泰国首次报告 Colletotrichum siamense 在菠萝上引起叶炭疽病。

IF 4.4 2区 农林科学 Q1 PLANT SCIENCES
Surapong Khuna, Jaturong Kumla, Tanapol Thitla, Chanokned Senwanna, Nakarin Suwannarach
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The internal transcribed spacer (ITS) region, actin (act), β-tubulin (tub2), calmodulin (CAL), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified using primer pairs ITS5/ITS4, ACT-512F/ACT-783R, T1/T22, CL1C/CL2C, and GDF1/GDR1, respectively (White et al. 1990; Weir et al. 2012). Sequences were deposited in GenBank (ITS: PP068858, PP068859, PP446789, PP446790; act: PP079636, PP079637, PP460760, PP460761; tub2: PP079638, PP079639, PP460762, PP460763; CAL: PP079634, PP079635, PP460758, PP460759; GAPDH: PP079632, PP079633, PP460756, PP460757). Maximum likelihood phylogenetic analyses of the concatenated five genes identified all isolates as C. siamense. To pathogenicity test, the mature leaves of a healthy plant were surface disinfested using 0.1% NaClO for 3 min, rinsed three times with sterile water, and wounded. 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引用次数: 0

摘要

菠萝蜜(Artocarpus heterophyllus Lam.)2023 年 6 月,在泰国清迈府 Chai Prakan 县(北纬 19°42'24",东经 99°01'59")的一块田地里观察到这种植物的叶炭疽病,在 1000 平方米的种植区中发病率约为 25%。初期症状为带黄晕的褐色斑点,扩大、拉长,直径 0.2 至 2 厘米,不规则,凹陷,褐色,带深褐色晕,叶片枯萎干枯。在高湿度条件下,病斑上长出淡黄色的分生孢子器。通过单孢子分离法(Tovar-Pedraza et al.)获得了四种形态相似的真菌分离物(SDBR-CMU492 至 SDBR-CMU495)。马铃薯葡萄糖琼脂(PDA)上的菌落直径为 70 至 85 毫米,白色至灰白色,带有棉状菌丝,在 25°C 下培养 1 周后,菌落呈淡黄色。所有分离物均产生无性结构。刚毛呈棕色,有 1 至 3 个隔膜,长 40 至 100 × 2.2 至 4.0 µm,基部呈圆柱形,顶端渐尖。分生孢子梗呈透明至淡褐色,有隔膜,有分枝。分生孢子细胞呈透明至淡褐色,圆柱形至瓶形, 7.4 - 27.2 × 2.0 - 4.5 µm。分生孢子单细胞,透明,壁光滑,圆柱形,末端圆形,具沟, 11.1-15.7 × 3.4-6.1 µm。外稃呈深褐色至黑色,椭圆形至不规则形,8.8 至 24.9 × 3.6 至 10 微米。从形态上看,所有分离物都与 Colletotrichum gloeosporioides 种群相似(Weir 等人,2012 年)。使用引物对 ITS5/ITS4、ACT-512F/ACT-783R、T1/T22、CL1C/CL2C 和 GDF1/GDR1 分别扩增了内部转录间隔区(ITS)、肌动蛋白(act)、β-微管蛋白(tub2)、钙调蛋白(CAL)和甘油醛-3-磷酸脱氢酶(GAPDH)基因(White 等,1990 年;Weir 等,2012 年)。序列已存入 GenBank(ITS:PP068858, PP068859, PP446789, PP446790; act:PP079636, PP079637, PP460760, PP460761; tub2: PP079638, PP079639, PP460762, PP460763; CAL:PP079634、PP079635、PP460758、PP460759;GAPDH:PP079632、PP079633、PP460756、PP460757)。通过对五个基因的最大似然系统发生分析,确定所有分离物均为 C. siamense。在致病性试验中,用 0.1% 的 NaClO 对健康植物的成熟叶片进行表面消毒 3 分钟,然后用无菌水冲洗 3 次,并进行伤口处理。在 25°C 的 PDA 上生长 2 周的每种分离株的分生孢子悬浮液(15 µl 含 1 × 106 个分生孢子/ml)被用来按附着法接种受伤和未受伤的样本。对照叶片用无菌蒸馏水模拟接种。每种处理进行 10 次重复,重复两次。将植物置于温度为 25 至 30°C、相对湿度为 80 至 90% 的温室中。7 天后,所有接种的叶片都出现褐色病斑,而对照叶片则没有症状。从 PDA 上的接种组织中重新分离出了暹罗毛霉菌(Colletotrichum siamense),从而完成了科赫假说(Koch's postulates)。在本研究之前,C. fructicola 和 C. gloeosporioides 在全球范围内引起了菠萝的叶炭疽病(Sangchote 等人,2003 年;Chitambar,2016 年)。澳大利亚(James 等人,2014 年)和 Bazil(Borges 等人,2023 年)报道了由 C. siamense 引起的胡柚叶炭疽病。在泰国,Bhunjun 等人(2019 年)报道了 C. artocarpicola 在菠萝中引起的叶炭疽病。因此,这是泰国首次报道 C. siamense 在菠萝上引起叶炭疽病。这一发现将为流行病学调查和未来管理这种疾病的方法提供信息。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
First Report of Colletotrichum siamense Causing Leaf Anthracnose on Jackfruit in Thailand.

