荔枝露疫菌粒線體核酸探針之選殖與分析

陳隆鐘; 賴學珍; 李崇正; 鍾依紋; 葉瑩; 安寶貞; Chen, L.C.; Lai, S.C; Lee, C.C.; Chung, Y.W.; Yeh Y.; Ann, P.J.

荔枝露疫菌粒線體核酸探針之選殖與分析

Other Title

Cloning and analysis of mtDNA probes of Peronophythora litchii

Journal

植物保護學會會刊

Journal Volume

40

Journal Issue

2

Pages

95-108

Date Issued

1998-06

Date

1998-06

Author(s)

陳隆鐘

賴學珍

李崇正

鍾依紋

葉瑩

安寶貞

Chen, L.C.

Lai, S.C

Lee, C.C.

Chung, Y.W.

Yeh Y.

Ann, P.J.

URI

https://scholars.tari.gov.tw/handle/123456789/4565

https://tpl.ncl.edu.tw/NclService/JournalContentDetail?SysId=A98015631

Abstract

利用三種內限制酵素EcoR I 、Hinc II 及(EcoR I + Sac I)剪切荔枝露疫菌PL024菌株之粒線體核酸，將所得核酸片段與pUC 18載體黏合並進行選殖，結果選殖到4個選殖株A2-3、HC3、HC9及ES6，其插入片段大小各為0.6 kb、0.4 kb、0.65 kb及0.6 kb，分別製備為核酸探針。並以墨點漬染法與25株荔枝露疫菌粒線體核酸進行雜合反應，均有雜合交互作用。另以17株Phytophthora菌株及Pythium splendens的全量核酸與核酸探針A2-3、HC3、HC9及ES6進行雜合反應，結果HC3、HC9、ES6核酸探針與Pythium splendens皆無雜合交互作用；ES6核酸探針與17株Phytophthora菌株沒有任何雜合交互作用，A2-3 核酸探針除Phytophthora cryptogea外都有雜合交互作用，顯示ES6核酸探針比A2-3 、HC3 及HC9核酸探針對荔枝露疫菌較具有專一性。經墨點潰染測試，ES6核酸探針的靈敏度可測到100 fg濃度的露疫菌粒線體核酸。 The mtDNA of Peronophythora litchii PL024 was digested with EcoR I, Hinc II and EcoR I / Sac I, and the DNA fragments were ligated with vector pUCI8. And then the recombinated plasmids were transformated into E. coli DH5α. There were four recombinat clones, designed as A2-3, HC3, HC9 and ES6 respectively. Their sizes are 0.6, 0.4, 0.65 and 0.6 kb. Probes A2-3, HC3, HC9 and ES6 showed strong signals in dot blot hybridization with 25 P. litchii isolates. Four probes hybridized with 17 Phytophthora isolates and Pythium splendens, and none of any hybridization signals were detected except probe A2-3. None of any hybridization signals was detected as ES6 probe hybridized with 17 Phytophthora isolates. Probe A2-3 hybridized with 17 Phytophthora isolates except Phytophthora cηptogea. Specificity of probe ES6 to P. litchii was higher than A2-3, HC3, HC9. In dot blotting analysis, a weak hybridization signals were shown when these four probes hybridized with 16 Pythium isolates, and ES6 probe was sensitive to mtDNA of Peronophythora litchii in concentration of 100 fg.

Subjects

荔枝露疫菌

粒線體核酸

核酸探針

墨點漬染法

Peronophythora litchii

mtDNA

DNA probes

dot blot hybridization

Publisher

中華植物保護學會

Type

journal article

File(s)

Name

40-2-95-108.pdf

Size

1.12 MB

Format

Adobe PDF

Checksum

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