Anthracnose Caused by Colletotrichum spaethianum on Polygonatum odoratum var. pluriflorum in Korea.

IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Plant disease Pub Date : 2024-10-11 DOI:10.1094/PDIS-05-24-0991-PDN
Joon-Ho Choi, Hyeon-Dong Shin, Young-Joon Choi
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As the disease progressed, brown necrotic lesions developed, covered with small, dark dots, indicating the presence of conidiomata of Colletotrichum. The disease often led to leaf blight, resulting in concentric lesions of a periodic and concentric arrangement of conidiomata, forming circular patterns within the affected tissue. It expanded rapidly during the rainy season producing water-soaked lesions but nearly slowed down during the dry season, leaving the plant with a ragged appearance. Monoconidial isolates were obtained from five fresh samples collected in Wanju, of which a representative isolate was deposited in the Korean Agricultural Culture Collection (KACC410442), and the dried specimen was housed at the herbarium of Jeonbuk National University (JBNU0135). Morphological characteristics of the fungus were examined with fresh and naturally infected leaves. Setae (n = 30) were dark brown to nearly blackish, 45 to 172 um long, 4.1 to 6.1 µm wide at the base, and becoming narrower upwards, apex pointed, 2 to 4-septate. Conidia were 1-celled, sometimes uniseptate before germination, slightly curved, with an acute or rounded apex and a more or less truncate base, and 11.4 to 23.2 × 3.1 to 4.2 µm. Appressoria were single or in groups, dark brown, lobate, irregularly shaped, and 6.0 to 11.9 × 7.0 to 14.8 µm. Two-week-old colonies grown on PDA at 25 ℃ reached 30-35 mm in diam., initially white, turning gray with age, with cottony aerial mycelia. These morphological characteristics were in good agreement with those of C. spaethianum (Damm et al. 2009; Liu et al. 2020). To confirm the morphology-based determination, genomic DNA was extracted from KACC410442, and multi-loci sequences of the internal transcribed spacer region (ITS) rDNA, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), actin (actA), chitin synthase (CHS1), histone (HIS3) and tubulin (TUB2) genes were determined, as outlined by Cannon et al. (2012) and Damm et al. (2009). The resulting sequences obtained in this study were registered to GenBank (PP739190 for ITS, PP741985 for GAPDH, PP741986 for actA, PP741987 for CHS1, PP741988 for HIS3, and PP741989 for TUB2) and compared with other sequences on GenBank using the BLASTn search tool. The results showed 100% identity to other C. spaethianum sequences (e.g., MT611068 for ITS, MN400659 for actA, OP965007 for CHS1, OR060963 for GAPDH, PP480643 for HIS3, and OQ613727 for TUB2). In a maximum parsimony tree, the Korean isolate formed a well-supported clade (100% bootstrap value) with reference sequences of C. spaethianum. Pathogenicity was tested by spraying a conidial suspension (1×104 conidia/ml) from a two-week-old culture onto ten leaves of three potted plants, without wounding. Ten water-sprayed leaves were served as controls. The plants were maintained in a dew chamber and later moved to a greenhouse at 25 to 30 °C. After two weeks, all inoculated plants developed anthracnose symptoms, while the control plants remained symptomless. The fungus isolated from the inoculated plants was morphologically identical to that initially observed, fulfilling Koch's postulates. C. spaethianum has been known on various plant species, but mostly on the Asparagaceae, including Anemarrhena (Okorley et al. 2019), Hosta (Damm et al. 2009; Cheon and Jeon 2016), and Polygonatum (Liu et al. 2020; Ma et al. 2021). 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引用次数: 0

Abstract

