圖一、臺灣十字花科根瘤病樣品採集分布圖
Fig. 1. Locations of brassica clubroot sample collected for this study.
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A B
C D
E F
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圖二、臺灣地區十字花科根瘤病之田間發病情形。A. 根瘤病菌感染後期，根部
嚴重腐敗（大同鄉南山村）；B. 高麗菜因根瘤病菌感染所生之巨大根瘤(仁愛鄉
梅峰)；C. 田間群集發病狀態（顏色較淺的區塊；仁愛鄉靜觀）；D. 高麗菜因根
瘤病造成地上部萎凋(阿里山鄉十字村)；E. 仁愛鄉華崗之病田（照片前方即為病
株聚集處）；F. 因嚴重根瘤病而遭廢棄之田(尖石鄉新光)。
Fig. 2. Clubroot disease condition in fields of Taiwan. A. Root rot in late stage of
disease (Datong Township). B. Large clubroot after infection (Ren’ai Township). C.
Group incidence in fields (shallow color; Ren’ai Township). D. Cabbages wilt after
infection (Alishan Township). E. Disease field (Ren’ai Township). F. Abandoned field
beacuase of serious clubroot disease (Jianshi Township).
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圖三、Plasmodiophora brassicae rDNA ITS 區域 PCR 增幅引子及增幅區域示意
圖。
Fig. 3. The primer set for amplification of ribosomal ITS region of Plasmodiophora
brassicae.
18S rDNA ITS 1 ITS 2 28S rDNA 5.8S rDNA
WC 682
WC 35
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圖四、十字花科根瘤病菌 ITS 區域 PCR 增幅結果之電泳分析圖：M：1 kb marker;；
1-3 為五峰鄉樣本；4-6：為尖石鄉鎮西堡樣本；7-9：為尖石鄉新光樣本；10:正
對照組；11:負對照組。
Fig. 4. The results of P. brassicae ITS region amplification. Lane M. 1 kb marker.
Lane 1-3. Samples from Wufeng Township. Lane 4-6. Samples from Smangus, Jianshi
Township. Lane 7-9. Samples from Cinsibu, Jianshi Township. Lane 10. Positive
control. Lane 11. Negative Control.
M 1 2 3 4 5 6 7 8 9 10 11
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圖五、臺灣地區十字花科根瘤病菌與其他國家病原菌之 ITS 親緣樹狀圖；梅山
(MS)、鹿谷(LG)、北投(BT)、復興(FS)、蘆竹(LJ)、武陵(WL)、五峰(WF)與尖石
鎮西堡和新光(CSB、SG)。其他國家的分析序列包括澳洲(AF231027.1)、韓國
(AF353998.1) 、 瑞 士 (EF195335) 、 英 國 (Y12831.1) 、 日 本 長 野 縣 七 個 樣 品
(AB094984.1 、 AB094980.1 、 AB094982.1 、 AB094977.1 、 AB094978.1 、
AB094981.1、AB094983.1)，以及日本名古屋之樣品（AB526843.1）。
Fig. 5. Phylogenic tree of ITS region of in Taiwan and other countries.
Australia(AF231027.1) 、 Korea(AF353998.1) 、 Switzerland(EF195335) 、 United
Kingdom(Y12831.1)、MS: Meishan. LG: Lugu. BT: Beitou. FS: Fushin. LJ: Luzhu.
WL: Wuling. WF: Wufeng. CSB: Cinsibu. SG: Smangus. Kuk (AB094984.1), Sak2
(AB094980.1), Sto (AB094982.1), DaiF (AB094977.1), Dail (AB094978.1), Sak3
(AB094981.1), and Yuk (AB094983.1) are from Nagano, Japan. AB526843.1 is from
Nagoya, Japan.
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圖六、P. brassicae RAPD 分子標記 PCR 增幅產物的電泳分析結果；1. 北投；2. 志
良；3. 新光；4. 鎮西堡；5. 五峰；6. 大同山；7. 榮興；8. 十字路；9. 利稻；
10. 關原；11. 洛韶；12. 蘆竹；13. 梅山；14. 春陽；15. 福壽山；16. 北東眼
山；17. 鹿谷。M: 1 kb DNA ladder，N: PCR 增幅負對照組樣品
Fig. 6. The results of P. brassicae RAPD marker amplification in Taiwan. 1. Beitou；
2. Zhiliang；3. Cinsibu；4. Smangus；5. Wufeng；6. Tatung mountain；7. Rongxing；
8. Shizilu；9. Lidao；10. Guanyuan；11. Luoshao；12. Luzhu；13. Meishan；14.
Chunyang；15. Fushou mountain；16. North Donyan mountain；17. Lugu. M: 1 kb
DNA ladder；N: negative control.
