薇属植物茉莉酸(JA)抗病途径相关基因'i>COI1?i>OPR3?i>MYC2?i>JAR1)的表达水平进行定量分析、/sec>
结果?(i>UBC可作?种蔷薇属植物共同适用的内参基因,可用于后续分析JA抗病途径相关基因的表达水平。(2)内源JA含量?种蔷薇属植物响应蔷薇盘二孢侵染的过程中存在差异。在黑斑病高抗植物受侵染0 ~ 4 d间JA含量下调? ~ 8 d间上调。在黑斑病易感植物受侵染0 ~ 8 d间,内源JA含量呈下调趋势。(3)JA合成相关基因
OPR3叉i>JAR1表达量在受侵染初期表达量趋势存在差异。在除荷花蔷薇外的其?种植物中+i>OPR3在侵染初期(0 ~ 0.5 d)表达下调,
JAR1在侵染初期表达上调、i>OPR3咋i>JAR1在侵染后期均表达上调,黑斑病易感材料的上调程度高于高抗材料。(4)JA信号传导相关基因
COI1叉i>MYC2在受侵染初期表达量趋势同样存在差异、i>COI1在黑斑病高抗材料受侵染初期上调表达,在黑斑病易感材料下调表达+i>MYC2?种植物受侵染0 ~ 2 d中均下调表达、i>COI1叉i>MYC2表达量在受侵? d后均上调表达,且在黑斑病易感植物中的上调程度大于黑斑病高抗材料、/sec>
结论与JA信号传导相关皃i>MYC2?i>COI1在蔷薇属植物抵御黑斑病病菌入侵过程中发挥负调控作用,且由JA通路介导的抵御死体营养型病原菌的侵染在后期发挥了作用、/sec>
Abstract:
ObjectivesIn order to select the appropriate reference gene and analyze the role of jasmonic acid in
Rosaspecies and cultivars responding to
Marssonina rosae.
Methods
R. multifloraf.
carnea,
R. xanthinaf.
spontanea,
R.glauca,
R. rugosa,
R. hybrida‘Porcelina and
R12-26 (hybrid of
R. rugosaand
R. hybrida‘Porcelina? were used as material, qRT-PCR technology, geNorm, NormFinder, BestKeeper were used to evaluate the expression stability of nine candidate reference genes (
ACT,
GAPDH,
PP2
A,
RCl2,
SAND,
TIP,
TUA,
TUBand
UBC). The expression levels of jasmonic acid resistance pathway related genes (
COI1,
OPR3,
MYC2 and
JAR1) in six
Rosaspecies and cultivars were quantitatively analyzed.
Result(1)
UBCwas the common reference gene of six
Rosaspecies and cultivars responding to
M.
rosae, and can be used for analysis the related genes expression levels to JA resistance pathway. (2) There were differences in JA content among the six plants responding to
M.
rosae. In disease-resistant plants, JA concentration decreased at 0? d and increased at 4? d. In disease- susceptible plants, JA concentration decreased at 0? d and increased significantly at 8?0 d. (3) The expression levels of JA synthesis-related genes
OPR3 and
JAR1 were different at the early stage of infection. The expression level of
OPR3 was down-regulated and
JAR1 up-regulated in five plants except
R. multifloraf.
carnea.
OPR3 and
JAR1 were all up-regulated in the late stage of infection, however the up-regulation degree in black-spot susceptible species was significantly higher than that of resistant species. (4) There were also differences in the expression levels of JA signaling related gene
COI1 and
MYC2 at the early stage of infection. The expression of
COI1 was up-regulated in disease-resistant plants and down-regulated in disease-susceptible plants at the early stage of infection,
MYC2 was down-regulated in 6 plants infected 0? days.
COI1 and
MYC2 were up-regulated after 2 days of infection, and the up-regulation degree in black-spot susceptible species was significantly higher than that of resistant species.
ConclusionsWe believe that
MYC2 and
COI1, which are related to JA signal transmissions, play a negative role in regulating black spot resistance in
Rosa, SA pathway mediated resistance to necrotrophic fungi played a role in the late stage of infection.
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