Polygonatum odoratum var. pluriflorum (Miq.) Ohwi (Asparagaceae), commonly known as Lesser Solomon's seal, is a perennial herbaceous plant widely distributed in the temperate regions of the Northern Hemisphere. It is known for tea and contains various pharmacologically active compounds (Chang et al. 2007). In September 2021, previously unseen anthracnose was encountered on Polygonatum odoratum var. pluriflorum in Chuncheon (37°45'07''N; 127°47'45''E) and Wanju (35°38'47''N; 127°31'16''E), Korea, with a disease incidence of 20 to 30% of the inspected plants. The disease symptoms begin with leaf yellowing (chlorosis) along the margins, progressing toward the central part of the leaf. As the disease progressed, brown necrotic lesions developed, covered with small, dark dots, indicating the presence of conidiomata of Colletotrichum. The disease often led to leaf blight, resulting in concentric lesions of a periodic and concentric arrangement of conidiomata, forming circular patterns within the affected tissue. It expanded rapidly during the rainy season producing water-soaked lesions but nearly slowed down during the dry season, leaving the plant with a ragged appearance. Monoconidial isolates were obtained from five fresh samples collected in Wanju, of which a representative isolate was deposited in the Korean Agricultural Culture Collection (KACC410442), and the dried specimen was housed at the herbarium of Jeonbuk National University (JBNU0135). Morphological characteristics of the fungus were examined with fresh and naturally infected leaves. Setae (n = 30) were dark brown to nearly blackish, 45 to 172 um long, 4.1 to 6.1 µm wide at the base, and becoming narrower upwards, apex pointed, 2 to 4-septate. Conidia were 1-celled, sometimes uniseptate before germination, slightly curved, with an acute or rounded apex and a more or less truncate base, and 11.4 to 23.2 × 3.1 to 4.2 µm. Appressoria were single or in groups, dark brown, lobate, irregularly shaped, and 6.0 to 11.9 × 7.0 to 14.8 µm. Two-week-old colonies grown on PDA at 25 ℃ reached 30-35 mm in diam., initially white, turning gray with age, with cottony aerial mycelia. These morphological characteristics were in good agreement with those of C. spaethianum (Damm et al. 2009; Liu et al. 2020). To confirm the morphology-based determination, genomic DNA was extracted from KACC410442, and multi-loci sequences of the internal transcribed spacer region (ITS) rDNA, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), actin (actA), chitin synthase (CHS1), histone (HIS3) and tubulin (TUB2) genes were determined, as outlined by Cannon et al. (2012) and Damm et al. (2009). The resulting sequences obtained in this study were registered to GenBank (PP739190 for ITS, PP741985 for GAPDH, PP741986 for actA, PP741987 for CHS1, PP741988 for HIS3, and PP741989 for TUB2) and compared with other sequences on GenBank using the BLASTn search tool. The results showed 100% identity to other C. spaethianum sequences (e.g., MT611068 for ITS, MN400659 for actA, OP965007 for CHS1, OR060963 for GAPDH, PP480643 for HIS3, and OQ613727 for TUB2). In a maximum parsimony tree, the Korean isolate formed a well-supported clade (100% bootstrap value) with reference sequences of C. spaethianum. Pathogenicity was tested by spraying a conidial suspension (1×104 conidia/ml) from a two-week-old culture onto ten leaves of three potted plants, without wounding. Ten water-sprayed leaves were served as controls. The plants were maintained in a dew chamber and later moved to a greenhouse at 25 to 30 °C. After two weeks, all inoculated plants developed anthracnose symptoms, while the control plants remained symptomless. The fungus isolated from the inoculated plants was morphologically identical to that initially observed, fulfilling Koch's postulates. C. spaethianum has been known on various plant species, but mostly on the Asparagaceae, including Anemarrhena (Okorley et al. 2019), Hosta (Damm et al. 2009; Cheon and Jeon 2016), and Polygonatum (Liu et al. 2020; Ma et al. 2021). To our knowledge, this is the first report of anthracnose caused by C. spaethianum on P. odoratum var. pluriflorum in Korea, highlighting the continuous need for disease management and control in the cultivation of this plant.

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韩国由 Colletotrichum spaethianum 在 Polygonatum odoratum var.
