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Article

Identification of New Sources of Resistance to Anthracnose Caused by Colletotrichum horii among Persimmon Germplasms

State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling 712100, China
*
Author to whom correspondence should be addressed.
Horticulturae 2022, 8(2), 180; https://doi.org/10.3390/horticulturae8020180
Submission received: 31 December 2021 / Revised: 12 February 2022 / Accepted: 18 February 2022 / Published: 21 February 2022
(This article belongs to the Special Issue Genetics and Breeding of Fruit Trees)

Abstract

:
Persimmon (Diospyros kaki) anthracnose, predominantly caused by Colletotrichum horii, is a destructive disease of persimmon. Thus, the evaluation of resistance resources is imperative for persimmon breeding and resistant variety deployment. In this study, the isolate from persimmon branches was identified as C. horii by using molecular and morphological characteristics. A total of 142 varieties were tested for anthracnose disease response by inoculation with the conidial suspension of C. horii. A significant variability was observed among the studied accessions. Only two accessions, which had a hypersensitive reaction with a slight lesion at the infection site at 14 days post inoculation, were highly resistant to anthracnose. A total of 7 and 24 accessions exhibited resistant and susceptible responses, respectively, to anthracnose. A total of 109 varieties with the highest proportion (76.76%) of accessions showed high susceptibility to C. horii. A total of 43 persimmon main cultivars were selected for the re-evaluation of the resistance level by counting the natural disease incidence of branches and fruits that showed similar resistant response. Moreover, nine of highly resistant or resistant accessions possessed desirable agronomic characters, including high fruit weight and strong growth potential. These resistant accessions could be used in the breeding of anthracnose-resistant persimmon varieties.

1. Introduction

Persimmon (Diospyros kaki) is regarded to originate from China [1] and is predominantly cultivated in East Asia, including China, Japan, and Korea. China, which has the largest area and production harvest in the world, produces 3.427 million tons, accounting for 75% among all the countries (The Food and Agriculture Organization of the United Nations, FAO, 2021). The persimmon fruit is rich in vitamins, antioxidants, and trace elements that are vital for human health and has been used in various medicinal and chemical industries and for commercial fruit consumption [2,3]. Currently, the cultivation of the persimmon crop is rapidly expanding in Israel, Italy, and Spain, suggesting that persimmon is becoming one of the most popular fruits worldwide.
The collection and evaluation of persimmon germplasms support the safe preservation of genetic diversity and facilitate the selection and breeding of excellent resistant varieties of persimmon [4]. The National Field Genebank for Persimmon (NFGP) in China began to collect persimmon germplasms in 1962 and was first built in 1987 in Meixian County, Shannxi, China [5]. Currently, the NFGP saves more than 1000 persimmon resources from different regions in China and a relatively high number of varieties with a high genetic diversity from other countries [6,7].
Persimmon anthracnose is a severe fungal disease, causing branches and leaves to wither, fruit to rot, and possibly the loss of the entire plant, gravely jeopardizing the development of the persimmon industry [8,9]. Previously, the pathogen that causes anthracnose was reported as Gloeosporium kaki by Shotaro Hori and Seiya Ito [10,11]. Maffei [12] described the leaf spot pathogen of persimmon from a specimen collected in Italy as Colletotrichum kaki. Von [13] later reported that the fungi previously described are the same and synonymous with C. gloeosporioides. However, in 2010, after examining the molecular and morphological characteristics of specimens from China, Japan, and New Zealand, Weir and Johnston renamed the persimmon pathogen as C. horii [14]. Additionally, the persimmon anthracnose can be caused by C. siamense, C. karstii, C. fructicola, C. nymphaeae, and C. melonis [15,16,17,18]. Persimmon anthracnose is a dangerous disease in China and in other nations across the world [15,16,17,18]. Carraro reported that C. fructicola, C. nymphaeae, and C. melonis cause persimmon anthracnose in Brazil [16]. Moreover, C. siamense and C. nymphaeae cause persimmon anthracnose in Korea [15,18]. With increased rain, the anthracnose in persimmon orchards becomes serious because many main varieties are susceptible to anthracnose. In addition to chemical control and cultivation management, breeding anthracnose-resistant varieties based on resistance evaluation is the most effective method to control anthracnose. Thus, screening resistance resources is imperative for breeding resistant varieties.
Abundant Diospyros germplasm resources are in the NFGP of China, and the evaluation and identification of persimmon germplasm resistance to anthracnose can provide new sources for resistance breeding programs. In the present study, we aim to (a) isolate and identify the pathogen of persimmon anthracnose, (b) evaluate the resistance level of 142 persimmon varieties in the NFGP, and (c) identify some anthracnose-resistant varieties for the future application of persimmon breeding.

2. Materials and Methods

2.1. Sample Collection and Pathogen Fungus Isolation

A total of 142 persimmon germplasm resources without infections and insect pests were preserved in the NFGP, Yangling, Shaanxi, China (34°16′56.24″ N, 108°04′27.95″ E; 420 m altitude).
For candidate pathogen isolation, symptomatic twigs of ‘Fuping Jianshi’ persimmon were randomly collected from NFGP at Yangling, China (34°17′52.55″ N, 108°04′05.58″ E) and brought to the laboratory for the isolation of putative causal agents. Samples were surface disinfected for 90 s with 70% ethanol and 1% sodium hypochlorite, rinsed twice with sterile distilled water. Sterilized small branch pieces (5 mm × 5 mm) were placed on PDA plates and then incubated at 25 °C with a photoperiod of 12 h until fungal growth was observed. Pathogen isolates were purified with single-spore culturing prior to use in subsequent experiments. The isolated pathogen fungus was named ‘FJ 3’.
For the microscopic morphology observation of the incidence of tissue samples after infection, the collected branches and fruit samples with different incidence degrees in different diseased parts were cut into small pieces of tissue (5–8 mm) at the junction of disease and health. After cutting, samples were quickly put into FAA fixative. Paraffin sections were prepared in accordance with the reported method [19].

