Taiwan Agricultural Research Institute Council of Agriculture, Executive Yuan
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(68(1):16-27)Establishment and Application of Multiplex Polymerase Chain Reaction for Simultaneous Detection of Transgenic Potatoes
No full text馬鈴薯 (Solanum tuberosum L.) 為我國常見的食用及加工作物，其塊莖在工業和飼料上具有很高的應用價值。本試驗以馬鈴薯物種專一性 (taxon-specific) 內生性基因尿苷二磷酸葡萄糖焦磷酸化酶 (UDP-glucosepyrophosphorylase; UGPase) 序列與基因轉殖馬鈴薯品項之專一性序列 (event-specific sequence) 建立多重聚合酶連鎖反應 (multiplex polymerase chain reaction; mPCR) 檢測技術。結果顯示，經多重聚合酶連鎖反應可正確增幅出目標片段，且混合2 或3 個不同轉殖品項之DNA 樣本，亦能檢出專一性目標片段；單一基因轉殖品項之稀釋樣本檢測其最低檢測極限 (limit of detection; LOD)，本方法之檢測靈敏度可達0.5%。將本方法送至4 個不同的試驗單位進行能力試驗，結果亦顯示具備良好穩定性與再顯性。另外，本試驗以快速馬鈴薯核酸萃取方法並利用毛細管電泳分析目標片段，可大幅縮減檢測流程所耗時間、人力成本，期望能應用於馬鈴薯檢測技術開發與基因改造 (基改) 馬鈴薯轉殖品項檢驗方法之參考。
Potato (Solanum tuberosum L.) is known as one of the most important food crops in the world and has high application in industry and feed. In this study, we present a multiplex polymerase chain reaction (mPCR) method for simultaneous detection of genetically modified (GM) potatoes. Quadruplex PCR assay targeting three event-specific sequences and the taxon-specific endogenous reference gene (UDP-glucose pyrophosphorylase; UGPase) were established. All potato DNA samples generated the expected PCR products, and the detection method performed robustly and specifically with various combinations of mixed GM potato events. The limit of detection (LOD) of each single GM sample in our mPCR detection assay was 0.5%. Furthermore, a proficiency test returned from four research sectors also indicated that our detection method had good reproducibility and repeatability. We also developed a simple, rapid and cost-efficient nucleic acid extraction method coupled with multiplex PCR and followed capillary electrophoresis. These results suggest that our methods are suitable for detecting multiplex PCR targets and have the potential for screening unknown genetically modified potato samples through a simple procedure
(68(1):59-68)Evaluation of Real-Time PCR Methods for the Detection of Banana bunchy top virus
No full text香蕉萎縮病毒 (Banana bunchy top virus; BBTV) 為一種危害香蕉之病原，它的偵測與確認已是台灣香蕉產區及世界重要課題。本研究使用即時螢光定量polymerase chain reaction (PCR) 技術，應用於鑑定香蕉感染BBTV 與否之檢測。本技術發展之即時螢光定量PCR 技術，同時利用含有BBTV-CP 基因之引子和探針，以real-time PCR (qPCR) 法與TaqMan® 螢光探針鑑定BBTV 外，並區分香蕉其他病毒 (香蕉條斑病毒及胡瓜嵌紋病毒)，且不與健康組織有反應。本技術將可應用在植株感染病毒早期檢測、篩選抗BBTV 之香蕉品系、田間植株病害檢測之判定，以及作為健康香蕉種苗生產與驗證。
Banana bunchy top virus (BBTV) is a damaging pathogen for banana, its detection being of critical importance to certification programs for Taiwan and worldwide. Here we report a quantitative real-time polymerase chain reaction assay (qPCR) based on TaqMan® chemistry to improve the diagnosis of this pathogen. Real-time PCR, targeting the amplification of CP gene fragment, was highly specific and capable of distinguishing BBTV from non-target banana viruses, Cucumber mosaic virus and Banana steak virus, which are two other common virus species in banana plants. The protocol was also evaluated for its specificity using healthy banana and water controls. None of the non-target or healthy samples reacted in qPCR. The developed technique is sensitive and reliable and allows rapid detection and quantitation of BBTV in both field and experimental material used for the surveillance and specific diagnosis of BBTV
(68(1):78-89)A Preliminary Study on Control Efficacy of Chemical Insecticides and Colored Sticky Cards against the Sweet Potato Flea Beetle (Chaetocnema confinis Crotch)
