玉米弯孢菌叶斑病菌检测技术及遗传多样性研究
|
摘要
玉米弯孢菌叶斑病是近几年在我国发展起来的一种对玉米危害很大的新病害，关于病菌致病性分化及遗传方面的报道较少，目前还不清楚病菌的生理分化情况，也少有关于病原菌遗传背景分析的研究，更缺乏病原菌快速鉴定和带菌种子检测技术的研究，本研究以病菌孢子形态学和可溶蛋白质多样性比较为基础，建立了两种免疫学的鉴定技术，并将该技术用于带菌玉米种子和幼苗的检测和诊断；同时，在分子水平用AFLP标记技术对该菌的遗传多样性用进行了分析，同时与多种相似病菌的AFLP分子标记进行了比较研究，确定了代表Curvularia lunata的特异性分子标记，为下一步的克隆、测序和建立分子生物学鉴定方法提供了捷径，也为植物病原真菌鉴定的分子标记筛选提供了范例。
首先，搜集国内外和分离四川主要玉米产区的玉米弯孢菌叶斑病样品，共获得73个纯化的相关菌株，其中包括C. lunata菌株34个，Cochlibolus carbonum菌株3个，Co. heterostrophus菌株13个，Alternaria alternata菌株8个。对其中的12个C. lunata菌株和6个相似或分离过程中容易混淆的菌株在分生孢子形态、菌落型态和可溶性蛋白质电泳图谱进行了比较研究。结果表明分生孢子形态是病原菌形态鉴定的主要依据，但在C. lunata和C. inaequalis之间孢子形态并不易区分；菌落型态在不同种之间存在明显的差异，同种之间也存在一定差异，但不稳定和存在变异。18个菌株SDS聚丙烯酰胺凝胶电泳图谱在种间存在明显的差异，C. lunata种内蛋白带也存在多样性。图谱显示Curvularia在Rf值为0.177处有一条该属的特征蛋白带，C. lunata在Rf值为0.225处有一条该种的特征蛋白带。聚类分析将未能确定种的13号菌株确定为C. inaequalis。
在以上蛋白质分析的基础上建立了C. lunata菌株鉴定和种子的免疫检
四川大学博土学位论文
测方法。用于 I．ELISA、ABC－ELISA和 DOt－ELISA三种方法比较研究了不
同抗血清的检测灵敏度和特异世，结果IELISA方便快捷，首选使用；
D。t－ELISA灵敏度稍低适用于基层检验。用 C 菌株混合的可溶性蛋白
质作为免疫抗原，获得一抗兔抗 C Iunata抗血清（PAhs）；抗血清经近似菌
可溶性蛋白质吸附后获得对C lunata菌株可溶性蛋白质具有特异性的抗血
清（APAbs卜 可应用于 C lunata菌株的鉴定；用玉米种子可溶性蛋白质对
APAs进行进一步吸附纯化，获得了与玉米种子蛋白质无交叉反应的特异性
抗血清（DAPAbS人可用于对带菌玉米种子的检测鉴定，并制定了相应的应
用程序。几种抗血清分别用于间接ELISA和ABC＊LISA兔疫分析，以比较
其检测灵敏度和特异性。
在免疫检测的基础上，为了从分子水平进一步了解新月弯抱菌的遗传背
景和多样性，以寻找能够作为 C Iunata种的分子标记，从而为建立该菌的分
子检测技术奠定基础，本研究首次用 AFLP指纹技术对C Iunata不同菌株及
相似菌株进行了遗传多样性研究。用改进的氯化书法获得了高质量的供试菌
株DNA，并优化了AFLP各步的试验条件。利用AFLP技术，分析了5个
C lunata菌株和5个不同种的近似菌株AFLP的DNA指纹特征，并进行了
不同引物的聚类分析。分别采用两对具有E．ZM．l 的选择性引物
EcoRJ－AC／Mel．A和EcoAI－AC／Mel．C，以及两对具有E－ZM－3的选择性引
物EcoXi叭 cTG和EcORI叭 （TC对供试菌进行选择性扩增。
结果供试菌株的AFLP指纹图谱具有很高的多态性，反映了菌株间存在丰富
的遗传多样性；四对引物共得到 11 69条扩增条带，其中多态性条带 778条，
占67％。根据不同引物的不同指纹图谱得到的聚类结果反映了 C lunata菌株
间以及近似菌株间种、属的遗传距离和亲缘关系；四对引物分别得到了
944hp、229hp、254hp、130hp、1129hp、296hp六条属于 C lunata的稳定的
特异性片段，这些特异性片段都有希望成为鉴定 C Iunata有效的分子标记，
为进一步确定用于鉴定c1U——的有效的分子标记奠定了基础。研究表明，
利用 AFLP不同的引物组合，不仅能稳定地检出 C Iunata和近似菌株丰富的
多态性，同时也是寻找特异性分子标记的理想方法。
Recenily there have been many reports all over the world about fungi disease
caused by Curvularia lunata [Cochliobolus Iunana]. The economic importance
and the wide range of its hosts can be found from these reports. The biggest
comznedty of the host of C.lunata is gramineous plants such as rice, malze,
wheat and sorghurn. Othedrise, cotton, cowpea,sunflower, onion etc could be
infected by C lunata too. C lunata is one of the importan seed-bome mycelial
fungus pathogen. Microscopic examination showed C.lunata could be deected
in all components of maize seeds.
Maize is the third importan food crop in addition to rice and wheat in China.
Com northem leaf blight (Helminthosporium turcicum) and com southern leaf
blight (Cochliobolus heterostrophus) were the main maize diseases in China in
the past. But recentiy a new disease named maize cedaria leaf spot or malze
yellow spot caused by C.lunata (infrequentiy by C hoequais), is becoming
prevalent in north China and is spreadin to south China.The disease is
charaterized by chlorotic spot symptoms on leaves: chlorotic pin points appeared
in the beginning, then yellow spots with a white center and a chlorotic halo
followed,and finaly leaves bligh when a lot of spotS joined togetheL Size of the
spot usually is 1 × 2mm-2mm. Maize cedaria leaf spot is easy to cothee with
com southem leaf bligh and com helrninthosporium leaf spot (Co. carbonum) in
symptoms. C lunata can be found on the sdse of seeds, the spores and
mycelium were also found on the buttrss roots, and on the soil surface but not
at
4
dePths of 5 or 10crn. So isolation of the causal agent C lunata is the main method
fOr diagnoses at presen though there are variabilihes of cultural cforteristics of
C lUnata isolates and coWons by other sdriilar fungi occur frequently in
the Process of isolation. There are some reportS on the variabi1ity growh and
sPOrulation of isolates from differen geograPhical locations, and on the
ulbetrUM of C IUnaha. So far there is still laCk of a raPid, sPecific and
sensitive tecboque for the diagnosis of C lunata and there are few stUdies on the
genetic baskground of Ans pathogen.
ln order to exPlore the differences betWeen C Iunata and otlier sbolar fungi
and establish a specific and sensihve diagnostic method fOr C lunata, this stUdy
has compared the moboometrics including the fungi colony and in medium and
the sPores of C Iunata in maize and Othr swhlar forgi aPPearing bequently by
isolation. We have compared their protein SDS polyacrytalde gel
electrophoresis pattems too. An immunological detection tecboque of C lunata
frOm cultUre and on the seed based on the assays of Proteins is established. We
also study the genetic diversihes using amPlified fragYnent 1ength polyInorphism
and comPared the molecular markers of C Iunata, the sbolar strains. Six specific
POlymorphic frgnents of AFLP have been found srochronously. They wiIl be
the twrtan molecular markers for idenopg C lunata. The main resultS are
followed.
First we collected 73 pure isolates of C IunaIa and their sindlar stains from
abroad, our councy and the main areas of maize in Sichuan, including 34 isolates
of C Iunata, 3 isolates of CO. Carbonum, l3 isolates of CO. heterosmphus, and 8
isolates of A. alternata. TWlve isolates of C Iunata and six sbolar sPecie strains
from haze or other hoSt plants wer exaInined for the analysis of mycelioid
colony morphometrics, sPores morphometrics and SDS polyaCryamide gel
eleCtrOPhoresis pattemS of their soluble proteins. The diffeences of colony
morphometrics of l8 isolates existed arnong differen SPeCies and the same specie
bu they wer no W and special. Th differences of SDS POlyaCrylndde gel
eleCtrphoresis pattems ekisted sighficantiy among differen Species and the
5
saxne sPecie. There was a sPecific protein band for CimIaria at relative mobility
value 0.l77 W0.177) and a sPeCial PrOtein band for C bo at Rf0.225.
