Exploitation of Techniques for Detecting Harmful Organisms on Imported Wood

長喙殼菌類真菌為世界上最重要之林木病害來源，包括了Ophiostoma及Ceratocysts屬及其無性繁殖態，為防範此類病原真菌藉由木材輸入而危害我國之森林原生樹種，並評估此類病原之風險，本計劃將針對進口木材及本土林木中長喙殼菌類真菌進行偵測，建立菌種形態特性及DNA序列資料庫，提供快速且正確檢測木材中此類有害真菌之基礎。除此之外，櫟樹猝死病菌亦是當前世界重要之森林病菌，本研究將針對其可能藉木材輸入之傳播途徑進行監測，以杜絕其危害。潛在病原菌之偵測及鑑定包括形態及分子序列方式，依據不同之菌種類別之特性，以特有之方式進行菌種分離，針對特定rDNA片段進行解序，建立形態及分子鑑定標準，有助於發展快速之檢測方法，攔截有害病原之輸入。Ophiostomatoid fungi are the most important forest pathogens in the world. Species of Ophiostoma and Ceratocystis and their anamorph genera are collectively referred to as the ophiostomatoid fungi. To prevent the introduction of these pathogens from plaguing our natural trees, and to evaluate their risk, the aims of this project is to inspect ophiostomatoid fungi on wood which is from importation or from indigenous plants. Morphological characteristics and DNA sequences of these fungi can be established to provide the basis for their accurate and quick detection. Additionally, Sudden Oak Death, caused by Phytophthora ramorum, is currently a serious problem in many countries. This pathogen will also be the major target in our inpection. Both methods of morphology identification and DNA sequence comparison will be applied. Specific approaches of isolation will be adopted to obtain specific groups of fungi. Specific primers are used to amplify specific DNA fragments for comparisons with DNA sequences deposited in GenBank. Based on the morphological and molecular data acquired, it can be expected that these potential pathogens can be warded off from our country

Systematic Studies on Pleosporales in Taiwan

隔孢菌目子囊菌為微小卻普遍存在之真菌，有重要的植物病原菌種類、亦有重要分解者之腐生性種類，當人們現今轉向生物資源尋找食品、工業或醫藥之可利用成分時，此類真菌必也佔有其潛在有用之角色。而唯有當正確的物種分類鑑定，及建立基礎的物種資料庫後，才有可能對目標生物做進一步的利用，甚至族群遺傳研究、生理生化成分分析及生態調查。位處亞熱帶的台灣具有多樣之植物相與生態環境，意味著正有無數之未知種類待我們來發掘及紀錄，本研究除了將修訂既有的紀錄種類外，亦將致力於探索不同之受質及生態環境，以期得到最多隔孢菌目子囊菌之種類，並以台灣之物種為起點，對重要及常見之屬進行系統之分類整理，在世界性隔孢菌目之分類亟需重整之際，此台灣種類之加入將能釐清許多分類上的問題。所有採集得到之菌株將進行純化培養並保存，以為永續使用之資源、探索有性無性世代聯結、及做分子系統分類之比對來佐證形態分類之結果。所有處理之菌種將以清晰之圖鑑呈現，以供分類鑑定之參考

Systematic Studies on Pleosporales in Taiwan (II)

生物之多樣性除了能維繫生態系之永續發展外，也提供了人類未來應用之潛在資源，而生物多樣性研究之基礎則在於物種之正確鑑定，這即需要一洽當而可遵行之分類系統。隔孢菌目子囊菌為微小卻普遍存在之真菌，有重要的植物病原菌種類、亦有重要分解者之腐生性種類，當人們現今轉向生物資源尋找食品、工業或醫藥之可利用成分時，此類真菌必也佔有其潛在有用之角色。然而此類真菌之鑑定主要依據少數之形態特徵，先前則因過度強調部分之分類特徵而導致了有爭議之結果，近年來之研究證據已說明了需對其進行分類修正之必要。本研究首先即是對台灣隔孢菌目子囊菌做廣泛之調查，相信在台灣的多樣植物相與生態環境中，必有無數之未知種類待我們來發掘，研究將致力於探索不同之受質，以期得到最多隔孢菌目子囊菌之種類。其次，隨著收集菌株的增加，將深入探討隔孢菌目之系統分類，結合分子及形態之特徵來釐清其中界定不清之屬間之關係，主要針對常見之四屬Astrosphaeriella、Lophiostoma、Massarina 及Trematosphaeria，而分子親緣關係樹之建構將使用ITS/5.8S、28S rDNA、與GPD 序列。所有採集得到之菌株將進行純化培養並保存，除提供進一步探索有性無性世代聯結外，更可為未來使用之資源。所有處理之菌種將以清晰之圖鑑呈現，以供分類鑑定之參考

