First report of basal rot of yellow dragon fruit (Selenicereus megalanthus) caused by Fusarium oxysporum in Peru.

IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Plant disease Pub Date : 2024-08-15 DOI:10.1094/PDIS-05-24-1060-PDN
Angel Fernando Huamán-Pilco, Marielita Arce-Inga, Jorge Huamán-Pilco, Vilma Aguilar-Rafael, Segundo Manuel Oliva-Cruz, Elgar Hernández-Diaz, Ysadora Fernández, Terry Jarianna Torres-Cruz, Jorge Ronny Díaz-Valderrama
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引用次数: 0

Abstract

Cultivation of yellow dragon fruit (Selenicereus megalanthus) in Peru has recently expanded (Verona-Ruiz et al. 2020). In August 2021, approximately 170 of 1,110 dragon fruit cuttings (15.3%) in the university's nursery (6°26'10'' S; 77°31'25'' W) showed basal rot symptoms. Initial symptoms included small brown spots on the base of stems, expanding towards the top that became soft and watery. All symptomatic plants eventually died, i.e., a severity of 100%. The disease was more prevalent on cuttings during the rooting phase than on well-established cuttings. We collected five symptomatic cuttings from throughout the nursery. Four sections of 1 × 1 cm2 of tissue adjacent to the diseased area were excised from each cutting, immersed for 1 min in 2% NaClO, rinsed twice with sterile distilled water, placed on potato dextrose agar (PDA) medium (four sections per Petri plate, five plates), and incubated at 25°C for 7 days. Morphologically similar mycelia grew from all sections, and five monosporic isolates were obtained, one per plate. Colonies grew fast, reaching 60 to 64 mm in 7 days, and produced violet-white cottony aerial mycelia with orange sporodochia on PDA, and abundant macro- and microconidia on synthetic nutrient-poor agar. Macroconidia were straight to slightly curved, typically with 2 to 3 septa, 16.6 to 23.3 × 1.7 to 3.7 µm (n = 30); microconidia were oval or kidney-shaped, and commonly hyaline, 6.7 to 16.4 × 2.5 to 4.7 µm (n = 40). Genomic DNA was extracted from isolate AFHP-100, then the ITS region and the TEF1 and RPB2 partial genes were amplified and sequenced (Accession numbers PP977433, OR437358, PP537149) following Gardes and Bruns (1993) and O'Donnell et al. (1998). We conducted a BLASTn search of ITS sequence against the NCBI "nr" database and local 'megablast' searches of TEF1 and RPB2 sequences against FUSARIUM-ID v.3.0 (Torres-Cruz et al. 2022). We found 100%, 98.19 to 99.84%, and 98.81 to 99.76% identities in ITS, TEF1, and RPB2 sequences, respectively, to the ex-epitype and other reference strains of Fusarium oxysporum (CBS 144134, NRRL26406, among others). A maximum likelihood phylogenetic analysis with a TEF1-RPB2 concatenated dataset with FUSARIUM-ID sequences also showed isolate AFHP-100 was F. oxysporum. A pathogenicity test was carried out by inoculating wounded healthy roots of three cuttings with submersion in a 5 × 106 conidia/ml suspension for 25 min. Then, the inoculated plants were planted in sterile soil. One cutting with wounded roots submerged in sterile water served as a control. In parallel, sterile soil was inoculated with 20 mL of the conidial suspension, and another three healthy cuttings were planted. A cutting planted in noninoculated soil also served as a control. Basal rot symptoms developed in all inoculated plants after 25 days. After re-isolation, the same fungus, corroborated based on micromorphology and TEF1 sequence (PP335689), was recovered, fulfilling Koch's postulates. The isolate was deposited in the KUELAP Herbarium (voucher KUELAP-3214), located and administered by the National University Toribio Rodriguez de Mendoza de Amazonas, in Chachapoyas, Peru. Fusarium oxysporum has been reported to cause basal stem rot in Bangladesh and Argentina (Mahmud et al. 2021; Wright et al. 2007), and stem blight in Malaysia (Mohd Hafifi et al. 2019) on dragon fruit. This is the first report of F. oxysporum causing basal rot in S. megalanthus in Peru. This fungus is among the most destructive plant pathogens, and the rapid expansion of the crop in Peru requires a comprehensive knowledge of the biotic factors influencing production. Therefore, this report is foundational to implementing proper control strategies.

