Rosskopf E N, Hong J, Kokalis-Burelle N
USDA-ARS, United States Horticultural Research Laboratory, Fort Pierce, FL 34945.
Plant Dis. 2014 Jan;98(1):161. doi: 10.1094/PDIS-06-13-0672-PDN.
During the 2013 winter cut flower production season, a severe anthracnose epidemic was observed on Lupinus mutabilis (syn L. cruckshanksii) on a commercial flower farm in Martin County, FL. Approximately 50% of the crop was lost to the disease. Symptoms included dark brown, irregularly shaped leaf spots, but more typically, there was a single severe twist in the stem, forming a distinctive necrotic crook. Margins of necrotic lesions were excised and surface sterilized by immersion in 1% sodium hypochlorite for 90 s, rinsed in sterile deionized water, and plated onto potato dextrose agar (PDA). Plates were incubated at approximately 27°C with cycles of 12 h light/12 h darkness. Infected tissue consistently produced colonies that were typical of the genus Colletotrichum. Conidia were primarily oval, with one rounded end and one pointed end, and were highly variable in size, ranging from 10 to 15 μm in length and 3.5 to 5.5 μm in width. Cultures were gray with orange spots, and no setae were observed. These morphological characteristics are consistent with those of Colletotrichum lupini (2). Identification of this species was confirmed by performing a BLASTn search with ITS sequence data (primers ITS4 and ITS5), which shared 99% identity with GenBank submission AJ301968, C. lupini var. setosum strain BBA 71310, isolated from L. luteus in Poland. Inoculum was produced by flooding PDA cultures with sterile deionized water, scraping with a rubber policeman, and passing the suspension through four layers of sterile cheesecloth. This preparation was used to inoculate 10 L. mutabilis and 10 L. hartwegii plants by injecting 10 μl of a suspension of 10 conidia/ml into the stem using a hypodermic needle (1). Ten additional plants were injected with sterile deionized water and maintained with the inoculated plants in the greenhouse for 4 weeks. All of the inoculated plants developed the previously-observed necrotic crook in the stem, whereas control plants developed no symptoms. The same organism was isolated from all inoculated plants. The ITS region was again sequenced, and the Polish strain was the closest match. The Floridian isolate sequence was deposited in GenBank (KF207599). Epidemics of anthracnose on ornamental lupins are common in most areas in which they are grown. In 1939, research plots of L. angustifolius were found with symptoms of anthracnose caused by Glomerella cingulata (3). Although it is not possible to determine if this isolate would be redefined as C. lupini, it does not seem likely since pathogenicity was confirmed on L. angustifolius and L. albus, but it did not cause infection on L. luteus (3) as has been reported for C. lupini (2). The finding of a lupin anthracnose in southeastern Florida is important to both the cut flower producers as well as vegetable producers who might consider some species of Lupinus as potential green manure crops. To the best of our knowledge, this is the first report of C. lupini or any Colletotrichum species on L. hartwegii and L. mutabilis in Florida. References: (1) W. H. Elmer et al. Plant Dis 85:216, 2001. (2) H. I. Nirenger et al. Mycologia 94:307, 2002. (3) J. L. Weimer. Phytopathology 43:249, 1943.
在2013年冬季切花生产季节,佛罗里达州马丁县的一个商业花卉农场里,羽扇豆(Lupinus mutabilis,同物异名L. cruckshanksii)上出现了严重的炭疽病疫情。约50%的作物因病损失。症状包括深褐色、形状不规则的叶斑,但更典型的是,茎部有一处严重扭曲,形成一个独特的坏死弯钩。切除坏死病斑边缘,浸入1%次氯酸钠中表面消毒90秒,用无菌去离子水冲洗,然后接种到马铃薯葡萄糖琼脂(PDA)平板上。平板在约27°C下培养,光照/黑暗周期为12小时/12小时。感染组织持续产生典型炭疽菌属的菌落。分生孢子主要为椭圆形,一端圆形,一端尖形,大小差异很大,长度为10至15μm,宽度为3.5至5.5μm。培养物呈灰色并有橙色斑点,未观察到刚毛。这些形态特征与羽扇豆炭疽菌(Colletotrichum lupini)一致(2)。通过对ITS序列数据(引物ITS4和ITS5)进行BLASTn搜索,确认了该物种的鉴定,其与GenBank提交的AJ301968、羽扇豆炭疽菌变种刚毛刺孢菌株BBA 71310(从波兰的窄叶羽扇豆中分离)有99%的同一性。通过用无菌去离子水淹没PDA培养物,用橡皮刮铲刮取,然后将悬浮液通过四层无菌粗棉布来制备接种物。使用皮下注射针将10μl浓度为10个分生孢子/ml的悬浮液注射到茎中,以此对10株多变色羽扇豆和10株哈氏羽扇豆进行接种(1)。另外10株植物注射无菌去离子水,并与接种的植物一起在温室中培养4周。所有接种的植物在茎部都出现了之前观察到的坏死弯钩,而对照植物没有症状。从所有接种的植物中都分离出了相同的生物体。再次对ITS区域进行测序,发现与波兰菌株最匹配。佛罗里达分离株的序列已存入GenBank(KF207599)。观赏羽扇豆上的炭疽病疫情在其种植的大多数地区都很常见。1939年,在窄叶羽扇豆的研究地块上发现了由围小丛壳(Glomerella cingulata)引起的炭疽病症状(3)。虽然无法确定该分离株是否会被重新定义为羽扇豆炭疽菌,但似乎不太可能,因为其对窄叶羽扇豆和白羽扇豆的致病性已得到证实,但对黄羽扇豆没有引起感染(3),而羽扇豆炭疽菌对此有报道(2)。在佛罗里达州东南部发现羽扇豆炭疽病,对切花生产者以及可能将某些羽扇豆物种视为潜在绿肥作物的蔬菜生产者都很重要。据我们所知,这是佛罗里达州关于哈氏羽扇豆和多变色羽扇豆上羽扇豆炭疽菌或任何炭疽菌属物种的首次报道。参考文献:(1)W. H. Elmer等人,《植物病害》85:216,2001年。(2)H. I. Nirenger等人,《真菌学》94:307,2002年。(3)J. L. Weimer,《植物病理学》43:249,1943年。