Systematic Studies on Massarina and Morphologically Similar Genera

Massarina 屬真菌之定義在於具有雙層壁之子囊，其內含有無色、具隔膜、紡錘形之孢子。而相同之孢子形態亦用來定義Lophiostoma 及Lophiotrema 二屬，導致此三屬之分類一直紛擾不清。以致於當真菌學者遇到此無色具隔膜孢子及雙層壁子囊之真菌時，多僅能以Massarina-like 真菌稱呼之，因為幾乎無法鑑定到種(甚至屬名)，而Massarina-like 真菌之鑑定問題也就一直困擾著真菌學者。此類真菌廣泛分布於世界各地，數量亦非常龐大，已含有750 個種名，而台灣僅有3 種之文獻記載。近來研究得到18 種之Massarina-like 真菌，但幾乎皆無法鑑定，其皆很可能為新種，然而在Massrina-like 真菌之分類問題獲得解決之前，這些種類的鑑定是難以得到答案的。所以本研究即是希望能解決此群真菌之分類問題，包含了兩大目標，一是更廣泛的收集Massarina-like 真菌之菌種，其二是運用所收集之菌種進行形態與分子親緣之分析，並整合二者之結果。在分子親緣分析方面，將使用不同之基因片段，包含ITS, LSU rDNA,SSU rDNA 以及partial RPB2，預期研究結果將能使此複雜之Massarina-like 真菌做更明確之分群，而也會有新屬之建立來對應。研究結果也將提供各物種之完整圖鑑，將使真菌學者不再受Massarina-like 真菌之鑑定問題困擾。The genus Massarina is characterized by bitunicate asci with hyaline, septate andfusiform to ellipsoid ascospores surrounded by a mucilaginous sheath. Lophiostoma andLophiotrema possess the same definition of ascospores. Consequently, species in thesegenera are actually in a mixture. However, mycologists, who encounter ascomycetes withbitunicate asci and hyaline septate ascospores, usually can not find the names for them andmerely attribute them as Massarina-like ascomycetes, because it is difficult to identify themto species, even genus level. Identification of Massarina-like fungi always frustratesmycologists. Species of Massarina-like species are ubiquitous and mainly saprophytic onwood. There are 18 Massarina-like species obtained in Tawian, most of which can not beassigned to any extant species, and are considered to be new species. These species isunlikely to have names before the classification of Massarina-like fungi is clarified. To solvethe problems exist in the identification of Massarina-like fungi. There are two aims of thisproject. The first aim is to conduct further comprehensive survey of Massarina-like fungi inTaiwan. The second aim is to investigate the systematics of Massarina-like fungi with thecombination of morphology and molecular phylogeny To investigate the phylogeneticrelationships of Massarina-like fungi, four different genes (ITS, LSU rDNA, SSU rDNA andpartial RPB2) will be sequenced and analyzed. It is expected that Massarina-like fungi canbe further divided into more subgroups, which merit new genera. Afterwards, by providingdetail and clear photos of these fungi, the problems in identifying Massarina-like fungi willnever confuse mycologists

Development of the Detection and Treatment Techniques for Imported Plant Products