Second, we have set uP the bological detection tecboque to ide
首先，搜集国内外和分离四川主要玉米产区的玉米弯孢菌叶斑病样品，共获得73个纯化的相关菌株，其中包括C. lunata菌株34个，Cochlibolus carbonum菌株3个，Co. heterostrophus菌株13个，Alternaria alternata菌株8个。对其中的12个C. lunata菌株和6个相似或分离过程中容易混淆的菌株在分生孢子形态、菌落型态和可溶性蛋白质电泳图谱进行了比较研究。结果表明分生孢子形态是病原菌形态鉴定的主要依据，但在C. lunata和C. inaequalis之间孢子形态并不易区分；菌落型态在不同种之间存在明显的差异，同种之间也存在一定差异，但不稳定和存在变异。18个菌株SDS聚丙烯酰胺凝胶电泳图谱在种间存在明显的差异，C. lunata种内蛋白带也存在多样性。图谱显示Curvularia在Rf值为0.177处有一条该属的特征蛋白带，C. lunata在Rf值为0.225处有一条该种的特征蛋白带。聚类分析将未能确定种的13号菌株确定为C. inaequalis。
在以上蛋白质分析的基础上建立了C. lunata菌株鉴定和种子的免疫检
四川大学博土学位论文
测方法。用于 I．ELISA、ABC－ELISA和 DOt－ELISA三种方法比较研究了不
同抗血清的检测灵敏度和特异世，结果IELISA方便快捷，首选使用；
D。t－ELISA灵敏度稍低适用于基层检验。用 C 菌株混合的可溶性蛋白
质作为免疫抗原，获得一抗兔抗 C Iunata抗血清（PAhs）；抗血清经近似菌
可溶性蛋白质吸附后获得对C lunata菌株可溶性蛋白质具有特异性的抗血
清（APAbs卜 可应用于 C lunata菌株的鉴定；用玉米种子可溶性蛋白质对
APAs进行进一步吸附纯化，获得了与玉米种子蛋白质无交叉反应的特异性
抗血清（DAPAbS人可用于对带菌玉米种子的检测鉴定，并制定了相应的应
用程序。几种抗血清分别用于间接ELISA和ABC＊LISA兔疫分析，以比较
其检测灵敏度和特异性。
在免疫检测的基础上，为了从分子水平进一步了解新月弯抱菌的遗传背
景和多样性，以寻找能够作为 C Iunata种的分子标记，从而为建立该菌的分
子检测技术奠定基础，本研究首次用 AFLP指纹技术对C Iunata不同菌株及
相似菌株进行了遗传多样性研究。用改进的氯化书法获得了高质量的供试菌
株DNA，并优化了AFLP各步的试验条件。利用AFLP技术，分析了5个
C lunata菌株和5个不同种的近似菌株AFLP的DNA指纹特征，并进行了
不同引物的聚类分析。分别采用两对具有E．ZM．l 的选择性引物
EcoRJ－AC／Mel．A和EcoAI－AC／Mel．C，以及两对具有E－ZM－3的选择性引
物EcoXi叭 cTG和EcORI叭 （TC对供试菌进行选择性扩增。
结果供试菌株的AFLP指纹图谱具有很高的多态性，反映了菌株间存在丰富
的遗传多样性；四对引物共得到 11 69条扩增条带，其中多态性条带 778条，
占67％。根据不同引物的不同指纹图谱得到的聚类结果反映了 C lunata菌株
间以及近似菌株间种、属的遗传距离和亲缘关系；四对引物分别得到了
944hp、229hp、254hp、130hp、1129hp、296hp六条属于 C lunata的稳定的
特异性片段，这些特异性片段都有希望成为鉴定 C Iunata有效的分子标记，
为进一步确定用于鉴定c1U——的有效的分子标记奠定了基础。研究表明，
利用 AFLP不同的引物组合，不仅能稳定地检出 C Iunata和近似菌株丰富的
多态性，同时也是寻找特异性分子标记的理想方法。
Recenily there have been many reports all over the world about fungi disease
caused by Curvularia lunata [Cochliobolus Iunana]. The economic importance
and the wide range of its hosts can be found from these reports. The biggest
comznedty of the host of C.lunata is gramineous plants such as rice, malze,
wheat and sorghurn. Othedrise, cotton, cowpea,sunflower, onion etc could be
infected by C lunata too. C lunata is one of the importan seed-bome mycelial
fungus pathogen. Microscopic examination showed C.lunata could be deected
in all components of maize seeds.
Maize is the third importan food crop in addition to rice and wheat in China.
Com northem leaf blight (Helminthosporium turcicum) and com southern leaf
blight (Cochliobolus heterostrophus) were the main maize diseases in China in
the past. But recentiy a new disease named maize cedaria leaf spot or malze
yellow spot caused by C.lunata (infrequentiy by C hoequais), is becoming
prevalent in north China and is spreadin to south China.The disease is
charaterized by chlorotic spot symptoms on leaves: chlorotic pin points appeared
in the beginning, then yellow spots with a white center and a chlorotic halo
followed,and finaly leaves bligh when a lot of spotS joined togetheL Size of the
spot usually is 1 × 2mm-2mm. Maize cedaria leaf spot is easy to cothee with
com southem leaf bligh and com helrninthosporium leaf spot (Co. carbonum) in
symptoms. C lunata can be found on the sdse of seeds, the spores and
mycelium were also found on the buttrss roots, and on the soil surface but not
at
4
dePths of 5 or 10crn. So isolation of the causal agent C lunata is the main method
fOr diagnoses at presen though there are variabilihes of cultural cforteristics of
C lUnata isolates and coWons by other sdriilar fungi occur frequently in
the Process of isolation. There are some reportS on the variabi1ity growh and
sPOrulation of isolates from differen geograPhical locations, and on the
ulbetrUM of C IUnaha. So far there is still laCk of a raPid, sPecific and
sensitive tecboque for the diagnosis of C lunata and there are few stUdies on the
genetic baskground of Ans pathogen.
ln order to exPlore the differences betWeen C Iunata and otlier sbolar fungi
and establish a specific and sensihve diagnostic method fOr C lunata, this stUdy
has compared the moboometrics including the fungi colony and in medium and
the sPores of C Iunata in maize and Othr swhlar forgi aPPearing bequently by
isolation. We have compared their protein SDS polyacrytalde gel
electrophoresis pattems too. An immunological detection tecboque of C lunata
frOm cultUre and on the seed based on the assays of Proteins is established. We
also study the genetic diversihes using amPlified fragYnent 1ength polyInorphism
and comPared the molecular markers of C Iunata, the sbolar strains. Six specific
POlymorphic frgnents of AFLP have been found srochronously. They wiIl be
the twrtan molecular markers for idenopg C lunata. The main resultS are
followed.
First we collected 73 pure isolates of C IunaIa and their sindlar stains from
abroad, our councy and the main areas of maize in Sichuan, including 34 isolates
of C Iunata, 3 isolates of CO. Carbonum, l3 isolates of CO. heterosmphus, and 8
isolates of A. alternata. TWlve isolates of C Iunata and six sbolar sPecie strains
from haze or other hoSt plants wer exaInined for the analysis of mycelioid
colony morphometrics, sPores morphometrics and SDS polyaCryamide gel
eleCtrOPhoresis pattemS of their soluble proteins. The diffeences of colony
morphometrics of l8 isolates existed arnong differen SPeCies and the same specie
bu they wer no W and special. Th differences of SDS POlyaCrylndde gel
eleCtrphoresis pattems ekisted sighficantiy among differen Species and the
5
saxne sPecie. There was a sPecific protein band for CimIaria at relative mobility
value 0.l77 W0.177) and a sPeCial PrOtein band for C bo at Rf0.225.
