Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (4): 761-772.doi: 10.16420/j.issn.0513-353x.2022-1220
• Genetic & Breeding · Germplasm Resources · Molecular Biology • Previous Articles Next Articles
HOU Xue, WANG Jiaojiao, ZHANG Wenwen, ZHAO Jianlong*(), MAO Zhenchuan*(
)
Received:
2023-10-07
Revised:
2024-02-28
Online:
2024-04-25
Published:
2024-04-26
Contact:
ZHAO Jianlong, MAO Zhenchuan
HOU Xue, WANG Jiaojiao, ZHANG Wenwen, ZHAO Jianlong, MAO Zhenchuan. Cloning of Pepper Ethylene-Responsive Proteinase Inhibitor Cacl-6468 and Its Effect on Resistance to Meloidogyne incognita[J]. Acta Horticulturae Sinica, 2024, 51(4): 761-772.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2022-1220
用途 Usage | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence |
---|---|---|
3′RACE克隆 3′RACE clone | 3′RACE | ACCTAGTAGATATACGTACA |
5′RACE克隆 5′RACE clone | 5′RACE | CTTAGTTTCAAGATCTACGA |
Cacl-6468基因克隆 Cacl-6468 clone | Cacl-6468-F | ATGGATAAGTTAGCTCATT |
Cacl-6468-R | TCAACCAACTTGGGGAGTTT | |
VIGS载体构建 VIGS vector construction | pTV-Cacl-6468-F | AGAAGGCCTCCATGGGGATCCGAAAAAGGACAAGTGGCCAGAA |
pTV-Cacl-6468-R | GAGACGCGTGAGCTCGGTACCAGACGAACTCGATCACATCTAAAATC | |
pTV-PDS-F | GTGAGTAAGGTTACCGAATTCTAGGGTGACGGATGAGGTGTTC | |
pTV-PDS-R | GAGACGCGTGAGCTCGGTACCTTCCAGTCTTCAGGCAAAAGAAG | |
qRT-PCR | β-Tubulin-F | GAGGGTGAGTGAGCAGTTC |
β-Tubulin-R | CTTCATCGTCATCTGCTGTC | |
qCacl-6468-F | AAAAGGACAAGTGGCCAGAA | |
qCacl-6468-R | TTGTGTAACTGGAGAACCATTCA | |
qCaERF66-F | AGAAGGTCGAAAGAGGAT | |
qCaERF66-R | GCTGAGGAATTAGGGTTA | |
qCaPTI1-F | GGTTCCAACTCACCAAAGTGATC | |
qCaPTI1-R | TTCTTGCCTATGGTTTGTGATGG | |
qCaPR1-F | GCCGTGAAGATGTGGGTCAATGA | |
qCaPR1-R | TGAGTTACGCCAGACTACCTGAGTA | |
qCaNPR1-F | ACTTCTTCGCCGACGCCAAG | |
qCaNPR1-R | GCCAACACATTCACCAGAGCATC |
Table 1 related genes and their qRT-PCR primer sequences
用途 Usage | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence |
---|---|---|
3′RACE克隆 3′RACE clone | 3′RACE | ACCTAGTAGATATACGTACA |
5′RACE克隆 5′RACE clone | 5′RACE | CTTAGTTTCAAGATCTACGA |
Cacl-6468基因克隆 Cacl-6468 clone | Cacl-6468-F | ATGGATAAGTTAGCTCATT |
Cacl-6468-R | TCAACCAACTTGGGGAGTTT | |
VIGS载体构建 VIGS vector construction | pTV-Cacl-6468-F | AGAAGGCCTCCATGGGGATCCGAAAAAGGACAAGTGGCCAGAA |
pTV-Cacl-6468-R | GAGACGCGTGAGCTCGGTACCAGACGAACTCGATCACATCTAAAATC | |
pTV-PDS-F | GTGAGTAAGGTTACCGAATTCTAGGGTGACGGATGAGGTGTTC | |
pTV-PDS-R | GAGACGCGTGAGCTCGGTACCTTCCAGTCTTCAGGCAAAAGAAG | |
qRT-PCR | β-Tubulin-F | GAGGGTGAGTGAGCAGTTC |
β-Tubulin-R | CTTCATCGTCATCTGCTGTC | |
qCacl-6468-F | AAAAGGACAAGTGGCCAGAA | |
qCacl-6468-R | TTGTGTAACTGGAGAACCATTCA | |
qCaERF66-F | AGAAGGTCGAAAGAGGAT | |
qCaERF66-R | GCTGAGGAATTAGGGTTA | |
qCaPTI1-F | GGTTCCAACTCACCAAAGTGATC | |
qCaPTI1-R | TTCTTGCCTATGGTTTGTGATGG | |
qCaPR1-F | GCCGTGAAGATGTGGGTCAATGA | |
qCaPR1-R | TGAGTTACGCCAGACTACCTGAGTA | |
qCaNPR1-F | ACTTCTTCGCCGACGCCAAG | |
qCaNPR1-R | GCCAACACATTCACCAGAGCATC |
Fig. 1 The evolutionary tree of the conserved domain amino acid sequence of the capsicum-ethylene response inhibitor gene Cacl-6468 and its sequence in other species
Fig. 2 Changes in the expression of resistance gene Cacl-6468 in roots of HDA149 pepper after inoculation with vir and avir population of Meloidogyne incognita * P < 0.05,** P < 0.01,**** P < 0.0001.
Fig. 3 Changes in root knot and egg mass number in HDA149 capsicum plants after inoculation with vir and avir population of Meloidogyne incognita after gene silencing and control plants H2O:Water control plant;pTV-PDS:Positive control plant;VIGS:Nematode resistance gene Cacl-6468 silencing plants. ** P < 0.01,**** P < 0.0001.
Fig. 4 Effects of HDA149 pepper nematode resistant gene silencing plants(VIGS)inoculated with avir population of Meloidogyne incognita on gene expression in salicylic acid and ethylene signaling pathways * P < 0.05,**** P < 0.0001.
Fig. 5 Staining of root tissue and the number of second-instar larvae in roots after inoculation with Meloidogyne incognita with HDA149 pepper H2O:Water control plant;pTV-PDS:Positive control plant;VIGS:Nematode resistance gene Cacl-6468 silencing plants. ** P < 0.01.
Fig. 6 Distribution of second-instar nematodes around the root tip of pepper in Meloidogyne incognita tropism test H2O:Water control;pTV-PDS:Positive control;VIGS:Anti-nematode gene Cacl-6468 silencing treatment. * P < 0.05.
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