Polygonatum odoratum var.它以茶闻名,含有多种药理活性化合物(Chang 等人,2007 年)。2021 年 9 月,在韩国春川(北纬 37°45'07'';东经 127°47'45'')和万州(北纬 35°38'47'';东经 127°31'16'')的玉竹变种 pluriflorum 上发现了以前从未见过的炭疽病,发病率占被检植株的 20% 至 30%。病害症状首先是叶片边缘发黄(萎黄),然后向叶片中央发展。随着病害的发展,叶片出现褐色坏死病斑,病斑上布满了小黑点,表明叶片上有 Colletotrichum 的分生孢子器。这种病通常会导致叶枯病,病斑上的分生孢子器呈周期性同心排列,在受害组织内形成环状图案。它在雨季迅速扩展,产生水渍状病斑,但在旱季几乎减缓,使植株外观破烂不堪。从万州采集的五个新鲜样本中获得了单孢子分离物,其中一个代表性分离物保存在韩国农业培养物保藏中心(KACC410442),干燥样本保存在全北国立大学标本室(JBNU0135)。用新鲜和自然感染的叶片检验真菌的形态特征。刚毛(n = 30)呈深褐色至近黑色,长 45 至 172 微米,基部宽 4.1 至 6.1 微米,向上变窄,先端尖,2 至 4 孔。分生孢子为单细胞,有时在发芽前为单隔膜,稍弯曲,先端锐尖或圆形,基部或多或少截形,直径 11.4-23.2 × 3.1-4.2 µm。外稃为单个或成群,深褐色,裂片状,形状不规则, 6.0-11.9 × 7.0-14.8 µm。在 25 ℃ 的 PDA 上生长两周的菌落直径达 30-35 毫米,最初为白色,随着时间的推移变为灰色,气生菌丝为棉状。这些形态特征与 C. spaethianum 的形态特征十分吻合(Damm 等,2009 年;Liu 等,2020 年)。为证实基于形态学的判定,从 KACC410442 提取了基因组 DNA,并按照 Cannon 等人(2012 年)和 Damm 等人(2009 年)的方法测定了内部转录间隔区(ITS)rDNA、3-磷酸甘油醛脱氢酶(GAPDH)、肌动蛋白(actA)、几丁质合成酶(CHS1)、组蛋白(HIS3)和微管蛋白(TUB2)基因的多序列。本研究获得的序列已登录到 GenBank(ITS 为 PP739190,GAPDH 为 PP741985,actA 为 PP741986,CHS1 为 PP741987,HIS3 为 PP741988,TUB2 为 PP741989),并使用 BLASTn 搜索工具与 GenBank 上的其他序列进行了比较。结果显示,与其他 C. spaethianum 序列(例如,ITS 为 MT611068,actA 为 MN400659,CHS1 为 OP965007,GAPDH 为 OR060963,HIS3 为 PP480643,TUB2 为 OQ613727)的同一性为 100%。在最大解析树中,韩国分离物与 C. spaethianum 的参考序列形成了一个支持良好的支系(100% 引导值)。致病性的测试方法是将两周培养的分生孢子悬浮液(1×104 个分生孢子/毫升)喷洒到三株盆栽植物的十片叶子上,不进行伤口处理。十片喷水的叶子作为对照。植物在露水室中养护,随后移至 25 至 30 °C 的温室中。两周后,所有接种的植株都出现了炭疽病症状,而对照植株仍无症状。从接种植物中分离出的真菌在形态上与最初观察到的真菌完全相同,符合科赫假说。已知 C. spaethianum 在多种植物物种上发生,但主要是在天门冬科植物上,包括 Anemarrhena(Okorley 等人,2019 年)、玉簪(Damm 等人,2009 年;Cheon 和 Jeon,2016 年)和 Polygonatum(Liu 等人,2020 年;Ma 等人,2021 年)。据我们所知,这是韩国首次报道由 C. spaethianum 在 P. odoratum var.
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来源期刊
Plant disease
Plant disease 农林科学-植物科学
CiteScore
5.10
自引率
13.30%
发文量
1993
审稿时长
2 months
期刊介绍: Plant Disease is the leading international journal for rapid reporting of research on new, emerging, and established plant diseases. The journal publishes papers that describe basic and applied research focusing on practical aspects of disease diagnosis, development, and management.
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