Second, we have set uP the bological detection tecboque to ide
引文
[1]白元俊等，1998，玉米弯孢菌叶斑病菌的生物学特性研究，辽宁农业科学，5：9-13
[2]曹以勤，彭金火，1990。几种禾本科植物上的弯孢菌，南京农业大学学报，13(4)：12-16。
[3]陈福生，罗信昌，周启，1999。酶联免疫技术检测植物病原真菌。植物检疫，13(1)：33-35。
[4]陈万全，冯洁，秦庆明等，1999。DNA分子标记在植物真菌病害研究中的应用。植物保护学报，26(3)：278-282。
[5]戴法超，高卫东，王晓鸣．1997，玉米弯孢菌叶斑病的初步研究简报．植物保护，28(1)：75-76．
[6]戴法超，王晓鸣，朱振东，等．1998，玉米弯孢菌叶斑病研究．植物病理学报，28(2)：123-129．
[7]戴芳澜，1979，中国真菌总汇。北京：科学出版社，936
[8]窦坦德，沈崇荛，2000。植物病原真菌检测技术研究进展。植物检疫，14(1)：31-33。
[9]窦坦德，沈崇尧，1997。大豆疫霉菌的血清学研究，菌物系统，16(2)：157-160
[10]窦坦德，沈崇尧，1997。快速检测大豆疫霉菌的酶联免疫吸附技术研究。植物检疫，11(3)：138-142
[11]傅俊范，白元俊等，1999，玉米弯孢菌叶斑病流行动态及产量损失测定，沈阳农业大学学报，30(3)：204-207．
[12]甘贤友等，1995，玉米弯孢霉叶斑病初步研究，植物保护，21(5)：24～25
[13]洪华珠，杨红等。1994，三株球孢白僵菌血清学性质的研究。真菌学报，13(1)：77-79。
[14]黄如貘等，应用裂解气相色谱法鉴定酵母菌的初步研究。见：程光胜等主编，分析微生物学专辑，北京：科学出版社，1988．194～197
[15]姜远良，1995。化学分类法在细菌和真菌系统分类学中的应用和发展。辽宁师范大学学报，18(2)，152-155。
[16]贾建航，李传友，伏健民等，1999，AFLP技术在玉米自交系类群分析研究中的应用。高技术通讯，4：43-46。
[17]李成文，1992。现代免疫化学技术．上海：上海科学技术出版社
[18]刘学敏，赵骞，曲金玲等。运用随机扩增多态性标记检测中国东北大豆灰斑病菌菌株遗传变异，植物病理学报，1998，28(1)：43～48
[19]卢乡怀，李明霞。1990，裂殖酵母属的酶及可溶性蛋白聚丙烯酰胺凝胶电泳。真菌学报，9(1)：50—55
[20]吕国忠，刘志恒，何富刚，等．1997，辽宁省爆发一种新病害—玉米弯孢菌叶斑病．沈阳农业大学学报，28(1)：75-76．
[21]吕国忠，薛玉梅，1999，玉米弯孢菌叶斑病菌毒素的生物活性测定及对玉米叶片超微结构的影响．沈阳农业大学学报，30(3)：212-214．
[22]蔺瑞明，高增贵，崔明珠，等．1999，玉米弯孢菌叶斑病苗期抗病性鉴定及遗传初步分析．植物保护，25(5)：1-3．
[23]戚佩坤．1978．玉米、高粱、谷子病原手册[M]北京：科学出版社，168-169．
[24]单卫星，陈受宜，康振生等，1995。DNA指纹技术分析在植病真菌群体遗传研究中的应用。西北农业大学学报，23(6)：84-89。
[25]魏景超，1975，水稻病原手册，科学出版社，198～204。
[26]徐大雅、黄河、王春平．1982，疫霉蛋白质的聚丙烯酰胺凝胶盘状电泳。真菌学报，1(1)：40—47
[27]徐德坤，王琪，殷秀东，等．1995，玉米弯孢菌叶斑病调查初报[J]．山东农业科学，6：38
[28]徐敬友，陆家云等，1993。六种疫霉菌体可溶性蛋白质和酯酶的聚丙烯酰胺凝胶平板电泳研究。真菌学报，12(1)：54-64。
[29]鄢洪海，高增贵，陈捷等，1999。玉米弯孢菌叶斑病研究进展。沈阳农业大学学报，30(3)：369-371．
[30]鄢洪海，高增贵等，2000。玉米弯孢菌叶斑病菌生理分化鉴定技术的研究，沈阳农业大学学报，31(5)：472-475．
[31]叶华智，张敏，秦芸等，2000。首次在四川发现玉米弯孢菌叶斑病[J]．四川植保，2：75-76．
[32]张定法，徐瑞富，张希福，等，1998。玉米弯孢霉叶斑病菌生物学特性的研究．河南职技师院学报，26(1)：18-21．
[33]张小平，陈强等，1999。用AFLP技术研究花生根瘤菌的遗传多样性。微生物学报，39(6)：483-488。
[34]赵玉锦，1988。植物生物化学技术与方法。北京：农业出版社，93-95。
[35]郑文明，刘峰，康振生等，2000。中国小麦条锈菌主要流行菌系的AFLP指纹分析。自然科学进展，10(6)：532-537。
[36]周志伟，黄河，毛霉目真菌的DNA提取及其GC含量的测定，微生物学通报，1991，18(5)：275～279
[37]Adiver-SS,Anahosur-KH. 1994. Association of Curvularia and Exserohilum species as sorghum moulds and their effect on seed germination and seedling growth. Current-Research-University-of-Agricultural-Sciences-Bangalore, 23(3-4):38-40.
[38]Alcom JL 1990, Additions to Bipolaris, Cochliobolus and Curvularia. Mycotaxon. 39,361-392.
[39]Alcorn JL 1991, New combinations and synonymy in Bipolaris and Curvularia and a new species of Exserohilum. Mycotaxon. 41,329-344.
[40]Anderson J B, Bailey S S, Pukila P J. 1989. Variation in ribosomal DNA among biological special of Armillaria, a genus of hoot infecting fungi. Evolution, 43:1652-1662
[41]Barrett B A, Kidwell K K. 1998. AFLP-based genetic diversity assessment among wheat cultivars from the Pacific Northwest. Crop Science, 38(5): 1261～1270
[42]Berbee M.L, Mona Pirseyedi, Hubbard S. 1999, Cochliobolus phylogenetics and the origin of known, highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase gene sequences. Mycologia 91(6), 964-977.
[43]Becket J et al. 1995. Com bined mapping of AFLP and RFLP markers in barley, Mol Gen Genet, 249:65-73
[44]Brill L.M, McClary R.D, Sinclair J.B 1994. Analysis of two ELISA Formats and Antigen Preparations Using Polyclonal Antibodies Against Phomopsis longicolla. Phytopathology. 84, 173-179.
[45]Burdon JJ,Marshall D R.Isozyme variation between species andforrnae speciales of the genus Puccinia Pers. Can J Bot, 1981,59:2628～2634
[46]Burns DT, et al.Pyrolysis gas chromatography as an aid to the identification of Penicillium species.Jchromatography, 1976, 116:107～115
[47]Burrell R G, Clayten C W, Galleyly M F, et al. 1966. Factors affecting the antigenicity of the mycelium of three species of Phytophthora. Phytopathology, 56:421-426
[48]Chen-WeiQun. Chen-FaJun,Zhang-TianYu, etc., 2000, Some saprotrophic deuteromycetes in Xisha Islands[J]. Mycosystema19(3):428-430.
[49]Chu Xining, Bai Yuming, and Yuan Jingming 1994: A comparative study of electrophoretic patterns of soluble proteins and zymograms from nine strains of Rhizopus. Acta Mycologica Sinica. 13(2), 121-126.
[50]Clark M F, Adams A N. 1977. Characteristics of the microplate method of Enzyme-Linked Immunosorbent Assay for the detection of plant vinuses J Gert Virology. 34:475-483
[51]Crowhust RN. Differentiation of Fusanumsolani f.sp.cucurbitae race 1 and 2 by random amplification of polymorphic DNA. Curr Cenet, 1991,20:391～396
[52]Damjana Rozman,Kristijan Jezernik,Radovan Komel. 1994, Ultrastructure and genotypic characterization of the filamentous fungus Cochliobotus lunata in comparaisom to the anamorphic strain Curvlaria lunata[J]. FEMS Microbiology Letters, 117:35-40.
[53]De Vallavieille C, Erselius LJ 1984, Variation in protein profiles in Phytophthora: survey of a compositive population of three species on citrus. Trans br Mycol Soc 83:473—479
[54]Deshmukh RN,Raut JG. 1993, Sites of Curvularia lunata infection in sorghum seed. Indian-Phytopathology, 46(3):251-252.
[55]Dewey F M. Detection of plant-invading fungus by monoclonal antibodies, In techniques for the rapid detection of plant pathogens pp47-62, eds J. M. Duncan and L. Tonance Uk: Blackwell Scientific Publication.
[56]Dorey-SE,Ayliffe-WH,Edrich-C,etc. 1997, Fungal keratitis caused by Curvularia lunata, with successful medical treatment[J]. Eye-London, 11:5,754-755.
[57]Gunasekaran M,et al.Differentiation of races of TiUetia caries and T.foetida by pyrolysis-gas-hiquid chromatography. My-cologia. 1979,71:1066～1071
[58]Ellis JJ. Species and varieties in the Rhizopus arrhizus-Rhizopus oryzae group as incated by their DNA comple mentarity Mycolgia
[59]Ellis M B. Dematiaceous Hyphomycetes. 1966. VII: Curvularia, Brachysporium etc. Mycological Papers, 106:1～57
[60]Ellis M B. 1971, Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, UK: 452～495
[61]Ellis M B. 1976, More Decatiaceous Hyphomyvetes. Commonwealth Mycological Institute, Kew, UK. 404～406
[62]Erselius LJ, Vallavieille CD 1984. Variation in protein profiles of Phytophihora: comparison of six species. Tran Br Mycol Soc 83: 463—472
[63]Fargues J, Duriez T, Andrieu S et al, 1975. Etude immunologique compree di souches de Metarrhizium anisopliae(Metsch.)sor. Champignon Hyphomycete entomopathogene. C. R. Acad. Sci Paris ser. D. 281:1781—1783
[64]Fargues J, Duriez T, Popeye R et al, 1981. Immunological characterization of the entomopathogenic hyphoycetes Beawveria and Metarhizium. Mycopathologia. 75:101—108
[65]Forster H, Kinscherf T G, Lang S A., et al. 1989. Restriction fragment length polymorphisms of the mitochondrial DNA of Phyphthora megasperma isolated from soybea, alfalfa and fruit trees. Canadian Journal Botany, 67:529-537
[66]Fred Ausubel et al. 1995. Short Protocols in Molecular Biology. John Wiley and Sons, Inc.,
[67]Gardes M,fortin JA,Mueller GM RFLP in nuclear ribosomal DNA of for Laccata spp:L.bicolor,L.laccata,Lproximn and L.anethstina. Phytopathol, 1990,80:1312～1317
[68]Gieason, M. L. Ghabral. S. A. and Ferriss. R. S.1987. Serological detecnon of Phomopsts longicolla in soybean seeds. Phyopathology 77:371-375
[69]Gill H S,Zentmyer G A.Identification of Phytophthora species by disc electrophoresis.Phytopathology, 1978,68:163～167
[70]Goodwin P H. et al. 1990. Detection of Phytophthora parasitica from soil and host tissue with a species-specific DNA probe. Phytopathology, 80:277-281
[71]Grajal, Martic M J, Simon C J, Muehlbauer F J et al. 1993. Use of random amplified polymorphic DNA (RAPD)to characterize race 2 of Fusarium oxysporum f. sp. pisi.Phytopathology, 83(6):612-614
[72]Hajek AE, Butt TM et al, 1991. Detection of Entomophaga maimaiga(Zygomycetes:Entomophthorales)using Enzyme-Linked Immunosorbent Assay. J. Invertebr. Pathol. 58:1—9
[73]Hamison J G. 1990. Quantification of Phytophthora infestans mycelium in potato leaves by ELISA. Inproceeding of the fourth COST-88 workshop pp 18-19 Invergowire Dundee Scotland 25-27 Sept.