No full text蕹菜小金花蟲 (Chaetocnema confinis Crotch) 為甘藷 [Ipomoea batatas (L.) Lam.] 常見之害蟲，其幼、成蟲分別為害藷塊及藷葉，影響產量及品質。本研究針對蕹菜小金花蟲成蟲，初步於室內評估現行登記及延伸使用殺蟲劑的藥效，以及有色黏紙誘捕效果。經圓葉法檢測9 種殺蟲劑對蕹菜小金花蟲之致死性，結果顯示20% 亞滅培 (acetamiprid) 可濕性粉劑、20% 達特南 (dinotefuran) 水溶性粒劑、20% 芬化利 (fenvalerate) 乳劑及43% 佈飛松 (profenofos) 乳劑之殺蟲效果較佳，處理後第3 日的死亡率分別可達96、82、82 及82%。關於有色黏紙的田間誘捕效果，5 種顏色 (螢光黃色、黃色、綠色、藍色及白色) 以螢光黃色所誘捕蟲數最多 (21.8 隻/黏紙)，藍色黏紙的效果最差 (1.4 隻/黏紙)。另根據各黏紙光譜相對反射率曲線，推測波長落在530 nm 左右之光線對蕹菜小金花蟲較具吸引力。不同高度下，擺放位置低於甘藷植冠的效果較佳，且捕捉蟲數隨擺放高度增加而顯著減少。此外，黏紙誘引效果亦受設置方位影響，本次試驗中面朝南方的黏紙捕捉蟲數最多，平均為38.8 隻/黏紙，而朝北面的蟲數則最少。
The sweet potato flea beetle (Chaetocnema confinis Crotch) is a common pest of sweet potato. The larvae and adults feed on storage roots and leaves, respectively, reducing the quality of both parts. Because of the lack of research on its control techniques, this study focused on C. confinis adults, evaluating the efficacy of currently recommended pesticides and sticky traps of various colors, heights, and orientations. The efficacy of 9 pesticides against C. confinis was assessed by the leaf disc method, and the results showed that acetamiprid (20% wettable powders; WP), dinotefuran (20% water soluble granules; SG), fenvalerate (20% emulsifiable concentrates; EC) and profenofos (43% EC) caused the greatest mortality, which reached 96, 82, 82 and 82% on the third day, respectively. In the sticky trap experiments, fluorescent yellow colored traps attracted the most C. confinis (21.8 beetles trap-1) among 5 colors (fluorescent yellow, yellow, green, blue and white), while blue was the least attractive (1.4 beetles trap-1). According to the spectral reflectance curves of the colored sticky cards, the green light with wavelengths around 530 nm might be more attractive to C. confinis. Besides, the traps hung lower than the sweet potato leaf canopy captured the most C. confinis, and the number captured decreased as the height increased. Lastly, trapping effectiveness was also affected by trap orientation. The sticky cards oriented to south captured the most C. confinis (38.8 beetles trap-1), while those facing the north captured the least in this single experiment
Wheat Yields and Cost-effectiveness of Various Cultivation Methods for Rice-Wheat Rotation System in Southern Asia and Taiwan
No full text稻麥輪作是南亞地區重要糧食生產栽培系統，種植小麥前的耕作模式可分整地、低整地與不整地，透過改變小麥播種前的耕作方式，可降低小麥延遲播種造成直接或間接對產量的影響，其中不整地雖可減少小麥田區準備的時間而不至於延後播種，但可能因土壤結構不利小麥根系生長造成減產。為了解決整地可能造成晚播的原因，較整地減少耕犁次數的低整地耕作可能是較為可行的替代方式，除了減少因多次耕犁而影響小麥播種外，低整地使土壤結構較為鬆散而具有高的水分滲透性，有助於小麥根系發展及截取水分，促進小麥生長發育，同時具較高的氮素利用效率，皆有助提升穀粒產量。臺灣小麥耕作方式有別於印度地區稻麥輪作，屬更集約的水稻－水稻－小麥輪作系統，耕作方式可分不整地撒播(no-till/broadcast sowing)、低整地撒播(reduced tillage/broadcast sowing)、低整地條播(reduced tillage/strip till)等三種，依據近年來於小麥主要產地調查得知，每公頃平均產量以低整地條播3,787 kg最高，較不整地撒播增產23.7%，造成產量差異主要係因低整地條播單位面積穗數顯著高於其他兩種。分析上述三種小麥播種方式之投入成本，結果顯示，雖然整地條播投入耕作成本較不整地撒播高，但因其產量最高，扣除投入耕作成本外仍有最佳之收益，應可推薦作為農民及農企業栽培管理的參考，將有助於提升臺灣小麥的產業競爭力及增進農民收益。
Rice-wheat rotation is an essential cropping system in southern Asia. Preparation prior to wheat sowing can be divided into tillage, reduced tillage, and no-tillage. Changing the tillage method alleviates the direct and indirect effects of delayed sowing on crop yields. In particular, no-tillage can reduce the preparation time and therefore avoid delayed sowing; however, this method leads to soil structure unconducive to wheat growths and yield losses. To address problems concerning delayed sowing due to tillage, reduced tillage that lowers the number of ploughing has been adopted as a viable alternative. This method creates loose soil structure with high water