Abstract
Main conclusion
Identification of MAPKKK genes in pear and functional characterization of PbrMAPKKK82 in response to pear black spot.
Abstract
Mitogen-activated protein kinase kinase kinase (MAPKKK) is located upstream of the MAPK cascade pathway. This region senses extracellular stimuli via the signaling molecule or by themselves and is activated by phosphorylation. In this study, we identified 108 PbrMAPKKK genes from the pear genome. The genes were divided into three subfamilies and contained the conserved domain. Except for chromosome 7, there were 93 PbrMAPKKK genes randomly distributed on 16 out of the 17 chromosomes, while 15 PbrMAPKKK genes were detected on unknown chromosomes. They largely originated from whole-genome duplication (WGD) and dispersed events. In the expression analysis of PbrMAPKKK genes in seven pear tissue types by using a database, 20 PbrMAPKKK genes were selected to verify the expression associated with different resistance in two varieties by quantitative real-time PCR (qRT-PCR). The results showed that PbrMAPKKK12, PbrMAPKKK13, PbrMAPKKK53, PbrMAPKKK60, PbrMAPKKK65, PbrMAPKKK82, PbrMAPKKK83, and PbrMAPKKK96 were correlated with black spot resistance. PbrMAPKKK3, PbrMAPKKK9, PbrMAPKKK11, PbrMAPKKK34, PbrMAPKKK80, PbrMAPKKK81, PbrMAPKKK99, and PbrMAPKKK100 were correlated with black spot susceptibility, while the PbrMAPKKK gene positively responded to the life process of pear resistance to black spot. Furthermore, virus-induced gene silencing (VIGS) indicated that the PbrMAPKKK82 gene enhanced resistance to pear black spot disease.
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Abbreviations
- CAT:
-
Catalase
- CHI:
-
Chitinase
- MAPKKK:
-
Mitogen-activated protein kinase kinase kinase
- PAL:
-
Phenylalanine ammonia-lyase
- PPO:
-
Polyphenol oxidase
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Acknowledgements
This work has been supported by the National Key Research and Development Program of China (2018YFD1000300), the National Science Foundation of China (31872070; 32072538), Fundamental Research Funds for the Central Universities (JCQY201901), the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, the Earmarked Fund for China Agriculture Research System (CARS-28), the Jiangsu Agriculture Science and Technology Innovation Fund (SCX(22)3215).
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425_2022_4020_MOESM1_ESM.pdf
Supplementary file1 Alignments analysis of MEKK subfamily from Pyrus bretchneideri. The typical conserved domain (G (T/S) PX(W/Y/F)MAPEV) was marked with red line. Different colors indicate different similarities (mazarine: 100%, magenta: ≥ 75%, cyan ≥ 50%) (PDF 177 KB)
425_2022_4020_MOESM2_ESM.pdf
Supplementary file2 Alignments analysis of ZIK subfamily from Pyrus bretchneideri. The typical conserved domain (GTPEFMAPE (L/V/M) (Y/F/L)) was marked with red line (PDF 160 KB)
425_2022_4020_MOESM3_ESM.pdf
Supplementary file3 Alignments analysis of RAF subfamily from Pyrus bretchneideri. The typical conserved domain (GTXX (W/Y) MAPE) was marked with red line (PDF 181 KB)
425_2022_4020_MOESM4_ESM.pdf
Supplementary file4 Phylogenetic tree of MAPKKK family genes from Pyrus bretchneideri and three Rosaceae species. The phylogenetic tree was generated based on the amino acid sequences of all MAPKKKs from four Rosaceae species by the NJ method using MEGA7.0. Bootstrap supports from 1000 replicates are indicated at each branch. The shapes and colors of nodes represent different sources. Gene name prefix ‘Pbr’ represents Chinese white pear, ‘Md’ represents Apple, ‘gene’ represents Strawberry, ‘ppa’ represents Peach, ‘AT’ represents Arabidopsis. The blue branch represents the Raf-like group, green branch represents the MEKK-like group, red branch represents the ZIK-like group (PDF 498 KB)
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Zhang, Z., Jia, L., Chen, Q. et al. Genome-wide identification of the mitogen-activated protein kinase kinase kinase (MAPKKK) in pear (Pyrus bretschneideri) and their functional analysis in response to black spot. Planta 257, 5 (2023). https://doi.org/10.1007/s00425-022-04020-z
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DOI: https://doi.org/10.1007/s00425-022-04020-z