[74]Hawksworth DL,Pitt J I. A new taxonomy for Monascus species based on cultural and microscopical characters. Australian Journal of Botany, 1983,31:51～61
[75]He G, Prakash C S. 1997. Indentification of polymorphic DNA markers in cultivated peanut(Arachis hypogaea L.). Euphytica, 2:143-149
[76]Hearn. V M. and Mackenzic. D.W. R, 1980. The preparation and partial purification of fractions from mycelial fungi withantigenic activity Mol. Immunol 17:1097-1103
[77]Hearn. V. M. Wilson. E. V. and Mackcnzie D. W. R. 1988. Characterization of purified wall antigens obtained from Aspergillus species. Pages 355-365 in: Fungal Antigens Isolation. Purification and Detection. E. Drouhet. G.T. Cole, L. de Repentigny, J, P. Large and B DuPont eds. Plenum Press New York.
[78]Hill M, Witsenbore H, Zabeau M. et al, 1996. PCR-based fingerprinting using AFLPs as a tool for studying genetic relationships in Lactuca spp. Theor Appl Genet., 8:1202-1210
[79]Ho HH, Zhuang NT, Liang ZR, Yu YN, 1983. Phytophthora cinnamomi on black locust(Robinia pseudoacacia)in Jiangsu Province of China, Mycologia 75:881—886
[80]Inam-ul-Haq; Sultan-Mahmood-Khan; 1999, Riaz-Ahmad.Physiological studies on six fungal isolates from rotted roots of cotton. Pakistan-Journal-of-Phytopathology. 11: 2,173-177; 14 ref.
[81]Irey MS, Comstock JC, 1991. Use of an enzyme-linked immunosorbent assay to detect leaf scald pathogen, Xantbomonas albilineans, in sugarcane. Journal of the American Society of Sugar Cane Technologists 11, 48-52
[82]Ito. 1979, Maize leaf spot caused by Curvulria lunata(Wakker)Boed. Summa-phytopathologica,5(3-4):181—184
[83]Javed-MS:Sheikh-AW; Idrees-M; Saleem-A. 1997, Fungi recorded from the onion seeds and control of Fusarium solani by chemicals. Pakistan-Journal-of-Phytopathology. 9: 1,93-96; 13 ref.
[84]Kaosiri T, Zentmyer GA 1980. Protein, esterase and peroxidase patterns in the Phytophthora palmivora complex from cacao. Mycologia 72:988—1000
[85]Kaur. 1973, A Curvularia leaf disease of maize. Kasetsart-Journal, 7(2)76—80
[86]Kistler H C., Benny N. 1989. The mitochontrial genome of Frsarium axysporum. Plasmid, 22:86-89
[87]Kohn L M. Pestsche D M., Bailey S R. et al. 1988. Restriction fragment length polymerphisms in the nuclear and mitochondrial DNA of Sclerotinia species. Phytopathology, 78:1047-1051
[88]Kozlowski M,Stepien pp.Restriction enzyme analysis of mitochondrial DNA of meders of fhe genus Aspergillus as an aidin toxonoomy. Mcrobiol, 1982,128:471～476
[89]Lacourt,Duncan. Specific detection of Phytophthora nicotianae using the polymerase chain reaction and primers basedon the DNA sepuence of its elictin gene parAI.Eur Jourplant pathol, 1997,103:73～83
[90]Lind V. 1990. Isolation of antigens for serlolgical identification of Pseudocercosporella herpotrichoides. Jormal of Plant Disease and Protection. 97(5):490-501
[91]Lind V. Isolation of antigens for serlolgical identification of Pseudocercosporella herpotrichoides. Jormal of Plant Disease and Protection. 1990, 97(5):490-501
[92]Lopes-JO. Jobim-NM. 1998, Dermatomycosis of the toe web caused by Curvularia lunata[J]. Revista-do-Instituto-de-Medicina-Tropical-de-Sao-Paula, 40: 5,327-328.
[93]Macri. 1976, Isolation and partial characeterization of phytotoxins from Curvularia lunata(Wakker)Boed. Physiogical-plant-pathology, 8(3):325—331
[94]Mandokhot, 1979. Chemical changes in maize leaves in response to leaf spot pathogens. Indian phytopatholigy, 1979, 32(4):658-660
[95]Manoj-Kumar; Agarwal-VK; Kumar-M. 1998, Location of seedborne fungi associated with discoloured maize seeds. Indian-Phytopathology, 51(3):247-250.
[96]Manoj-Kumar, Agarwal-VK, Kumar-M. 1999, Effect of fungicidal seed treatment on seedborne fungi, germination and seedling vigour of Maize. Seed-Research, 26(2):147-151.
[97]Mandokhot, 1979, Chemical changes in maize leaves in response to leaf spot pathogens. Indian phytopathology, 32(4): 658—660
[98]Manicom B Q, Bar-Joseph M, Rosner A, et al. 1987. Potential applications of random DNA probes and restriction fragment length polymorphisms in the taxonomy of the Fusaria. Phytopatholohy, 77(5):669-672
[99]Miller, S. A., Rittenburg, J. H., Petersen, F.P and Grothaus, G.D. 1988. Application of rapid, field-usable immunoassay for the diagnosis and monitoring of fungal pathogens in plants. Br. Crop Prot. Conf. Pests Dis. 2:795-803
[100]Moricca, Ragazzi, Kasuga. Detection of Fusaniumoxysporum f.sp. Vasinfectum in cotton tissue by polymerase chain reaction. Plant patholo, 1998,47:486～494
[101]Moudallal, Z., Altschuh, D., briand, J. P, and Van regenmortel, M. H. V. 1984. Comparative sensitivity of different ELISA procedures for detecting monoclonal antibodies. J. Immunol. Methods 68:35-43
[102]Moukhamedov R Hu X, Nazar R N, and Robb J. 1994. Use of PCR ribosomal intergenic sequences for the diagnosis of Verticillium tricorpus. Phytopathology, 84:256-259
[103]Mullis KB, Faloona F, Scharf Scharf SF et al. Specific emzymaticamplification of DNA in uitro:the plomerase chain reaction. Cold Spring Harbor Symposium on Quantitative Biolo, 1968,51:263～267
[104]Musgrave, D. R. Grose, T A., latch, G. C. M., and Christensen, M. J. 1986. Purification and characterization of the antigens of endophytic fungi isolated from Lolium perenne and Festuca arundinacea in New Zealand. N. Z. J. Agric. Res. 29:121-128
[105]Nameth, S. T, Shane, W. W., and Stier, J, C. 1990. Development of a monoclonal antibody for detection of Leptosphaeria korrae, the causal agent of necrotic ringspot disease of turfgrass. Phytopathology 80:1208-1211
[106]Newton A C, Regliski T. 1993. An ELISA for quantifying midew biomass. Joumai of Plant Diseases and Protection 100:176-179
[107]Niu YC.Comparisn of four formae speciales of Puccinia striiformis West. Proceedings of the Second China-Japan International Congress of Mycology, Beijing, 1992,119-120
[108] Nwachukwu-EO. 1995, Evaluation of seed health testing methods for the detection of seed-borne fungi of African yam bean. Agrosearch, 1(2): 123-127.
[109] Olufolaji, 1985. Comparative studies of the effect of Helminthosporium maydis and Curvularia pallescens infected on total nitrogen of maize leaves. Cryptogamie-Mycologie, 6(3): 197-200
[110] Olufolaji, 1984. Sporulation and growth of Curvularia pallescens as affected by media temperature and nitrogen sources. Phytopathology, 74(3): 260-263 Curvularia pallescens
[111] Osiewacz H.D,Ridder. R. 1991, Genome analusis of imperfect fungi:electrophoretic and characterization of the nuclear gene coding for gpd of Curvlaria lunata[J].Curr. Genet, 20:151-155.
