Stanosz G R, Kimbler D L
Departments of Plant Pathology and Forestry, University of Wisconsin, Madison 53706.
Plant Dis. 1997 Mar;81(3):311. doi: 10.1094/PDIS.1997.81.3.311A.
Shoot blight was observed on lodgepole pine (Pinus contorta Douglas ex Loud.) seedlings in fall 1994 at the USDA Forest Service Bessey Tree Nursery, Halsey, NE. Shoots were stunted, cankered, and sometimes curled at the tips, and bore brown needles that often had been killed before full elongation. Pycnidia were present on necrotic needles and stems. Conidia from these pycnidia produced pure cultures of Sphaeropsis sapinea (Fr.:Fr.) Dyko & Sutton in Sutton. The fungus was identified based on cultural, pycnidial, and conidial characteristics and comparison with known isolates. Also, random amplified polymorphic DNA markers of isolates were consistent with those of the A morphotype of S. sapinea (1). The ability of each of the two recognized morphotypes of S. sapinea to cause shoot blight of lodgepole pine was tested by inoculation of potted, 2-year-old seedlings in a greenhouse. Elongating terminal shoots were wounded by removing a needle pair approximately 1 cm below the shoot apex. A plug cut from an actively growing culture on water agar (WA) was placed fungus-side-down on the wound. The two isolates used (128, A morphotype; and 124, B morphotype) are representative of larger collections of these two morphotypes for which aggressiveness has been compared (2). Noncolonized WA plugs were placed on similarly wounded control seedlings. Nonwounded control seedlings also were used. Parafilm was wrapped around the shoots to hold the agar plugs in place and was removed 1 week later. Each treatment was applied to 10 seedlings in each of five completely randomized replicates. After 4 weeks, the condition (living or dead) of shoot tips was recorded, and the length of necrotic stem (canker) below the point of inoculation was measured to the nearest 0.25 cm. Segments of shoots were harvested, surface sterilized, and incubated to determine the presence of the pathogen. Analyses of variance were performed with Minitab for Windows version 10.1 software (Minitab Inc., State College, PA). Seedlings inoculated with either morphotype developed symptoms resembling those observed in the nursery. Neither incidence of shoot tip death nor severity (as indicated by canker length) differed greatly according to isolate morphotype. The means for number of shoot tips killed by the A and B isolates were 8.8 and 6.4 (out of 10), respectively (P = 0.07). The means for lengths of cankers produced by the A and B isolates were 4.9 and 3.8 cm, respectively (P = 0.10). Among controls, only one wounded seedling developed any symptoms. The pathogen was identified on incubated shoot segments from inoculated seedlings, but not those of control seedlings. This is the first report of S. sapinea as a cause of shoot blight of lodgepole pine in the U.S., and it associates the A morphotype with damage observed in the Bessey nursery. Further, the ability of the B morphotype to cause disease of lodgepole pine is established. Results of inoculations contrast with those obtained with red pine (Pinus resinosa Aiton) seedlings, on which the B morphotype was not aggressive (2). Susceptibility of lodgepole pine seedlings to both A and B morphotypes of S. sapinea, however, is similar to that exhibited by the related species, jack pine (P. banksiana Lamb.) (2). These species are classified in subsection "contortae" of the genus Pinus. Both morphotypes of S. sapinea should be considered as potential causes of shoot blight encountered on lodgepole pine. References: (1) G. R. Stanosz et al. Plant Dis. 80:1175, 1996. (2) J. T. Blodgett and G. R. Stanosz. Plant Dis. 81:143, 1997.
1994年秋季,在美国农业部林业局位于内布拉斯加州哈尔西的贝西树木苗圃中,黑松(Pinus contorta Douglas ex Loud.)幼苗上出现了梢枯病症状。病枝发育受阻,出现溃疡,梢尖有时卷曲,针叶呈褐色,且往往在完全伸长之前就已枯死。坏死的针叶和茎上出现了分生孢子器。从这些分生孢子器中分离得到的分生孢子在萨顿的培养基上培养出了纯的松球壳孢菌(Sphaeropsis sapinea (Fr.:Fr.) Dyko & Sutton)。该真菌是根据培养特征、分生孢子器特征和分生孢子特征,并与已知分离株进行比较而鉴定出来的。此外,分离株的随机扩增多态性DNA标记与松球壳孢菌A形态型的标记一致(1)。通过在温室中接种盆栽2年生幼苗,测试了松球壳孢菌两种公认形态型引起黑松梢枯病的能力。在伸长的顶梢上,于梢尖下方约1厘米处去除一对针叶造成伤口。从水琼脂(WA)上活跃生长的培养物上切下的菌块,菌面朝下放置在伤口上。所用的两个分离株(128,A形态型;124,B形态型)代表了这两种形态型的大量菌株收集,且已对其侵袭性进行了比较(2)。未接种的WA菌块放置在同样造成伤口的对照幼苗上。也使用了未受伤的对照幼苗。用Parafilm包裹梢部以固定琼脂菌块,1周后去除。每个处理在5个完全随机重复中,每个重复处理10株幼苗。4周后,记录梢尖的状况(存活或死亡),并测量接种点下方坏死茎(溃疡)的长度,精确到0.25厘米。采集梢段,进行表面消毒并培养,以确定病原体的存在。使用Minitab for Windows版本10.1软件(Minitab公司,宾夕法尼亚州州立学院)进行方差分析。接种任何一种形态型的幼苗都出现了与苗圃中观察到的症状相似的症状。根据分离株形态型,梢尖死亡发生率和严重程度(以溃疡长度表示)差异均不大。A和B分离株导致死亡的梢尖平均数分别为8.8和6.4(共10个)(P = 0.07)。A和B分离株产生的溃疡平均长度分别为4.9厘米和3.8厘米(P = 0.10)。在对照中,只有一株受伤的幼苗出现了任何症状。在接种幼苗的培养梢段上鉴定出了病原体,而对照幼苗的梢段上未鉴定出。这是美国关于松球壳孢菌作为黑松梢枯病病因的首次报道,并且将A形态型与在贝西苗圃中观察到的损害联系起来。此外,确定了B形态型引起黑松病害的能力。接种结果与红松(Pinus resinosa Aiton)幼苗的接种结果形成对比,在红松幼苗上B形态型没有侵袭性(2)。然而,黑松幼苗对松球壳孢菌A和B两种形态型的易感性与相关物种短叶松(P. banksiana Lamb.)相似(2)。这些物种都属于松属“contortae”亚组。松球壳孢菌的两种形态型都应被视为黑松上遇到的梢枯病的潜在病因。参考文献:(1)G. R. Stanosz等人,《植物病害》80:1175,1996年。(2)J. T. Blodgett和G. R. Stanosz,《植物病害》81:143,1997年。