41 research outputs found
基因轉殖作物品種檢測技術之開發-利用35S啟動子分子標誌檢測轉基因玉米及大豆。
[[abstract]]本研究係分別以轉抗殺草劑基因之大豆以及轉抗蟲基因之玉米作為模式植物,建立基因轉殖植物在DNA層次之初步檢測分析。研究中基因轉殖與非基因轉殖之玉米及大豆共十七個樣品,經比較不同DNA萃取成本、DNA純度以及聚合酵素連鎖反應結果,以 WizardTM Genomic DNA Purification Kit 萃取結果較佳。另比較利用巢式聚合酵素連鎖反應 (Nested Polymerase Chain Reaction) 以及一般聚合酵素連鎖反應,進行基因轉殖大豆及玉米之鑑定,另以歐盟認可之基因轉植產品偵測試劑組 (Biosmart Allin 1.0 GMO Screening System) 為對照,經依作物調整其聚合酵素連鎖反應條件,結果可正確測得基因轉殖玉米及大豆樣品中的CAMV 355 啟動子特有之150bp DNA片段;此外,依CAMV 35S啟動子序列,所合成之35S 1 / 35S 2 引子以及35S-F / 35S-R引子,亦可分別用以測得基因轉殖樣品中CAMV 35S啟動子特有之195bp DNA片段與207bp之DNA片段
Identifications of capsicum spp. by isozymes and randomly amplified polymorphic DNA (RAPD) markers
本試驗以番椒 (Capsicum spp) 為材料,探討利用植物性狀分類、同功異
構酵素及逢機增殖多型性 DNA 標誌 (RAPD marker) 三種方法,分析不同
自交系、雜交種及栽培種間的種原鑑別。目的為:(1)建立區別番椒品系
之分子標誌,(2) 建立品系間遺傳連鎖群之關係,(3) 建立快速及正確的
甜椒種子純度的鑑定技術。番椒品系之植物性狀鑑別,採用 IBPGR
(International Board for Plant Genetic Resources) 番椒性狀分類標
準,調查結果顯示由花冠、花萼及每節位花朵數可區別番椒 (Capsicum)
屬 C. annuum L. 及 C. chinense Jacquin 二個種,其中供試番椒品系
中除 P1697 為 C. chinense Jacquin 外其餘均為 C. annuum L。調查果
實性狀則又可區分 C. annuum 中的 7品系。唯甜椒種內的F1雜交種無法
依外表性狀區別。同功異構酵素分析的結果顯示C. annuum L.與C.
chinense Jacquin二個種間之 EST、GOT、PGI、PGM、PRX、SkDH 及 SOD
等 7種酵素圖譜明顯不同。其中在 C. annuum L.種內之 GOT、PRX、PGI
、SOD 則具有不同的多型性酵素圖譜,唯僅能清楚識別 1 ~ 3 個品系,
尚無法區分所有 C. annuum L. 中之品系。此外萃取緩衝液與取樣部位都
會影響同功異構酵素圖譜的解析度;以萃取過程不易褐化的緩衝液所得之
酵素解析力較高。不同取樣部位則視不同酵素而有不同表現,其中 PRX、
SOD、PGI 以下位葉之多型性表現較清晰, GOT、MDH、PGM、SkDH 則為中
位葉解析較好,而 EST則以上位葉的活性及解析度較高。自300種逢機引
子 (random primer) 中篩選具有多型性 DNA 標誌,獲得10種可產生多型
性的引子。其中有 6個引子 UBC313,UBC327,UBC346,UBC457 UBC483及
UBC484 產生的 RAPD 圖譜可以清晰的將供試的番椒 8品系加以區別,以
此引子的特定DNA片段的標誌可有效的用來鑑別甜椒品系之F1雜交種,
及12個甜椒栽培品種。連鎖群分析的結果顯示12個甜椒栽培品種的遺傳背
景很相似,可能近源於甜椒P852品系,5個甜椒自交系中以P1717及P852較
相似,P1657及P1740次之,而P859較遠;3個辣椒及5個甜椒自交系屬於不
同族群。本試驗結果顯示RAPD的分子標誌可有效的區別番椒種原及F1雜交
種,估算親緣關係及鑑定種子純度;尤其是親源接近的品系,更可增加鑑
定的準確性及敏感度。另外利用這些特定的多型性 DNA 標誌片段,可進
行種子純度檢測,以區別出混雜的異品種個體。
Morphological traits,isozymes and randomly amplified polymor-
phic DNA (RAPD) markers were used to study the identification
of Capsicum species.The purposes of this study were to
establish molecular markers for classification of the Capsicum
spp., to establish the genetic linkage relationships and a
quick, technique for the identification of seed purity of
Capsicum spp. The results of isozyme analysis indicated that
there were significantly different in the zymograms of EST,
GOT, PGI, PGM, PRX, SkDH and SOD between C. annuum L. and C.
chinense Jacquin. Distinctly polymorphic patterns were
exhibited among the C. annuum L., but only three lines can be
distinguished. Besides, the isozyme patterns might vary with
different extracting buffers and sampling positions. For
example, the expressions of PRX, SOD and PGI were clearer in
lower leaf, GOT, PGM and SkDH in middle leaf and EST in upper
leaf. A total of 10 primers was selected from 300 random
primers that showed the polymorphic patterns in Capsicum spp..
Among , the RAPD patterns of six primers,i.e."UBC 313","UBC
327" , "UBC 346", "UBC 457","UBC 483" and "UBC 484", could
clearly eight tested Capsicum lines and might be used as an
effective for the identification of F1 hybrids and cultivated
varieties of Capsicum spp.. The results of linkage cluster
analysis revealed that the genetic background of 12 cultivars
was very similar, and might originate from P852. Among the
five inbred lines of sweet pepper, P1717 had a closer relation
with P852, next by P1657 and P1740, but P 859 was farthest.The
three hot pepper lines belonged to different population, when
compared with these five inbred lines of sweet pepper. The
results suggested that the RAPD markers is preciser and more
sensitive than morphological and isozyme identifications and
can be used as an effective tool to distinguish plant
germplasms
設施甜椒關鍵害蟲管理及安全生產模式之研發與應用
本研究為強化設施農業全方位服務,提升設施蔬果產值能並達到不用農藥或農藥使用最少化的蔬菜生產目標。針對設施栽培甜椒,進行試驗模擬其安全生產模式之應用及評估效治效益,藉以擬定以生物防治為基礎的病蟲害綜合管理及安全生產策略,以黃(藍色)黏板監測關鍵病蟲害,掌握發生密度,以適時防治。於甜椒栽培初期以每株釋放4-7 隻南方小黑花椿象(Oriusstrigicollis (Poppius),防治薊馬及其他小型害蟲,搭配以植物油混方及石灰硫磺合劑,可有效壓制蚜蟲及細蟎等小型害蟲的發生密度,提高甜椒良果率,全程不施用農藥達到優質安全生產目標