[112] Oyekan. 1997, Effect of planting date on the incidence and severity of common fumgal diseases of maize in western stata. Nigerian-Journal-of-plant protection, 3:11-15
[113] Patchara PP, Osborn TC,Williams PH et al. Genetic analysis and identification of amplified fragment length polymorphism market linded to the alml avirulence gene of Lepeosphaeria mucnlans. Phytopatho, 1998,88:1068～1072
[114] Peter B, Marianne, Marga. Detection and identification of Phytophthora fragariae Hickmae by the polymerase chain reaction.Eur Jour plant pathol, 1997,103:345～355
[115] Rataj-Guranowska M, Wolk o B. 1991. Com parison of Fusarium oxysporum and Fusarium oxysporum var. reclons by analuyzing the isoenzyme and serological patterns. J Pythopathol, 132:289-293
[116] Reddy M N, Stahmann M A, Isozyme patterns of F usarium species and their significance in taxonomy. Phytopathol Z, 1972,74:115～125
[117] Ricker, R. W., Marois, J. J., Dlott, J. W., Bostock, R. M and Morrison, J. C. 1991 Immunodetection and quantification of botrytis cinerea on harvested wine grapes. Phytopathology 81:404-411
[118] Rott P, Davis Md, Baudin P, 1994. Serological variability in Xantbomonas albilineans causal agent of leaf scald disease of sugarcane. Plant Patbology 43,344-9
[119] Rott P, Chatenet M, Granier M, Baudin P, 1988. Recognition and detection of Xantbomonas and Clavibacter xyli xubsp. Xyli by indirect immunmofluorescence and enzyme immunoassays. Agronomia Tropicale 43,158
[120] Rozman,-D; Jezemik,-K; Komel,-R. 1994, Ultrastructure and genotypic characterization of the filamentous fungus Cochliobolus lunatus in comparison to the anamorphic strain Curvularia lunata. FEMS-Microbiology-Letters. 117:1,35-40; 17 ref.
[121] Irey MS, Comstock JC, 1991.Use of an enzyme-linked immunosorbent assay to detect leaf scald pathogen, Xantbomonas albilineans, in sugarcane. Journal of the American Society of Sugar Cane Technologists 11, 48-52
[122] Sambrook J, Fritish EF, Maniatis T. Moleculer cloing:alabonatory manual(2nd ed).Cold spring harbor Laboratory,New York:Cold spring Harbor, 1989
[123] Savage, S. D., and Sail, M. A. 1981. Radioimmunosorbent assay for Botrytis cinerea. Phytopathology 71:411-415
[124] Scheffer RP 1997. The nature of disease in plants. Cambridge University Press, New York.
[125] Shelby, R. 1985. Use of enzyme-linked immunosorbent assay(ELISA) as an aid in the detection of Diaporthe phaseolorum var. caulivora. Proc. South. Soybean Dis. Work.12:70
[126] Shashi-Sharma, Gupta-RBL, Yadava-CPS,etc. 1999, Effect of certain soil fungi on Metarhizium and Beauveria spp. and their pathogenicity against Holotrichia consanguinea[J]. Indian-Phytopathology, 52(2): 196-197.
[127] Shimizu S, Aizawa K, 1988. Serological classification of Beawveria bassiana. J. Invertebr. Pahol. 52:348—353
[128] Shimizu S, Aizawa K, 1992. Antigenic structures of blastospores of three entomopathogenic Paecilomyces Spieces and Nomuraea rileyi. J. Invertebr. Pathol. 59:106—108
[129] Sinclair, J. B. 1991. Latent infection of soybean plants and seeds by fungi. Plant Dis. 75:220-224
[130] Sivanesan A. 1984. The Bitunicate Ascomycetes and their anamorphs, Vaduz[C]. Cramer
[131] Sivanesan A. 1987, Graminicolous of Bipolaris, Curvlaria, Drechslera, Exserohilum and their telemorphs[J]. Mycol Pap, 158:261.
[132] Solanke-RB, Kore-SS,Sudewad-SM. 1997, Detection of soybean seed horne pathogens and effect of fungicides. Journal-of-Maharashtra-Agricultural-Universities,22(2):168-170.
[133] Solanke-RB; Hussaini-MM; Jawale-LN; Bonde-VJ. 1997, Effect of fungicidal seed treatment on seed health of sunflower under storage conditions. Journal-of-Maharashtra-Agricultural-Universities. 22: 3,349-350; 3 ref.
[134] Soviur R J, Pethica L M, Soddell J A. 1985, Electrophoretic patterns of sporangiospore proteins of Rhizopus isolates as taxonomic characters. Trans Brit Mycol Soc, 84: 701—708
[135] Somani-RB; Ingle-RW; Wanjari-SS. 1994, Variability in Curvularia lunata (Wakker)Boedijn causing grain mold in sorghum. International-Sorghum-and-Millets-Newsletter. No. 35, 106-107; 1 ref.
[136] Solanke-RB, Kore-SS,Sudewad-SM. 1997. Detection of soybean seed borne pathogens and effect of fungicides. Journal-of-Maharashtra Agricultural Universities, 22(2):168-170.
[137] Sundaram. S., Plasencia, J., and Banttari, E. E 1991. Enzyme-linked immunosorbent assay for detection of Verlicillium spp. Using antiscra produced to V. dahliae from potato. Phytopathology 81:1485-1489
[138] Tae-Ho Hah Msc. 1998. Heredity and phylogeny in the Alstroemeria using molecular markers (AFLP). From Wageningen Plant Breeding Research Theme 1
[139] Takenaka. S. 1992. Use of immunological methods with antiribosome serums to detect snow mold fungi in whcat plants. Phytopathology 82. 896-901
[140] Thomas C M et al. 1995. Identification of amplified restriction fragment polymorphism (AFLP) markers tightly linked to the tomato Cf-9 gene for resistance to Cladosporium fuivum, The Plant journal, 8(5):785-794
[141] Tijssen, P. 1990. Practice and Theory of Enzyme Immunoassays. Elsevier, New York 549 pp.
[142] Tsuda M, Ueyama A. 1977, Pseudocochliobolus nisikadoi, the perfect state of Helminthosporium coicis[J]. Mycologia, 69:1109-1120.
[143] Tsuda M, Ueyama A. 1982, Pseudocochliobolus verruculosus and variability of conidium morphology[J]. Mycologia, 74:563-568.
[144] Tsuda M, Ueyama A. 1985, Two new Pseudocochliobolus and a new specise of Curvlaria[J]. Trans Mycol Soc Japan, 26:321-330.
[145] Ushamalini-C; Rajappan-K; Manickam-K. 1999, Suppression of phenol and defense related enzymes in cowpea seed by seed-borne fungi. Acta-Phytopathologica-et-Entomologica-Hungarica. 34: 1-2, 51-55; 17 ref.
[146] Velicheti, R. K. Lamison, C., Brill, L. M. and Sinclair, J. B. 1993. Immunodetection of Phomopsis species in asymptomatic soybean plants. Plant Dis. 77:70-73
[147] Vilgalgse R, Gonzalez D. 1990. Ribobomal DNA restriction length polymorphisms in Rhizoctonia solani, Phytopathology, 80:151-158
[148] Vos P et al. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research, 23(21):4407-4414
[149] Wang Z. K, Comstock J. C. et. 1999. Comparison of PCR, DIA, ELISA and isolation on semiselective medium for detection of Xanthomonas albilineans, the causal agent of leaf scald of sugarcane. Plant Pathology, 48,245-252.
[150] Wong A L,Wiletts HJ. Electrophoretic studies of Australasian,North American and European isolates of Sclerotinia sclerotiorumand related species. J Gen Micro, 1975,90:355～359
[151] WycoffK L, Ayers A R. 1990. Monoclonal antibodies tosurface and extracellular antigens of a fungal plant pathogen, phytophthona megaspemaf sp. glycinea, recognize carbohydrate epitopes. Physiological and Molecular Plant Pathology. 37(1):55-79
[152] WycoffK L, jellison J and Ayers A R. 1987. Monoclonal antibodies to glycoprotein antigens of a fungal plant pathogen, Phytopathora megasperma f. sp. glycinea. Plant Physiology,,85:508-515
[153] WycoffK L, Ayers A R. 1991. Localization of carbonlydrate antigens in the walls of Phytophthora mesgasperma f. sp. glycinea by monoclonal antibodies, Physiological and Molecular Plant Pathology, 39:95-109
[154] Yao-CL,Magill-CW, Frederiksen, etc. 1991, Detection and identification of Peronosclerospora sacchari in maize by DNA hybridization[J]. Phytopathology, 81(8):901-905.
