Bioinformatics,Subcellular Localization and Expression Analysis of SsRPM1 Gene in Wild Solanum sisymbriifolium Lam.

doi:10.7668/hbnxb.20195481

Abstract

Abstract:

In order to explore the role of NBS-LRR gene RPM1 in the resistance to Verticillium wilt in Solanum,it took the wild Solanum sisymbriifolium Lam.as material,and cloned the homologous sequence of RPM1 gene on the basis of its transcriptional sequencing.The physicochemical properties and molecular structure of the sequence encoded protein were analyzed,the evolutionary relationship tree was constructed,and subcellular localization was performed in tobacco.At the same time,the relative expression level of RPM1 gene in different parts of Solanum sisymbriifolium Lam.was detected,as well as the relative expression level at for time points(0,24,48 and 72 h)after inoculation with Verticillium dahliae(a pathogen of Verticillium wilt).The results showed that the total length of RPM1 gene(SsRPM1)was 2 772 bp,encoding 924 amino acids.SsRPM1 protein,with a total molecular weight of 105.99 ku,was an alkaline hydrophilic protein without transmembrane structure.SsRPM1 protein was mainly composed of α helix and random coil,including LRR,NBC and CC domains.Solanum dulcamara RPM1 protein had the closest relationship with it.Subcellular localization in tobacco found that the protein was located on the cell membrane.SsRPM1 gene was expressed in different organs(root,stem and leaf),among which the stem had the highest relative expression,followed by leaf and root.After inoculation with V.dahliae,in general,SsRPM1 gene expression in both control group and inoculation group showed a trend of first increasing and then decreasing.The relative expression level of SsRPM1 gene was the highest at 24 h after inoculation.Compared with the control group,the relative expression level of SsRPM1 gene in inoculation treatment was lower.It is suggested that SsRPM1 is a negative regulatory gene in response to Verticillium wilt stress.

Key words: Wild eggplant, RPM1 gene, Verticillium wilt, Bioinformatics, Subcellular localization, Gene expression

CLC Number:

HU Yuying, SUN Mao, WANG Qian, LI Zhibin, BAO Rui, GUI Min, ZHONG Qiuyue, DU Guanghui, WU Liyan. Bioinformatics,Subcellular Localization and Expression Analysis of SsRPM1 Gene in Wild Solanum sisymbriifolium Lam.[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 27-33. doi: 10.7668/hbnxb.20195481.

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Figures/Tables 10

Fig.1 1% agarose gel electrophoresis of PCR amplication products M.5 000 bp DNA Marker;1—2.PCR products.

Fig.2 Prediction result of hydrophobicity of SsRPM1 protein

Fig.3 Prediction of secondary structure(A) and tertiary structure(B) of SsRPM1 protein

Fig.4 Prediction result of phosphoric acid sites in SsRPM1 protein

Fig.5 Domain analysis of SsRPM1 protein

Fig.6 Amino acid sequences alignment of RPM1 proteins in 5 different species

Fig.7 Phylogenetic tree of RPM1 proteins of different plant species

Fig.8 Subcellular localization analysis in tobacco cells of SsRPM1 protein

Fig.9 Relative expression level of SsRPM1 gene in different organs of S.sisymbriifolium Different letters indicate significant difference between different organs(P<0.05).

Fig.10 Relative expression level of SsRPM1 gene after S.sisymbriifolium inoculation with V.dahliae Different letters indicate significant differences between all the treatments and times(P<0.05).

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