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北美首次报道芍药生盘多毛孢引起牡丹枝枯病

First Report of Pestalotiopsis paeoniicola Causing Twig Blight on Paeonia suffruticosa in North America.

作者信息

Marra R E, Li D-W

机构信息

Department of Plant Pathology and Ecology, Connecticut Agricultural Experiment Station, 123 Huntington Street, New Haven.

Valley Laboratory, Connecticut Agricultural Experiment Station, 153 Cook Hill Road, Windsor.

出版信息

Plant Dis. 2009 Sep;93(9):966. doi: 10.1094/PDIS-93-9-0966C.

Abstract

Native to China, tree peony (Paeonia suffruticosa Andrews) is a perennial valued for its showy, often fragrant flowers. In May 2007, samples received from two field-grown tree peonies from Torrington, CT exhibited twig blight characterized by small, black spots on the bark of living or dead branches, and associated with subsequent death and loss of the branches. Colonies on potato dextrose agar (PDA) attained 5.5 to 6.5 cm in diameter in 9 days at 25°C and produced abundant, white, aerial mycelium, later turning pink and forming scattered, wet, black, acervular conidiomata containing abundant conidia that were five-celled, fusiform, smooth, straight, or slightly curved, and 24.0 ± 1.8 × 7.1 ± 0.5 μm (n = 20); three intermediate cells were dark brown and end cells were hyaline; two to four hyaline whip-like appendages on the apical cell, 26.6 ± 4.1 μm long, and one appendage on the basal cell, 6.7 ± 1.1 μm long. We identified the fungus as Pestalotiopsis paeoniicola (Tsukam. & T. Hino) J.G. Wei & T. Xu (= Pestalotia paeoniicola Tsukam. & T. Hino) based on morphology and host and have deposited a culture with CBS (CBS 124745). Originally described from Paeonia suffruticosa in Japan (1,3), the fungus is also found in China on Paeonia lactiflora (Pall.) (2). To confirm pathogenicity, two 5-year-old potted plants of Paeonia suffruticosa cv. Shichifukujin were inoculated with the fungus as follows: a 2-week-old PDA culture was used to produce a conidial spore suspension in sterile water; incisions (5 mm) were made with a sterile scalpel; 4 μl of either the conidial suspension or water were applied; and the wounds were wrapped with Parafilm. Each plant received three replicates each of the treatment and the control. Plants were loosely covered with plastic bags, kept on laboratory benches with ambient light for 1 month, and transferred to a greenhouse for an additional 2 months. Subsequent inspection revealed irregular or elongate, grayish brown lesions at the treatment inoculations, while the controls remained symptom free. Lesions gradually girdled the branches and the infected cortex turned dark brown. At advanced stages, small, black acervuli developed. Treatment and control tissues were cut into four pieces, surface sterilized, and placed on malt extract agar in petri dishes, four per dish. These were incubated for 9 days at 25°C under ambient light. Reisolation of P. paeoniicola only from tissues that had been treated with the pathogen, not from control inoculations, confirmed that the causal agent was P. paeoniicola. DNA sequences were obtained from the β-tubulin gene (1,512 bp) and the internal transcribed spacer (ITS1 and ITS2) and 5.8S regions of the rDNA (550 bp) and were deposited in GenBank (Nos. FJ975603 and FJ997645, respectively). Neither ITS nor β-tubulin sequences distinguished this P. paeoniicola isolate from Pestalotiopsis spp. whose sequences are deposited in GenBank. Morphological characteristics identify the fungus as P. paeoniicola, constituting, to our knowledge, the first report of this pathogen in North America. Discovery of P. paeoniicola in field-grown plants warrants further monitoring by growers because of the uncertain level of threat this pathogen may pose to the tree peony industry. References: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia: 224. Harvard University Press, Cambridge, MA, 1961. (2) F. Tai. Sylloge Fungorum Sinicorum: 1021. Sci. Acad. Sin, Peking, 1979. (3) E. Tsukamoto et al. Ann. Phytopathol. Soc. Jpn. 4:183, 1956.

摘要

牡丹(Paeonia suffruticosa Andrews)原产于中国,是一种多年生植物,因其艳丽且常具芳香的花朵而备受珍视。2007年5月,从美国康涅狄格州托灵顿市两棵田间种植的牡丹上采集的样本表现出嫩枝枯萎症状,其特征为活枝或死枝树皮上出现小黑斑,并伴随枝条随后死亡和脱落。在马铃薯葡萄糖琼脂(PDA)培养基上,菌落于25°C条件下9天内直径达到5.5至6.5厘米,产生大量白色气生菌丝,随后变为粉红色,并形成分散的、湿润的、黑色的分生孢子盘,其中含有大量分生孢子。分生孢子为五细胞,梭形,表面光滑,直或稍弯曲,大小为24.0 ± 1.8 × 7.1 ± 0.5 μm(n = 20);三个中间细胞为深褐色,两端细胞透明;顶端细胞有两到四条透明鞭状附属物,长26.6 ± 4.1 μm,基部细胞有一条附属物,长6.7 ± 1.1 μm。基于形态学和寄主,我们将该真菌鉴定为芍药拟盘多毛孢(Pestalotiopsis paeoniicola (Tsukam. & T. Hino) J.G. Wei & T. Xu = Pestalotia paeoniicola Tsukam. & T. Hino),并已将一份培养物保藏于荷兰真菌生物多样性中心(CBS 124745)。该真菌最初从日本的牡丹(Paeonia suffruticosa)上描述(1,3),在中国也发现于芍药(Paeonia lactiflora (Pall.))上(2)。为确认致病性,对两株5年生盆栽牡丹品种“七福神”进行如下接种:用两周龄的PDA培养物在无菌水中制备分生孢子悬浮液;用无菌手术刀切割(5毫米);分别接种4微升分生孢子悬浮液或水;并用石蜡膜包裹伤口。每株植物的处理组和对照组均设三个重复。植株用塑料袋松散覆盖,置于实验室台面上,在自然光下放置1个月,然后转移至温室再放置2个月。随后检查发现,处理接种部位出现不规则或细长的灰褐色病斑,而对照组无症状。病斑逐渐环绕枝条,受感染的皮层变为深褐色。在后期,形成小的黑色分生孢子盘。将处理组和对照组组织切成四块,表面消毒后置于培养皿中的麦芽提取物琼脂上,每皿四块。在25°C、自然光下培养9天。仅从接种病原菌的组织中重新分离到芍药拟盘多毛孢,而未从对照接种中分离到,这证实病原菌为芍药拟盘多毛孢。从β-微管蛋白基因(1,512 bp)以及核糖体DNA的内部转录间隔区(ITS1和ITS2)和5.8S区域(550 bp)获得了DNA序列,并分别保藏于GenBank(登录号分别为FJ975603和FJ997645)。ITS和β-微管蛋白序列均未将该芍药拟盘多毛孢分离株与保藏于GenBank中的拟盘多毛孢属其他种的序列区分开来。形态特征将该真菌鉴定为芍药拟盘多毛孢,据我们所知,这是该病原菌在北美的首次报道。由于该病原菌对牡丹产业可能构成的威胁程度尚不确定,种植者应对田间种植的牡丹植株中芍药拟盘多毛孢的发现进行进一步监测。参考文献:(1)E. F. Guba。《单毛孢属和盘多毛孢属专论》:224。哈佛大学出版社,马萨诸塞州剑桥,1961年。(2)戴芳澜。《中国真菌总汇》:1021。科学出版社,北京,1979年。(3)E. Tsukamoto等人。《日本植物病理学会会报》4:183,1956年。

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