[2]曹以勤，彭金火，1990。几种禾本科植物上的弯孢菌，南京农业大学学报，13(4)：12-16。
[3]陈福生，罗信昌，周启，1999。酶联免疫技术检测植物病原真菌。植物检疫，13(1)：33-35。
[4]陈万全，冯洁，秦庆明等，1999。DNA分子标记在植物真菌病害研究中的应用。植物保护学报，26(3)：278-282。
[5]戴法超，高卫东，王晓鸣．1997，玉米弯孢菌叶斑病的初步研究简报．植物保护，28(1)：75-76．
[6]戴法超，王晓鸣，朱振东，等．1998，玉米弯孢菌叶斑病研究．植物病理学报，28(2)：123-129．
[7]戴芳澜，1979，中国真菌总汇。北京：科学出版社，936
[8]窦坦德，沈崇荛，2000。植物病原真菌检测技术研究进展。植物检疫，14(1)：31-33。
[9]窦坦德，沈崇尧，1997。大豆疫霉菌的血清学研究，菌物系统，16(2)：157-160
[10]窦坦德，沈崇尧，1997。快速检测大豆疫霉菌的酶联免疫吸附技术研究。植物检疫，11(3)：138-142
[11]傅俊范，白元俊等，1999，玉米弯孢菌叶斑病流行动态及产量损失测定，沈阳农业大学学报，30(3)：204-207．
[12]甘贤友等，1995，玉米弯孢霉叶斑病初步研究，植物保护，21(5)：24～25
[13]洪华珠，杨红等。1994，三株球孢白僵菌血清学性质的研究。真菌学报，13(1)：77-79。
[14]黄如貘等，应用裂解气相色谱法鉴定酵母菌的初步研究。见：程光胜等主编，分析微生物学专辑，北京：科学出版社，1988．194～197
[15]姜远良，1995。化学分类法在细菌和真菌系统分类学中的应用和发展。辽宁师范大学学报，18(2)，152-155。
[16]贾建航，李传友，伏健民等，1999，AFLP技术在玉米自交系类群分析研究中的应用。高技术通讯，4：43-46。
[17]李成文，1992。现代免疫化学技术．上海：上海科学技术出版社
[18]刘学敏，赵骞，曲金玲等。运用随机扩增多态性标记检测中国东北大豆灰斑病菌菌株遗传变异，植物病理学报，1998，28(1)：43～48
[19]卢乡怀，李明霞。1990，裂殖酵母属的酶及可溶性蛋白聚丙烯酰胺凝胶电泳。真菌学报，9(1)：50—55
[20]吕国忠，刘志恒，何富刚，等．1997，辽宁省爆发一种新病害—玉米弯孢菌叶斑病．沈阳农业大学学报，28(1)：75-76．
[21]吕国忠，薛玉梅，1999，玉米弯孢菌叶斑病菌毒素的生物活性测定及对玉米叶片超微结构的影响．沈阳农业大学学报，30(3)：212-214．
[22]蔺瑞明，高增贵，崔明珠，等．1999，玉米弯孢菌叶斑病苗期抗病性鉴定及遗传初步分析．植物保护，25(5)：1-3．
[23]戚佩坤．1978．玉米、高粱、谷子病原手册[M]北京：科学出版社，168-169．
[24]单卫星，陈受宜，康振生等，1995。DNA指纹技术分析在植病真菌群体遗传研究中的应用。西北农业大学学报，23(6)：84-89。
[25]魏景超，1975，水稻病原手册，科学出版社，198～204。
[26]徐大雅、黄河、王春平．1982，疫霉蛋白质的聚丙烯酰胺凝胶盘状电泳。真菌学报，1(1)：40—47
[27]徐德坤，王琪，殷秀东，等．1995，玉米弯孢菌叶斑病调查初报[J]．山东农业科学，6：38
[28]徐敬友，陆家云等，1993。六种疫霉菌体可溶性蛋白质和酯酶的聚丙烯酰胺凝胶平板电泳研究。真菌学报，12(1)：54-64。
[29]鄢洪海，高增贵，陈捷等，1999。玉米弯孢菌叶斑病研究进展。沈阳农业大学学报，30(3)：369-371．
[30]鄢洪海，高增贵等，2000。玉米弯孢菌叶斑病菌生理分化鉴定技术的研究，沈阳农业大学学报，31(5)：472-475．
[31]叶华智，张敏，秦芸等，2000。首次在四川发现玉米弯孢菌叶斑病[J]．四川植保，2：75-76．
[32]张定法，徐瑞富，张希福，等，1998。玉米弯孢霉叶斑病菌生物学特性的研究．河南职技师院学报，26(1)：18-21．
[33]张小平，陈强等，1999。用AFLP技术研究花生根瘤菌的遗传多样性。微生物学报，39(6)：483-488。
[34]赵玉锦，1988。植物生物化学技术与方法。北京：农业出版社，93-95。
[35]郑文明，刘峰，康振生等，2000。中国小麦条锈菌主要流行菌系的AFLP指纹分析。自然科学进展，10(6)：532-537。
[36]周志伟，黄河，毛霉目真菌的DNA提取及其GC含量的测定，微生物学通报，1991，18(5)：275～279
[37]Adiver-SS,Anahosur-KH. 1994. Association of Curvularia and Exserohilum species as sorghum moulds and their effect on seed germination and seedling growth. Current-Research-University-of-Agricultural-Sciences-Bangalore, 23(3-4):38-40.
[38]Alcom JL 1990, Additions to Bipolaris, Cochliobolus and Curvularia. Mycotaxon. 39,361-392.
[39]Alcorn JL 1991, New combinations and synonymy in Bipolaris and Curvularia and a new species of Exserohilum. Mycotaxon. 41,329-344.
[40]Anderson J B, Bailey S S, Pukila P J. 1989. Variation in ribosomal DNA among biological special of Armillaria, a genus of hoot infecting fungi. Evolution, 43:1652-1662
[41]Barrett B A, Kidwell K K. 1998. AFLP-based genetic diversity assessment among wheat cultivars from the Pacific Northwest. Crop Science, 38(5): 1261～1270
[42]Berbee M.L, Mona Pirseyedi, Hubbard S. 1999, Cochliobolus phylogenetics and the origin of known, highly virulent pathogens, inferred from ITS and glyceraldehyde-3-phosphate dehydrogenase gene sequences. Mycologia 91(6), 964-977.
[43]Becket J et al. 1995. Com bined mapping of AFLP and RFLP markers in barley, Mol Gen Genet, 249:65-73
[44]Brill L.M, McClary R.D, Sinclair J.B 1994. Analysis of two ELISA Formats and Antigen Preparations Using Polyclonal Antibodies Against Phomopsis longicolla. Phytopathology. 84, 173-179.
[45]Burdon JJ,Marshall D R.Isozyme variation between species andforrnae speciales of the genus Puccinia Pers. Can J Bot, 1981,59:2628～2634
[46]Burns DT, et al.Pyrolysis gas chromatography as an aid to the identification of Penicillium species.Jchromatography, 1976, 116:107～115
[47]Burrell R G, Clayten C W, Galleyly M F, et al. 1966. Factors affecting the antigenicity of the mycelium of three species of Phytophthora. Phytopathology, 56:421-426
[48]Chen-WeiQun. Chen-FaJun,Zhang-TianYu, etc., 2000, Some saprotrophic deuteromycetes in Xisha Islands[J]. Mycosystema19(3):428-430.
[49]Chu Xining, Bai Yuming, and Yuan Jingming 1994: A comparative study of electrophoretic patterns of soluble proteins and zymograms from nine strains of Rhizopus. Acta Mycologica Sinica. 13(2), 121-126.
[50]Clark M F, Adams A N. 1977. Characteristics of the microplate method of Enzyme-Linked Immunosorbent Assay for the detection of plant vinuses J Gert Virology. 34:475-483
[51]Crowhust RN. Differentiation of Fusanumsolani f.sp.cucurbitae race 1 and 2 by random amplification of polymorphic DNA. Curr Cenet, 1991,20:391～396
[52]Damjana Rozman,Kristijan Jezernik,Radovan Komel. 1994, Ultrastructure and genotypic characterization of the filamentous fungus Cochliobotus lunata in comparaisom to the anamorphic strain Curvlaria lunata[J]. FEMS Microbiology Letters, 117:35-40.
[53]De Vallavieille C, Erselius LJ 1984, Variation in protein profiles in Phytophthora: survey of a compositive population of three species on citrus. Trans br Mycol Soc 83:473—479
[54]Deshmukh RN,Raut JG. 1993, Sites of Curvularia lunata infection in sorghum seed. Indian-Phytopathology, 46(3):251-252.
[55]Dewey F M. Detection of plant-invading fungus by monoclonal antibodies, In techniques for the rapid detection of plant pathogens pp47-62, eds J. M. Duncan and L. Tonance Uk: Blackwell Scientific Publication.
[56]Dorey-SE,Ayliffe-WH,Edrich-C,etc. 1997, Fungal keratitis caused by Curvularia lunata, with successful medical treatment[J]. Eye-London, 11:5,754-755.
