Yang S Y, Su S C, Liu T, Fan G, Wang J, Leng P S
Laboratory for Silviculture and Conservation Ministry, Beijing Forestry University, Bejing, 100083, China.
Department of Landscape, Beijing Agricultural College, Beijing, 102206, China.
Plant Dis. 2011 Oct;95(10):1314. doi: 10.1094/PDIS-04-11-0269.
In the 1990s, pistachio (Pistacia vera L. 'Kerman' and 'Peters') was introduced in China. They are found in many orchards in Beijing and Gansu and Hebei provinces, northern China. In 2009, a new disease was observed on leaves, stems, and fruits in pistachio orchards in Gansu Province. Disease incidence in 8- to 12-year-old orchards was 30%. Yield losses reached 25%. Symptoms began as discrete, sunken, black spots, approximately 10 mm in diameter, followed by circular lesions that eventually coalesced with tissue death recorded and orange fructifications developed on lesions. Pieces of diseased leaves, stems, and fruits were surfaced sterilized and placed on 2% potato dextrose agar (PDA) at 25°C. A fungus was consistently isolated. After 10 days, cultures on PDA showed aerial, white mycelium that turned gray to grayish black with a salmon-to-orange conidial mass at 25°C and a 12-h photoperiod. Brown, 80 to 120 μm long setae were observed in the acervulus. Conidia were hyaline, fusiform to nearly straight, and averaged 12 to 18 × 3 to 5 μm. On the basis of morphological characteristics, the fungus was identified as Colletotrichum gloeosporioides (Penz.) Sacc. (2). On PDA, 0.5 μg/ml of benomyl was applied for the sensitivity test (3). Benomyl completely inhibited the growth of the fungus. Mycelial DNA was extracted, PCR amplified using ITS1 and ITS4 primers for the ribosomal DNA internal transcribed spacers 1 and 2, and sequenced. The DNA sequence was recorded in GenBank as No. HQ631378. The DNA sequence was blasted showing 99% identity with Accession Nos. GQ144454 and GU004376, for C. gloeosporioides. Pathogenicity tests were conducted under greenhouse conditions at 25°C. Three replicates of 2-year-old 'Kerman' plants were inoculated with mycelial PDA plugs placed on 0.5-cm stem wounds and then wrapped with Parafilm. Controls were inoculated with PDA plugs without the fungus. After 3 weeks, stem cankers were observed on inoculated plants. Control plants remained healthy. Pathogenicity was also tested on injured leaves and fruits. A 10-μl drop of a spore suspension of 10 conidia/ml was applied on 'Kerman' and 'Peters' leaves and 'Kerman' fruits and placed on plates with a wet filter paper at 25°C. Small, black lesions were observed at 2 days after inoculation. At 7 days, necrotic lesions covered the entire surface. C. gloeosporioides was reisolated from necrotic lesions. Controls did not develop symptoms. C. acutatum has been reported on pistachio in Australia (1), but to our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on pistachio. References: (1) G. J. Ash and V. M. Lanoiselet. Australas. Plant Pathol. 30:365, 2001. (2) J. Y. Lu. Plant Pathogenic Mycology. China Agricultural Press, Beijing, 2001. (3) N. A. R. Peres et al. Plant Dis. 86:620, 2002.
20世纪90年代,阿月浑子(黄连木属‘克尔曼’和‘彼得斯’品种)被引入中国。在中国北方的北京、甘肃和河北省的许多果园都有种植。2009年,在甘肃省的阿月浑子果园中,叶片、茎干和果实上发现了一种新病害。8至12年生果园的发病率为30%。产量损失达25%。症状开始表现为离散的、凹陷的黑色斑点,直径约10毫米,随后形成圆形病斑,最终病斑合并导致组织死亡,并在病斑上产生橙色分生孢子盘。将病叶、病茎和病果的组织进行表面消毒后,置于25℃的2%马铃薯葡萄糖琼脂(PDA)培养基上。持续分离出一种真菌。10天后,PDA培养基上的培养物在25℃、12小时光照周期条件下,长出气生白色菌丝体,后变为灰色至灰黑色,带有鲑鱼色至橙色的分生孢子团。在分生孢子盘中观察到棕色、长80至120微米的刚毛。分生孢子无色透明,梭形至近直形,平均大小为12至18×3至5微米。根据形态特征,该真菌被鉴定为胶孢炭疽菌(Penz.)Sacc.(2)。在PDA培养基上,用0.5微克/毫升的苯菌灵进行敏感性测试(3)。苯菌灵完全抑制了该真菌的生长。提取菌丝体DNA,使用核糖体DNA内部转录间隔区1和2的ITS1和ITS4引物进行PCR扩增并测序。该DNA序列在GenBank中记录为编号HQ631378。经比对,该DNA序列与胶孢炭疽菌的登录号GQ144454和GU004376的序列一致性达99%。在25℃的温室条件下进行致病性测试。将2年生的‘克尔曼’植株分成3组,每组接种放置在0.5厘米茎伤口上的PDA培养基菌丝块,然后用Parafilm包裹。对照组接种不含真菌的PDA培养基块。3周后,在接种植株上观察到茎溃疡。对照植株保持健康。还对受伤的叶片和果实进行了致病性测试。将10个分生孢子/毫升的孢子悬浮液10微升滴在‘克尔曼’和‘彼得斯’叶片以及‘克尔曼’果实上,并置于25℃、铺有湿滤纸的培养皿中。接种2天后观察到小的黑色病斑。7天后,坏死病斑覆盖整个表面。从坏死病斑中再次分离出胶孢炭疽菌。对照未出现症状。在澳大利亚,曾报道过阿月浑子上的尖孢炭疽菌(1),但据我们所知,这是首次报道胶孢炭疽菌引起的阿月浑子炭疽病。参考文献:(1)G. J. Ash和V. M. Lanoiselet。《澳大利亚植物病理学》30:365,2001。(2)J. Y. Lu。《植物病原真菌学》。中国农业出版社,北京,2001。(3)N. A. R. Peres等人。《植物病害》86:620,2002。