從進口木材及原始林木中檢測出有害昆蟲及真菌，且探討昆蟲與其傳播之真菌病害間之關聯，建立檢疫檢測昆蟲及真菌之模式，獲得進口原木及我國原始林木中昆蟲及真菌種類之資料。利用穩定性同位素技術，分析不同地區來源及不同部位之碳與氮穩定同位素含量，以開發檢測梨接穂來源之技術，期能有效查緝未經檢疫之走私接穂，杜絕外來病蟲害污染。釐清「榴槤夜蛾」(Durian borer)相關昆蟲之系統分類，並確認各種取食榴槤夜蛾成蟲與幼蟲之正確鑑別方式，以助於鑑定並評估其對台灣農業與生態環境之風險，及評估相關昆蟲種類是否可能對台灣的農業與生態系造成危害。針對寄主範圍相當廣泛之翠菊黃萎病菌質體（aster yellows phytoplasma group ,AY group)，以PCR技術，發展篩檢健康活植物苗之技術，以防堵此一病原菌之入侵，俾維護台灣之農業產業。Comprehensive survey of harmful insects and fungi from imported wood and indigenous forest is conducted, and the acquired organisms are documented. Information regarding the connection between insect vectors and fungi can therefore be established. The model of detecting harmful organisms can therefore be obtained, which is very helpful in operating quarantine insepection. Isotope technique, including carbon and nitrogen isotope composition analyses, is applied to identify the source of pear scion for quarantine purpose. Pear twigs and grafting scions from different areas are assayed in an attempt to establish a technique to track out the sources, and then recognize those imported from legal or illegal ways. The result will not only provide an standard for quarantine operation, but also reduce the impact of introduced harmful organisms. Clarification of the systematic positions of durian borers and their relevant species is undertaken. Correct methods to identify adults and larvae of different durian borer species will subsequently be established. The proper information can therefore be provided to assess the risk of impact of these insects on the agriculture of Taiwan. To prevent the introduction of Aster yellows (AY) phytoplasmas into Taiwan, a accurate and sensitive approach is needed to detect the AY phytoplasma from imported seedlings. A PCR-based detection system will be developed in this project to coincide with the quarantine requirements

Studies on Biodiversity and Systematics of Loculoascomycetes in Taiwan

和歐美溫帶國家相比，地處亞熱帶地區且高山林立之台灣擁有無盡之菌類資源，然而我們對其研究仍明顯不足，當現今人們越來越重視生態之保育與永續利用時，對我國本土菌類資源之調查就更顯重要。普遍存在之腔室子囊菌綱之真菌是亟待開發之領域，不僅在於其種類繁多可豐富我國之生物多樣性，更在於其存在許多分類之問題有待解決，所以本研究即是先針對台灣之腔室子囊菌進行調查，然後有系統的累積許多常見屬之資料，如Astrosphaeriella、Byssosphaeria、Didymosphaeria、Herpotrichia、Kirschsteiniothelia、Massarina、Melanomma、Trematosphaeria 等屬，並整合國外種類之標本觀察，及分子親緣分析之結果，建立各屬之正確定義。對於各屬收集之種類都進行照相及描述，以出版專屬專論供學界參考為目標，而培養之菌株皆進行保存，以為永續利用之資源。在採集腐生種類之腔室子囊菌之同時，亦將收集屬於絕對寄生菌之星盾炱科菌，該科之種類為熱帶地區所特有，除了對其多樣性進行調查外，亦將研究其與寄主間之共同演化關係。此計劃希望藉由逐一的對腔室子囊菌綱之重要屬之釐清，建構出完善之腔室子囊菌綱之系統分類。Compared with temperate countries, Taiwan, situated in subtropics and covered withmany high elevation mountain areas, certainly embraces highly diverse fungal species.However, the survey of fungal diversity is still far from enough in Taiwan. While the naturalconservation and sustainability are attracting public attention, the intense survey of nativefungal flora becomes a crucial issue. The microscopic loculoascomycetes are ubiquitous. Notonly for the biodiversity, but also the systematics, there is plenty of work remaining to bedone on loculoascomycetes. The aim of this project is to comprehensively surveyloculoascomycetous fungi in Taiwan. Systemically gathered are materials belonging to somecommon genera, such as Astrosphaeriella, Byssosphaeria, Didymosphaeria, Herpotrichia,Kirschsteiniothelia, Massarina, Melanomma and Trematosphaeria. Specimens are lent fromforeign herbariums for morphologically comparing with Taiwan materials. Morphologicalobservations integrating with molecular phylogenetic analyses will shape the precisedefinition of many genera. The fungi obtained are isolated in pure culture and maintained inculture collection center in an attempt to be screened for useful secondary metabolites in thefuture. Genus monographs are expected to be issued after adequate data are acquired. Inaddition to the saprobic loculoascomycetes, biotrophic Asterincaeae is also involved in thisstudy. This family exclusively occurs in tropic areas. Taiwan undoubtedly harbors numerousspecies of this family. All the need is to survey their diversity. The coevolution betweenAsterinaceae species with their host will be investigated. Conclusively and hopefully asound systematics of the Loculoascomycetes can be gradually achieved after more and moreloculoascomycetous genera are studies

