Mauzey S J, Koike S T, Bull C T
Hartnell Community College, Salinas, CA and California State University, Monterey Bay, Undergraduate Research Opportunities Center, Seaside 93955.
University of California Cooperative Extension, Salinas 93901.
Plant Dis. 2011 Jan;95(1):71. doi: 10.1094/PDIS-09-10-0642.
In 2008, field-grown cabbage (Brassica oleracea var. capitata L., cv. Grenader) from Monterey County, California showed symptoms on the wrapper leaves of immature plants that had formed heads. Initial symptoms consisted of small, brown, water-soaked flecks surrounded by chlorotic haloes. These flecks later coalesced into large, irregularly shaped, brown-black, necrotic lesions with chlorotic haloes visible on both adaxial and abaxial surfaces of the leaf. This disease resulted in lower quality and reduced marketability of the cabbage. Five gram-negative, blue-green fluorescing bacteria were isolated from separate lesions on different plants on King's medium B agar. The isolates were positive for levan formation and negative for oxidase and arginine dihydrolase. The isolates did not cause soft rot on potato slices but did induce a hypersensitive reaction in tobacco (Nicotiana tabacum L. cv. Samsun). These data indicated that the bacteria belonged to Lelliot's LOPAT group 1 (2). Repetitive extragenic palindromic sequence (rep)-PCR using the BOXA1R primer resulted in identical DNA fragment banding patterns for the cabbage isolates and the pathotype of Pseudomonas cannabina pv. alisalensis (formerly P. syringae pv. alisalensis). Additionally, both P. cannabina pv. alisalensis and the five cabbage isolates were sensitive to bacteriophage PBS1 while the pathotype strain of P. syringae pv. maculicola was not (1). Pathogenicity of the five cabbage isolates was evaluated in two independent experiments. Inoculum was prepared by growing the bacteria on nutrient agar for 48 h (27°C), suspending the bacteria in 0.01 M phosphate buffer (pH 7.0), adjusting each suspension to 0.6 OD at 600 nm (approximately 10 CFU/ml), and adding three to five drops of Tween 20. In each experiment, two cabbage, broccoli raab (Brassica rapa subsp. rapa cv. Sorrento), and oat (Avena sativa cv. Montezuma) plants were inoculated for each isolate by spraying until runoff. Positive control plants were inoculated with the pathotype strain of P. cannabina pv. alisalensis and negative control plants were treated with sterile 0.01 M phosphate buffer. The plants were placed in a mist chamber for 48 to 72 h and then in a greenhouse (20 to 25°C). After 7 to 10 days, foliar symptoms similar to symptoms observed on the original diseased cabbage plants developed on all inoculated plants including the positive control plants inoculated with P. cannabina pv. alisalensis. Additionally severe symptoms on broccoli raab and minor symptoms on oats developed on plants inoculated with cabbage strains or P. cannabina pv. alisalensis. For each experiment, bacteria reisolated from symptomatic tissue were identical to the bacteria used to inoculate the plants and to P. cannabina pv. alsialensis for rep-PCR DNA fragment banding pattern and sensitivity to phage PBS1. There were no symptoms on any of the cabbage and oat negative controls. Additionally, there were no symptoms on any broccoli raab negative controls in the first experiment; however, in the second experiment, a small (<1 mm) lesion was detected on one leaf of one plant. To our knowledge, this is the first report of P. cannabina pv. alisalensis causing bacterial blight of cabbage in California. This disease may have significant impact because of the large acreage of cabbage grown in California (approximately 5,666 ha annually). References: (1) C. T. Bull et al. Syst. Appl. Microbiol. 33:105, 2010. (2) R. A. Lelliott. J. Appl. Bacteriol. 29:470, 1966.
2008年,加利福尼亚州蒙特雷县田间种植的甘蓝(Brassica oleracea var. capitata L.,品种Grenader)在已形成叶球的未成熟植株的外层叶片上出现症状。最初症状为小的、褐色的、水渍状斑点,周围有褪绿晕圈。这些斑点随后融合成大的、形状不规则的、棕黑色坏死斑,叶片的正面和背面均可见褪绿晕圈。这种病害导致甘蓝品质下降,市场适销性降低。从不同植株上的单独病斑中,在King氏培养基B琼脂上分离出5株革兰氏阴性、蓝绿色荧光细菌。这些分离菌株产果聚糖呈阳性,氧化酶和精氨酸双水解酶呈阴性。这些分离菌株在土豆片上不引起软腐,但在烟草(Nicotiana tabacum L.,品种Samsun)上能引发过敏反应。这些数据表明这些细菌属于Lelliot的LOPAT第1组(2)。使用BOXA1R引物进行的重复外显子回文序列(rep)-PCR,甘蓝分离菌株和大麻假单胞菌丁香致病变种(Pseudomonas cannabina pv. alisalensis,以前称为丁香假单胞菌丁香致病变种P. syringae pv. alisalensis)的致病型产生相同的DNA片段条带模式。此外,大麻假单胞菌丁香致病变种和5株甘蓝分离菌株对噬菌体PBS1敏感,而丁香假单胞菌斑点致病变种的致病型菌株则不敏感(1)。在两项独立实验中评估了5株甘蓝分离菌株的致病性。通过在营养琼脂上于27°C培养细菌48小时来制备接种物,将细菌悬浮于0.01M磷酸盐缓冲液(pH 7.0)中,将每种悬浮液在600nm处调整至0.6 OD(约10CFU/ml),并加入三至五滴吐温20。在每个实验中,通过喷雾直至径流,对每种分离菌株接种两株甘蓝、两株抱子芥(Brassica rapa subsp. rapa,品种Sorrento)和两株燕麦(Avena sativa,品种Montezuma)植株。阳性对照植株接种大麻假单胞菌丁香致病变种的致病型菌株,阴性对照植株用无菌0.01M磷酸盐缓冲液处理。将植株置于雾室中48至72小时,然后置于温室(20至25°C)。7至10天后,在所有接种植株上,包括接种了大麻假单胞菌丁香致病变种的阳性对照植株上,出现了与原始患病甘蓝植株上观察到的症状相似的叶部症状。此外,接种甘蓝菌株或大麻假单胞菌丁香致病变种的植株上,抱子芥出现严重症状,燕麦出现轻微症状。对于每个实验,从有症状组织重新分离出的细菌,在rep-PCR DNA片段条带模式和对噬菌体PBS1的敏感性方面,与用于接种植株的细菌以及大麻假单胞菌丁香致病变种相同。所有甘蓝和燕麦阴性对照均无症状。此外,在第一个实验中,所有抱子芥阴性对照均无症状;然而,在第二个实验中,在一株植物的一片叶子上检测到一个小的(<1mm)病斑。据我们所知,这是大麻假单胞菌丁香致病变种在加利福尼亚州引起甘蓝细菌性叶枯病的首次报道。由于加利福尼亚州种植的甘蓝面积较大(每年约5666公顷),这种病害可能具有重大影响。参考文献:(1)C. T. Bull等人,《系统与应用微生物学》33:105,2010年。(2)R. A. Lelliott,《应用细菌学杂志》29:470,1966年。