permeability, thereby facilitates the development and water acquisition of wheat roots. Accordingly, it is conducive to wheat growth, engenders high nitrogen utilization rates, and improves crop yields. Wheat cropping in Taiwan differs from that in India in that Taiwanese farmers generally adopt a rice- rice-wheat rotation system, the cultivation methods of which are divided into no-tillage with broadcast sowing, reduced tillage with broadcast sowing, and reduced tillage with strip till. Recent surveys on main wheat production regions in Taiwan have reported that reduced tillage with strip till leads to the highest mean yield at 3,787 kg per hectare, which is 23.7% higher than that of no-tillage with broadcast sowing. The difference in yields is mainly attributable to the notably higher panicle number per unit area of reduced tillage with strip till compared with other tillage methods. This study analyzed the costs of the three cultivation methods. The results indicate that despite the higher cost of reduced tillage with strip till compared with other methods, it leads to the highest yield and is therefore the most cost-effective after the cultivation cost is accounted for. This study can serve as a reference for farmers and agricultural businesses to manage their cultivation, helping the Taiwanese wheat industry improve its competitiveness and profits
A Study of Equilibrium Relative Humidity Properties for Soybeans and Beans
No full text此研究在不同處理之樣本中量測五種溫度下大豆，紅豆與綠豆之平衡相對濕度。由非線性迴歸技術可知修正Halsey式是最適用於此類農產品之模式。大豆之ERH物性具有遲滯性但不顯著，在同一相對濕度下其對含水率影響之量化差距約0.5%以內。乾燥處理溫度減少了持水能力。紅豆的ERH物性也受到遲滯性與乾燥處理溫度之影響。以此研究結果與各文獻資料之比較可知測定技術與品種對上述產品之ERH物性無顯著影響。針對高蛋白質，高油量之農產品建立其單－ERH模式乃是可行。
The equilibrium relative humidity (ERR) properties of soybeans, red beans and mung beans were determined at temperatures ranging from 5 to 50℃. The Modified-Halsey equation is the only adequate model to fit these ERH data. Hysteresis effect was found for the ERR properties of soybeans, however, the difference of moisture content between desorption and adsorption were within 0.5% at the same RH condition. The drying temperatare reduces the water hold ability of soybeans. The ERH/EMC relationships of redbeans were affected by the hysteresis and the drying temperature. Comparing these results of this research with the previously published data, the effects of the determinating technique, and varieties were insignificant for soybeans and redbeans. It is available to establish a specific ERH/EMC model for each high protein and oily products
Antifungal effect and chitinase activities of the froth of spittlebug Poophilus costalis (Walker) (Hemiptera: Cercopoidea: Aphrophoridae)
No full textNymphs of xylophagous spittlebug Poophilus costalis (Walker, 1851) produce froth that covers their entire body surface. However, no mold has ever been found on this froth under natural conditions. This study has shown that the froth of P. costalis can repress the growth of Fusarium oxysporum f. sp. pisi. Three chitinases, i.e., beta-N-acetylglucosaminidase, chitobiosidase, and endochitinase, with specific activities of 1.759, 0.365, and 1.172 U/mg protein, respectively, and a protease with minimal activity have been detected in this froth. beta-N-actylglucosaminidase activity was also detected in the source of the froth, the nymphal Malpighian tubules and the host plant Commelina diffusa Burm. f. These findings suggest that the antifungal effect of the froth from P. costalis may be attributed to the chitinase produced by its Malpighian tubules and/or its host plant
