Cloning and Expression of a Pathogenesis Related Protein Gene EjPR1 from Loquat Induced by Colletotrichum gloeosporioides

doi:10.16420/j.issn.0513-353x.2020-0141

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (2): 367-376.doi: 10.16420/j.issn.0513-353x.2020-0141

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Cloning and Expression of a Pathogenesis Related Protein Gene EjPR1 from Loquat Induced by Colletotrichum gloeosporioides

ZOU Hui¹^,², ZHAO Liang¹, YUAN Ting¹^,², GUO Qigao¹, WU Di²^,^*(), LIANG Guolu¹^,^*()

¹Key Laboratory of Horticulture Science for Southern Mountains Regions,Ministry of Education,College of Horticulture and Landscape Architecture;Southwest University,Chongqing 400715,China
²Academy of Agricultural Sciences of Southwest University;State Cultivation Base of Crop Stress Biology for Southern Mountainous Land of Southwest University,Chongqing 400715,China

Received:2020-05-29 Revised:2020-09-06 Online:2021-02-25 Published:2021-03-09
Contact: WU Di,LIANG Guolu E-mail:wudisuper610@126.com;lianggl@swu.edu.cn

Abstract

Abstract:

A full length cDNA of pathogenesis-related protein 1 gene named EjPR1 was obtained from the leaves of‘Peluches’which was a loquat cultivar with high resistance to anthracnose treated by Colletotrichum gloeosporioides using RT-PCR and RACE technology. Bioinformatics analysis indicated that the full length of EjPR1 was 731 bp（GenBank number：MN92775）,the length of 5′ UTR and 3′ UTR were 79 bp and 91 bp,respectively. The length of open reading frame（ORF）was 561 bp encoding 186 amino acids with isoelectric point 5.63 and molecular weight 21 485.34 D which was consistent with the results of prokaryotic expression. Multiple alignment analysis based on the amino acids encoded by different PR1 genes from six other species of Rosaceae showed that EjPR1 was conserved among them with the similarity of 76.41%-94.36%. The results of time series expression analysis after inoculation with anthrax indicated that EjPR1 expression was up-regulated by infection and the highest expression was observed 96 h after inoculation in highly resistant cultivar while there was no significant difference in highly susceptible cultivar. It was speculated that EjPR1 might be involved in disease defense of loquat.

Key words: Eriobotrya japonica, EjPR1, gene clone, prokaryotic expression, expression analysis

CLC Number:

S667.3

ZOU Hui, ZHAO Liang, YUAN Ting, GUO Qigao, WU Di, LIANG Guolu. Cloning and Expression of a Pathogenesis Related Protein Gene EjPR1 from Loquat Induced by Colletotrichum gloeosporioides[J]. Acta Horticulturae Sinica, 2021, 48(2): 367-376.

Figures/Tables 7

Table 1 The sequences of primer used in this study

用途
Primer funtion

序列
Sequence

EjPR1
3′RACE
5′RACE
EjPR1-CDs
pET-EjPR1
EjPR1D
EjActin

F：GGTTGCATAGGAGAGTCCTTG;R：GCACATTTGACCAGCCTCGC
Outer：GATGTGGGACTTTCAACTCG;Inner：CAAGCGACGCAGTGAAGACA
Long：TGTCTTCACTGCGTCGCTTG;Short：CGAGTTGAAAGTCCCACATC
F：ATGAAGGCACAATTCCTCTTTATCC;R：CTAGTATGGCCTCTCACCAATATAA
F：CGGGATCCATGAAGGCACAATTCCTCTTTATCC;R：CGAGCTCCTAGTATGGCCTCTCACCAATATAA
F：AAGCGACGCAGTGAAGACA;R：ACAACCCTAGCACACCCGA
F：AATGGAACTGGAATGGTCAAGGC;R：TGCCAGATCTTCTCCATGTCATCCCA

Fig. 1 Result of amplification product of EjPR1 from loquat

Fig. 2 The full length of EjPR1 ORF and deduced amino acid sequences

Fig. 3 Multiple sequence alignment of EjPR1 protein

Fig. 4 Phylogenetic tree of PR1 protein from plants The number at the branch of evolutionary tree indicate the confidence of the branch.

Fig. 5 Analysis of EjPR1 gene expression in‘Peluches’and‘Senwei Zaosheng’at different hpi * shows that there are significant differences between different treatments at the same time point（P < 0.05）.

Fig. 6 SDS-PAGE analysis of protein expressed in Escherichia coli by pET-EjPR1

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Cloning and Expression of a Pathogenesis Related Protein Gene EjPR1 from Loquat Induced by Colletotrichum gloeosporioides

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