Identification and Functional Analysis of Walnut JrPR1 and JrGLU in Relation to Anthracnose Resistance

doi:10.16420/j.issn.0513-353x.2024-0558

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (7): 1696-1706.doi: 10.16420/j.issn.0513-353x.2024-0558

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Identification and Functional Analysis of Walnut JrPR1 and JrGLU in Relation to Anthracnose Resistance

XU Haifeng, WANG Guifang, XIANG Kun, SHEN Guangning, ZHANG Meiyong, and CHEN Xin^*()

Key Laboratory of Huanghuai Facility Horticulture Engineering，Ministry of Agriculture and Rural Affairs，National Germplasm Repository of Walnut and Chestnut，Shandong Institute of Pomology，Tai’an，Shandong 271000，China

Received:2025-04-16 Revised:2025-06-20 Online:2025-07-23 Published:2025-07-23
Contact: and CHEN Xin

Abstract

Abstract:

Pathogenesis-related genes and β-1,3-glucanase genes play important roles in plant disease resistance.‘Lukang 8’and‘Xiangling’walnut cultivars，which exhibit significant differences in anthracnose resistance，were used as experimental materials. 10 days after in vitro inoculation with Colletotrichum gloeosporioides，the husk lesion on‘Xiangling’significantly enlarged，while‘Lukang 8’showed no obvious changes. Transcriptome analysis revealed that the differentially expressed genes in the husks of the two cultivars were enriched in disease resistance-related pathways such as plant-pathogen interaction，MAPK signaling pathway，and plant hormone signal transduction. The preliminarily identified that JrPR1，JrPR5，JrGLU and JrGLU5 are highly expressed in the disease-resistant cultivar‘Lukang 8’. The tissue-specific analysis showed that JrPR5 and JrGLU5 are highly expressed in buds and leaves，respectively，while JrPR1 and JrGLU are highly expressed in fruits，suggesting they are candidate genes for regulating fruit anthracnose resistance. Transient expression in tobacco leaves revealed that JrPR1 is localized to the cell wall and intercellular spaces，while JrGLU is localized to the cell wall. Additionally，the physicochemical properties，secondary，and tertiary structures of their encoded proteins were analyzed. Transient expression in walnut fruits revealed that both JrPR1 and JrGLU can significantly inhibit the growth of C. gloeosporioides and enhance anthracnose resistance in the fruits.

Key words: walnut, pathogenesis-related gene, β-1,3-glucanase, anthracnose

XU Haifeng, WANG Guifang, XIANG Kun, SHEN Guangning, ZHANG Meiyong, and CHEN Xin. Identification and Functional Analysis of Walnut JrPR1 and JrGLU in Relation to Anthracnose Resistance[J]. Acta Horticulturae Sinica, 2025, 52(7): 1696-1706.

Figures/Tables 8

Fig. 1 Analysis of anthracnose resistance in‘Lukang 8’and‘Xiangling’walnuts

Fig. 2 Gene enrichment analysis of‘Lukang 8’and‘Xiangling’walnuts after inoculation with Colletotrichum gloeosporioides A：Principal component analysis（PCA）；B：KEGG analysis. T：Lukang 8；X：Xiangling. T0 and T10，X0 and X10 represent the 0 days and 10 days post-infection with Colletotrichum gloeosporioides，respectively. The same below

Fig. 3 Heatmap of PR and GLU genes between‘Lukang 8’and‘Xiangling’walnut’s husks on day 10 post-inoculation with Colletotrichum gloeosporioides Colors ranging from blue to red indicate higher levels of gene expression

Fig. 4 Tissue-specific expression of PR and GLU genes Different lowercase letters indicate significant differences between tissues，P < 0.01

Fig. 5 Subcellular localization analysis of PR1 and GLU in tobacco leaves

Table 1 Physicochemical properties and secondary structure of PR1 and GLU proteins

基因名 Gene name	染色体定位 Chromosomal location	分子量/kD Molecular weight	等电点 Isoelectric point	平均亲水性 Average hydrophilicity	不稳定指数 Instability index	α螺旋/% Alpha helix	延伸链/% Extended strand	β转角/% Beta turn	无规则卷曲/% Random coil
PR1	NC_049913.1	22.10	9.51	-0.06	31.99	40.40	17.68	3.03	38.89
GLU	NC_049905.1	37.80	4.80	-0.10	34.49	35.63	20.11	5.75	38.51

Fig. 6 Structural analysis of PR1 and GLU proteins A：Secondary structure arrangement of PR1 and GLU proteins；B：Tertiary structure models of PR1 and GLU proteins

Fig. 7 Resistance analysis of walnut fruits after transient expression of PR1 and GLU and inoculation with Colletotrichum gloeosporioides for 6 d pIR：IL60-1 + IL60-2；pIR-PR1：IL60-1 + PR1-IL60-2；pIR-GLU：IL60-1+ GLU-IL60-2. Different lowercase letters indicate significant differences，P < 0.01

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Identification and Functional Analysis of Walnut JrPR1 and JrGLU in Relation to Anthracnose Resistance

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