1 2 3 4 5 6 7 8 M 9 10 11 12 13 14 15 16 M 17 N
1kb
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圖七、具芥子酶活性的真菌其生長及產孢構造圖：A. 左為 Fusarium oxysporum
(桃園)分離株，右為 Humicola fuscoatra (仁愛)分離株；B. Aspergillus japonicus var.
aculeatus 的產孢構造；C. Aspergillus japonicus var. aculeatus(北投)菌落。
Fig. 7. Growth of myrosinase producing fungi. A. Fusarisum oxysporum from
Taoyuan (left). Humicola fuscoatra from Ren'ai, Nantou (right). B. The sporulation
structure of Aspergillus japonicus var. aculeatus. C. Aspergillus japonicus var.
aculeatus from Beitou.
C
A B
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Humicola fuscoatra strain NRRL 25765 internal transcribed spacer 1, partial sequence;
5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence
圖八、自桃園分離之 Humicola fuscoatra 之 ITS-5.8S rDNA 序列在 NCBI BLAST
比對結果。
Fig. 8. The ITS-5.8S rDNA sequence of Humicola fuscoatra isolated from Taoyuan
was BLASTed against NCBI database
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Humicola fuscoatra strain NRRL 25765 internal transcribed spacer 1, partial sequence;
5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence
圖九、自仁愛鄉分離之 Humicola fuscoatra 之 ITS-5.8S rDNA 序列在 NCBI BLAST
比對結果。
Fig. 9. The ITS-5.8S rDNA sequence of Humicola fuscoatra isolated from Ren’ai was
BLASTed against NCBI database.
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Humicola fuscoatra strain NRRL 25765 internal transcribed spacer 1, partial sequence;
5.8S ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence
圖十、自仁愛鄉分離之另一株 Humicola fuscoatra 之 ITS-5.8S rDNA 序列在 NCBI
BLAST 比對結果。
Fig. 10. The ITS-5.8S rDNA sequence of Humicola fuscoatra isolated from Ren’ai
was BLASTed against NCBI database.
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Fusarium oxysporum isolate FoxySIN9 18S ribosomal RNA gene, partial sequence;
internal transcribed spacer 1, 5.8S ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and 28Sribosomal RNA gene, partial sequence
圖十一、自桃園分離之 Fusarium oxysporum 之 ITS-5.8S rDNA 序列在 NCBI
BLAST 比對結果。
Fig. 11. The ITS-5.8S rDNA sequence of Fusarium oxysporum isolated from Taoyuan
was BLASTed against NCBI database.
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Aspergillus aculeatus strain DQ2 internal transcribed spacer 1, partial sequence; 5.8S
ribosomal RNA gene and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence
圖十二、自北投分離之 Aspergillus japonicus var. aculeatus (同種異名: Aspergillus
aculeatus)之 ITS-5.8S rDNA 序列在 NCBI BLAST 比對結果。
Fig. 12. The ITS-5.8S rDNA sequence of Aspergillus japonicus var. aculeatus isolated
from Beitou was BLASTed against NCBI database.
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圖十三、利用 sinigrin-barium agar 鑑別具有 myrosinase 活性之真菌菌株。A.以
Aspergillus oryzae 作測詴，左盤為 SBA，右盤則未加 sinigrin。左盤可清楚見到
白色霧狀物；B.田間分離菌株實際的測詴情況。
Fig. 13. Identification of myrosinase-producing fungi with sinigrin-barium agar. A.
Aspergillus oryzae for control test. Left, colony on SBA.Right, colony on medium
without sinigrin. B. The test of fungi isolated from fields.