2.2. Molecular Characterization of the Fungus

DNA was extracted from the isolated pathogen, and the genomic DNA sequences of the ITS region of rDNA, beta-tubulin (TUB2) gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS-1), and a partial sequence of the actin (ACT) were amplified with primers of ITS1F/ITS4, T1/Bt2b, GDF1/GDRl, ACT-512F/ACT-783R, and CHS-79F/CHS-345R, respectively [20] (Table S1). The amplified reaction system contained a total volume of 25 μL, including 2.5 μL of 2 × Reaction Mix, 0.2 μL Golden DNA Polymerase, 1 μL template DNA, 1 μL of 10 μmol/L primers, and 9.3 μL ddH2O. PCR conditions were as follows: pre-denaturation at 95 °C for 3 min, denaturation at 95 °C for 30 s, annealing at 56 °C for 30 s, and extension at 72 °C for 1 min for 42 cycles. The last extension was at 72 °C for 5 min. PCR products were detected by 1% agarose gel electrophoresis. Subsequently, products were cloned into the T-Vector pMD19 (TaKaRa, Dalian, China), and three positive clones for each fragment were sequenced (Aoke Technology Co., Ltd, Beijing, China). A phylogenetic tree was obtained using the MEGA5 software [21].

2.3. Artificial Inoculation and Natural Disease Incidence

After 18 days of culturing for purified pathogen isolation, conidia suspensions were prepared by flooding the culture plates and removing the fungal mycelium. The number of conidia was determined using a hemocytometer, and the conidia suspension was adjusted to a 1 × 106/mL concentration as a backup.
For artificial inoculation, an inoculum concentration of 1.0 × 106 conidia/mL for the ‘FJ 3’ fungus was used to infect the uninjured young branches (n = 3) by 5 μL suspensions with a short interval of 1.5 cm (Figure 1). The growth of diseased spots was recorded using a Vernier caliper for 15 consecutive days. The experiment was conducted twice in 2019 and 2020.
For natural disease incidence in field, diseased fruits (≥20) and branches (≥20) with uniform growth potential were recorded in four different directions of the persimmon tree. The surface or sepals of the fruit exhibiting typical black spots were counted, and the incidence was presented as percent anthracnose infection. Branches showing typical anthracnose lesions were evaluated as infected.

2.4. Disease Resistance Criterion

For the artificial inoculation, the standard of disease classification is shown in Table 1. Disease index (DI) = Σ (number of branches infected in the rank × corresponding disease rank) × 100 / total number of infected branches × highest rank. On the basis of the range of DI, persimmon accessions were classified into highly resistant (0 < DI ≤ 10.0), resistant (10.0 < DI ≤ 30.0), susceptible (30.0 < DI ≤ 50.0), and highly susceptible (50.0 < DI ≤ 100.0, Table 2). If the disease severity varied between two years, a high disease grade of the year should be used as a rating scale of the accession resistance.
Under natural field conditions, the severity of fruits and branches was categorized using a 1–4 scale, where highly resistant ≤ 2%, 2% ≤ resistant < 5%, 5% ≤ susceptible < 20%, and highly susceptible ≥ 20%. Fruit and branch severity values were presented as percent infected samples. At least 20 samples were measured for each persimmon variety.

3. Results

3.1. Symptoms of Persimmon Anthracnose Disease

The anthracnose fungus may attack the fruits, twigs, and leaf petioles in susceptible persimmon. When the fruit is infected with anthracnose, black spots appear on the surface of the fruit first and diseased spots slowly expand and dent downwards, finally causing the fruit to soften (Figure 1A). The lesion location had a modest number of stone cells, the peel was softened, and the infection gradually spread throughout the flesh.
Anthracnose symptoms on young twigs first appeared in dark spots, and minute spots developed into dark brown specks, showing a clear dividing line between diseased and symptomless tissues. The anthracnose fungus still attacked the xylem of branches and led to collapse. With the spread of disease spots, adjacent lesions connected until the entire twig was infected (Figure 1B).
The fungus infected petioles and caused similar dark brown spots (Figure 1C). With the development of the disease, leaves gradually fell off.

3.2. Morphological and Molecular Characterization of C. horri

For fungal isolation, symptomatic twigs of persimmon were randomly collected and brought to the laboratory for the isolation of putative causal agents. Samples were surface disinfected for 90 s with 70% ethanol and 1% sodium hypochlorite, then rinsed twice with sterile distilled water. Sterilized small branch pieces (5 mm × 5 mm) were placed on PDA plates. Representative purified isolates of the ‘FJ 3’ fungus colonies resembling Colletotrichum spp. were selected for further phylogenetic and morphological analyses. The ‘FJ 3’ fungal strain grew rapidly on PDA, thereby filling the entire Petri dish (90 mm) after 11 days at 25 °C and 12 h photoperiod. Colonies of ‘FJ 3’ were initially white and gradually became grayish on PDA with dark concentric zonation and regular margins in reverse (Figure 2). Few conidia (no conidial mass) were observed across the colony after 15 days of incubation at 25 °C on PDA. Conidia were hyaline, single-celled, straight, cylindrical apices and measured up to 10.5 to 17.1 μm × 4.0 to 5.56 μm (n = 100, average = 13.6 μm × 4.6 μm, Figure 2). The ‘FJ 3’ isolate showed similar morphological traits with the previously reported C. horri [22].
Five isolates obtained from the ‘Fuping Jianshi’ persimmon were identified as the same fungal species on the basis of multigene phylogenetic and morphological analyses (Figure 3). The persimmon anthracnose ‘FJ 3’ fungus was selected for further phylogenetic analysis. Then, genomic DNA sequences were amplified with ITS1F/ITS4, T1/Bt2b, GDF1/GDRl, ACT-512F/ACT-783R, and CHS-79F/CHS-345R (Weir et al., 2012). These amplicons were submitted to GenBank with accession numbers of OL347726 for ITS, OL364188 for GAPDH, OL364190 for CHS-1, OL364191 for TUB2, and OL364189 for ACT. The BLAST search in GenBank revealed that sequences showed high similarity to those of C. horii. Amplification sequences from the ‘FJ 3’ fungus were identical to JX010450 for CHS-1, 98.34% to GQ329690 for ITS, 98.34% to JX010450 for GAPDH, 99.29% to JX009438 for ACT, and 99.74% to JX010450 for TUB2. The maximum likelihood tree was obtained from a concatenated dataset of ITS, ACT, CHS, GAPDH, and TUB2 gene sequences of the C. gloeosporioides species complex with 1000 bootstrapping, showing that the present ‘FJ 3’ isolate clustered with the reference isolate C. horii (NBRC 7478) with high bootstrap support. Thus, the current ‘FJ 3’ isolate was identified as C. horii on the basis of cultural characteristics and sequence similarity data (Figure 3).