Jackfruit (Artocarpus heterophyllus Lam.) is commonly grown in Thailand. In June 2023, leaf anthracnose on this plant was observed at a field in Chai Prakan District (19°42'24"N, 99°01'59"E), Chiang Mai Province, Thailand, with ~25% disease incidence in a 1000-m2 plantation area. The initial symptom had brown spots with a yellow halo, enlarged, elongated, 0.2 to 2 cm in diameter, irregular, sunken, brown, with a dark brown halo, and leaves withered and dried. Pale yellow conidiomata developed on the lesions in high humidity. Ten symptomatic leaves were used to isolate the fungal causal agents through a single spore isolation method (Tovar-Pedraza et al. 2020). Four fungal isolates (SDBR-CMU492 to SDBR-CMU495) with similar morphology were obtained. Colonies on potato dextrose agar (PDA) were 70 to 85 mm in diameter, white to grayish white with cottony mycelia, the reverse pale yellow after incubation at 25°C for 1 week. All isolates produced asexual structures. Setae were brown with 1 to 3 septa, 40 to 100 × 2.2 to 4.0 µm, a cylindrical base, and acuminate tip. Conidiophores were hyaline to pale brown, septate, and branched. Conidiogenous cells were hyaline to pale brown, cylindrical to ampulliform, 7.4 to 27.2 × 2.0 to 4.5 µm. Conidia were one celled, hyaline, smooth walled, cylindrical, ends rounded, guttulate, 11.1 to 15.7 × 3.4 to 6.1 µm. Appressoria were dark brown to black, oval to irregular, 8.8 to 24.9 × 3.6 to 10 µm. Morphologically, all isolates resembled the Colletotrichum gloeosporioides species complex (Weir et al. 2012). The internal transcribed spacer (ITS) region, actin (act), β-tubulin (tub2), calmodulin (CAL), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified using primer pairs ITS5/ITS4, ACT-512F/ACT-783R, T1/T22, CL1C/CL2C, and GDF1/GDR1, respectively (White et al. 1990; Weir et al. 2012). Sequences were deposited in GenBank (ITS: PP068858, PP068859, PP446789, PP446790; act: PP079636, PP079637, PP460760, PP460761; tub2: PP079638, PP079639, PP460762, PP460763; CAL: PP079634, PP079635, PP460758, PP460759; GAPDH: PP079632, PP079633, PP460756, PP460757). Maximum likelihood phylogenetic analyses of the concatenated five genes identified all isolates as C. siamense. To pathogenicity test, the mature leaves of a healthy plant were surface disinfested using 0.1% NaClO for 3 min, rinsed three times with sterile water, and wounded. Conidia suspensions (15 µl of 1 × 106 conidia/ml) of each isolate grown on PDA at 25°C for 2 weeks were used to inoculate wounded and unwounded samples by the attached method. Control leaves were mock inoculated with sterile distilled water. Ten replications were conducted for each treatment and repeated twice. Plants were placed in a greenhouse at 25 to 30°C and 80 to 90% relative humidity. After 7 days, all inoculated leaves displayed brown lesions, while control leaves had no symptoms. Colletotrichum siamense was reisolated from inoculated tissues on PDA to complete Koch's postulates. Prior to this study, C. fructicola and C. gloeosporioides caused leaf anthracnose on jackfruit worldwide (Sangchote et al. 2003; Chitambar 2016). Leaf anthracnose on jackfruit caused by C. siamense has been reported from Australia (James et al. 2014) and Bazil (Borges et al. 2023). In Thailand, Bhunjun et al. (2019) reported that C. artocarpicola causes leaf anthracnose in jackfruit. Therefore, this is first report of C. siamense causing leaf anthracnose on jackfruit in Thailand. The finding will inform epidemiological investigations and future approaches to managing this disease.

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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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