台灣葡萄晚腐病發生調查及其主要病原Colletotrichum viniferum之病原性與親緣關係

Occurence of Grape Ripe Rot in Taiwan and the Pathogenicity and Phylogenetic Relationship of Its Primary Causal Agent Colletotrichum viniferum

林筑蘋(Chu-Ping Lin) ； 王智立(Chih-Li Wang) ； 蔡志濃(Jyh-Nong Tsai) ； 戴裕綸(Yu-Lun Dai) ； 安寶貞(Pao-Jan Ann) ； 詹月梅(Yue-Mei Zhan) ； 黃順源(Shun-Yuan Huang)

《台灣農業研究》 71卷2期 (2022/06) Pp. 135-157

https://doi.org/10.6156/JTAR.202206_71(2).0004

摘要

由Colletotrichum spp.引起之葡萄晚腐病（grape ripe rot）為影響台灣葡萄產業最重要的病害之一，為更瞭解引發台灣葡萄晚腐病之病原菌的主要種類、親緣關係、病徵及田間感染分布狀況，本研究自2012-2020年間於葡萄主要產區採集呈現典型葡萄晚腐病徵之葡萄果實、具壞疽病斑之枝條或葉片，藉由組織分離得Colletotrichum spp.分離株共40株。利用internal transcribed spacer（ITS）、lyceraldehyde-3-phosphatedehydrogenase（GAPDH）、actin（ACT）、beta-tubulin 2 （TUB2）以及chitin synthase 1（CHS-1）等5個基因建立多重序列親緣關係樹，結果顯示自罹病果實分離之28個分離株主要為Colletotrichum viniferum，並可分成兩個次支序群（subclade），屬於次支序群T菌株分離比例高達85.7%，屬於次支序群C的菌株僅占3.6%；其他菌株分別為Colletotrichum fructicola（7.1%）及Colletotrichum tropicale（3.6%）。C. viniferum次支序群T菌株分布範圍包括苗栗縣卓蘭鎮、台中市新社區與霧峰區、彰化縣大村鄉以及南投縣信義鄉、國姓鄉、水里鄉與埔里鎮等葡萄主要產地；調查結果也顯示可自田間罹病葉片上分離到C. viniferum。田間葡萄幼果接種試驗結果顯示C. viniferum次支序群T之菌株可自幼果期潛伏感染果實，並於果實轉色初期至成熟時產生典型晚腐病徵；溫室葡萄盆栽接種試驗證明C. viniferum次支序群T之菌株可引起葉片壞疽，病徵多自葉緣一端開始發展，葉片壞疽周圍常伴隨大範圍黃暈，嚴重時造成落葉，部分菌株可引起葉脈褐化。此外，藉人工接種試驗亦觀察到葡萄卷鬚壞疽與花穗壞疽等病徵。從田間調查與病原性試驗結果可確認C. viniferum次支序群T之菌株可於葡萄果實、葉片、卷鬚與花穗等組織造成多型態的病徵。

關鍵字

次支序群 ； 葉部病徵 ； 多基因序列親緣關係 ； 葡萄晚腐病 ； Colletotrichum viniferum

並列摘要

Grape ripe rot caused by Colletotrichum spp. is one of the most devastating diseases for grape production in Taiwan. To reveal population structures, phylogenetic relationships, symptoms, and geographic distribution of the causal organisms in the field, grape samples with the typical ripe rot symptoms on fruits, and necrotic lesions on twigs and leaves were collected and investigated. Moreover, symptoms from field samples and inoculated plants were also recorded. From 2012 to 2020, a total of 40 Colletotrichum isolates were isolated and identified with the phylogenetic analysis based on the concatenated sequences of internal transcribed spacer (ITS), lyceraldehyde-3-phosphatedehydrogenase (GAPDH), actin (ACT), beta-tubulin 2 (TUB2), and chitin synthase 1 (CHS-1) genes. The phylogenetic tree showed that grape ripe rot in Taiwan was mainly caused by Colletotrichum viniferum, which was divided into two subclades, subclade T and subclade C. Among 28 isolates that were isolated from diseased fruits of grape ripe rot, 85.7% of these isolates belonged to C. viniferum subclade T, 3.6% to C. viniferum subclade C, 7.1% to Colletotrichum fructicola, and 3.6% to Colletotrichum tropicale. The results also showed that C. viniferum subclade T could be isolated not only from diseased fruits but also from symptomatic leaves. The pathogens are distributed in most major grape-growing areas, including Zhuolan in Miaoli County, Xinshe and Wufeng in Taichung City, Dacun in Changhua County, and Xinyi, Guoxing, Shuili and Puli in Nantou County. When inoculated on young fruits in planta in the field, the isolates of C. viniferum subclade T induced the typical ripe rot symptoms on grapefruits after veraison, suggesting that the pathogens were able to establish a latent infection. When inoculated on whole potted plants in a greenhouse, isolates of C. viniferum subclade T caused various symptoms on leave, such as necrotic lesions with a large area of yellow halo on the leave margin, defoliation, and browning leaf veins. Furthermore, necrosis on shoots and blossoms was also observed. The results from field investigations and artificial inoculations indicate that the isolates of C. viniferum subclade T could cause various symptoms on fruits, leaves, shoots, and blossoms of grapes.

並列關鍵字

Subclade ； Foliar symptoms ； Multilocus phylogeny