New genus, new species and new records of ascomycetes in Taiwan

由全省各地採集，共整理了四十種子囊菌，其中包括兩個新屬、十個新種、一個新 組合種、 二十六個新記錄種及三種新寄主的記錄， 新屬為 Urosporellopsis Svanesania， 新種為 Acantharia chaetomides、 Drmatodothis euonymi、 Dermatodothis symploci、 Gnomonia appendiculate、 Micropeltis symploci、 Physalospora erycibe、 Rosenscheldiella perseae、 Sivanesanirubicola、 Urosporellopsis taiwanensis、Vestergrenia clematidis，新組合種為 Coleroa deadii， 新記錄種為 Acanthari aterrima、Acanthonitschkda argentinensis、 Acanthonitschka pulchella、 Anthostomella caulicola、Apiospora montagnei 、 Astrosphaeriella trochus、 Brtia moriformis、 Capronia pilosella、 Cucurbidothis pithyophila、 Datodothis javanica、Fracchiaea broomeiana、 Gnomoniella sp.、 Herpotrichia schiedermayerna、Leptosphaeria doliolum、 Melanomma pulvis-pyrius、Ophiovals femoralis、Paraphaeosphaeria michotii 、Parmulina japonica、Phyllachora bauhiniae var. longispora、Phyllachora cinnamcmi、 Phyllachorella micheliae、 Pseudothis coccodes、 Thyridium vertitum、Tubeufia sp.、Viadracula annuliformis、 Yoshinagaia quercus 等 ， 而有新寄主記載的為 Aphisphaerella petrakii 及 Amphisphaeria saccharicola、Rosenscheldiella orbis。 #9201878 #920187

Taxonomic Studies on Some Common Genera of the Loculoascomycetes

本研究選擇腔室子囊菌綱中五個常見的屬進行形態之分類研究，並重新評估其屬的界定，此五屬為Leptosphaeria，Byssosphaeria，Herpotrichia，Astrosphaeriella及Kirschsteiniothelia。而特別強調的是Leptosphaeria屬，因為長久以來此屬之鑑定一直是分類學家之一大難題，此研究即主要針對歐洲之種類，以模式標本為主，以求對此屬有更清晰之了解。並依現在一般接受的有關此屬觀念，將此種類繁多的屬細分為三個分類群，分別為doliolum群、maculans群及dryadis群，以各群之代表種類之小種名命名，各群間之差異主要在於孢子之型態，doliolum群之孢子深褐色具三隔膜、有疣突、有外套膜，及孢子兩端之細胞壁有明顯之加厚; maculans群之孢子淡褐色具三至多個隔膜、較細長且孢子兩端常有附屬絲; dryadis群之孢子淡至深褐色具三隔膜。亦並從演化觀點來探討區分此三分類群之各特徵，各分類群以主要代表性種類為例，詳細描述以供比較。重新研究歐洲標本所歸納之分類特徵來區分Byssosphaeria及Herpotrichia屬，其中Byssosphaeria屬子囊短至長柄、孢子褐色，而Herpotrichia屬子囊幾乎無柄、孢子無色，並以台灣之原生種類為例來說明此二屬之差異，包括了兩個新種。對於Astrosphaeriella屬，由於缺乏明確之定義，以致於囊括了非常異質性的種類，而其亦可區分為三個界定清晰之分類群，分別為包含模式種之典型Astrosphaeriella群，和Trematosphaeria屬相似之Trematosphaeria-like群及和Massarina相似之Massarina-like群，本研究以台灣之種類為例說明典型Astrosphaeriella群及Trematosphaeria-like群之特徵，並描述兩個新種。最後對Kirschsteiniothelia屬之模式種進行研究，由於之前對該模式種之觀念認定有誤，包含了兩個不同的種，在此對其提出修正，釐清了該屬之正確觀念，並建立一新組合種。以上是針對這幾個常見屬之主要問題進行形態上的探討，冀望能在其混沌不清之分類現象下提供一新的歸類依據及思考方向，最終當然是希望能使整個腔室子囊菌綱之分類系統能臻至完善。The generic limits of some selected genera in the Loculoascomycetes are reassessed morphologically. They are Leptosphaeria, Byssosphaeria, Herpotrichia, Astrosphaeriella and Kirschsteiniothelia. Especially emphasis is placed on the Leptosphaeria, because it is a historically intractable genus for mycologists. Three groups of Leptosphaeria species, doilolum group, maculans group and dryadis group, are proposed, each of which is desiginated by the epithet of the species most exemplifying it. These three groups are separated mainly on the basis of ascospore morphology. Ascospores in doliolum group are ellipsoid, 3-septate, dark brown, verrucose, surrounded by a sheath, in dryadis group are ellipsoid, 3-septate, pale to dark brown, smooth without sheath or appendage, and in maculans group are elongated ellipsoid to cylindrical, 3 to multiseptate, yellowish brown, usually with an appendage at each end. Characters used in the distinction of these three groups are evaluated from the aspects of evolution. Representative species are described and illustrated to elucidate respective groups. The relationship between Byssosphaeria and Herpotrichia is evaluated with an account on the taxonomic value of characters. The two genera are distinguishable. Asci in Byssosphaeria are short to long-stalked, while in Herpotrichia asci are sessile to short-stalked. Ascospores of Byssosphaeria are pale to dark brown, while those of Herpotrichia are hyaline Taiwan materials which are referred to both genera, including two new species, are presented to provide a clear comparison between them. With regard to Astrosphaeriella, due to its heterogeneity, it can be subdivided into three groups of species. Individuals groups comprise typical Astrosphaeriella species, Trematosphaeria-like species and Massarina-like species respectively. The group constituted by typical Astrosphaeriella species and the group by Trematosphaeria-like species are included in this study, and they are represented by materials from Taiwan. Two new species are introduced. Finally the generic definition of Kirschsteiniothelia is clarified by redescribing the generic type and segregating a species previously synonymized under this type species. A new combination is therefore proposed. This study provides a new insight toward a completely clarification of the genera involved. Hopefully a sound classification of the Loculoascomycetes can be gradually approached.Contents 中文摘要-------------------------------------------------------1 Abstract ------------------------------------------------------2 Introduction --------------------------------------------------3 Chapter 1 : Morphological delimitation of the doliolum-group in Leptosphaeria, with an evalution of the current generic definition-----------------------------------------------------7 Chapter 2 : Delimitation of the maculans-group in Leptosphaeria --30 Chapter 3 : Delimitation of the dryadis-group in Leptosphaeria -----57 Chapter 4 : Byssosphaeria and Herpotrichia from Taiwan, with notes on the taxonomic relationship between these two genera -------71 Chapter 5 : Astrosphaeriella from Taiwan, including two new specie--------------------------------------------------------87 Chapter 6 : On the type species of the genus Kirschsteiniothelia, K. aethiops ----------------------------102 Conclusion---------------------------------------------------111 附錄 : 中文大綱----------------------------------------------11

