Cheng B P, Lu L M, Peng A T, Song X B, Ling J F, Chen X
Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences and Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Guangzhou 510640, China.
Zhejiang Citrus Research Institute, Taizhou 318020, China.
Plant Dis. 2014 Jun;98(6):845. doi: 10.1094/PDIS-09-13-0951-PDN.
Citrus reticulata Blanco cv. Nian Ju, an important ornamental plant, is traditionally displayed during the Chinese Spring Festival because its golden fruits are a symbol of auspiciousness. In the spring of 2012, foliar blight was observed on 10 to 30% of the Nian Ju plants at four nurseries in Yangjiang, Guangdong Province, China. Initial symptoms appeared as brown to black foliar lesions, followed by expansion of spots into blight. Some young branches also had necrosis. During frequent rainfall and prolonged wet periods at 22°C to 30°C, white and dense mycelia and sporangia were observed on the infected seedlings. To isolate the causal organism, leaves and stems were cut into sections. Each section included some partial lesion and adjacent asymptomatic tissues. They were surface-disinfested in 1% sodium hypochlorite for 60 s, rinsed three times with sterile water, and placed on V8 juice agar (V8A) at 25°C. After 3 days, 10 isolates were obtained and purified by single-zoospore method. These isolates were identified to species level by sequencing the rRNA internal transcribed spacer (ITS) region. Four representative isolates had an identical ITS sequence (GenBank Accession No. KF750568), which had 99% homology with Phytophthora capsici sequences in GenBank. In addition, all recovered isolates were identical in morphological characteristics. They produced caducous, papillate, and ovoid to ellipsoid sporangia (Length × width = 46.2 ± 7.7 × 23.6 ± 11.3 μm), often with a tapered base. The average length of pedicels was 33.3 ± 4.5 μm. All isolates are A2 mating type. They produced gametangia when paired with an A1 tester of P. capsici isolated from pepper on V8A. Plerotic oospores were 25.3 ± 2.1 μm in diameter. Amphigynous antheridia were 13.6 ± 2.8 μm long and 11.2 ± 0.9 μm wide. Oogonia were 27.4 ± 3.2 μm in diameter. To determine the pathogenicity, three 3-year-old potted C. reticulata cv. Nian Ju plants were sprayed with 20 ml of zoospore suspension from one representative isolate at 10 per ml. Two control plants were sprayed with 20 ml distilled water. All plants were then maintained at 90% relative humidity at 25°C with a 12-h photoperiod. Symptoms similar to those observed in the nurseries developed on all inoculated plants but not on any control plants after 10 days. The pathogenicity test was repeated once and similar results were obtained. P. capsici was recovered from all inoculated plants and resultant isolates had identical morphology to that of the isolates used for inoculation. P. capsici has a relatively broad host range including pumpkins, cucumbers, peppers, beans, squashes, and spinach (1,2). To our knowledge, this is the first report of foliar blight of C. reticulata cv. Nian Ju caused by P. capsici. This study indicates that P. capsici is potentially an important pathogen of C. reticulata cv. Nian Ju plants and further investigations into its epidemiology and development of site-specific integrated management programs for this new disease are warranted. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. American Phytopathological Society, St. Paul, MN, 1996. (2) D. Tian and M. Babadoost. Plant Dis. 88:485. 2004.
年桔(Citrus reticulata Blanco cv. Nian Ju)是一种重要的观赏植物,传统上在中国春节期间摆放,因为其金黄色的果实是吉祥的象征。2012年春季,在中国广东省阳江市的4个苗圃中,10%至30%的年桔植株出现了叶枯病。最初的症状表现为叶片上出现褐色至黑色的病斑,随后病斑扩大形成叶枯。一些嫩枝也出现坏死。在22°C至30°C的频繁降雨和长时间潮湿条件下,在受感染的幼苗上观察到白色且密集的菌丝体和孢子囊。为分离致病生物,将叶片和茎切成小段。每段包括一些部分病斑和相邻的无症状组织。将它们在1%次氯酸钠中进行表面消毒60秒,用无菌水冲洗3次,然后置于25°C的V8汁琼脂(V8A)上。3天后,获得了10个分离物,并通过单游动孢子法进行纯化。通过对核糖体RNA内部转录间隔区(ITS)区域进行测序,将这些分离物鉴定到种的水平。4个代表性分离物具有相同的ITS序列(GenBank登录号KF750568),与GenBank中辣椒疫霉(Phytophthora capsici)的序列具有99%的同源性。此外,所有回收的分离物在形态特征上是相同的。它们产生脱落性、具乳突、卵形至椭圆形的孢子囊(长×宽 = 46.2 ± 7.7 × 23.6 ± 11.3 μm),基部通常渐细。梗的平均长度为33.3 ± 4.5 μm。所有分离物均为A2交配型。当与从辣椒中分离的辣椒疫霉A1测试菌株在V8A上配对时,它们产生配子囊。满器卵孢子直径为25.3 ± 2.1 μm。雌雄同丝雄器长13.6 ± 2.8 μm,宽11.2 ± 0.9 μm。藏卵器直径为27.4 ± 3.2 μm。为确定致病性,对3株3年生盆栽年桔植株喷洒来自一个代表性分离物的20 ml游动孢子悬浮液,每毫升含10个游动孢子。2株对照植株喷洒20 ml蒸馏水。然后将所有植株在25°C、相对湿度90%、光周期为12小时的条件下培养。10天后,所有接种植株都出现了与苗圃中观察到的类似症状,而对照植株未出现任何症状。致病性测试重复了一次,获得了类似的结果。从所有接种植株中都分离到了辣椒疫霉,所得分离物的形态与用于接种的分离物相同。辣椒疫霉具有相对广泛的寄主范围,包括南瓜、黄瓜、辣椒、豆类、南瓜和菠菜(1,2)。据我们所知,这是关于辣椒疫霉引起年桔叶枯病的首次报道。本研究表明,辣椒疫霉可能是年桔植株的一种重要病原菌,有必要对其流行病学以及针对这种新病害制定特定地点的综合管理方案进行进一步研究。参考文献:(1)D. C. Erwin和O. K. Ribeiro。《世界疫霉病》。美国植物病理学会,明尼苏达州圣保罗,1996年。(2)D. Tian和M. Babadoost。《植物病害》88:485。2004年。