3.3. Evaluation of Anthracnose Resistance among Persimmon Germplasms by Artificial Infection

Persimmon accessions were grown in NFGP at Yangling, China (34°17′52.55″ N, 108°04′05.58″ E). In 2019 and 2020, this area received total rainfall values of 656 and 683 mm, respectively, and average temperatures of 13.0 °C and 13.2 °C, respectively. Rainfall varied between months of the same year, whereas the temperature of the area fluctuated little, especially between different years. The rainfall from July to September accounts for nearly half of the annual rainfall, and this period represents the hottest three months (highest average temperature (26.1–26.3 °C) is observed in July). The temperature and relative humidity from July to September are conducive to the occurrence of the persimmon anthracnose disease.
The resistance level was defined in four categories, including highly resistant, resistant, susceptible, and highly susceptible, on the basis of the DI of inoculated branches in 2019 and 2020 (Figure 4 and Figure 5). A significant variability was observed among the studied accessions. Only two accessions, which had a hypersensitive reaction with a slight lesion at the injection site at 15 days postinoculation, were highly resistant to anthracnose. Seven cultivars exhibited a resistant response to anthracnose and accounted for 4.93% of the 142 accessions (Figure 4). The susceptible response was observed in 24 accessions, which accounted for 16.90% of the total. A total of 109 varieties with the highest proportion (76.76%) of the accessions showed a highly susceptible response to C. horii ‘FJ 3’ (Figure 4 and Table 3).

3.4. Evaluation of the Anthracnose Resistance for Selected Persimmon Accessions by Natural Disease Incidence

A total of 43 persimmon main cultivars were selected for the re-evaluation of anthracnose resistance in the nature field in 2018 and 2019 (Table 4). Four accessions were highly resistant and had no symptomatic anthracnose in branches and fruits, and nine accessions were resistant with average percent disease incidence ≤ 5. In 34 remaining accessions, 5 and 29 were susceptible and highly susceptible, respectively, with percent disease incidence values of 5–20% and 20–100%, respectively. The data of repeated evaluation experiments showed that persimmons performed consistent resistance by artificial infection and natural disease incidence, except for two accessions, i.e., ‘Zaozhuang Ehuangshi’ and ‘Ningbo Tongpenshi’.

3.5. Agronomic Characteristics of Blast Resistant Finger Millet Genotypes

The agronomic traits of nine anthracnose-resistant germplasms were analyzed, and results showed that highly resistant genotypes, i.e., ‘Kangbing Jianshi’ and ‘Yuanqu Bayuehuang’, had strong growth potential, orange-yellow peel color, glutinous texture, and none had a cross-concave form on the fruit apex (Table S2). ‘Hyakume’, which could naturally remove astringency on the tree during the ripening stage, was classified under the pollination-variant nonastringent type, whereas others were classified under the pollination-constant astringent type. Moreover, ‘Hyakume’ had the highest average fruit weight of 192 g and maximum fruit weight of 386 g. ‘Zhaoan Yuanxiaoshi’ and ‘Baoshan Dafangshi’ had the most days of maintaining fruit crispness, with 27 days. All varieties could be used as female parent materials for resistance breeding. In addition, the ‘Hyakume’ persimmon, with occasional male flowers, could be used as a male parent for the breeding program (Table S2).

