Guan Y M, Lu B H, Wang Y, Gao J, Wu L J
Laboratory of Plant Pathology, Department of Agronomy, Jilin Agricultural University, Changchun, 130118, Jilin, China, and Institute of Special Wild Economic Animal and Plant Science, CAAS, Changchun, 130112, Jilin, China.
Laboratory of Plant Pathology, Department of Agronomy, Jilin Agricultural University, Changchun, 130118, Jilin, China.
Plant Dis. 2014 Jun;98(6):844. doi: 10.1094/PDIS-09-13-0922-PDN.
Ginseng (Panax ginseng C.A. Meyer) is a treasure of traditional Chinese medicine resources and has been used in food, health care products, and medicines in China for over 4,000 years. Jilin Province is the largest producing region in the world. Ginseng root rot disease caused by Fusarium spp. is the chief disease of ginseng with more than 25% incidence. Infected ginseng roots could not be used, which brought economic losses, and the species of the pathogens were not clear. The above-ground leaves of infected ginseng plants became red in the early stages and later wilted or died. All parts of root including rhizome, taproot, and fibrous root can be infected. Brown lesions were mainly formed on the surface and expanded gradually. Finally, multiple lesions converged and caused root rot. Diseased roots collected in June 2012 from 4-year-old ginseng (cv. Damaya) from Jingyu County and Jiaohe City, Jilin Province. Six isolates were obtained by cutting the roots into 5 mm pieces, disinfecting in 3% NaOCl for 3 min, and isolating on half-strength acidified potato dextrose agar (PDA). All isolates were purified from single spore. They produced white and cottony aerial mycelium cultured on PDA, microconidia were in the majority, oval or reniform, and length ranged from 5.97 to 12.01 μm. Macroconidia were in the minority, fusiform to conical, usually 3 to 5 septa, length ranged from 13.99 to 26.50 μm, thin-walled, with a hooked apical cell and a foot-shaped basal cell. A variety of structures of chlamydospores were produced, in chains or in pairs, basidixed or in the middle of the hyphae. Preliminary conclusions were made according to the morphological characteristics, which were consistent with the descriptions of Fusarium redolens (2). All the isolates of Fusarium spp. were accurately identified based on molecular biological methods of rDNA internal transcribed spacer (ITS) and the elongation factor (EF1-α) gene sequences analysis. ITS sequences analysis (GenBank Accession No. KF499035) resulted in a 100% match for one accession of F. redolens (X94169) by BLAST in the NCBI nucleotide database, and EF1-α gene (KF499036) had 100% similarity with 12 accessions of F. redolens (e.g., AF324319) by BLAST in the FUSARIUM-ID database (1). All isolates of F. redolens were tested for pathogenicity on ginseng root in vitro and healthy plants in a greenhouse, respectively. The surface of healthy ginseng roots were washed and disinfested in 70% alcohol for 3 min and plant in flower pots with sterile sand and sorghum grain infested with 14-day-old isolates and no inoculation as the control. Each experiment was repeated three times. Root rots were assessed 21 days after planting and all the isolates consistently caused root rot on inoculated plants, taproot and fibrous root exhibited brown to black root rot and plant wilting, whereas no root rot was observed in the controls and the pathogen was successfully re-isolated. The results were confirmed by the in vitro and healthy plant inoculation of ginseng root. F. redolens was identified previously as F. oxysporum, but recent molecular studies have shown that they are distant species in phylogenetic analysis (3). To our knowledge, this is the first report of F. redolens causing root rot of ginseng in China. References: (1) M. D. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) K. O'Donnell et al. Mycologia 90:465,1998.
人参(Panax ginseng C.A. Meyer)是中国传统医药资源中的瑰宝,在中国用于食品、保健品和药品已有4000多年历史。吉林省是世界上最大的人参产区。由镰刀菌属(Fusarium spp.)引起的人参根腐病是人参的主要病害,发病率超过25%。受感染的人参根无法使用,造成了经济损失,且病原菌种类尚不明确。受感染人参植株的地上叶片早期变红,随后枯萎或死亡。根的所有部分,包括根茎、主根和须根均可被感染。根表面主要形成褐色病斑,并逐渐扩展。最终,多个病斑融合导致根腐。2012年6月从吉林省靖宇县和蛟河市4年生人参(品种大马牙)上采集病根。将根切成5毫米小段,在3%次氯酸钠中消毒3分钟后,接种于半强度酸化马铃薯葡萄糖琼脂(PDA)上,获得6个分离株。所有分离株均经单孢纯化。在PDA上培养时,它们产生白色棉絮状气生菌丝,多数为小型分生孢子,椭圆形或肾形,长度为5.97至12.01微米。大型分生孢子较少,梭形至锥形,通常有3至5个隔膜,长度为13.99至26.50微米,薄壁,顶端细胞呈钩状,基部细胞呈足形。产生了多种结构的厚垣孢子,成链状或成对,着生于担子上或菌丝中部。根据形态特征初步判断,与变红镰刀菌(Fusarium redolens)的描述一致(2)。基于核糖体DNA内转录间隔区(ITS)和延伸因子(EF1-α)基因序列分析的分子生物学方法,对所有镰刀菌属分离株进行了精确鉴定。ITS序列分析(GenBank登录号KF499035)在NCBI核苷酸数据库中通过BLAST与变红镰刀菌(X94169)的一个登录号100%匹配,EF1-α基因(KF499036)在FUSARIUM-ID数据库中通过BLAST与12个变红镰刀菌登录号(如AF324319)100%相似(1)。分别对所有变红镰刀菌分离株进行了人参根的体外致病性测试和温室健康植株致病性测试。将健康人参根表面洗净,在70%乙醇中消毒3分钟,种植于装有无菌沙子和接种14日龄分离株的高粱粒的花盆中,不接种作为对照。每个实验重复三次。种植21天后评估根腐情况,所有分离株均在接种植株上一致引起根腐,主根和须根表现出褐色至黑色根腐以及植株枯萎,而对照中未观察到根腐,且病原菌成功再分离。通过人参根的体外和健康植株接种证实了结果。变红镰刀菌以前被鉴定为尖孢镰刀菌(F. oxysporum),但最近的分子研究表明,在系统发育分析中它们是远缘物种(3)。据我们所知,这是中国关于变红镰刀菌引起人参根腐病的首次报道。参考文献:(1)M. D. Geiser等人,《欧洲植物病理学杂志》110:473,2004年。(2)J. F. Leslie和B. A. Summerell,《镰刀菌实验室手册》,Blackwell出版社,艾姆斯,爱荷华州,2006年。(3)K. O'Donnell等人,《真菌学》90:465,1998年。