A
B
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A B
C D
E
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圖十四、產生芥子酶真菌在 10、15、20、25 及 30℃下生長速度的分析結果，各
溫度代表圖依序為 A、B、C、D、E。縱座標-菌落直徑大小(單位
cm)，橫坐標-天數；桃一、仁一、仁五皆為 Humicola fuscoatra，Fusarium 為從桃園分得之 F.
oxysporum，Aspergillus 為從北投分得之 Aspergillus japonicus var. aculeatus。
Fig. 14. The growth speed of myrosinase-producing fungi under different
temperatures . 3 isolates of Humicola fuscoatra are from Taoyuan and Ren’ai.
Fusarium oxysporum is isolated from Taoyuan. Aspergillus japonicus var. aculeatus is
isolated from Beitou. Y-axis: diameter of colonies. X-axis: days.
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A B
C.
C D
E
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圖十五、利用穴植管進行產生芥子酶真菌根圈定棲之測詴。A. 詴驗進行示意圖；
B. 定 棲 測 詴 分 離 得 到 的 Aspergillus japonicus var. aculeatus 菌 落 在 Rose
Bengal-PDA 培養基上的生長狀況，每盤中的各點代表穴植管內不同深度的土壤
或根段；C. 自未接種真菌之穴植管土壤或根段分離真菌；D、E. 自培養菌落挑
取孢子及產孢構造，於 40x 物鏡下之鏡檢圖，比例尺表示 10 μm。
Fig.15. Dibbling-tubes planting for the colonization test of myrosinase-producing
fungi. A. The schematic diagram of experiment. B. Conolies of Aspergillus japonicus
var. aculeatus from different soil depth on Rose Bengal-PDA. C. Fugi isolated from
non- inoculated soil and roots in the dibbling-tube. D & E. Spores and sporulation
structure of Aspergillus japonicus var. aculeatus. Bar: 10 μm.
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圖十六、溫室防治測詴。A、C-為僅於土中添加菜子粕者；B、D-為 Aspergillus
japonicus var. aculeatus 和菜子粕組合之防治測詴。
Fig. 16. Results of greenhouse prevention test. A & C. Canola meal. B & D.
Aspergillus japonicus var. aculeatus with canola meal.
A B
C D
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圖十七、北投田間測驗採得之樣本圖；圖中根瘤分別代表:左-未處理；中-Humicola
sp.加菜子粕；右-Aspergillus japonicus var. aculeatus 加菜子粕。
Fig. 17. Samples collected from Beitou. The roots of non-treated (left), treated with
Humicola sp. with rape seed cake (middle), and Aspergillus japonicus var. aculeatus
with rape seed cake (right).
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A
C D
E F
G H
B
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圖十八、梅峰農場田間測詴之實況。A、B 為測詴前施用製劑後敷設塑膠布進行
燻蒸；C、D 為兩重複區；E-H 為各處理植株生長狀況，依序為處理 1: Aspergillus
japonicus var. aculeatus 製劑及菜子粕、處理 2:僅有 Aspergillus japonicus var.
aculeatus 製劑、處理 3:僅有菜子粕、處理 4:未施任何處理。詴驗以 RCBD 設計，
每個處理兩重複。
Fig. 18. Condition of field test in Meifeng farm. A & B. The plastic sheetings are
covered on soil before the test. C & D. The areas of two repeats. E. Treatment 1. The
formulation of Aspergillus japonicus var. aculeatus with canola meal. F. Treatment 2.
Only the formulation of Aspergillus japonicus var. aculeatus. G. Treatment 3. Only
canola meal. H. Treatment 4. Non-treated. The experiment is designed with RCBD
method. Every treatment was repeated twice.
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圖十九、地上部之呈現。A、B 為第一和第二重複之代表植株比較，由左至右依 序為處理 1、2、3、4；C 為所有得到之 13 顆甘藍結球；D、E 為第一和第二重 複之外葉大小之比較，由左至右依序為處理 1、2、3、4。
Fig.19. Above-ground parts of cabbage in Meifeng farm. A & B. Plants of the two
repeats. The treatments are 1, 2, 3,and 4 from left to right in the figure. C. All
cabbages in the test. D & E. Leaves of two repeats. The treatments are 1, 2, 3, and 4
from left to right in the figure.
A B
C D
E
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lower right one is the non-germinated spore (Black bar is 1 μm.)
0.00%