4. Discussion

The C. gloeosporioides complex is the most common and polyphagous species complex of the genus. Weir et al. [20] reported that C. gloeosporioides is a complex consisting of 22 species. To date, 57 species have been described [23]. The genus has been voted as one of the top 10 fungal plant pathogens in molecular plant pathology [24]. On the basis of multigene phylogenetic analysis and morphological characteristics, the pathogen causing persimmon anthracnose has been renamed as C. horii, which also belongs to the C. gloeosporioides species complex [14]. To the best of our knowledge, most individual species in the C. gloeosporioides species complex, such as C. horii, C. fructicola, and C. siamense, may cause persimmon anthracnose [15,16,25] (Figure 1). Moreover, some different plants can be infected by the same anthrax species [14,26], C. acutatum from pepper can cause typical persimmon anthracnose symptoms, showing black spot disease in Korea [27,28]. Thus, these species (C. gloeosporioides is a species complex) are extremely difficult to distinguish on the basis of morphological characteristics only. Thus, pathogenic strains from persimmon anthracnose should be identified on the basis of the multigene phylogenetic analysis. In the present study, the ‘FJ 3’ isolate from persimmon was identified as C. horri by the combined analysis of molecular and morphological characteristics. Moreover, the ‘FJ 3’ isolate was tested for pathogenicity on persimmon twigs in the laboratory and nature field to confirm the isolate’s ability to cause the anthracnose disease. Therefore, the identification of the anthracnose pathogen in the national persimmon germplasm resource nursery is helpful to further carry out and deepen the research on the pathogenesis and defense mechanism of this anthracnose and provide a basis for the prevention and control of persimmon anthracnose.
Persimmon anthracnose, a serious disease occurring in many countries where persimmon is cultivated, is predominantly caused by C. gloeosporioides [29]. Persimmon anthracnose symptoms first appear in the spring as spots which then develop into dark lesions. Under high temperature and humidity conditions, adjacent lesions may integrate together, increasing in size until the entire twig is infected [30]. In the present study, the anthracnose pathogen can result in dark lesions in the fruit, calyx, branches, and leaf of persimmon, showing similar results with previous reports [9,15,22,30]. In addition, the C. gloeosporioides species complex consists of important plant pathogens that can cause anthracnose in economic crops, such as chili [31], mango [32], strawberry [33], apples [34], grapes [35], bananas [36,37], and tomato [38]. As a destructive disease of persimmon trees, C. horii can cause leaf defoliation, fruit rot, and even the death of the whole plant [39]. Unfortunately, most persimmon varieties are not resistant to anthracnose. For example, ‘Gongcheng Shuishi’ and ‘Fuping Jianshi’, which have the largest cultivation area in China, are susceptible to persimmon anthracnose. In some persimmon-growing areas in China, anthracnose can reduce the yield of persimmon by more than 50% [40].
Anthracnose-resistant persimmon varieties are important for the breeding of new resistant cultivars and the development of the persimmon industry. However, the evaluation and identification of persimmon germplasms with anthracnose resistance remain unknown. Persimmon is believed to originate in China and has abundant genetic diversity, with at least 1000 varieties [2,7]. Therefore, the identification of excellent persimmon resources resistant to anthracnose is necessary for resistance breeding. On the basis of the artificial infection, two accessions (i.e., ‘Kangbing Jianshi’ and ‘Quyuan Bayuehuang’), which had a hypersensitive reaction with a slight lesion at the injection site at 14 days post inoculation, were highly resistant to anthracnose, and six cultivars (i.e., ‘Yeshi’, ‘Pucheng Fangshi’, ‘Bayuehuang’, ‘Lushi 5’, and ‘ Junqianzi) exhibited a resistant response to anthracnose. Furthermore, 43 main cultivars of persimmon were selected for the re-evaluation of anthracnose resistance in the nature field in 2018 and 2019. The resistance level of germplasm resources was consistent with the previous evaluation results of artificial infection. Moreover, through the evaluation of agronomic characters, the above eight resistance resources showed excellent characters and could be used in resistance breeding in the future. For example, ‘Kangbing Jianshi’ is suitable for making dried persimmon due to the heart-shaped fruit, and ‘Quyuan Bayuehuang’ is early maturing with high yield and strong growth potential.

5. Conclusions

Persimmon anthracnose, a destructive disease for persimmon, is predominantly caused by C. horii pathogen, resulting in shoot wilting, fruit rotting, and even death of the whole plant. In this study, the isolate ‘FJ 3’ from persimmon trees in the NFGP of China was further identified as C. horii by using morphological characteristics and multigene phylogenetic analysis. A total of 142 persimmon varieties were tested for anthracnose disease response by inoculation with the conidial suspension of C. horii in the NFGP. Resistance level was defined in four categories, including highly resistant, resistant, susceptible, and highly susceptible, on the basis of the DI of the inoculated branch. A significant variability was observed among the studied accessions. Only two varieties, which had a hypersensitive reaction with a slight lesion at the injection site at 14 days post inoculation, were highly resistant to anthracnose. Seven cultivars exhibited a resistant response to anthracnose and accounted for 4.93% of the 142 accessions. A susceptible response was observed for 24 accessions, which accounted for 16.90% of the total. A total of 109 varieties with the highest proportion (76.76%) of the accessions showed a highly susceptible response to C. horii. A total of 43 persimmon main cultivars were selected for the re-evaluation of anthracnose resistance by natural disease incidence in 2018 and 2019 and showed similar resistance levels. Moreover, eight highly resistant or resistant accessions possessed desirable agronomic characters. The use of these resistance sources in persimmon breeding should aid in expanding the genetic diversity and in the development of new resistant varieties.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/horticulturae8020180/s1. These amplicons have been submitted to GenBank with accession numbers of OL347726 for ITS, OL364188 for GAPDH, OL364190 for CHS-1, OL364191 for TUB2, and OL364189 for ACT. Table S1: Primers for pathogen identification; Table S2: The agronomic characteristics of anthracnose resistant persimmon genotypes.

