The Molecular Regulatory Mechanism of StCAT1 and StCAT3 in Resistance to Early Blight in Potatoes

doi:10.7668/hbnxb.20195821

Abstract

Abstract:

Catalase(CAT)plays a critical role in plant resistance against biotic stresses.To investigate the functions of StCAT1 and StCAT3 in potato defense against Alternaria solani infection,this study cloned the cDNA sequences of StCAT1 and StCAT3 via RT-PCR.Sequencing results demonstrated that both genes were 1 479 bp in length,encoding 493 amino acids.Phylogenetic tree construction revealed that StCAT1 shared high homology with SlCAT1 from tomato,while StCAT3 exhibited high homology with both SlCAT3(tomato)and SpCAT3 from wild tomato.Following A.solani infection for 6 days,the expression of StCAT1 and StCAT3 was significantly upregulated by approximately 3.9-fold and 8.7-fold,respectively.A co-silencing vector targeting both StCAT1 and StCAT3 was constructed,and transient co-silencing was achieved using virus-induced gene silencing(VIGS).qRT-PCR screening identified an effective co-silenced potato line(VIGS-3)with suppressed StCAT1 and StCAT3 expression.Upon A.solani inoculation,the silenced line exhibited 42% larger lesion areas compared to the control,indicating that co-silencing StCAT1/StCAT3 compromised potato resistance to early blight.DAB(3,3'-diaminobenzidine)staining further demonstrated that H₂O₂ accumulation in co-silenced leaves was significantly elevated,with staining intensity being 3.8-fold higher than that in the control.These results demonstrate that StCAT1 and StCAT3 regulate H₂O₂ homeostasis to mediate potato resistance against A.solani,providing a theoretical foundation for elucidating the molecular mechanisms of CAT genes in potato-pathogen interactions.

Key words: Early Blight in Potatoes, StCAT1, StCAT3, VIGS, H₂O₂

CLC Number:

LIU Xin, LI Qian, LIU Fangming, DING Mingya, YANG Zhihui, ZHU Jiehua. The Molecular Regulatory Mechanism of StCAT1 and StCAT3 in Resistance to Early Blight in Potatoes[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(5): 171-178. doi: 10.7668/hbnxb.20195821.

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Figures/Tables 8

Tab.1 Primers used in this study

引物名称 Primer name	序列(5'—3') Sequence(5'—3')
StCAT1-F	ATGGATCCCTCTAAGTATCGC
StCAT1-R	TCACATTGTGGGCTTCACA
StCAT3-F	ATGGATCCTTACAAGTATCGTCC
StCAT3-R	TCACATTGTAGGCCTTATATTGAGAC
V-CAT1-F	CG GAATTCCTGGAAACTATCCTGAGTGG
V-CAT1-R	CACCGTATTGGACCAAACTATAGGTACCCC
V-CAT3-F	TATTGGACCAAACTATAATGGATCC
	TTACAAGTATCGTCCTTCAAG
V-CAT3-R	CTCCTGGGGTCCAGACACCTGTTATT GGTACCCC
qStCAT1-F	CCGTGATGGAATGAAGTT
qStCAT1-R	GCCTGTAATCTTGTGGAA
qStCAT3-F	CCTTCTTCTACGATGATG
qStCAT3-R	TACACAAGACTCTAATAG

Fig.1 PCR amplification of the StCAT1 and StCAT3 genes M.DL2000 DNA Marker;1.PCR product of StCAT1; 2.PCR product of StCAT3.

Fig.2 Construction of the phylogenetic tree of StCAT1 and StCAT3

Fig.3 Analysis of the expression patterns of StCAT1 and StCAT3 during the early blight infection period Different lowercase letters indicate significant differences (P<0.05,Student's t-test).The same as Fig.5—7.

Fig.4 Construction of the co-silencing vector for StCAT1 and StCAT3 A.PCR amplification of the silenced gene sequences of StCAT1 and StCAT3:M.DL2000 DNA Marker,1.StCAT1 silenced fragment,2.StCAT3 silenced fragment;B.PCR amplification of the silenced fusion fragments of StCAT1 and StCAT3:M.DL5000 DNA Marker;C.Double enzyme digestion verification of the co-silencing vector of StCAT1 and StCAT3 genes:M.DL5000 DNA Marker.

Fig.5 qRT-PCR identification of positive potato plants with co-silenced StCAT1 and StCAT3 genes

Fig.6 Resistance evaluation of co-silenced StCAT1 and StCAT3 plants to early blight disease A.Phenotype of early blight resistance evaluation.

Fig.7 DAB staining and intensity quantification of infected leaf tissue A.DAB staining of leaves from silenced and control plants after inoculation with early blight.

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