[57]Gunasekaran M,et al.Differentiation of races of TiUetia caries and T.foetida by pyrolysis-gas-hiquid chromatography. My-cologia. 1979,71:1066～1071
[58]Ellis JJ. Species and varieties in the Rhizopus arrhizus-Rhizopus oryzae group as incated by their DNA comple mentarity Mycolgia
[59]Ellis M B. Dematiaceous Hyphomycetes. 1966. VII: Curvularia, Brachysporium etc. Mycological Papers, 106:1～57
[60]Ellis M B. 1971, Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, UK: 452～495
[61]Ellis M B. 1976, More Decatiaceous Hyphomyvetes. Commonwealth Mycological Institute, Kew, UK. 404～406
[62]Erselius LJ, Vallavieille CD 1984. Variation in protein profiles of Phytophihora: comparison of six species. Tran Br Mycol Soc 83: 463—472
[63]Fargues J, Duriez T, Andrieu S et al, 1975. Etude immunologique compree di souches de Metarrhizium anisopliae(Metsch.)sor. Champignon Hyphomycete entomopathogene. C. R. Acad. Sci Paris ser. D. 281:1781—1783
[64]Fargues J, Duriez T, Popeye R et al, 1981. Immunological characterization of the entomopathogenic hyphoycetes Beawveria and Metarhizium. Mycopathologia. 75:101—108
[65]Forster H, Kinscherf T G, Lang S A., et al. 1989. Restriction fragment length polymorphisms of the mitochondrial DNA of Phyphthora megasperma isolated from soybea, alfalfa and fruit trees. Canadian Journal Botany, 67:529-537
[66]Fred Ausubel et al. 1995. Short Protocols in Molecular Biology. John Wiley and Sons, Inc.,
[67]Gardes M,fortin JA,Mueller GM RFLP in nuclear ribosomal DNA of for Laccata spp:L.bicolor,L.laccata,Lproximn and L.anethstina. Phytopathol, 1990,80:1312～1317
[68]Gieason, M. L. Ghabral. S. A. and Ferriss. R. S.1987. Serological detecnon of Phomopsts longicolla in soybean seeds. Phyopathology 77:371-375
[69]Gill H S,Zentmyer G A.Identification of Phytophthora species by disc electrophoresis.Phytopathology, 1978,68:163～167
[70]Goodwin P H. et al. 1990. Detection of Phytophthora parasitica from soil and host tissue with a species-specific DNA probe. Phytopathology, 80:277-281
[71]Grajal, Martic M J, Simon C J, Muehlbauer F J et al. 1993. Use of random amplified polymorphic DNA (RAPD)to characterize race 2 of Fusarium oxysporum f. sp. pisi.Phytopathology, 83(6):612-614
[72]Hajek AE, Butt TM et al, 1991. Detection of Entomophaga maimaiga(Zygomycetes:Entomophthorales)using Enzyme-Linked Immunosorbent Assay. J. Invertebr. Pathol. 58:1—9
[73]Hamison J G. 1990. Quantification of Phytophthora infestans mycelium in potato leaves by ELISA. Inproceeding of the fourth COST-88 workshop pp 18-19 Invergowire Dundee Scotland 25-27 Sept.
[74]Hawksworth DL,Pitt J I. A new taxonomy for Monascus species based on cultural and microscopical characters. Australian Journal of Botany, 1983,31:51～61
[75]He G, Prakash C S. 1997. Indentification of polymorphic DNA markers in cultivated peanut(Arachis hypogaea L.). Euphytica, 2:143-149
[76]Hearn. V M. and Mackenzic. D.W. R, 1980. The preparation and partial purification of fractions from mycelial fungi withantigenic activity Mol. Immunol 17:1097-1103
[77]Hearn. V. M. Wilson. E. V. and Mackcnzie D. W. R. 1988. Characterization of purified wall antigens obtained from Aspergillus species. Pages 355-365 in: Fungal Antigens Isolation. Purification and Detection. E. Drouhet. G.T. Cole, L. de Repentigny, J, P. Large and B DuPont eds. Plenum Press New York.
[78]Hill M, Witsenbore H, Zabeau M. et al, 1996. PCR-based fingerprinting using AFLPs as a tool for studying genetic relationships in Lactuca spp. Theor Appl Genet., 8:1202-1210
[79]Ho HH, Zhuang NT, Liang ZR, Yu YN, 1983. Phytophthora cinnamomi on black locust(Robinia pseudoacacia)in Jiangsu Province of China, Mycologia 75:881—886
[80]Inam-ul-Haq; Sultan-Mahmood-Khan; 1999, Riaz-Ahmad.Physiological studies on six fungal isolates from rotted roots of cotton. Pakistan-Journal-of-Phytopathology. 11: 2,173-177; 14 ref.
[81]Irey MS, Comstock JC, 1991. Use of an enzyme-linked immunosorbent assay to detect leaf scald pathogen, Xantbomonas albilineans, in sugarcane. Journal of the American Society of Sugar Cane Technologists 11, 48-52
[82]Ito. 1979, Maize leaf spot caused by Curvulria lunata(Wakker)Boed. Summa-phytopathologica,5(3-4):181—184
[83]Javed-MS:Sheikh-AW; Idrees-M; Saleem-A. 1997, Fungi recorded from the onion seeds and control of Fusarium solani by chemicals. Pakistan-Journal-of-Phytopathology. 9: 1,93-96; 13 ref.
[84]Kaosiri T, Zentmyer GA 1980. Protein, esterase and peroxidase patterns in the Phytophthora palmivora complex from cacao. Mycologia 72:988—1000
[85]Kaur. 1973, A Curvularia leaf disease of maize. Kasetsart-Journal, 7(2)76—80
[86]Kistler H C., Benny N. 1989. The mitochontrial genome of Frsarium axysporum. Plasmid, 22:86-89
[87]Kohn L M. Pestsche D M., Bailey S R. et al. 1988. Restriction fragment length polymerphisms in the nuclear and mitochondrial DNA of Sclerotinia species. Phytopathology, 78:1047-1051
[88]Kozlowski M,Stepien pp.Restriction enzyme analysis of mitochondrial DNA of meders of fhe genus Aspergillus as an aidin toxonoomy. Mcrobiol, 1982,128:471～476
[89]Lacourt,Duncan. Specific detection of Phytophthora nicotianae using the polymerase chain reaction and primers basedon the DNA sepuence of its elictin gene parAI.Eur Jourplant pathol, 1997,103:73～83
[90]Lind V. 1990. Isolation of antigens for serlolgical identification of Pseudocercosporella herpotrichoides. Jormal of Plant Disease and Protection. 97(5):490-501
[91]Lind V. Isolation of antigens for serlolgical identification of Pseudocercosporella herpotrichoides. Jormal of Plant Disease and Protection. 1990, 97(5):490-501
[92]Lopes-JO. Jobim-NM. 1998, Dermatomycosis of the toe web caused by Curvularia lunata[J]. Revista-do-Instituto-de-Medicina-Tropical-de-Sao-Paula, 40: 5,327-328.
[93]Macri. 1976, Isolation and partial characeterization of phytotoxins from Curvularia lunata(Wakker)Boed. Physiogical-plant-pathology, 8(3):325—331
[94]Mandokhot, 1979. Chemical changes in maize leaves in response to leaf spot pathogens. Indian phytopatholigy, 1979, 32(4):658-660
[95]Manoj-Kumar; Agarwal-VK; Kumar-M. 1998, Location of seedborne fungi associated with discoloured maize seeds. Indian-Phytopathology, 51(3):247-250.
[96]Manoj-Kumar, Agarwal-VK, Kumar-M. 1999, Effect of fungicidal seed treatment on seedborne fungi, germination and seedling vigour of Maize. Seed-Research, 26(2):147-151.
[97]Mandokhot, 1979, Chemical changes in maize leaves in response to leaf spot pathogens. Indian phytopathology, 32(4): 658—660
[98]Manicom B Q, Bar-Joseph M, Rosner A, et al. 1987. Potential applications of random DNA probes and restriction fragment length polymorphisms in the taxonomy of the Fusaria. Phytopatholohy, 77(5):669-672
[99]Miller, S. A., Rittenburg, J. H., Petersen, F.P and Grothaus, G.D. 1988. Application of rapid, field-usable immunoassay for the diagnosis and monitoring of fungal pathogens in plants. Br. Crop Prot. Conf. Pests Dis. 2:795-803
[100]Moricca, Ragazzi, Kasuga. Detection of Fusaniumoxysporum f.sp. Vasinfectum in cotton tissue by polymerase chain reaction. Plant patholo, 1998,47:486～494
[101]Moudallal, Z., Altschuh, D., briand, J. P, and Van regenmortel, M. H. V. 1984. Comparative sensitivity of different ELISA procedures for detecting monoclonal antibodies. J. Immunol. Methods 68:35-43
[102]Moukhamedov R Hu X, Nazar R N, and Robb J. 1994. Use of PCR ribosomal intergenic sequences for the diagnosis of Verticillium tricorpus. Phytopathology, 84:256-259
[103]Mullis KB, Faloona F, Scharf Scharf SF et al. Specific emzymaticamplification of DNA in uitro:the plomerase chain reaction. Cold Spring Harbor Symposium on Quantitative Biolo, 1968,51:263～267
[104]Musgrave, D. R. Grose, T A., latch, G. C. M., and Christensen, M. J. 1986. Purification and characterization of the antigens of endophytic fungi isolated from Lolium perenne and Festuca arundinacea in New Zealand. N. Z. J. Agric. Res. 29:121-128
[105]Nameth, S. T, Shane, W. W., and Stier, J, C. 1990. Development of a monoclonal antibody for detection of Leptosphaeria korrae, the causal agent of necrotic ringspot disease of turfgrass. Phytopathology 80:1208-1211
[106]Newton A C, Regliski T. 1993. An ELISA for quantifying midew biomass. Joumai of Plant Diseases and Protection 100:176-179
[107]Niu YC.Comparisn of four formae speciales of Puccinia striiformis West. Proceedings of the Second China-Japan International Congress of Mycology, Beijing, 1992,119-120
[108] Nwachukwu-EO. 1995, Evaluation of seed health testing methods for the detection of seed-borne fungi of African yam bean. Agrosearch, 1(2): 123-127.