Author Contributions

Conceptualization, C.G. and Y.Y.; methodology, Q.C. and Y.L.; software, Q.C. and J.H.; validation, C.G., Y.Y., and J.H.; writing—original draft preparation, C.G. and J.H.; writing—review and editing, C.G.; visualization, Y.Y.; supervision, Y.Y.; funding acquisition, Y.Y. and C.G. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by National Key Research and Development Program of China (2019YFD1000600) and National Horticulture Germplasm Resources Center (NHGRC2020-NH06).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data are contained within the article.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Anthracnose symptoms and plant tissue sectioning caused by C. horri on persimmon. Disease lesions on (A) fruit, (B) branch, and (C) petiole. The red ellipse indicates the infection site of anthracnose, “a” indicates the location of the disease, and “×” means magnification.
Figure 1. Anthracnose symptoms and plant tissue sectioning caused by C. horri on persimmon. Disease lesions on (A) fruit, (B) branch, and (C) petiole. The red ellipse indicates the infection site of anthracnose, “a” indicates the location of the disease, and “×” means magnification.
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Figure 2. Cultural and morphological characteristics of the C. horii ‘FJ 3’ isolation growing on PDA: (A) the upper surface, (B) the lower surface, (C) conidia (bar = 20 μm), and (D) mycelial growth (bar = 50 μm) of C. horii.
Figure 2. Cultural and morphological characteristics of the C. horii ‘FJ 3’ isolation growing on PDA: (A) the upper surface, (B) the lower surface, (C) conidia (bar = 20 μm), and (D) mycelial growth (bar = 50 μm) of C. horii.
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Figure 3. Phylogenetic analysis using the neighbor-joining method through comparative studies of nucleotide sequences of the internal transcribed spacer (ITS) region of rDNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a partial sequence of the actin (ACT), chitin synthase (CHS-1), and beta-tubulin (TUB2) genes from the present isolates with those of other Colletotrichum spp. in the C. gloeosporioides species complex retrieved from GenBank. The ‘FJ 3’ isolate is emphasized in the red circle. Numbers above the branches represent bootstrap values.
Figure 3. Phylogenetic analysis using the neighbor-joining method through comparative studies of nucleotide sequences of the internal transcribed spacer (ITS) region of rDNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a partial sequence of the actin (ACT), chitin synthase (CHS-1), and beta-tubulin (TUB2) genes from the present isolates with those of other Colletotrichum spp. in the C. gloeosporioides species complex retrieved from GenBank. The ‘FJ 3’ isolate is emphasized in the red circle. Numbers above the branches represent bootstrap values.
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Figure 4. Representative varieties of ‘Qianxian Muwashi’ and ‘Hebei Lianhuashi’ exhibiting high susceptibility and susceptibility, respectively, to C. horii after artificial infection in (A) 2019 and (B) 2020.
Figure 4. Representative varieties of ‘Qianxian Muwashi’ and ‘Hebei Lianhuashi’ exhibiting high susceptibility and susceptibility, respectively, to C. horii after artificial infection in (A) 2019 and (B) 2020.
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Figure 5. Representative varieties of ‘Chengsushi’ and ‘Yuanqu Bayuehong’ exhibiting resistant and highly resistant responses, respectively, to C. horii after artificial infection in (A) 2019 and (B) 2020.
Figure 5. Representative varieties of ‘Chengsushi’ and ‘Yuanqu Bayuehong’ exhibiting resistant and highly resistant responses, respectively, to C. horii after artificial infection in (A) 2019 and (B) 2020.
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Table 1. Branch lesion standard for persimmon anthracnose.
Table 1. Branch lesion standard for persimmon anthracnose.