輸出入木材及木質包裝材有害生物檢測技術之研發

The aims of this project is to inspect detrimental fungi, particularly ophiostomatoid fungi, on wood which is from importation or from indigenous plants. Morphological characteristics and DNA sequences of these fungi can be established to provide the basis for their accurate and quick detection. Both methods of morphology identification and DNA sequence comparison will be applied. Specific approaches of isolation will be adopted to obtain specific groups of fungi. Specific primers are used to amplify specific DNA fragments for comparisons with DNA sequences deposited in GenBank. Based on the morphological and molecular data acquired, it can be expected that these potential pathogens can be warded off from our country. As to the imported / exported wood packaging materials, a standard sampling procedure of a fast screening process and a monitoring system will be set up for these materials. In this project, three tasks will be included, (i) literature survey of important pests and pathogens for wood (ii) establishing a standard procedures for inspecting and identifying of pests and plant pathogens and (iii) continuous monitoring important pests and pathogens information of imported / exported wood packaging materials, are proposed to accomplish our goal. The success of this project would help quarantine officers monitor and control important imported / exported wood packaging materials importing from wood packaging material.本計劃將針對進口原木材及本土林木中重要病原(特別是長喙殼菌類真菌)進行偵測，建立菌種形態特性及DNA序列資料庫，提供快速且正確檢測木材中此類有害真菌之基礎。潛在病原菌之偵測及鑑定包括形態及分子序列方式，依據不同之菌種類別之特性，以特有之方式進行菌種分離，針對特定rDNA片段進行解序，建立形態及分子鑑定標準，有助於發展快速之檢測方法，攔截有害病原之輸入。對於木質包裝材，建立重要病蟲害資訊庫及建立進出口木質包裝材之標準採樣流程、快速病蟲害鑑定技術及監測系統。重要工作項目包括收集世界各國經由木質包裝材輸入之重要病蟲害文獻資料、建立進出口木質包裝材之標準採樣流程及病蟲害檢測程序，持續監測經木質包裝材輸出入之重要病蟲害等四大項。預期收集經由進口木質包裝材輸入之重要病蟲害資訊並建立資料庫，以及建立有效取樣標準流程，進而協助檢疫人員監測與管制經木質包裝材輸入之重要病蟲害