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由铁杉球座菌引起的北美乔松在佐治亚州的枝枯病

Shoot Blight Caused by Sirococcus tsugae on Eastern Hemlock (Tsuga canadensis) in Georgia.

作者信息

Stanosz G R, Smith D R, Sullivan J P, Mech A M, Gandhi K J K, Dalusky M J, Mayfield A E, Fraedrich S W

机构信息

Department of Plant Pathology, University of Wisconsin-Madison 53706.

Georgia Forestry Commission, Gainesville 30507.

出版信息

Plant Dis. 2011 May;95(5):612. doi: 10.1094/PDIS-11-10-0833.

Abstract

Eastern hemlock (Tsuga canadensis) is an ecologically and economically important conifer from the north-central United States to the east coast of North America to the southern Appalachian Mountains. In early spring 2010, blighted shoot tips of eastern hemlock were observed at widely separated locations in the Chattahoochee National Forest in north Georgia. Damage did not appear to be directly related to hemlock woolly adelgid (Adelges tsugae) activity, which was sporadic or absent in some areas where symptoms were observed. A preliminary survey in March 2010 revealed that incidence of blighted shoots on individual trees varied, but was as high as 70%. Stems of shoots produced the previous year were frequently necrotic, had lost needles, and bore pycnidia with hyaline, two-celled conidia consistent with those of Sirococcus tsugae (2,3). Later in the spring and summer, shoots of the current year's growth became blighted, with sporulation of S. tsugae also on dead and dying needles. While S. tsugae previously has been reported on T. heterophylla, T. mertensiana, Cedrus atlantica, and C. deodara in western North America, it has only recently been reported on eastern hemlock (1), and its ability to induce shoot blight has not been proven. Pure cultures (2,3) were obtained on streptomycin-amended potato dextrose agar (PDA) and their identity was confirmed by species-specific PCR primers (4). Nuclear rDNA internal transcribed spacer sequence (554 nucleotides) also was obtained for isolate 10-05 and deposited in GenBank (Accession No. HQ256769). This sequence was found to be identical to sequences previously deposited for S. tsugae isolates. Isolate 10-05 and a second isolate (10-06) were used to inoculate potted 2-year-old eastern hemlock seedlings in a growth chamber at 20°C with a 16-h photoperiod. Conidia were collected by flooding 1-month-old colonies on PDA with sterile water. Expanding shoots on one branch of each seedling were wounded using scissors to cut the tips off needles and stems, while another branch remained nonwounded. Ten seedlings per isolate were inoculated by spraying to runoff with a suspension of 5 × 10 conidia ml in sterile water, and five similarly treated control seedlings were sprayed with sterile water. Seedlings were covered with plastic bags to maintain high humidity for 4 days. Germination of conidia of each isolate incubated on water agar in this growth chamber was >80% after 24 h. Symptoms were evaluated and reisolation was attempted on streptomycin-amended PDA 2 months after inoculation. Symptoms of seedlings inoculated with either isolate included chlorotic and necrotic needle spots, browning of cut edges of needles, browning and death of needle tips and entire needles, death of stem tips with retention of dead needles, and needle loss. Symptoms of control seedlings were limited to slight browning of cut edges of needles. The fungus was reisolated from wounded shoots of 17 of 20 inoculated seedlings and nonwounded shoots of 5 of 20 inoculated seedlings and was not cultured from control seedlings. To our knowledge, this is the first report of S. tsugae in Georgia and also the first demonstration of its ability to produce symptoms that have been attributed to it on any tree species. References: (1) M. Miller-Weeks and W. Ostrofsky. USDA. Forest Service. Online publication. NA-PR-01-10, 2010. (2) A. Y. Rossman et al. For. Pathol. 38:47. (3) D. R. Smith et al. For. Pathol. 33:141, 2003. (4) D. R. Smith and G. R. Stanosz. For. Pathol. 38:156, 2008.

摘要

东部铁杉(加拿大铁杉)是一种在生态和经济方面都很重要的针叶树,其分布范围从美国中北部延伸至北美东海岸,直至南部的阿巴拉契亚山脉。2010年早春,在佐治亚州北部查塔胡奇国家森林中相距甚远的不同地点,观察到了东部铁杉受枯萎的梢尖。损害似乎与铁杉球蚜(铁杉球蚜)的活动没有直接关系,在一些观察到症状的地区,铁杉球蚜的活动是零星的或不存在的。2010年3月的初步调查显示,单株树木上枯萎梢的发生率各不相同,但高达70%。前一年长出的嫩枝茎干常常坏死,针叶脱落,并带有分生孢子器,其中有透明的、双细胞的分生孢子,与云杉球座菌的分生孢子一致(2,3)。在春季和夏季后期,当年生长的嫩枝也出现枯萎,在枯死和即将枯死的针叶上也有云杉球座菌的孢子形成。虽然之前在北美西部的异叶铁杉、墨氏铁杉、大西洋雪松和喜马拉雅雪松上已报道过云杉球座菌,但它最近才在东部铁杉上被报道(1),而且其诱发嫩枝枯萎的能力尚未得到证实。在添加链霉素的马铃薯葡萄糖琼脂(PDA)上获得了纯培养物(2,3),并通过物种特异性PCR引物(4)确认了其身份。还获得了分离株10 - 05的核糖体DNA内部转录间隔区序列(554个核苷酸),并保存在GenBank中(登录号HQ256769)。发现该序列与之前为云杉球座菌分离株保存的序列相同。分离株10 - 05和第二个分离株(10 - 06)被用于在生长室中对盆栽的2年生东部铁杉幼苗进行接种,生长室温度为20°C,光周期为16小时。通过用无菌水淹没PDA上1个月大的菌落来收集分生孢子。用剪刀剪掉每株幼苗一个枝条上正在伸展的嫩枝梢尖和茎尖使其受伤,而另一个枝条保持未受伤。每个分离株接种10株幼苗,用5×10个分生孢子/毫升的无菌水悬浮液喷雾至径流,另外5株经过类似处理的对照幼苗用无菌水喷雾。用塑料袋覆盖幼苗以保持高湿度4天。在该生长室中,接种在水琼脂上的每个分离株的分生孢子在24小时后萌发率>80%。接种2个月后,在添加链霉素的PDA上评估症状并尝试进行再分离。接种任何一个分离株的幼苗症状包括褪绿和坏死的针斑、针叶切口边缘变褐、针叶尖端和整个针叶变褐和死亡、茎尖死亡但枯死的针叶仍保留以及针叶脱落。对照幼苗的症状仅限于针叶切口边缘轻微变褐。从20株接种幼苗中的17株受伤嫩枝和20株接种幼苗中的5株未受伤嫩枝上重新分离到了该真菌,对照幼苗上未培养出该真菌。据我们所知,这是云杉球座菌在佐治亚州的首次报道,也是其在任何树种上产生已归因于它的症状的能力的首次证明。参考文献:(1)M. Miller - Weeks和W. Ostrofsky。美国农业部。森林服务局。在线出版物。NA - PR - 01 - 10,2010年。(2)A. Y. Rossman等人。《森林病理学》38:47。(3)D. R. Smith等人。《森林病理学》33:141,2003年。(4)D. R. Smith和G. R. Stanosz。《森林病理学》38:156,2008年。

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