RankDescription
0No lesion on branches
10% < Ratio of total lesion diameter to total branch length ≤ 20%
220% < Ratio of total lesion diameter to total branch length ≤ 40%
340% < Ratio of total lesion diameter to total branch length ≤ 60%
460% < Ratio of total lesion diameter to total branch length ≤ 80%
580% < Ratio of total lesion diameter to total branch length ≤ 100%
Table 2. Levels and types of resistance to persimmon anthracnose.
Table 2. Levels and types of resistance to persimmon anthracnose.
Resistance LevelsDisease Index (DI)Resistance
10 < DI ≤ 10.0Highly Resistant (HR)
310.0 < DI ≤ 30.0Resistant (R)
530.0 < DI ≤ 50.0Susceptible (S)
750.0 < DI ≤ 100.0Highly Susceptible (HS)
Table 3. Anthracnose resistance level based on the average lesions of persimmon germplasm inoculated with C. horri isolate.
Table 3. Anthracnose resistance level based on the average lesions of persimmon germplasm inoculated with C. horri isolate.
Group20192020
NumberVarietyAverage Lesions (mm)Disease IndexResistance LevelResistanceAverage Lesions (mm)Disease IndexResistance LevelResistanceIdentification Result
1Xiangfen Qiyuehong14.32 ± 0.61007HS14.36 ± 0.231007HSHS
2Jishan Hanbanjin13.25 ± 0.98967HS11.94 ± 1.19887HSHS
3Jishan Banshi8.54 ± 1.69687HS4.67 ± 0.4405SHS
4Yongji Qingshi11.67 ± 0.77807HS6.52 ± 1.38527HSHS
5Baishi11.97 ± 1.99847HS8.42 ± 1.05687HSHS
6Meixian Qinghuamanaitou11.85 ± 3.14887HS13.99 ± 0.531007HSHS
7Qingxuanshi14.88 ± 0.241007HS7.87 ± 1.91567HSHS
8Yaoxian Wuhuashi6.84 ± 0.66567HS6.71 ± 1.79567HSHS
9Wenxi Pingdingshi14.03 ± 1.52967HS14.65 ± 0.711007HSHS
10Lintong Jiandingshi13.47 ± 1.62927HS11.46 ± 1.14847HSHS
11Lintong Fangshi5.99 ± 1.4445S12.55 ± 0.69967HSHS
12Lintong Huojing8.89 ± 2.63727HS9.5 ± 2.58727HSHS
13Chengou Huishi6.66 ± 4.94567HS5.9 ± 4.54485SHS
14Weiboshi6.45 ± 1.98567HS5.52 ± 3.12445SHS
15Boai Bayuehuang11.18 ± 1.08847HS12.01 ± 2.17887HSHS
16Shiyangshi14.45 ± 0.461007HS10.28 ± 3.01847HSHS
17Wanghoushi12.63 ± 2.37927HS13.64 ± 11007HSHS
18Meipishi10.33 ± 2.46767HS8.54 ± 1.69687HSHS
19Ernizi11.86 ± 0.34807HS12.92 ± 1.63927HSHS
20Hongxuanshi12.39 ± 2.12887HS13.09 ± 2.23927HSHS
21Meixian Niuxinshi13.71 ± 0.881007HS12.09 ± 1.67967HSHS
22Yichuanling11.37 ± 1.94807HS10.19 ± 1.45767HSHS
23Xunyang Huoshi7.37 ± 1.13567HS7.28 ± 1.08567HSHS
24Xunyang Guanguanshi10.24 ± 3.31807HS8.53 ± 2.46647HSHS
25Nanzhang Jianshi14.19 ± 0.241007HS11.09 ± 2.89847HSHS
26Luoyang Guilianqing14.87 ± 0.191007HS6.77 ± 3.78527HSHS
27Liuba Huoshi9.63 ± 2.65767HS8.19 ± 3.07607HSHS
28Binxian Shuishi7.13 ± 3.27527HS12.97 ± 1967HSHS
29Huaitaishi6.99 ± 0.69607HS8.02 ± 2.11647HSHS
30Heixinshi14.35 ± 0.371007HS13.94 ± 0.751007HSHS
31Denglongshi10.85 ± 3.02807HS3.9 ± 4.37325SHS
32Yidu Tuoshi11.44 ± 2.7847HS7.26 ± 3.37567HSHS
33Zhouqu Huoshi7.95 ± 1.89647HS4.69 ± 1.81405SHS
34Changan Fudingjian13.95 ± 0.81007HS14.32 ± 0.821007HSHS
35Shuhuangshi7.59 ± 2.22647HS10.97 ± 0.58807HSHS
36Jincheng Gaishi9.6 ± 3.42727HS9.47 ± 3.52727HSHS
37Tongguan Lianhuashi10.15 ± 3.06767HS11.1 ± 1.56847HSHS
38Licheng Mianshi12.36 ± 2.28727HS14.43 ± 0.181007HSHS
39Matian FangSs11.95 ± 0.95927HS14.07 ± 0.611007HSHS
40Heishi10.49 ± 0.62807HS11.23 ± 3.34887HSHS
41Xiaoyi Niuxinshi13.16 ± 2.22967HS6.01 ± 0.76485SHS
42Xiaoerzao13.53 ± 1.15967HS11.89 ± 1887HSHS
43Cangshan Niuxinshi6.38 ± 2.97485S7.51 ± 1.27647HSHS
44Mianrangshi14.81 ± 0.11007HS14.09 ± 1.091007HSHS
45Xingyang Shuishi9.98 ± 2.11767HS12.05 ± 3.56887HSHS
46Suxian Niuxinshi9.13 ± 1.41727HS11.77 ± 2.76887HSHS
47Qujing Shuishi11.42 ± 1.27887HS6.9 ± 1.38607HSHS
48Qujing Xiaoshuishi13.1 ± 2.35927HS12.55 ± 2.04927HSHS
49Biantashi12.91 ± 1.52927HS12.01 ± 2.84887HSHS
50Fuzhengbing13.49 ± 1.45967HS12.97 ± 1.88967HSHS
51Qiuzhengbing11.04 ± 1.42847HS10.04 ± 2.08807HSHS
52Chikelang11.62 ± 1.12887HS9.12 ± 1.59687HSHS
53Hongmiandan12.91 ± 0.871007HS15 ± 01007HSHS
54Huxian Huangmiandan15 ± 01007HS14.87 ± 0.271007HSHS
55Xichou Huoshi15 ± 01007HS15 ± 01007HSHS
56Jiro8.27 ± 1.52647HS9.93 ± 0.22807HSHS