[109] Olufolaji, 1985. Comparative studies of the effect of Helminthosporium maydis and Curvularia pallescens infected on total nitrogen of maize leaves. Cryptogamie-Mycologie, 6(3): 197-200
[110] Olufolaji, 1984. Sporulation and growth of Curvularia pallescens as affected by media temperature and nitrogen sources. Phytopathology, 74(3): 260-263 Curvularia pallescens
[111] Osiewacz H.D,Ridder. R. 1991, Genome analusis of imperfect fungi:electrophoretic and characterization of the nuclear gene coding for gpd of Curvlaria lunata[J].Curr. Genet, 20:151-155.
[112] Oyekan. 1997, Effect of planting date on the incidence and severity of common fumgal diseases of maize in western stata. Nigerian-Journal-of-plant protection, 3:11-15
[113] Patchara PP, Osborn TC,Williams PH et al. Genetic analysis and identification of amplified fragment length polymorphism market linded to the alml avirulence gene of Lepeosphaeria mucnlans. Phytopatho, 1998,88:1068～1072
[114] Peter B, Marianne, Marga. Detection and identification of Phytophthora fragariae Hickmae by the polymerase chain reaction.Eur Jour plant pathol, 1997,103:345～355
[115] Rataj-Guranowska M, Wolk o B. 1991. Com parison of Fusarium oxysporum and Fusarium oxysporum var. reclons by analuyzing the isoenzyme and serological patterns. J Pythopathol, 132:289-293
[116] Reddy M N, Stahmann M A, Isozyme patterns of F usarium species and their significance in taxonomy. Phytopathol Z, 1972,74:115～125
[117] Ricker, R. W., Marois, J. J., Dlott, J. W., Bostock, R. M and Morrison, J. C. 1991 Immunodetection and quantification of botrytis cinerea on harvested wine grapes. Phytopathology 81:404-411
[118] Rott P, Davis Md, Baudin P, 1994. Serological variability in Xantbomonas albilineans causal agent of leaf scald disease of sugarcane. Plant Patbology 43,344-9
[119] Rott P, Chatenet M, Granier M, Baudin P, 1988. Recognition and detection of Xantbomonas and Clavibacter xyli xubsp. Xyli by indirect immunmofluorescence and enzyme immunoassays. Agronomia Tropicale 43,158
[120] Rozman,-D; Jezemik,-K; Komel,-R. 1994, Ultrastructure and genotypic characterization of the filamentous fungus Cochliobolus lunatus in comparison to the anamorphic strain Curvularia lunata. FEMS-Microbiology-Letters. 117:1,35-40; 17 ref.
[121] Irey MS, Comstock JC, 1991.Use of an enzyme-linked immunosorbent assay to detect leaf scald pathogen, Xantbomonas albilineans, in sugarcane. Journal of the American Society of Sugar Cane Technologists 11, 48-52
[122] Sambrook J, Fritish EF, Maniatis T. Moleculer cloing:alabonatory manual(2nd ed).Cold spring harbor Laboratory,New York:Cold spring Harbor, 1989
[123] Savage, S. D., and Sail, M. A. 1981. Radioimmunosorbent assay for Botrytis cinerea. Phytopathology 71:411-415
[124] Scheffer RP 1997. The nature of disease in plants. Cambridge University Press, New York.
[125] Shelby, R. 1985. Use of enzyme-linked immunosorbent assay(ELISA) as an aid in the detection of Diaporthe phaseolorum var. caulivora. Proc. South. Soybean Dis. Work.12:70
[126] Shashi-Sharma, Gupta-RBL, Yadava-CPS,etc. 1999, Effect of certain soil fungi on Metarhizium and Beauveria spp. and their pathogenicity against Holotrichia consanguinea[J]. Indian-Phytopathology, 52(2): 196-197.
[127] Shimizu S, Aizawa K, 1988. Serological classification of Beawveria bassiana. J. Invertebr. Pahol. 52:348—353
[128] Shimizu S, Aizawa K, 1992. Antigenic structures of blastospores of three entomopathogenic Paecilomyces Spieces and Nomuraea rileyi. J. Invertebr. Pathol. 59:106—108
[129] Sinclair, J. B. 1991. Latent infection of soybean plants and seeds by fungi. Plant Dis. 75:220-224
[130] Sivanesan A. 1984. The Bitunicate Ascomycetes and their anamorphs, Vaduz[C]. Cramer
[131] Sivanesan A. 1987, Graminicolous of Bipolaris, Curvlaria, Drechslera, Exserohilum and their telemorphs[J]. Mycol Pap, 158:261.
[132] Solanke-RB, Kore-SS,Sudewad-SM. 1997, Detection of soybean seed horne pathogens and effect of fungicides. Journal-of-Maharashtra-Agricultural-Universities,22(2):168-170.
[133] Solanke-RB; Hussaini-MM; Jawale-LN; Bonde-VJ. 1997, Effect of fungicidal seed treatment on seed health of sunflower under storage conditions. Journal-of-Maharashtra-Agricultural-Universities. 22: 3,349-350; 3 ref.
[134] Soviur R J, Pethica L M, Soddell J A. 1985, Electrophoretic patterns of sporangiospore proteins of Rhizopus isolates as taxonomic characters. Trans Brit Mycol Soc, 84: 701—708
[135] Somani-RB; Ingle-RW; Wanjari-SS. 1994, Variability in Curvularia lunata (Wakker)Boedijn causing grain mold in sorghum. International-Sorghum-and-Millets-Newsletter. No. 35, 106-107; 1 ref.
[136] Solanke-RB, Kore-SS,Sudewad-SM. 1997. Detection of soybean seed borne pathogens and effect of fungicides. Journal-of-Maharashtra Agricultural Universities, 22(2):168-170.
[137] Sundaram. S., Plasencia, J., and Banttari, E. E 1991. Enzyme-linked immunosorbent assay for detection of Verlicillium spp. Using antiscra produced to V. dahliae from potato. Phytopathology 81:1485-1489
[138] Tae-Ho Hah Msc. 1998. Heredity and phylogeny in the Alstroemeria using molecular markers (AFLP). From Wageningen Plant Breeding Research Theme 1
[139] Takenaka. S. 1992. Use of immunological methods with antiribosome serums to detect snow mold fungi in whcat plants. Phytopathology 82. 896-901
[140] Thomas C M et al. 1995. Identification of amplified restriction fragment polymorphism (AFLP) markers tightly linked to the tomato Cf-9 gene for resistance to Cladosporium fuivum, The Plant journal, 8(5):785-794
[141] Tijssen, P. 1990. Practice and Theory of Enzyme Immunoassays. Elsevier, New York 549 pp.
[142] Tsuda M, Ueyama A. 1977, Pseudocochliobolus nisikadoi, the perfect state of Helminthosporium coicis[J]. Mycologia, 69:1109-1120.
[143] Tsuda M, Ueyama A. 1982, Pseudocochliobolus verruculosus and variability of conidium morphology[J]. Mycologia, 74:563-568.
[144] Tsuda M, Ueyama A. 1985, Two new Pseudocochliobolus and a new specise of Curvlaria[J]. Trans Mycol Soc Japan, 26:321-330.
[145] Ushamalini-C; Rajappan-K; Manickam-K. 1999, Suppression of phenol and defense related enzymes in cowpea seed by seed-borne fungi. Acta-Phytopathologica-et-Entomologica-Hungarica. 34: 1-2, 51-55; 17 ref.
[146] Velicheti, R. K. Lamison, C., Brill, L. M. and Sinclair, J. B. 1993. Immunodetection of Phomopsis species in asymptomatic soybean plants. Plant Dis. 77:70-73
[147] Vilgalgse R, Gonzalez D. 1990. Ribobomal DNA restriction length polymorphisms in Rhizoctonia solani, Phytopathology, 80:151-158
[148] Vos P et al. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research, 23(21):4407-4414
[149] Wang Z. K, Comstock J. C. et. 1999. Comparison of PCR, DIA, ELISA and isolation on semiselective medium for detection of Xanthomonas albilineans, the causal agent of leaf scald of sugarcane. Plant Pathology, 48,245-252.
[150] Wong A L,Wiletts HJ. Electrophoretic studies of Australasian,North American and European isolates of Sclerotinia sclerotiorumand related species. J Gen Micro, 1975,90:355～359
[151] WycoffK L, Ayers A R. 1990. Monoclonal antibodies tosurface and extracellular antigens of a fungal plant pathogen, phytophthona megaspemaf sp. glycinea, recognize carbohydrate epitopes. Physiological and Molecular Plant Pathology. 37(1):55-79
[152] WycoffK L, jellison J and Ayers A R. 1987. Monoclonal antibodies to glycoprotein antigens of a fungal plant pathogen, Phytopathora megasperma f. sp. glycinea. Plant Physiology,,85:508-515
[153] WycoffK L, Ayers A R. 1991. Localization of carbonlydrate antigens in the walls of Phytophthora mesgasperma f. sp. glycinea by monoclonal antibodies, Physiological and Molecular Plant Pathology, 39:95-109
[154] Yao-CL,Magill-CW, Frederiksen, etc. 1991, Detection and identification of Peronosclerospora sacchari in maize by DNA hybridization[J]. Phytopathology, 81(8):901-905.