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中国由变红镰刀菌引起的西洋参根腐病。

American Ginseng Root Rot Caused by Fusarium redolens in China.

作者信息

Fan Sanhong, Zhao Fangjie, Zhang Jiguang, Shang Wenjing, Hu Xiaoping

机构信息

Northwest A&F University, 12469, College of Life Science, Yangling, Shaanxi, China;

Northwest A&F University, 12469, College of Plant Protection, Yangling, Shaanxi, China;

出版信息

Plant Dis. 2021 Jan 13. doi: 10.1094/PDIS-12-20-2600-PDN.

Abstract

American ginseng (Panax quinquefolium L.) originating from North America is one of important herbal medicine and economic crops . With the increasing market demand, China has become the third producer and the largest consumer country of American ginseng. However, continuous cropping obstacle has become the most serious problem for the production of American ginseng, and the continuous cropping of soils usually lead to accumulations of root fungal pathogens and increasing plant disease occurrence (1), root rot caused by the notorious soil-borne pathogenic fungi, Fusarium spp., results in a significant reduction of yield and quality of American ginseng. Investigation of American ginseng root rot was carried out in Liuba county, Shaanxi province, China from 2017 to 2019. About 20% of over 3-year-old American ginseng showed varied root rot symptoms in newly reclaimed fields, and more than 70% in continuous American ginseng planting fields. Among these root rot diseases, we found one kind of disease which shows symptoms of red leaves in initial stage and yellow or yellow brown lesions at the reed heads or taproots. The lesions mainly appear on the root surface; however, the vascular tissue has no discoloration. The aboveground parts become wilted and died, and the whole root appears dark brown rots. Fifteen Fusarium spp. isolates were obtained by cutting diseased rot roots into 5 × 5 mm2 pieces, disinfecting in 70% ethanol for 1 min, rinsing 2 ~ 3 times in sterile water for 1 min and isolating on PDA medium including 50 μg/mL streptomycin sulfate. All the isolates have identical morphological characteristics. The colony was white with curved and uplifted aerial hyphae in central region. The colony diameter was 48 ~51 mm after 6 days at room temperature. Microconidia were oval to cylindrical shape with 0 to 1 septa, ranged from 6.24 to10.09 μm long; the macroconidia were fusiform to conical with a hooked apical cell and a foot-shaped basal cell, usually 3 to 5 septa, ranged from 31.45 to 42.52 μm long. The chlamydospores were not found under our culture condition. Preliminary data analysis showed that the morphological characteristics of these isolates were consistent with the descriptions of Fusarium redolens (2). To clarify the fungus in the taxonomy , the rDNA internal transcribed spacer (ITS), the translation elongation factor 1 alpha (TEF1-α) and the RNA polymerase II subunit 1 (RPB1) fragments of two randomly selected isolates were amplified and sequenced. The sequences of the corresponding fragments of the two isolates were identical. The blast results in the GenBank and FUSARIUM-ID databases show the isolates belong to F. redolens (3). Previous study indicated F. redolens has an indistinguishable relative, F. hostae (4). Although the ITS sequence (MW331695) cannot provide enough information to distinguish them, the phylogenetic tree combined the sequence of TEF1-α (tempID: 2407237 ) and RPB1 (tempID: 2407229) clearly showed that the isolates are F. redolens. (Fig) The pathogenicity of a representative isolate, YP04, was tested on ginseng taproot by in vivo inoculation experiments with three replications. The taproot surface of 2-year-old healthy ginseng was washed and disinfested with 75% alcohol for 1 min and rinsed with sterile water, and dried. The surface of taproot was injured with sterilized steel needles and immersed in 1 × 106 /ml spore suspension (sterile water for control plants) for 30 min. The treatment and control plants were transplanted in 20 cm diameter flowerpots filled with sterilized humus and cultured in a greenhouse at 18-23°C. Six days after transplanting, the leaves began to turn red. The cortex of ginseng taproot showed yellow brown lesions and the vascular tissue turn to light yellow. Fifteen days after transplanting, the aboveground parts of treatment plants began to wilting and the taproots showed serious rots. no taproot rot was observed in the controls. The pathogen was re-isolated from the diseased taproots successfully. To our knowledge, this is the first report of F. redolens causing root rot of American ginseng in China.

摘要

原产于北美的西洋参(Panax quinquefolium L.)是重要的草药和经济作物之一。随着市场需求的增加,中国已成为西洋参的第三大生产国和最大消费国。然而,连作障碍已成为西洋参生产中最严重的问题,土壤连作通常会导致根际真菌病原体积累,植物病害发生率增加(1),由臭名昭著的土传致病真菌镰刀菌属引起的根腐病,导致西洋参产量和品质大幅下降。2017年至2019年在中国陕西省留坝县对西洋参根腐病进行了调查。在新开垦的田地中,超过3年生的西洋参约20%表现出不同程度的根腐症状,而在连作西洋参种植田中,这一比例超过70%。在这些根腐病中,我们发现了一种病害,其初期症状为红叶,芦头或主根出现黄色或黄褐色病斑。病斑主要出现在根表面;然而,维管束组织没有变色。地上部分枯萎死亡,整个根呈现深褐色腐烂。将患病腐烂根切成5×5 mm2的小块,在70%乙醇中消毒1分钟,在无菌水中冲洗23次,每次1分钟,然后在含有50μg/mL硫酸链霉素的PDA培养基上分离,获得了15株镰刀菌属分离物。所有分离物具有相同的形态特征。菌落白色,中部气生菌丝弯曲且隆起。室温下培养6天后,菌落直径为4851 mm。小型分生孢子椭圆形至圆柱形,有0至1个隔膜,长6.24至10.09μm;大型分生孢子梭形至圆锥形,顶端细胞钩状,基部细胞足形,通常有3至5个隔膜,长31.45至42.52μm。在我们的培养条件下未发现厚垣孢子。初步数据分析表明,这些分离物的形态特征与变红镰刀菌(Fusarium redolens)的描述一致(2)。为了在分类学上明确该真菌,对随机选择的两个分离物的核糖体DNA内部转录间隔区(ITS)、翻译延伸因子1α(TEF1-α)和RNA聚合酶II亚基1(RPB1)片段进行了扩增和测序。两个分离物相应片段的序列相同。在GenBank和FUSARIUM-ID数据库中的比对结果表明,这些分离物属于变红镰刀菌(3)。先前的研究表明,变红镰刀菌有一个难以区分的近缘种,即霍氏镰刀菌(F. hostae)(4)。虽然ITS序列(MW331695)不能提供足够的信息来区分它们,但结合TEF1-α序列(临时编号:2407237)和RPB1序列(临时编号:2407229)构建的系统发育树清楚地表明,这些分离物是变红镰刀菌。(图)通过体内接种实验对一个代表性分离物YP04在人参主根上的致病性进行了测试,重复三次。将2年生健康人参的主根表面洗净,用75%酒精消毒1分钟,用无菌水冲洗,然后晾干。用消毒后的钢针损伤主根表面,并将其浸入1×106 /ml的孢子悬浮液(对照植株用无菌水)中30分钟。处理植株和对照植株移栽到装满消毒腐殖质的直径20 cm的花盆中,在18-23°C的温室中培养。移栽6天后,叶片开始变红。人参主根皮层出现黄褐色病斑,维管束组织变为淡黄色。移栽15天后,处理植株的地上部分开始枯萎,主根出现严重腐烂。对照植株未观察到主根腐烂。从患病主根中成功重新分离出病原体。据我们所知,这是中国首次报道变红镰刀菌引起西洋参根腐病。

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