57Luoyang Shuiniuxin14.92 ± 0.141007HS15 ± 01007HSHS
58Lantian Shuishi13.17 ± 0.88967HS8.82 ± 5.08687HSHS
59Nanjing Gaozhuangshi7.49 ± 1.87607HS3.69 ± 3.65325SHS
60Lantian Dafangshi10.57 ± 1.28847HS9.7 ± 2.44727HSHS
61Binxian Jiandingshi14.28 ± 0.681007HS14.78 ± 0.471007HSHS
62Huxian Dashi13.71 ± 1.3967HS13.97 ± 0.61007HSHS
63Sifangshi14.76 ± 0.131007HS14.35 ± 0.351007HSHS
64Niutoushi12.42 ± 0.8927HS8.73 ± 0.33647HSHS
65Guyangshi8.84 ± 0.72687HS5.76 ± 0.4445SHS
66Yangshuo Niuxinshi8.14 ± 1.54647HS3.7 ± 2.52365SHS
67Shieryueshi12.15 ± 1.34927HS7.88 ± 2.7647HSHS
68Baoshan Dashuishi7.79 ± 1.25647HS4.64 ± 4.2365SHS
69Dafeng Niuxinshi11 ± 1.28887HS8.59 ± 1.61687HSHS
70Longhui Ruanzao10.24 ± 2.8767HS10.61 ± 3.81807HSHS
71Liuheshi13.63 ± 0.971007HS11.18 ± 1.74847HSHS
72Jurong Bianshi14.41 ± 0.571007HS13.13 ± 1.18967HSHS
73Suqian Bianshi6.63 ± 1.03527HS6.02 ± 1.23527HSHS
74Pixian Biangangshi11.77 ± 3.41847HS11.08 ± 2.23887HSHS
75Silenggaishi11.55 ± 2.3887HS10.15 ± 0.98767HSHS
76Huixian Dashi11.87 ± 1.7887HS13.06 ± 5.89647HSHS
77Huixian Shuishi11.94 ± 1.58927HS5.6 ± 2.83485SHS
78Pijianglou8.35 ± 3.61647HS6.9 ± 1.94567HSHS
79Ichidagaki5.91 ± 0.67527HS5.24 ± 1.98445SHS
80Shougatsu7.19 ± 1.76607HS4.16 ± 0.95365SHS
81Sakugosho12.02 ± 1.76887HS13.09 ± 1.23967HSHS
82Changan Shuishi10.89 ± 3.28807HS12.24 ± 2.15927HSHS
83Changan Shaoshi14.49 ± 0.281007HS14.19 ± 0.651007HSHS
84Huxian Tieguoshi6.94 ± 2.76567HS5.45 ± 0.74445SHS
85XiaoShuishi14.29 ± 0.871007HS11.66 ± 1.69927HSHS
86Xingyang Huoguan14.39 ± 0.651007HS13.76 ± 1.091007HSHS
87Qingchutou12.02 ± 2.03887HS8.84 ± 4.81767HSHS
88Yanshitiansheng-0112.27 ± 1.22927HS12.64 ± 1.72927HSHS
89Shahe Huoshi14.61 ± 0.341007HS13.85 ± 0.691007HSHS
90Jingjing Gaishi9.01 ± 2.77687HS10.63 ± 2.43807HSHS
91Pingshan Niujinshi13.65 ± 1.52967HS13.3 ± 2.19967HSHS
92Huaxian Baixuanshi14.09 ± 1.131007HS13.65 ± 0.981007HSHS
93Heibei Lianhuashi112.99 ± 2.35927HS12.29 ± 2.92927HSHS
94Shiyueshi14.54 ± 0.271007HS13.73 ± 1.061007HSHS
95Lintong Diaoshi8.9 ± 4.66647HS6.34 ± 1.46527HSHS
96Laoshigou11 ± 1.28887HS8.59 ± 1.61687HSHS
97Baokangshi10.24 ± 2.8767HS10.61 ± 3.81807HSHS
98Daezi13.63 ± 0.971007HS11.18 ± 1.74847HSHS
99Changsha Shuishi15 ± 01007HS8.96 ± 4.79717HSHS
100Fuping Jianshi15 ± 01007HS15 ± 01007HSHS
101Chaoyang Yuanxiaoshi8.46 ± 1.34667HS9.61 ± 0.92807HSHS
102Conghuashi14.31 ± 0.521007HS15 ± 01007HSHS
103Dabaoshi13.09 ± 0.76927HS8.79 ± 5.18687HSHS
104Dabiegaishi7.86 ± 1.78607HS3.71 ± 3.53325SHS
105Dali Daqiyuehuang10.66 ± 1.32857HS9.65 ± 2.32737HSHS
106Dali Qiyuehuang14.51 ± 0.761007HS14.85 ± 0.551007HSHS
107Damianhu13.73 ± 1.36977HS13.97 ± 0.61007HSHS
108Dangshan Ehuangshi14.76 ± 0.131007HS14.35 ± 0.451007HSHS
109Dangshan Niuxinshi13.12 ± 1.6917HS8.52 ± 0.95737HSHS
110Yongji Mushi3.57 ± 1.81325S3.46 ± 1.35283RS
111Hangzhou Niuxinshi5.52 ± 0.65445S4.79 ± 0.53405SS
112Chetoushi4.03 ± 0.99365S3.69 ± 1.04365SS
113Shagu23.89 ± 0.54325S3.51 ± 1.03283RS
114Fuyang Gongshi4.37 ± 0.93405S4.59 ± 0.46405SS
115Qianxian Muwashi5.94 ± 2.49485S4.86 ± 1.66405SS
116Putian Goushi2.37 ± 0.32203R5.09 ± 0.55405SS
117Xiaobaxianshi5.36 ± 0.39405S5.81 ± 2.21445SS
118Guangzhou Niuxinshi5.41 ± 0.78445S3.22 ± 0.82283RS
119Zhengyang Baheshi5.53 ± 0.93485S4.98 ± 0.52405SS
120Gongcheng Shuishi2.76 ± 1.16283R3.5 ± 0.78365SS
121Sibanmianshi3.21 ± 1.59283R5.89 ± 3.39485SS
122Zhoushan Changshi3.79 ± 0.34405S4.1 ± 1.88405SS
123Tangcunshi2.44 ± 0.47243R3.28 ± 0.6365SS
124Caojishi4.38 ± 1.83405S4.32 ± 2.51325SS
125Songyang Bianshi4.85 ± 0.42405S4.18 ± 1.01365SS
126Mancheng Niuxinshi6.1 ± 0.92485S3.34 ± 1.42325SS
127Taian Jingmianshi3.8 ± 3.45365S1.53 ± 3.32163RS
128Baoshan Hongshi3.76 ± 1.28385S3.53 ± 0.98375SS
129Baoshan Shuishi3.67 ± 1.71315S5.92 ± 3.49495SS
130Ganmaokui3.97 ± 0.56425S4.09 ± 1.91405SS
131Changan Fudingshi2.49 ± 0.66233R3.38 ± 0.66375SS
132Huxian Jiandingshi4.54 ± 1.68395S4.36 ± 2.29325SS
133Youhou3.11 ± 1.06293R4.24 ± 1.01375SS
134Ningbo Tongpenshi1.05 ± 0.33123R1.31 ± 1.22123RR
135Zhaoan Yuanxiaoshi1.87 ± 0.32163R2.34 ± 0.33243RR