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秘鲁首次报告由 Fusarium oxysporum 引起的黄火龙果(Selenicereus megalanthus)基腐病。
最近,秘鲁的黄火龙果(Selenicereus megalanthus)种植面积有所扩大(Verona-Ruiz 等人,2020 年)。2021 年 8 月,该大学苗圃(南纬 6°26'10'';西经 77°31'25'')中的 1,110 份火龙果插条中约有 170 份(15.3%)出现基腐症状。最初的症状包括茎基部出现褐色小斑点,并向顶部扩展,变得松软多水。所有出现症状的植株最终都死亡了,即严重程度达到 100%。这种病在生根期的插条上比在生长良好的插条上更为普遍。我们从整个苗圃中采集了五株有症状的插条。从每个插条上切除病变区域附近 1 × 1 cm2 的四片组织,在 2% 的 NaClO 中浸泡 1 分钟,用无菌蒸馏水冲洗两次,放在马铃薯葡萄糖琼脂(PDA)培养基上(每个培养皿四片,共五个培养皿),在 25°C 下培养 7 天。所有切片上都长出了形态相似的菌丝,并获得了五个单孢分离株,每个培养皿一个。菌落生长迅速,7 天内长到 60 至 64 毫米,在 PDA 上产生紫白色棉状气生菌丝和橙色孢子囊,在贫养分的合成琼脂上产生大量大孢子和小孢子。大锥体直到微弯,通常有 2 到 3 个隔膜,16.6 到 23.3 × 1.7 到 3.7 µm(n = 30);微锥体呈椭圆形或肾形,通常呈透明状,6.7 到 16.4 × 2.5 到 4.7 µm(n = 40)。从分离物 AFHP-100 中提取基因组 DNA,然后按照 Gardes 和 Bruns(1993 年)以及 O'Donnell 等人(1998 年)的方法扩增 ITS 区域以及 TEF1 和 RPB2 部分基因并进行测序(登录号 PP977433、OR437358 和 PP537149)。我们根据 NCBI "nr "数据库对 ITS 序列进行了 BLASTn 搜索,并根据 FUSARIUM-ID v.3.0 (Torres-Cruz 等,2022 年)对 TEF1 和 RPB2 序列进行了本地 "megablast "搜索。我们发现,ITS、TEF1 和 RPB2 序列分别与前表皮镰刀菌及其他 Fusarium oxysporum 参考菌株(CBS 144134、NRRL26406 等)具有 100%、98.19%-99.84% 和 98.81%-99.76%的相同性。利用 TEF1-RPB2 与 FUSARIUM-ID 序列的数据集进行的最大似然系统发生分析也表明,分离物 AFHP-100 是 F. oxysporum。致病性试验是通过将 5 × 106 个分生孢子/毫升的悬浮液浸泡 25 分钟后接种三株插条的健康伤根来进行的。然后,将接种的植株种植在无菌土壤中。将一根带伤的插条浸入无菌水中作为对照。同时,在无菌土壤中接种 20 毫升分生孢子悬浮液,并种植另外三株健康插条。在未接种土壤中种植的插条也作为对照。25 天后,所有接种的植株都出现了基腐病症状。重新分离后,根据微观形态学和 TEF1 序列(PP335689)证实,找到了相同的真菌,符合科赫假说。该分离物被存放在秘鲁查查波亚斯托里比奥-罗德里格斯-德-门多萨-德-亚马孙国立大学(National University Toribio Rodriguez de Mendoza de Amazonas)的 KUELAP 植物标本馆(凭证 KUELAP-3214)。据报道,氧孢镰刀菌(Fusarium oxysporum)在孟加拉国和阿根廷(Mahmud 等人,2021 年;Wright 等人,2007 年)可导致火龙果基部茎腐病,在马来西亚(Mohd Hafifi 等人,2019 年)可导致火龙果茎枯病。这是首次报道 F. oxysporum 在秘鲁导致 S. megalanthus 基腐病。这种真菌是最具破坏性的植物病原体之一,秘鲁火龙果作物的迅速发展需要全面了解影响生产的生物因素。因此,本报告对于实施适当的控制策略具有基础性意义。
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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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