136Chengsushi2.74 ± 1.09243R2 ± 0.36203RR
137Baoshan Dafangshi1.6 ± 3.89163R2.41 ± 3.31203RR
138Hyakume1.41 ± 0.51203R2.33 ± 1.33243RR
139Yanjing Yingshi1.36 ± 1.74163R3.31 ± 9.5243RR
140Zaozhuang Ehuangshi1.87 ± 0.71163MR0 ± 001HRR
141Kangbing Jianshi0 ± 001HR0 ± 001HRHR
142Yuanqu Bayuehong0.92 ± 0.2583HR0 ± 001HRHR
Table 4. Anthracnose resistance level for 43 selected persimmon germplasms based on the severity of fruits and branches in the nature field.
Table 4. Anthracnose resistance level for 43 selected persimmon germplasms based on the severity of fruits and branches in the nature field.
Group20182019
NumberVarietyInfected Fruits (%)Infected Braches (%)Average (%)ResistanceInfected Fruits (%)Infected Braches (%)Average (%)ResistanceIdentification Result
1Jishan Hanbanjin50.00%0.00%20.00%HS11.54%35.00%25.62%HSHS
2Yongji Qingshix24.29%24.29%HSx31.90%31.90%HSHS
3Lintong Jiandingshi12.09%26.00%20.44%HS40.00%16.46%25.87%HSHS
4Lintong Fangshi30.00%14.29%20.57%HS41.00%23.08%30.25%HSHS
5Lintong Huojing50.00%2.83%21.70%HS50.00%1.03%20.62%HSHS
6Boai Bayuehuang15.38%26.09%21.80%HS18.42%55.00%40.37%HSHS
7Meixian Niuxinshi30.20%7.00%16.28%S50.00%1.43%20.86%HSHS
8Luoyang Guilianqing57.89%1.85%24.27%HSx21.05%21.05%HSHS
9Denglongshi34.38%12.50%21.25%HS35.29%21.81%27.20%HSHS
10Yidu Tuoshi29.82%18.57%23.07%HS100.00%11.43%46.86%HSHS
11Zhouqu Huoshi33.33%11.76%20.39%HS62.50%4.44%27.70%HSHS
12Changan Fudingjianx16.00%16.00%S6.90%30.00%20.76%HSHS
13Shuhuangshi37.50%11.58%21.95%HSx52.50%31.50%HSHS
14Jincheng Gaishi40.00%10.00%22.00%HSx45.00%45.00%HSHS
15Tongguan Lianhuashi0.00%40.00%24.00%HSx32.50%32.50%HSHS
16Licheng Mianshi12.50%30.00%23.00%HS4.44%40.00%25.78%HSHS
17Matian FangSs56.00%29.05%39.83%HS100.00%0.00%40.00%HSHS
18Xingyang Shuishi33.33%17.14%23.62%HS70.00%21.00%40.60%HSHS
19Fuzhengbing100.00%8.33%45.00%HS75.00%17.50%40.50%HSHS
20Hongmiandan40.54%10.00%22.21%HS100.00%14.29%48.57%HSHS
21Lantian Shuishi50.00%2.56%21.54%HS58.10%61.00%59.84%HSHS
22Sifangshi18.50%17.14%17.68%S79.32%19.50%43.43%HSHS
23Gongcheng Shuishi21.02%12.50%15.91%S100.00%14.29%48.57%HSHS
24Baoshan Dashuishi60.00%1.67%25.00%HS90.00%18.33%47.00%HSHS
25Jurong Bianshi51.02%8.25%25.36%HSx50.00%50.00%HSHS
26Suqian Bianshi100.00%14.29%48.57%HS100.00%21.65%52.99%HSHS
27Huixian Dashi100.00%16.30%49.78%HS61.54%50.00%54.62%HSHS
28Huaxian Baixuanshix18.60%18.60%HSx55.00%55.00%HSHS
29Fuping Jianshix50.00%50.00%HSx60.00%60.00%HSHS
30Youhou7.55%3.03%4.84%R0.00%20.00%12.00%SS
31Putian Goushi8.33%5.80%6.81%S7.69%15.00%12.08%SS
32Sibanmianshi5.88%7.50%6.85%S8.46%14.74%12.23%SS
33Zhoushan Changshi14.29%4.79%8.59%S28.57%5.00%14.43%SS
34Tangcunshi0.00%8.33%5.00%R21.74%10.00%14.50%SS
35Ningbo Tongpenshix0.00%0.00%HR5.00%0.00%2.00%HRHR
36Zhaoan Yuanxiaoshix0.00%1.39%R5.06%0.00%2.03%RR
37Chengsushix2.33%0.00%R1.22%2.63%2.07%RR
38Baoshan Dafangshix0.55%1.23%R0.00%3.54%2.12%RR
39Hyakumex0.00%1.67%Rx2.13%2.13%RR
40Yanjing Yingshi0.00%0.00%0.00%HR1.37%2.63%2.13%RR
41Zaozhuang Ehuangshi0.00%0.00%0.00%HR0.00%1.67%1.00%HRHR
42Kangbing Jianshix0.00%0.00%HR0.00%0.00%0.00%HRHR
43Yuanqu Bayuehongx0.00%0.00%HR0.00%0.00%0.00%HRHR
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Guan, C.; Hu, J.; Li, Y.; Che, Q.; Yang, Y. Identification of New Sources of Resistance to Anthracnose Caused by Colletotrichum horii among Persimmon Germplasms. Horticulturae 2022, 8, 180. https://doi.org/10.3390/horticulturae8020180

AMA Style

Guan C, Hu J, Li Y, Che Q, Yang Y. Identification of New Sources of Resistance to Anthracnose Caused by Colletotrichum horii among Persimmon Germplasms. Horticulturae. 2022; 8(2):180. https://doi.org/10.3390/horticulturae8020180

Chicago/Turabian Style

Guan, Changfei, Jie Hu, Yongkuan Li, Qinghui Che, and Yong Yang. 2022. "Identification of New Sources of Resistance to Anthracnose Caused by Colletotrichum horii among Persimmon Germplasms" Horticulturae 8, no. 2: 180. https://doi.org/10.3390/horticulturae8020180

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